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Approximately one third of all mammalian genes are essential for life. Phenotypes resulting from mouse knockouts of these genes have provided tremendous insight into gene function and congenital disorders. As part of the International Mouse Phenotyping Consortium effort to generate and phenotypically characterize 5000 knockout mouse lines, we have identified 410 lethal genes during the production of the first 1751 unique gene knockouts. Using a standardised phenotyping platform that incorporates high-resolution 3D imaging, we identified novel 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 identified in our screen, thus providing a novel dataset that facilitates prioritization and validation of mutations identified in clinical sequencing efforts.

Present initial reliability results of the Mi Nino scale and validation of scale responses with mealtime observations to determine whether it captures parent food-related behaviors in low-literacy, Spanish-speaking parents of preschool children. Mi Niño was developed to capture "parent-centered" and "child-centered" food-related behaviors with items based on the previous English language My Child at Mealtime tool, and tailored through cognitive interviews for low-income Spanish audiences. Factor analysis will be conducted and reliability of the resulting scales tested. Mealtime observations will be used to assess the accuracy of item responses to observed mealtime behaviors. Mi Niño was completed by 205 Hispanic parents from WIC or Head Start. Exploratory factor analysis of the responses will be conducted to examine whether the responses correspond to the "parent-centered" and "child-centered" subscales of MCMT. A subsample (n = 60) completed mealtime videotaping in their homes. All videotapes were coded by two independent coders for parent food-related behaviors using Noldus v.10 for the following behaviors: physical manipulation, feeding child, verbal demands to eat, bargaining; eating statements, eating inquiries, requests to eat, preference statements about food, and general food statements. Inter-rater reliability was established between coders through double-coding of 40% of coded videotapes (Cohens kappa >.70). All discrepancies (coded outside of a five-second window of each other or application of different codes) between the two coders on the shared videotapes were resolved through discussion and consensus. The total number of occurrences of each code was exported to SPSS v.22 and then adjusted for the length of the meal, resulting in a rate per minute (RPM) for each behavior. Pearson's correlations will examine whether rate of the observed behaviors corresponds to the behavior rating on the Mi Ninos scale. Parent food-related behaviors are related to the development of obesogenic behaviors in children. Validation of a Spanish language self-assessment tool to capture these behaviors would be a valuable tool to help programs effectively target them for obesity prevention. [ABSTRACT FROM AUTHOR]

Copper (Cu) deficiency can result in embryonic and yolk sac developmental anomalies. Cu deficiency decreases superoxide dismutase activity, and increases superoxide anions, which can react with nitric oxide (NO) to form peroxynitrite. One consequence of peroxynitrite formation is a decrease in NO availability, which can adversely affect development. When gestation day (GD) 8.5 embryos from Cu adequate (Cu+) or Cu deficient (Cu-) dams were cultured for 48 h in Cu+ or Cu medium, respectively, and subsequently incubated in KRB buffer for 2 h, Cu-/Cu- embryo and yolk sac (YS) had low NO levels in the conditioned media compared to Cu+/Cu+ controls. Similar results were obtained from GD 10.5 Cu+ vs. Cu- embryos collected in vivo. In a dose response study, Cu-/Cu- embryo and YS had a blunted NO response to increasing doses of the NOS agonist, acetylcholine in KRB buffer compared to Cu+/Cu+ controls. Endothelial NOS (eNOS) mRNA expression was low in Cu-/Cu- YS compared to controls. Inducible NOS protein was not detected in YS of either group at this stage of gestation. While eNOS protein was similar, phosphoSer1177 eNOS was lower in Cu-/Cu- YS than Cu+/Cu+ YS. To test whether altering NO levels could modulate Cu deficiency-induced teratogenicity, embryos were cultured in the presence of the NO donor, DETA/NONOate (20 µM). The addition of the NO donor to Cu- media improved developmental score and decreased the incidence of developmental anomalies in Cu-/Cu-embryos and YS. These data support the concept that altered NO availability contributes to Cu deficiency-induced embryo abnormalities and impaired YS vessel development. [ABSTRACT FROM AUTHOR]

To validate two pediatric obesity risk assessment tools for low-income Spanish speaking parents and one diet quality tool; determine the feasibility of EFNEP embedded in a medical clinic; coordinate an AFRI obesity grant program journal supplement. The focus of year four (Y4) was to complete participant data collection and entry to validate assessment tools. Enrollment for the EFNEP intervention concluded with post surveys and in-person interviews administered after each of 21 intervention rounds. Hispanic (> 99%) parents (n = 239) from WIC or Head Start completed the five data collection points and a blood sample (n = 169 children). Children were classified as being overweight (10%) or obese (16%). Children in the highest BMI percentile-for-age quartile have significantly higher lipid (229 ± 82), and metabolic (118 ± 33) indices, and larger waist circumference (58.8 ± 5.7) than children in the lowest BMI percentile-for-age quartile (188 ± 95, 95 ± 38, 49.9 ± 2.6, respectively). A sample of 60 families were videotaped at mealtime and all of the videotapes have been coded for parent behaviors and 20% for child behaviors. One-hundred and five parents attended at least one EFNEP session who were referred by their child's pediatrician. All parents engaged in setting nutrition and parenting goals and 71 parents were interviewed to assess the relevance of Motivational Modeling & feasibility of attending EFNEP intervention. In addition, cultural adaptation and design were completed for Mis Vegetales, a vegetable variety tool to measure diet quality. The HK website was also updated to include Spanish language versions of Healthy Kids (HK) and My Child at Meal Time and the newly validated HK 19-item tool http://healthykids.ucdavis.edu/. Journal supplement planning began with communication to project directors and three journals were contacted for comparison bids. Products and findings can support efforts to reduce childhood obesity risk in low-income Spanish-speaking families by providing valid tools to identifying risky nutrition and parenting behaviors and illustrating their utility in expanding EFNEP's reach to medical clinics. [ABSTRACT FROM AUTHOR]

Despite a substantial body of research, we lack fundamental understanding of the pathophysiology of COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) including pulmonary and cardiovascular outcomes, in part due to limitations of murine models. Most models use transgenic mice (K18) that express the human (h) angiotensin converting enzyme 2 ( ACE2 ), ACE2 knock-in (KI) mice, or mouse-adapted strains of SARS-CoV-2. Further, many SARS-CoV-2 variants produce fatal neurologic disease in K18 mice and most murine studies focus only on acute disease in the first 14 days post inoculation (dpi). To better enable understanding of both acute (<14 dpi) and post-acute (>14 dpi) infection phases, we describe the development and characterization of a novel non-lethal KI mouse that expresses both the ACE2 and transmembrane serine protease 2 ( TMPRSS2 ) genes (h ACE2 /h TMPRSS2 ). The human genes were engineered to replace the orthologous mouse gene loci but remain under control of their respective murine promoters, resulting in expression of ACE2 and TMPRSS2 instead of their murine counterparts. After intranasal inoculation with an omicron strain of SARS-CoV-2, h ACE2 /h TMPRSS2 KI mice transiently lost weight but recovered by 7 dpi. Infectious SARS-CoV-2 was detected in nasopharyngeal swabs 1-2 dpi and in lung tissues 2-6 dpi, peaking 4 dpi. These outcomes were similar to those in K18 mice that were inoculated in parallel. To determine the extent to which h ACE2/ h TMPRSS2 KI mice are suitable to model pulmonary and cardiovascular outcomes, physiological assessments measuring locomotion, behavior and reflexes, biomonitoring to measure cardiac activity and respiration, and micro computed tomography to assess lung function were conducted frequently to 6 months post inoculation. Male but not female SARS-CoV-2 inoculated h ACE2/ h TMPRSS2 KI mice showed a transient reduction in locomotion compared to control saline treated mice. No significant changes in respiration, oxygen saturation, heart rate variability, or conductivity were detected in SARS-CoV-2 inoculated mice of either sex. When re-inoculated 6 months after the first inoculation, h ACE2/ h TMPRSS2 KI became re-infected with disease signs similar to after the first inoculation. Together these data show that a newly generated h ACE2/ h TMPRSS2 KI mouse can be used to study mild COVID-19., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Liu, Brostoff, Ramirez, Wong, Rowland, Heffner, Flores, Willis, Evans, Lanoue, Lloyd and Coffey.)

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., (© 2023 The Authors.)

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.