Docosahexaenoic acid supplementation during porcine oocyte in vitro maturation improves oocyte quality and embryonic development by enhancing the homeostasis of energy metabolism.

Although supplementation with docosahexaenoic acid (DHA) during porcine oocyte IVM is well-established, the available data are limited due to the lack of consistency. Moreover, to our knowledge, the anti-oxidant effects of DHA on porcine oocytes have not been reported. Hence, this study aimed to examine the effects of DHA supplementation on the regulation of energy metabolism during porcine oocyte maturation to improve oocyte maturation and embryonic development. By supplementing the IVM medium with various DHA concentrations, 25 µM DHA was identified as the optimal concentration which improved intraoocyte glutathione content and enhanced embryonic development after parthenogenesis. Compared to embryos derived from the control group, those derived from SCNT or IVF showed significantly improved blastocyst formation upon DHA supplementation during IVM. In addition, various transcription factors associated with oocyte development and apoptosis in mature oocytes were beneficially regulated in the DHA-treated oocytes. Moreover, DHA improved the AMP-activated protein kinase (AMPK)-regulatory ability of porcine oocytes and ameliorated nuclear maturation and embryonic development, which were decreased by artificially downregulating AMPK. To our knowledge, this is the first study to examine the effects of DHA as an AMPK regulator on oocyte maturation and embryo development in pigs. Furthermore, DHA addition to the IVM medium upregulated the relative expression of genes associated with mitochondrial potential and lipid metabolism. Therefore, the membrane potential of mitochondria (evaluated based on the JC-1 aggregate/JC-1 monomer ratio) and the levels of fatty acids and lipid droplets in matured oocytes increased, resulting in increased ATP synthesis. In conclusion, the DHA treatment of porcine oocytes with 25 µM DHA during IVM enhances the homeostasis of energy metabolism by improving mitochondrial function and lipid metabolism, leading to improved quality of matured oocytes and enhanced embryonic developmental potential of in vitro produced (IVP) embryos. Thus, 25 µM DHA supplementation could serve as a tool for improving the quality of IVP embryos. The study findings provide a basis for further research on improving the production efficiency of cloned animals by securing high-quality matured oocytes and enhancing energy metabolism in mammalian oocytes, including those of pigs.

Life-course trajectories of children through the U. S foster care system.

OBJECTIVE: Foster care is surprisingly commonplace in the United States, with more than one in twenty children experiencing placement in their lifetimes. Due to the complexity of foster care (e.g. age at contact, length of stay), we still lack a clear idea of how children move through the child welfare system. We pose the question, "do distinct typologies of system trajectories exist that may be used to inform policy analyses?" METHODS: We used child maltreatment reports and foster care case records for all U.S. children born in FY2006 and placed in care between birth and age 15. Using sequence analysis, we classified child-level system trajectories into distinct clusters. Then, we employed multilevel multinomial regression to explore child and system characteristics associated with each. RESULTS: Three typologies were identified: 1) early-entry-exit, 2) school-age-entry, and 3) late-entry. Early-entry-exit cases typically entered and left foster care before turning three, were frequently adopted, and had little ongoing system contact. School-age-entry children typically entered between ages 5-10, were in care for the shortest amount of time, and mostly exited to reunification. Late-entry children typically entered between ages 9-11, entered with substantial CPS history and remained in care into mid-adolescence. CONCLUSIONS: Our findings provide the first description of foster care trajectories in the US. Both practice and policy formulation can benefit from these empirically supported descriptions. Using such trajectory typologies, researchers can now explore how trajectories may predict wellbeing outcomes. We discuss how the differences among the typologies may inform identification of service needs and outcomes.

Evidence-Based Home Visiting Provisions and Child Maltreatment Report Rates: County-Level Analysis of US National Data From 2016 to 2018.

This observational ecological study examined county-level associations between evidence-based home visiting (EHV) provisions and child maltreatment report (CMR) rates, using national county-level data from 2016-2018. We found that longitudinal changes of EHV provisions were significantly negatively associated with county CMR rates while controlling for potential confounders. Our model estimated that after EHV provisions were launched in counties, their CMR rates decreased (or after they were ceased, rates increased) by 2.21 per 1000 children overall, 2.88 per 1000 children aged 0-5, 2.59 per 1000 children aged 6-11, 2.13 per 1000 male children, and 2.24 per 1000 female children. When limiting attention to EHV provisions funded by the Maternal, Infant and Early Childhood Home Visiting (MIECHV) program, we found no significant association perhaps because MIECHV-funded EHV provisions were a small subset of all EHV provisions. These findings propose potential protective impacts of county EHV provisions on overall county CMR rates. Yet, the small effect sizes suggest that EHV provisions should be considered as a part of a complete response to child maltreatment rather than in isolation. Given that EHV is provided to a very small part of the population, nevertheless, our findings suggest that expanding coverage would increase effect sizes.

Policies to Reduce Child Poverty and Child Maltreatment: A Scoping Review and Preliminary Estimates of Indirect Effects.

This study includes a scoping review of prior studies investigating the effects of policy changes on child poverty rates. It further conducts an empirical analysis to estimate the relationship between child poverty rates and child maltreatment report (CMR) rates, utilizing national county-level data. The study then calculates the indirect effects of policy changes on CMR rates, mediated through child poverty rates, by integrating information from previous studies with its own empirical findings. Among the policy changes explored in prior studies, those related to a child allowance and a fully refundable Child Tax Credit demonstrate the largest indirect effects but also the highest costs. The expansion of in-kinds and near-cash benefits, such as the Supplemental Nutrition Assistance Program benefits and housing vouchers, shows moderate effects with moderate costs. Tax credits like the Earned Income Tax Credit exhibit lower effects and costs when targeted at the lowest earners, and moderate effects and costs for broader expansion. Focused tax credits, such as the Child and Dependent Care Tax Credit, had lower effects and costs, even if made fully refundable. Despite certain limitations, the study's approach yields consistent estimates with a recent simulation study, indicating its potential validity. While some proposed policy changes may seem expensive, implementing them is anticipated to substantially reduce CMR rates, with the benefits outweighing the associated costs. Overall, the findings suggest that addressing child poverty to reduce CMRs is an attractive strategy with numerous potential benefits.

Identification of Novel Biomarkers for Early Diagnosis of Atherosclerosis Using High-Resolution Metabolomics.

Atherosclerosis (AS) is a metabolic disorder and the pre-stage of several cardiovascular diseases, including myocardial infarction, stroke, and angina pectoris. Early detection of AS can provide the opportunity for effective management and better clinical results, along with the prevention of further progression of the disease. In the current study, an untargeted and targeted metabolomic approach was used to identify possible metabolic signatures that have altered levels in AS patients. A total of 200 serum samples from individuals with AS and normal were analyzed via liquid chromatography-high-resolution mass spectrometry. Univariate and multivariate analysis approaches were used to identify differential metabolites. A group of metabolites associated with bile acids, amino acids, steroid hormones, and purine metabolism were identified that are capable of distinguishing AS-risk sera from normal. Further, the targeted metabolomics approach confirmed that six metabolites, namely taurocholic acid, cholic acid, cortisol, hypoxanthine, trimethylamine N-oxide (TMAO), and isoleucine, were found to be significantly upregulated, while the concentrations of glycoursodeoxycholic acid, glycocholic acid, testosterone, leucine, methionine, phenylalanine, tyrosine, and valine were found to be significantly downregulated in the AS-risk sera. The receiver operating characteristic curves of three metabolites, including cortisol, hypoxanthine, and isoleucine, showed high sensitivity and specificity. Taken together, these findings suggest cortisol, hypoxanthine, and isoleucine as novel biomarkers for the early and non-invasive detection of AS. Thus, this study provides new insights for further investigations into the prevention and management of AS.

RCKD: Response-Based Cross-Task Knowledge Distillation for Pathological Image Analysis.

We propose a novel transfer learning framework for pathological image analysis, the Response-based Cross-task Knowledge Distillation (RCKD), which improves the performance of the model by pretraining it on a large unlabeled dataset guided by a high-performance teacher model. RCKD first pretrains a student model to predict the nuclei segmentation results of the teacher model for unlabeled pathological images, and then fine-tunes the pretrained model for the downstream tasks, such as organ cancer sub-type classification and cancer region segmentation, using relatively small target datasets. Unlike conventional knowledge distillation, RCKD does not require that the target tasks of the teacher and student models be the same. Moreover, unlike conventional transfer learning, RCKD can transfer knowledge between models with different architectures. In addition, we propose a lightweight architecture, the Convolutional neural network with Spatial Attention by Transformers (CSAT), for processing high-resolution pathological images with limited memory and computation. CSAT exhibited a top-1 accuracy of 78.6% on ImageNet with only 3M parameters and 1.08 G multiply-accumulate (MAC) operations. When pretrained by RCKD, CSAT exhibited average classification and segmentation accuracies of 94.2% and 0.673 mIoU on six pathological image datasets, which is 4% and 0.043 mIoU higher than EfficientNet-B0, and 7.4% and 0.006 mIoU higher than ConvNextV2-Atto pretrained on ImageNet, respectively.

Evidence for the circulation of genotype 4 Eurasian avian-like lineage swine H1N1 influenza A viruses on Korean swine farms, obtained using a newly developed one-step multiplex RT-qPCR assay.

Genotype 4 (G4) Eurasian avian-like lineage swine H1N1 influenza A viruses, which are reassortants containing sequences from the pandemic 2009 H1N1 virus lineage, triple-reassortant-lineage internal genes, and EA-lineage external genes, have been reported in China since 2013. These have been predominant in pig populations since 2016 and have exhibited pandemic potential. In this study, we developed a one-step multiplex RT-qPCR assay targeting the M, HA1, and PB2 genes to detect G4 and related EA H1N1 viruses, with detection limits of 1.5 × 101 copies/µL and 1.15 × 10-2 ng/µL for the purified PCR products and RNA templates, respectively. The specificity of the detection method was confirmed using various influenza virus subtypes. When the one-step multiplex RT-qPCR assay was applied to swine respiratory samples collected between 2020 and 2022 in Korea, a virus related to G4 EA H1N1 strains was detected. Phylogenetic analysis based on portions of all eight genome segments showed that the positive sample contained HA, NA, PB2, NS, and NP genes closely related to those of G4 EA H1N1 viruses, confirming the ability of our assay to accurately detect G4 EA H1N1 viruses in the field.

Risk factors and neglect subtypes: Findings from a nationally representative data set.

Child neglect is a multidimensional concept encompassing various forms. Prior studies suggest that risk factors differ by neglect subtypes such as physical or supervisory neglect, but few studies address how risk factors vary between other neglect subtypes. This study aimed to examine how risk factors were related to neglect subtypes such as physical neglect, lack of supervision, exposure to domestic violence, substance-abusing parent, and mixed neglect. This study used secondary data from a nationally representative sample of children (National Survey of Child and Adolescent Well-Being-II, N = 5,872), and 786 children with a first-time child protective services investigation for neglect allegations alone were selected. Multinomial logistic regression analysis was used to explore how individual, family, and community risk factors may be associated with specific neglect subtypes. Five risk factors were able to discriminate between subtypes of neglect. For example, being a young child is associated with a greater risk of experiencing multiple forms of neglect. Caregiver's mental health problem is associated with a higher likelihood of being referred for multiple forms of neglect, particularly as compared with the risk of being referred for lack of supervision. Having poor social support is associated with a higher risk of physical neglect, and caregiver high stress is related to a higher risk of domestic violence. While most intervention programs target risk factors for overall child neglect, not specific neglect subtypes cases, our findings suggest that the intervention approach based on an understanding of the heterogeneity in risk factors between neglect subtypes is advisable. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Has the relationship between community poverty and child maltreatment report rates become stronger or weaker over time?

BACKGROUND: Poverty is among the most powerful predictors of child maltreatment risk and reporting. To date, however, there have been no studies assessing the stability of this relationship over time. OBJECTIVE: To examine whether the county-level relationship between child poverty rates and child maltreatment report (CMR) rates changed over time in the United States in 2009-2018, overall and across of child age, sex, race/ethnicity, and maltreatment type. PARTICIPANTS AND SETTING: U.S. Counties in 2009-2018. METHODS: Linear multilevel models estimated this relationship and its longitudinal change, while controlling for potential confounding variables. RESULTS: We found that the county-level relationship between child poverty rates and CMR rates had intensified almost linearly from 2009 to 2018. Per one-percentage-point increase in child poverty rates, CMR rates significantly increased by 1.26 per 1000 children in 2009 and by 1.74 per 1000 children in 2018, indicating an almost 40 % increase in the poverty-CMR relationship. This increasing trend was also found within all subgroups of child age and sex. This trend was found among White and Black children, but not among Latino children. This trend was strong among neglect reports, weaker among physical abuse reports, and not found among sexual abuse reports. CONCLUSIONS: Our findings highlight the continued, perhaps increasing importance of poverty as a predictor of CMR. To the degree that our findings can be replicated, they could be interpreted as supporting an increased emphasis on reducing child maltreatment incidents and reports through poverty amelioration efforts and the provision of material family supports.

Elevated risk of cervical cancer in elderly women with incident ulcerative colitis in South Korea.

The association between ulcerative colitis (UC) and uterine cervical cancer is still unclear. To investigate cervical cancer risk in South Korean women with UC, we analyzed the Korean National Health Insurance claims data. UC was defined using both ICD-10 codes and UC-specific prescriptions. We analyzed incident cases of UC diagnosed between 2006 and 2015. Age-matched women without UC (control group) were randomly selected from the general population (1:3 ratio). Hazard ratios were calculated using multivariate Cox proportional hazard regression, and the event was defined as occurrence of cervical cancer. A total of 12,632 women with UC and 36,797 women without UC were enrolled. The incidence of cervical cancer was 38.8 per 100,000 women per year in UC patients and 25.7 per 100,000 women per year in controls, respectively. The adjusted HR for cervical cancer was 1.56 (95% CI 0.97-2.50) in the UC group with reference to the control group. When stratified by age, the adjusted HR for cervical cancer was 3.65 (95% CI 1.54-8.66) in elderly UC patients (≥ 60 years) compared to elderly control group (≥ 60 years). Within UC patients, increased age (≥ 40 years) and low socioeconomic status were associated with an increased risk of cervical cancer. The incidence of cervical cancer was found to be higher among elderly patients (≥ 60 years) with newly diagnosed UC in South Korea, compared to age-matched controls. Therefore, regular cervical cancer screening is recommended for elderly patients who have recently been diagnosed with UC.

Longitudinal changes in the county-level relationship between opioid prescriptions and child maltreatment reports, United States, 2009-2018.

This article examines whether county opioid prescription rates were associated with county child maltreatment report (CMR) rates in the United States and whether this relationship changed over time. We linked multiple national data sets to assemble retail opioid prescription data, CMR data, rural-urban codes (to control for urbanicity), and census data (to control for other community characteristics, such as poverty rates) covering 2009-2018. Multilevel linear modeling analyzed the linked data. We found that the strength of the county-level relationship between opioid prescription rates and CMR rates increased almost linearly during the study period. The relationship was not significant in 2009-2011; it became significant in 2012 and grew stronger in the next 6 years. In 2012, there was one more CMR per 1,000 children in a county for every 14.3 more opioid prescriptions per 100 people. In 2018, the number of prescriptions related to this effect was 3.6. In other words, the county-level relationship between opioid prescriptions and CMRs was four times as strong in 2018 as it had been in 2012. This trend was also observed within all subgroups of child age and sex. By type, this trend was somewhat more pronounced for neglect, but somewhat less for sexual abuse. Our findings suggest a growing need for greater efforts to prevent child maltreatment in communities with high opioid prescription rates. Further research is warranted to reveal the underlying factors for this concerning trend. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

County-Level Relationships Between Foreign-Born Residents, Latinos, Immigration Enforcement, and Child Maltreatment Report Rates in the United States, 2015-2018.

U.S. Latino and foreign-born populations show lower child maltreatment report (CMR) rates despite their low socioeconomic positions, perhaps due to protective cultural factors within these populations. However, discriminatory Immigration and Customs Enforcement (ICE) activities may attenuate such protection. We examined how ethnic and foreign-born compositions and local ICE activities were associated with community CMR rates, overall and within racial/ethnic groups (i.e., White, Black, Latino), and how these associations changed over time. We used national county-level data linking multiple administrative/archival data sources (i.e., CMR, Census, and ICE data) longitudinally for 2015 to 2018 across the United States. Multilevel (county-years, counties, and states) models estimated how percentages of Latino, percentages of foreign-born, and ICE arrest rates were related to overall and race/ethnicity-specific CMR rates among counties while adjusting for a range of demographic, socioeconomic, child care burden, health insurance, residential mobility, and urbanicity factors. Higher percentages of foreign-born residents within counties were significantly associated with lower CMR rates, both overall and within all racial/ethnic groups. These protective associations became significantly stronger over the study period. Higher percentages of Latino residents were significantly associated with lower total and White CMR rates but not with Black or Latino CMR rates. The interaction between the percentage of Latino residents and year was not significant. ICE arrest rates showed no significant associations with CMR rates. Our findings suggest that communities with more foreign-born and Latino residents may be more protective against CMRs. While the foreign-born and Latino concentrations were both independently predictive of decreased CMR rates, the protective associations of the foreign-born concentration were more consistent within racial/ethnic groups and grew stronger over time. These findings suggest the need to investigate community-level protective mechanisms that may explain these results. The null findings for ICE activity also require further research with alternative measures of discriminatory state action.

Risk of Gastric Cancer among Patients with Newly Diagnosed Ulcerative Colitis: A Nationwide Population-Based Study.

BACKGROUND: Few studies have investigated the risk of gastric cancer (GC) in ulcerative colitis (UC), and the results have been inconsistent. This study aimed to assess the risk of gastric cancer in newly diagnosed UC patients. METHODS: Based on claims data from Korean National Health Insurance from January 2006 to December 2015, we identified 30,546 patients with UC and randomly selected 88,829 non-UC individuals as controls, who were matched by age and sex. Multivariate Cox proportional hazards regression was used to calculate adjusted hazard ratios (HRs) for gastric cancer events, with covariates taken into account. RESULTS: During the study period, a total of 77 (0.25%) patients with UC and 383 (0.43%) non-UC individuals were diagnosed with GC. After multivariable adjustment, the HR for GC was 0.60 (95% CI: 0.47-0.77) in patients with UC, using non-UC individuals as the reference group. When stratified by age, the adjusted HRs for GC in UC patients were 0.19 (95% CI: 0.04-0.98) for those aged 20-39 years at the time of UC diagnosis, 0.65 (95% CI: 0.45-0.94) for 40-59, and 0.60 (95% CI: 0.49-0.80) for ≥60 as compared to non-UC individuals in the corresponding age groups. When stratified by sex, the adjusted HR for GC was 0.54 (95% CI: 0.41-0.73) in male UC patients of all ages. Within UC patients, a multivariable analysis revealed that the HR for GC was 12.34 (95% CI: 2.23-68.16) for those aged ≥ 60 years at the time of diagnosis of UC. CONCLUSIONS: Patients with UC had a decreased GC risk compared with non-UC individuals in South Korea. Within the UC population, advancing age (≥60 years) was identified as a significant risk factor for GC.

Racial/Ethnic Differences in Child Protective Services Reporting, Substantiation and Placement, With Comparison to Non-CPS Risks and Outcomes: 2005-2019.

We used National Child Abuse and Neglect Data System and Census data to examine Black-White and Hispanic-White disparities in reporting, substantiation, and out-of-home placement both descriptively from 2005-2019 and in multivariate models from 2007-2017. We also tracked contemporaneous social risk (e.g., child poverty) and child harm (e.g., infant mortality) disparities using non-child protective services (CPS) sources and compared them to CPS reporting rate disparities. Black-White CPS reporting disparities were lower than found in non-CPS risk and harm benchmarks. Consistent with the Hispanic paradox, Hispanic-White CPS reporting disparities were lower than risk disparities but similar to harm disparities. Descriptive and multivariate analyses of data from the past several years indicated that Black children were less likely to be substantiated or placed into out-of-home care following a report than White children. Hispanic children were slightly more likely to be substantiated or placed in out-of-home care than White children overall, but this difference disappeared in multivariate models. Available data provide no evidence that Black children were overreported relative to observed risks and harms reflected in non-CPS data. Reducing reporting rates among Black children will require addressing broader conditions associated with maltreatment.

Community Food Insecurity and Child Maltreatment Reports: County-Level Analysis of U.S. National Data From 2009 to 2018.

Current literature suggests that food insecurity increases child maltreatment risk. Yet, existing evidence is limited to individual-level associations among low-income, high-risk populations based on local, mostly urban data. This study aims to generalize prior findings to community-level associations in general populations, using national data including all urban-rural areas. We examined, for the first time, if food insecurity rates increase child maltreatment report (CMR) rates at the county level after controlling for potential confounders. We examined both within-community longitudinal changes (i.e., within-effects) and inter-community differences (i.e., between-effects) of food insecurity rates and their associations with CMR rates. We also examined differences by age, sex, race/ethnicity, maltreatment type, and urbanicity. We constructed longitudinal county-level data by linking multiple national databases, including all substantiated and unsubstantiated CMR records, the Map the Meal Gap's community food insecurity estimates, and Census data. The data covered over 96% of U.S. counties from 2009 to 2018. For analysis, we used within-between random effects models. Regarding between-effects, we found that in inter-community comparisons, higher food insecurity rates were significantly associated with increased CMR rates. This association was consistent by age, sex, maltreatment type, and urbanicity. For within-effects, we found that the association between longitudinal changes of food insecurity rates and CMR rates significantly differed by urbanicity. Specifically, longitudinal increases of food insecurity rates significantly increased CMR rates among large urban counties, but not among small urban and rural counties. Study findings highlight the importance of conducting further research to better understand the mechanisms through which food insecurity impact child maltreatment at both individual and community levels. Our community-level findings from general populations especially have significant implications for community-based programs and large-scale policies to achieve population-level impact on child well-being.

Testing a QuantCrit-Informed Approach to the Empirical Study of Race/Ethnicity and Child Maltreatment.

The past several years have seen calls from QuantCrit scholars to "disaggregate" samples into same-race groups. To date, however, there has been no attempt to empirically evaluate the benefits of disaggregation within a child welfare sample. Using a child maltreatment dataset derived from the National Child Abuse and Neglect Data System and Census data, we empirically evaluate the utility of employing sample disaggregation (in which separate records are created for White, Black and Latino populations in each county) as well as variable creation disaggregation (in which we avoid using "full county" economic measures, but instead employ "same race/ethnicity" measures). Using model fit and convergence with findings from individual-level studies as evaluation metrics, we find that both kinds of disaggregation are demonstrably beneficial. We recommend that sample and variable disaggregation be considered by any future researchers using national geographically structured child maltreatment data.

Do county mental health, physical health, and care provider availability predict child maltreatment report rates?

BACKGROUND: Research on community-level relationships between mental/physical health and child maltreatment is sparse. OBJECTIVE: We examined how rates of mental distress, physical distress, mental health professionals, and primary care physicians were related to child maltreatment report rates at the county level. PARTICIPANTS AND SETTING: U.S. counties from 2014 to 2017. METHODS: Within-between random effects models estimated both within-effects (i.e., longitudinal changes) and between-effects (i.e., inter-county differences) of mental distress rates, physical distress rates, mental health professional rates, and primary care physician rates and their associations with overall and age-specific maltreatment report rates, while adjusting for potential confounders. RESULTS: Longitudinal increases of mental distress rates marginally significantly (p < .10) increased overall maltreatment report rates (ß = 0.50) and significantly (p < .05) increased age 0-5 maltreatment report rates (ß = 0.84). Conversely, longitudinal increases of mental health professional rates significantly decreased overall (ß = -0.38), age 0-5 (ß = -0.59), and age 6-11 (ß = -0.31) maltreatment report rates and marginally significantly decreased age 12-17 maltreatment report rates (ß = -0.13). Between-effects of metal distress rates and mental health professional rates were mostly not significant. Neither within-effects nor between-effects of physical distress rates and primary care physician rates were significant. CONCLUSIONS: Our findings suggest that community mental distress is a risk factor for child maltreatment reports and that community availability of mental health professionals is a protective factor. Community-based strategies to address mental distress and human resource approaches to supply sufficient mental health professionals in communities may help reduce maltreatment report rates in communities. Further research is required to confirm our findings and to better understand underlying mechanisms.

Community home visiting services and child maltreatment report rates, Illinois zip codes, 2011-2018.

BACKGROUND: Research is sparse on the community-level impacts of home visiting programs on child maltreatment. OBJECTIVE: To examine community-level associations between state-funded home visiting programs (i.e., IDHS-HV), federal-funded home visiting programs (i.e., MIECHV), and child maltreatment report (CMR) rates, overall and within subgroups of age, sex, and maltreatment type. PARTICIPANTS AND SETTING: 3824 zip code-years during 2011-2018 in Illinois for IDHS-HV/CMR associations and 1896 zip code-years during 2015-2018 for MIECHV/CMR associations. METHODS: We measured county-level IDHS-HV rates (per 1000 children aged 0-5) since data were only available at that level. MIECHV rates (per 1000 children aged 0-5), CMR rates (per 1000 children), and all controls were measured at the zip code level. We used spatial linear models to handle spatial autocorrelation. RESULTS: Adjusted for controls, longitudinal increases of IDHS-HV rates were significantly associated with decreased overall CMR rates (coefficient: -0.28; 95 % CI: -0.45, -0.11), age 0-5 CMR rates (-0.52; -0.82, -0.22), age 6-11 CMR rates (-0.31; -0.55, -0.06), male CMR rates (-0.25; -0.45, -0.05), female CMR rates (-0.29; -0.49, -0.08), and neglect report rates (-0.13; -0.24, -0.02). In contrast, longitudinal increases of MIECHV rates were significantly associated with increased CMR rates within several subgroups. CONCLUSIONS: Our findings suggest that increasing state-funded home visiting services in communities may have benefits in lowering their CMR rates. Given the very low MIECHV service rates and the federal policy that requires MIECHV to target at-risk communities, the significant positive MIECHV/CMR associations we found might indicate MIECHV programs are typically in higher risk communities.

A desirable transgenic strategy using GGTA1 endogenous promoter-mediated knock-in for xenotransplantation model.

Pig-to-human organ transplantation is a feasible solution to resolve the shortage of organ donors for patients that wait for transplantation. To overcome immunological rejection, which is the main hurdle in pig-to-human xenotransplantation, various engineered transgenic pigs have been developed. Ablation of xeno-reactive antigens, especially the 1,3-Gal epitope (GalT), which causes hyperacute rejection, and insertion of complement regulatory protein genes, such as hCD46, hCD55, and hCD59, and genes to regulate the coagulation pathway or immune cell-mediated rejection may be required for an ideal xenotransplantation model. However, the technique for stable and efficient expression of multi-transgenes has not yet been settled to develop a suitable xenotransplantation model. To develop a stable and efficient transgenic system, we knocked-in internal ribosome entry sites (IRES)-mediated transgenes into the α 1,3-galactosyltransferase (GGTA1) locus so that expression of these transgenes would be controlled by the GGTA1 endogenous promoter. We constructed an IRES-based polycistronic hCD55/hCD39 knock-in vector to target exon4 of the GGTA1 gene. The hCD55/hCD39 knock-in vector and CRISPR/Cas9 to target exon4 of the GGTA1 gene were co-transfected into white yucatan miniature pig fibroblasts. After transfection, hCD39 expressed cells were sorted by FACS. Targeted colonies were verified using targeting PCR and FACS analysis, and used as donors for somatic cell nuclear transfer. Expression of GalT, hCD55, and hCD39 was analyzed by FACS and western blotting. Human complement-mediated cytotoxicity and human antibody binding assays were conducted on peripheral blood mononuclear cells (PBMCs) and red blood cells (RBCs), and deposition of C3 by incubation with human complement serum and platelet aggregation were analyzed in GGTA1 knock-out (GTKO)/CD55/CD39 pig cells. We obtained six targeted colonies with high efficiency of targeting (42.8% of efficiency). Selected colony and transgenic pigs showed abundant expression of targeted genes (hCD55 and hCD39). Knocked-in transgenes were expressed in various cell types under the control of the GGTA1 endogenous promoter in GTKO/CD55/CD39 pig and IRES was sufficient to express downstream expression of the transgene. Human IgG and IgM binding decreased in GTKO/CD55/CD39 pig and GTKO compared to wild-type pig PBMCs and RBCs. The human complement-mediated cytotoxicity of RBCs and PBMCs decreased in GTKO/CD55/CD39 pig compared to cells from GTKO pig. C3 was also deposited less in GTKO/CD55/CD39 pig cells than wild-type pig cells. The platelet aggregation was delayed by hCD39 expression in GTKO/CD55/CD39 pig. In the current study, knock-in into the GGTA1 locus and GGTA1 endogenous promoter-mediated expression of transgenes are an appropriable strategy for effective and stable expression of multi-transgenes. The IRES-based polycistronic transgene vector system also caused sufficient expression of both hCD55 and hCD39. Furthermore, co-transfection of CRISPR/Cas9 and the knock-in vector not only increased the knock-in efficiency but also induced null for GalT by CRISPR/Cas9-mediated double-stranded break of the target site. As shown in human complement-mediated lysis and human antibody binding to GTKO/CD55/CD39 transgenic pig cells, expression of hCD55 and hCD39 with ablation of GalT prevents an effective immunological reaction in vitro. As a consequence, our technique to produce multi-transgenic pigs could improve the development of a suitable xenotransplantation model, and the GTKO/CD55/CD39 pig developed could prolong the survival of pig-to-primate xenotransplant recipients.