Obesity Risk Assessment for Spanish-Speaking Immigrant Families with Young Children in the United States: Reliability and Validity with Nutrient Values.

The purpose is to examine validity and reliability for an obesity risk assessment tool developed in Spanish for immigrant families with children, 3-5 years old using an 8-week cross-sectional design with data collected over 1 year at Head Start and Special Supplemental Nutrition Program for Women, Infants and Children [WIC]. Parent/child dyads (206) provided a child obesity risk assessment, three child modified 24 h dietary recalls, three child 36+ h activity logs and one parent food behavior checklist. Main outcome measures were convergent validity with nutrients, cup equivalents, and diet quality and three assessments of reliability that included item difficulty index, item discrimination index, and coefficient of variation. Validity was demonstrated for assessment tool, named Niños Sanos. Scales were significantly related to variables in direction hypothesized [p ≤ 0.05]: Healthy Eating Index, fruit/vegetable cup equivalents, folate, dairy cup equivalents, vitamins D, ß-carotene, fiber, saturated fat, sugar, time at screen/ sleep/physical activity and parent behaviors. Three measures of reliability were acceptable. The addition of nutrient values as an analytical validation approach adds strength and consistency to previously reported Niños Sanos validation results using children's blood biomarkers and body mass index. This tool can be used by health professionals as an assessment of obesity risk in several capacities: (1) screener for counseling in a clinic, (2) large survey, (3) guide for participant goal setting and tailoring interventions, and (4) evaluation.

Cross-Cultural Adaptation and Initial Validation of Mi Niño: A Tool to Measure Food-Related Parenting Practices of Spanish-Speaking Parents with Young Children.

Background: Accurate measurement of food-related parenting practices is necessary to inform related interventions and program evaluation. Valid tools reflect cultural attributes that affect household food environments and feeding practices. Simple, unidirectional language adaptation approaches are insufficient to capture these attributes in assessment tools. My Child at Mealtime (MCMT) is a 27-item, validated, visually enhanced self-assessment tool to measure food-related parenting practices of low-income English-speaking parents of preschoolers. Objectives: The aim of this study was to describe the cross-cultural adaptation of MCMT into its Spanish version Mi Niño a la Hora the Comer (Mi Niño) and to establish its face validity, factor structure, and internal consistency. Methods: MCMT was adapted into its Spanish version after an iterative process that triangulated cognitive interviews with verification of conceptual equivalence by content experts to establish face validity and semantic equivalence. The resulting tool underwent confirmatory factor analysis to determine whether internal consistency was equivalent across the 2 versions. Results: Four rounds of cognitive interviews (n = 5, n = 6, n = 2, and n = 4, respectively) with Spanish-speaking women caregivers of children aged 3-5 y recruited from Head Start were conducted. Ten items were modified throughout the adaptation process. Modifications included improved clarity (6 items), comprehension (7 items), appropriateness (4 items), suitability (4 items), and usefulness (2 items) of text and/or accompanying visuals. Confirmatory factor analysis with a sample of Spanish-speaking caregivers (n = 243) resulted in 2 reliable factors representing "child-centered" (α = 0.82) and "parent-centered" (α = 0.87) food-related parenting practices. Conclusions: Face validity, semantic equivalence, and internal consistency of Mi Niño were established. This tool can be used in community settings to inform program content and measure changes in food-related parenting practices of Spanish-speaking parents and assist in setting food-related parenting goals. The next steps include exploring the correspondence of Mi Nino with mealtime behaviors observed through video recording.

Genome-wide screening reveals the genetic basis of mammalian embryonic eye development.

BACKGROUND: Microphthalmia, anophthalmia, and coloboma (MAC) spectrum disease encompasses a group of eye malformations which play a role in childhood visual impairment. Although the predominant cause of eye malformations is known to be heritable in nature, with 80% of cases displaying loss-of-function mutations in the ocular developmental genes OTX2 or SOX2, the genetic abnormalities underlying the remaining cases of MAC are incompletely understood. This study intended to identify the novel genes and pathways required for early eye development. Additionally, pathways involved in eye formation during embryogenesis are also incompletely understood. This study aims to identify the novel genes and pathways required for early eye development through systematic forward screening of the mammalian genome. RESULTS: Query of the International Mouse Phenotyping Consortium (IMPC) database (data release 17.0, August 01, 2022) identified 74 unique knockout lines (genes) with genetically associated eye defects in mouse embryos. The vast majority of eye abnormalities were small or absent eyes, findings most relevant to MAC spectrum disease in humans. A literature search showed that 27 of the 74 lines had previously published knockout mouse models, of which only 15 had ocular defects identified in the original publications. These 12 previously published gene knockouts with no reported ocular abnormalities and the 47 unpublished knockouts with ocular abnormalities identified by the IMPC represent 59 genes not previously associated with early eye development in mice. Of these 59, we identified 19 genes with a reported human eye phenotype. Overall, mining of the IMPC data yielded 40 previously unimplicated genes linked to mammalian eye development. Bioinformatic analysis showed that several of the IMPC genes colocalized to several protein anabolic and pluripotency pathways in early eye development. Of note, our analysis suggests that the serine-glycine pathway producing glycine, a mitochondrial one-carbon donator to folate one-carbon metabolism (FOCM), is essential for eye formation. CONCLUSIONS: Using genome-wide phenotype screening of single-gene knockout mouse lines, STRING analysis, and bioinformatic methods, this study identified genes heretofore unassociated with MAC phenotypes providing models to research novel molecular and cellular mechanisms involved in eye development. These findings have the potential to hasten the diagnosis and treatment of this congenital blinding disease.

Analysis of genome-wide knockout mouse database identifies candidate ciliopathy genes.

We searched a database of single-gene knockout (KO) mice produced by the International Mouse Phenotyping Consortium (IMPC) to identify candidate ciliopathy genes. We first screened for phenotypes in mouse lines with both ocular and renal or reproductive trait abnormalities. The STRING protein interaction tool was used to identify interactions between known cilia gene products and those encoded by the genes in individual knockout mouse strains in order to generate a list of "candidate ciliopathy genes." From this list, 32 genes encoded proteins predicted to interact with known ciliopathy proteins. Of these, 25 had no previously described roles in ciliary pathobiology. Histological and morphological evidence of phenotypes found in ciliopathies in knockout mouse lines are presented as examples (genes Abi2, Wdr62, Ap4e1, Dync1li1, and Prkab1). Phenotyping data and descriptions generated on IMPC mouse line are useful for mechanistic studies, target discovery, rare disease diagnosis, and preclinical therapeutic development trials. Here we demonstrate the effective use of the IMPC phenotype data to uncover genes with no previous role in ciliary biology, which may be clinically relevant for identification of novel disease genes implicated in ciliopathies.

Mendelian gene identification through mouse embryo viability screening.

BACKGROUND: The diagnostic rate of Mendelian disorders in sequencing studies continues to increase, along with the pace of novel disease gene discovery. However, variant interpretation in novel genes not currently associated with disease is particularly challenging and strategies combining gene functional evidence with approaches that evaluate the phenotypic similarities between patients and model organisms have proven successful. A full spectrum of intolerance to loss-of-function variation has been previously described, providing evidence that gene essentiality should not be considered as a simple and fixed binary property. METHODS: Here we further dissected this spectrum by assessing the embryonic stage at which homozygous loss-of-function results in lethality in mice from the International Mouse Phenotyping Consortium, classifying the set of lethal genes into one of three windows of lethality: early, mid, or late gestation lethal. We studied the correlation between these windows of lethality and various gene features including expression across development, paralogy and constraint metrics together with human disease phenotypes. We explored a gene similarity approach for novel gene discovery and investigated unsolved cases from the 100,000 Genomes Project. RESULTS: We found that genes in the early gestation lethal category have distinct characteristics and are enriched for genes linked with recessive forms of inherited metabolic disease. We identified several genes sharing multiple features with known biallelic forms of inborn errors of the metabolism and found signs of enrichment of biallelic predicted pathogenic variants among early gestation lethal genes in patients recruited under this disease category. We highlight two novel gene candidates with phenotypic overlap between the patients and the mouse knockouts. CONCLUSIONS: Information on the developmental period at which embryonic lethality occurs in the knockout mouse may be used for novel disease gene discovery that helps to prioritise variants in unsolved rare disease cases.

A novel DPH5-related diphthamide-deficiency syndrome causing embryonic lethality or profound neurodevelopmental disorder.

PURPOSE: Diphthamide is a post-translationally modified histidine essential for messenger RNA translation and ribosomal protein synthesis. We present evidence for DPH5 as a novel cause of embryonic lethality and profound neurodevelopmental delays (NDDs). METHODS: Molecular testing was performed using exome or genome sequencing. A targeted Dph5 knockin mouse (C57BL/6Ncrl-Dph5em1Mbp/Mmucd) was created for a DPH5 p.His260Arg homozygous variant identified in 1 family. Adenosine diphosphate-ribosylation assays in DPH5-knockout human and yeast cells and in silico modeling were performed for the identified DPH5 potential pathogenic variants. RESULTS: DPH5 variants p.His260Arg (homozygous), p.Asn110Ser and p.Arg207Ter (heterozygous), and p.Asn174LysfsTer10 (homozygous) were identified in 3 unrelated families with distinct overlapping craniofacial features, profound NDDs, multisystem abnormalities, and miscarriages. Dph5 p.His260Arg homozygous knockin was embryonically lethal with only 1 subviable mouse exhibiting impaired growth, craniofacial dysmorphology, and multisystem dysfunction recapitulating the human phenotype. Adenosine diphosphate-ribosylation assays showed absent to decreased function in DPH5-knockout human and yeast cells. In silico modeling of the variants showed altered DPH5 structure and disruption of its interaction with eEF2. CONCLUSION: We provide strong clinical, biochemical, and functional evidence for DPH5 as a novel cause of embryonic lethality or profound NDDs with multisystem involvement and expand diphthamide-deficiency syndromes and ribosomopathies.

Supplier-origin mouse microbiomes significantly influence locomotor and anxiety-related behavior, body morphology, and metabolism.

The mouse is the most commonly used model species in biomedical research. Just as human physical and mental health are influenced by the commensal gut bacteria, mouse models of disease are influenced by the fecal microbiome (FM). The source of mice represents one of the strongest influences on the FM and can influence the phenotype of disease models. The FM influences behavior in mice leading to the hypothesis that mice of the same genetic background from different vendors, will have different behavioral phenotypes. To test this hypothesis, colonies of CD-1 mice, rederived via embryo transfer into surrogate dams from four different suppliers, were subjected to phenotyping assays assessing behavior and physiological parameters. Significant differences in behavior, growth rate, metabolism, and hematological parameters were observed. Collectively, these findings show the profound influence of supplier-origin FMs on host behavior and physiology in healthy, genetically similar, wild-type mice maintained in identical environments.

Feasibility of Colocating a Nutrition Education Program into a Medical Clinic Setting to Facilitate Pediatric Obesity Prevention.

PURPOSE: Within a medical clinic environment, pediatric obesity prevention education for families faces challenges. Existing long-term government-funded nutrition education programs have the expertise and staff to deliver. The purpose is to determine feasibility of colocating the Expanded Food and Nutrition Education Program (EFNEP) into a medical clinic setting to support pediatric obesity prevention. METHODS: Physicians from a large university teaching and research hospital (n = 73) and 4 small Medicaid-serving community clinics (n = 18) in the same geographic area in northern California were recruited and trained in the patient-referral protocol for a primary prevention intervention provided by EFNEP. The 8-week intervention deployed in the medical clinics, included general nutrition, physical activity and parenting topics anchored with guided goal setting and motivational modeling. Referral, enrollment, and attendance data were collected for 2 years. Parent and physician feasibility surveys, parent interviews and parent risk assessment tools were administered. Paired-sample t-test analysis was conducted. RESULTS: Twenty intervention series with parents of patients (n = 106) were conducted at 5 clinics. Physicians (n = 92) generated 686 referrals. Every 6 referrals generated 1 enrolled parent. Physicians (91%, n = 34) reported the intervention as useful to families. Parents (n = 82) reported improved child behaviors for sleep, screen time, physical activity, and food and beverage offerings (P < .0001) and at family mealtime (P < .001). Focus group interviews (n = 26) with 65 participants indicated that parents (97%) reacted positively to participating in the intervention with about a third indicating the classes were relevant to their needs. CONCLUSION: The intervention is a feasible strategy for the 5 medical clinics. Physicians referred and parents enrolled in the intervention with both physicians and parents indicating positive benefits. Feasibility is contingent upon physician awareness of the intervention and motivation to refer patients and additional EFNEP and clinic staff time to enroll and keep parents engaged.

Biallelic UBE4A loss-of-function variants cause intellectual disability and global developmental delay.

PURPOSE: To identify novel genes associated with intellectual disability (ID) in four unrelated families. METHODS: Here, through exome sequencing and international collaboration, we report eight individuals from four unrelated families of diverse geographic origin with biallelic loss-of-function variants in UBE4A. RESULTS: Eight evaluated individuals presented with syndromic intellectual disability and global developmental delay. Other clinical features included hypotonia, short stature, seizures, and behavior disorder. Characteristic features were appreciated in some individuals but not all; in some cases, features became more apparent with age. We demonstrated that UBE4A loss-of-function variants reduced RNA expression and protein levels in clinical samples. Mice generated to mimic patient-specific Ube4a loss-of-function variant exhibited muscular and neurological/behavioral abnormalities, some of which are suggestive of the clinical abnormalities seen in the affected individuals. CONCLUSION: These data indicate that biallelic loss-of-function variants in UBE4A cause a novel intellectual disability syndrome, suggesting that UBE4A enzyme activity is required for normal development and neurological function.

Obesity Risk Assessment Tool for Low-Income Spanish Speaking Immigrant Parents with Young Children: Validity with BMI and Biomarkers of Obesity.

Children of Hispanic origin bear a high risk of obesity. Child weight gain trajectories are influenced by the family environment, including parent feeding practices. Excessive body fat can result in unhealthful metabolic and lipid profiles and increased risk of metabolic diseases. The objective was to estimate criterion validity of an obesity risk assessment tool targeting Spanish-speaking families of Mexican origin using anthropometric measures and blood values of their young children. A cross-sectional study design with five data collection sessions was conducted over an eight-week period and involved 206 parent/child dyads recruited at Head Start and the Special Supplemental Nutrition Program for Women, Infants and Children in Northern California. Main outcome measures were criterion validity of Niños Sanos, a pediatric obesity risk assessment tool, using anthropometric measures and blood biomarkers. Niños Sanos scores were inversely related to child BMI-for-age percentiles (p = 0.02), waist-for-height ratios (p = 0.05) and inversely related to blood biomarkers for the metabolic index (p = 0.03) and lipid index (p = 0.05) and positively related to anti-inflammatory index (p = 0.047). Overall, children with higher Niños Sanos scores had more healthful lipid, metabolic and inflammatory profiles, as well as lower BMI-for-age percentiles and waist-to height ratios, providing evidence for the criterion validity of the tool. Niños Sanos can be used by child obesity researchers, by counselors and medical professionals during clinic visits as a screening tool and by educators as a tool to set goals for behavior change.

Obesity Risk Assessment Tool among 3-5 Year Olds: Validation with Biomarkers of Low-Grade Chronic Inflammation.

Background: Many families with young children practice nutrition, parenting, and lifestyle behaviors that set their children on trajectories for unhealthful weight gain. Potential adverse health effects of excessive body fat can result in the secretion of proinflammatory molecules and increased risk of inflammation and metabolic diseases. A pediatric obesity risk assessment tool named Healthy Kids (HK), demonstrated validity in a longitudinal study with child's measured BMI and 36-hour diet, screen, sleep, and activity logs. Our objective was to provide additional evidence of validity with low-income families with literacy issues using an inflammation index composed of four proinflammatory biomarkers. Methods: Parent/child pairs (n = 104) from Head Start and Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) provided HK, blood samples, and measured heights/weights. Select child inflammatory markers were discretized into two groups of HK scores. Data were analyzed with a mixed model adjusted for children's age and BMI. Results: A significant HK-time interaction effect was shown for the child inflammation index with two data collection points 1 year apart (pdid = 0.039). This index increased over 12 months in children with less healthful behaviors (p = 0.007), but not in children with more healthful profiles (p = 0.58). Conclusions: Children with less healthful HK scores had an elevated inflammation index indicating a low-grade chronic systemic inflammatory state. Taken together with our previously published findings, the HK tool has potential as a rapid and easy-to-administer assessment of the family environment and the child's obesity risk. HK can be useful for federal nutrition programs for evaluation, risk assessment, goal setting, and/or program planning in clinical and community environments.

An Obesity Risk Assessment Tool for Young Children: Validity With BMI and Nutrient Values.

OBJECTIVE: Demonstrate validity and reliability for an obesity risk assessment tool for young children targeting families' modifiable home environments. DESIGN: Longitudinal design with data collected over 100 weeks. SETTING: Head Start and the Special Supplemental Nutrition Program for Women, Infants, and Children. PARTICIPANTS: Parent-child pairs (n = 133) provided food behavior assessments; 3 child-modified, 24-hour dietary recalls; 3 ≥ 36-hour activity logs; and measured heights and weights. MAIN OUTCOME MEASURE: Five measures of validity and 5 of reliability. RESULTS: Validity was excellent for the assessment tool, named Healthy Kids, demonstrating an inverse relationship with child body mass index percentile-for-age (P = .02). Scales were significantly related to hypothesized variables (P ≤ .05): fruit or vegetable cup equivalents; folate; vitamins A, C, and D; ß-carotene; calcium; fiber; sugar; screen, sleep, and physical activity minutes; and parent behaviors. Measures of reliability were acceptable. CONCLUSIONS AND IMPLICATIONS: Overall, children with higher Healthy Kids scores had a more healthful profile as well as lower body mass index percentiles-for-age 1.5 years later. Healthy Kids has potential for use by nutrition professionals as a screening tool to identify young children most at risk for excess weight gain, as an evaluation to assess intervention impact, and as a counseling tool to tailor intervention efforts. Future research should include validation in other settings and with other populations.

Corrigendum: High-throughput discovery of novel developmental phenotypes.

This corrects the article DOI: 10.1038/nature19356.

High-throughput discovery of novel developmental phenotypes.

Approximately one-third of all mammalian genes are essential for life. Phenotypes resulting from knockouts of these genes in mice have provided tremendous insight into gene function and congenital disorders. As part of the International Mouse Phenotyping Consortium effort to generate and phenotypically characterize 5,000 knockout mouse lines, here we identify 410 lethal genes during the production of the first 1,751 unique gene knockouts. Using a standardized phenotyping platform that incorporates high-resolution 3D imaging, we identify phenotypes at multiple time points for previously uncharacterized genes and additional phenotypes for genes with previously reported mutant phenotypes. Unexpectedly, our analysis reveals that incomplete penetrance and variable expressivity are common even on a defined genetic background. In addition, we show that human disease genes are enriched for essential genes, thus providing a dataset that facilitates the prioritization and validation of mutations identified in clinical sequencing efforts.

Vegetable behavioral tool demonstrates validity with MyPlate vegetable cups and carotenoid and inflammatory biomarkers.

Young children are not meeting recommendations for vegetable intake. Our objective is to provide evidence of validity and reliability for a pictorial vegetable behavioral assessment for use by federally funded community nutrition programs. Parent/child pairs (n=133) from Head Start and the Special Supplemental Nutrition Program for Women, Infants and Children [WIC] provided parent-administered vegetable tools, three child 24-hour diet recalls, child blood sample and measured heights/weights. The 10-item Focus on Veggies scale, with an alpha of .83 and a stability reliability coefficient of .74, was positively related to vegetables in cup equivalents [p≤.05]; dietary intakes of folate, vitamin C, ß-carotene, potassium and magnesium [p≤.05-.01]; and soluble fiber [p≤.001]. The child vegetable scores were related to the parent's mediators [p≤.00001] and vegetable behaviors [p≤.00001]. Children's plasma inflammatory markers were negatively related to the 10 item scale [p≤.05] and are indicators of the child's health status. The positive relationship between the serum carotenoid index and a sub-scale of child vegetable behaviors offered additional support for criterion validity [p≤.05]. Finally, the inverse relationship of BMI-for-age percentile one year post baseline and a sub-scale of child vegetable behaviors supported the predictive validity [p≤.05]. Focus on Veggies, a simple assessment tool, can inform practitioners about the child's health status. A child with a high score, shows a healthful profile with a lower inflammation index, higher carotenoid index, lower BMI and higher vegetable intake. In conclusion, validity of Focus on Veggies has been demonstrated using vegetable cup equivalents and micronutrient intakes, anthropometry and blood biomarkers.

Influence of intracellular zinc on cultures of rat cardiac neural crest cells.

BACKGROUND: Developmental zinc (Zn) deficiency increases the incidence of heart anomalies in rat fetuses, in regions and structures derived from the outflow tract. Given that the development of the outflow tract requires the presence of cardiac neural crest cells (cNCC), we speculated that Zn deficiency selectively kills cNCC and could lead to heart malformations. METHODS: Cardiac NCC were isolated from E10.5 rat embryos and cultured in control media (CTRL), media containing 3 µM of the cell permeable metal chelator N, N, N', N'-tetrakis (2-pyridylmethyl) ethylene diamine (TPEN), or in TPEN-treated media supplemented with 3 µM Zn (TPEN + Zn). Cardiac NCC were collected after 6, 8, and 24 h of treatment to assess cell viability, proliferation, and apoptosis. RESULTS: The addition of TPEN to the culture media reduced free intracellular Zn pools and cell viability as assessed by low ATP production, compared to cells grown in control or Zn-supplemented media. There was an accumulation of reactive oxygen species, a release of mitochondrial cytochrome c into the cytoplasm, and an increased cellular expression of active caspase-3 in TPEN-treated cNCC compared to cNCC cultured in CTRL or TPEN + Zn media. CONCLUSION: Zn deficiency can result in oxidative stress in cNCC, and subsequent decreases in their population and metabolic activity. These data support the concept that Zn deficiency associated developmental heart defects may arise in part as a consequence of altered cNCC metabolism.

Reproductive and developmental outcomes, and influence on maternal and offspring tissue mineral concentrations, of (-)-epicatechin, (+)-catechin, and rutin ingestion prior to, and during pregnancy and lactation in C57BL/6J mice.

Maternal nutrition can have a significant effect on developmental processes during pregnancy and lactation. While certain flavonoids have been postulated to be beneficial for health, little is known about the effects of ingestion during pregnancy and lactation on the mother and progeny. We report on the effects of maternal consumption of high levels of certain flavonoids on reproductive and developmental outcomes in a mouse model. C57BL/6J female mice were fed a control diet (CT), the CT diet supplemented with 1% or 2% of a mix of epicatechin and catechin (EC1, EC2), or rutin (RU1, RU2) prior to, during pregnancy, and lactation. A subset of dams was killed on gestation day (GD) 18.5 to evaluate fetal outcomes and the remainder was allowed to deliver to evaluate offspring. Maternal food intake, body and tissue weight did not differ among groups. The number of resorptions, implantations, litter size, postnatal survival, body weight, and skeletal development were also similar. Alterations in maternal and offspring liver mineral concentrations were observed. The current results indicate that consumption of high amounts of epicatechin, catechin, and rutin during gestation and lactation is not associated with any marked developmental effects, although changes in liver mineral concentrations were noted.

Dietary factors and the risk for acute infant leukemia: evaluating the effects of cocoa-derived flavanols on DNA topoisomerase activity.

There is cumulative strong evidence that diets rich in flavanols can provide certain positive health benefits, particularly with respect to the cardiovascular system. Consequently, it has been suggested that increasing one's dietary intake of flavanols may be of benefit. Complicating this idea, there are reports that high intakes of certain flavonoids during pregnancy are associated with an increased risk for acute infant leukemia due to a poison effect of select polyphenolic compounds on DNA topoisomerase (topo) II activity that promotes aberrant chromosomal translocations. In the current study, we characterized the effects of select flavanols (epicatechin and catechin monomers), and select flavanol dimers and longer oligomers, on topo II activity, and on cellular toxicity in vitro. In contrast to the chemotherapeutic drug etoposide (VP16) and the flavonol quercetin, which strongly inhibited topo II activity and increased the formation of cleavage complexes demonstrating a poison effect, the flavanols epicatechin and catechin had little effect on topo II enzyme activity. Accordingly, several fold greater concentrations of the flavanols were required to achieve cellular toxicity similar to that of quercetin and VP16 in cultures of myeloid and lymphoid cells. Low cellular toxicity and limited topo II inhibition were also observed with a procyanidin-rich cocoa extract. Of all the flavanols tested, the dimers (B2, B5 and a mix of both) exerted the greatest inhibition of topo II and inhibited cellular proliferation rates at concentrations similar to quercetin. However, in contrast to quercetin, the dimers did not function as topo II poisons. Collectively, our in vitro data show that cocoa-derived flavanols have limited effects on topo II activity and cellular proliferation in cancer cell lines. We predict that these compounds are likely to have limited leukemogenic potential at physiological concentrations.

Influence of copper on early development: prenatal and postnatal considerations.

Copper (Cu) is an essential nutrient whose requirement is increased during pregnancy and lactation. These represent times of critical growth and development, and the fetus and neonate are particularly vulnerable to deficiencies of this nutrient. Genetic mutations that predispose the offspring to inadequate stores of Cu can be life threatening as is observed in children with Menkes disease. During the last decade, severe Cu deficiency, once thought to be a rare condition, has been reported in the literature at an increasing frequency. Secondary Cu deficiencies can be induced by a variety of ways such as excessive zinc or iron intake, certain drugs, and bariatric surgery. Premature and low birth weight infants can be born with low Cu stores. A number of mechanisms can contribute to the teratogenicity of Cu including decreased activity of select cuproenzymes, increased oxidative stress, decreased nitric oxide availability, altered iron metabolism, abnormal extracellular matrix protein crosslinking, decreased angiogenesis and altered cell signaling among others. The brain, heart, and vessels as well as tissues such as lung, skin and hair, and systems including the skeletal, immune, and blood systems, are negatively affected by suboptimal Cu during development. Additionally, persistent structural, biochemical, and functional adverse effects in the offspring are noted even when Cu supplementation is initiated after birth, supporting the concept that adequate Cu nutriture during pregnancy and lactation is critical for normal development. Although Cu-containing IUDs are an effective method for increasing intrauterine Cu concentrations and for reducing the risk of pregnancy, high amounts of dietary Cu are not thought to represent a direct developmental risk.