Development and Applications of a Zebrafish (Danio rerio) CYP1A-Targeted Monoclonal Antibody (CRC4) with Reactivity across Vertebrate Taxa: Evidence for a Conserved CYP1A Epitope.

CYP1A is a heme-thiolate enzyme associated with the cytochrome P4501A1 monooxygenase system and is inducible by a wide variety of xenobiotics and endogenous ligands that bind and activate the aryl hydrocarbon receptor (AHR). The AHR-CYP1A axis is important for detoxification of certain xenobiotics and for homeostatic balance of endogenous sex hormones, amine hormones, vitamins, fatty acids, and phospholipids. Herein, we generated and described applications of a zebrafish CYP1A-targeted monoclonal antibody (mAb CRC4) that fortuitously recognizes induced CYP1A across vertebrate taxa, including fish, chicken, mouse, rat, and human. We then demonstrated that mAb CRC4 targets a highly conserved epitope signature of vertebrate CYP1A. The unique complimentary determining region (CDR) sequences of heavy and light chains were determined, and these Ig sequences will allow for the expression of recombinant mAb CRC4, thus superseding the need for long-term hybridoma maintenance. This antibody works well for immunohistochemistry (IHC), as well as whole-mounted IHC in zebrafish embryos. Monoclonal antibody CRC4 may be particularly useful for studying the AHR-CYP1A axis in multiple vertebrate species and within the context of Oceans and Human Health research. By using archived samples, when possible, we actively promoted efforts to reduce, replace, and refine studies involving live animals.

Advancing health policy education in nursing: American Association of Colleges of Nursing Faculty Policy Think Tank.

Based upon an awareness of the need to increase its policy footprint, the American Association of Colleges of Nursing's (AACN) Board of Directors convened a think tank of nurse experts in the field of policy curriculum to advise the Board. The goal of the think tank was to review the current landscape, analyze trends, and create a set of recommendations for AACN's Board to consider. The Faculty Policy Think Tank (FPTT) met between 2016 and 2017 to review and reflect on methods to increase expertise of nursing faculty and students in health policy and make recommendations to the AACN Board to advance the agenda of the organization, member schools, and the profession. This article describes the methods and processes the Think Tank employed to develop a set of recommendations for the AACN Board of Directors.

Rating of the Effectiveness of 26 Psychiatric and Seizure Medications for Autism Spectrum Disorder: Results of a National Survey.

OBJECTIVE: The objective of this study was to provide an evaluation of the benefits and adverse effects (AEs) of psychiatric and seizure medications commonly used for individuals with autism spectrum disorder (ASD). METHODS: As part of the National Survey on Treatment Effectiveness for Autism, we report ratings of 26 psychiatric and seizure medications by 505 participants. Each medication was rated with a standardized scale for overall benefits, overall AEs, and specific symptoms affected. The frequency of use and net perceived benefit (overall benefit minus overall AE) are reported. RESULTS: Most medications were rated as having a slightly greater benefit than AE. Six medications (lamotrigine, oxcarbazepine, clonidine, guanfacine, buspirone, and sertraline) had benefit ratings that were more than twice their adverse rating. Conversely, some medications had slightly negative net benefit ratings (worse AEs than benefits on average), including Adderall, Paroxetine, Quetiapine, Olanzapine, and Topiramate. However, there were wide variations in individual ratings of benefit and AEs, suggesting that clinical response to medications was highly variable, so these scores simply represent averages. A ranking of the top medications (those with the highest net perceived benefit) for each of 18 different symptoms is provided, which may provide some clinical guidance as to which medications may be most worth considering for a given symptom. A comparison of the survey results with the results of clinical trials shows generally good agreement in terms of medication benefits with some differences; in some cases the differences are because the clinical trials did not assess all of the symptoms assessed by this survey. CONCLUSIONS: It is hoped that physicians and their patients will find the survey results useful in selecting the most promising medications for a given symptom, and also for monitoring for likely benefits and AEs, especially for medications for which few or no studies have been carried out in ASD populations.

Female Sex Offenders: Public Awareness and Attributions.

Traditional gender roles, sex scripts, and the way female sex offenders are portrayed in the media may lead to misconceptions about who can commit sexual offenses. Sexual crimes by women may go unnoticed or unreported if there is a general lack of awareness that females commit these crimes. Data from the 2012 Nebraska Annual Social Indicators Survey were used to determine whether the public perceives women as capable sex offenders and the perceived causes of female sex offending. The traditional focus on male sex offenders by researchers, media, and politicians, in addition to gender stereotypes, introduces the possibility of group differences (e.g., between men and women) in perceptions of female sex offenders. Consequently, two secondary analyses were conducted that tested for group differences in both the public's perception of whether females can commit sex offenses and the explanations selected for why females sexually offend. The findings suggest that the public does perceive women as capable sex offenders, although there were group differences in the causal attributions for female sex offending.

3,4-Dihydroxy-benzohydroxamic acid (Didox) suppresses pro-inflammatory profiles and oxidative stress in TLR4-activated RAW264.7 murine macrophages.

Didox (3,4-dihydroxy-benzohydroxamic acid), is a synthetic ribonucleotide reductase (RR) inhibitor derived from polyhydroxy-substituted benzohydroxamic acid, and originally developed as an anti-cancer agent. Some studies indicate that didox may have anti-oxidative stress-like properties, while other studies hint that didox may have anti-inflammatory properties. Using nitric oxide production in response to LPS treatment as a sensitive screening assay for anti-inflammatory compounds, we show that didox is very potent at levels as low as 6.25 µM, with maximal inhibition at 100 µM. A qRT-PCR array was then employed to screen didox for other potential anti-inflammatory and anti-oxidative stress-related properties. Didox was very potent in suppressing the expression of these arrayed mRNA in response to LPS, and in some cases didox alone suppressed expression. Using qRT-PCR as a follow up to the array, we demonstrated that didox suppresses LPS-induced mRNA levels of iNOS, IL-6, IL-1, TNF-α, NF-κß (p65), and p38-α, after 24h of treatment. Treatment with didox also suppresses the secretion of nitric oxide, IL-6, and IL-10. Furthermore, oxidative stress, as quantified by intracellular ROS levels in response to macrophage activators LPS and phorbol ester (PMA), and the glutathione depleting agent BSO, is reduced by treatment with didox. Moreover, we demonstrate that nuclear translocation of NF-κß (p65) in response to LPS is inhibited by didox. These findings were supported by qRT-PCR for oxidative stress genes SOD1 and catalase. Overall, this study supports the conclusion that didox may have a future role in managing acute and chronic inflammatory diseases and oxidative stress due to high production of ROS.

Functional plasticity before the cradle: a review of neural functional imaging in the human fetus.

The organization of the brain is highly plastic in fetal life. Establishment of healthy neural functional systems during the fetal period is essential to normal growth and development. Across the last several decades, remarkable progress has been made in understanding the development of human fetal functional brain systems. This is largely due to advances in imaging methodologies. Fetal neuroimaging began in the 1950-1970's with fetal electroencephalography (EEG) applied during labor. Later, in the 1980's, magnetoencephalography (MEG) emerged as an effective approach for investigating fetal brain function. Most recently, functional magnetic resonance imaging (fMRI) has arisen as an additional powerful approach for examining fetal brain function. This review will discuss major developmental findings from fetal imaging studies such as the maturation of prenatal sensory system functions, functional hemispheric asymmetry, and sensory-driven neurodevelopment. We describe how with improved imaging and analysis techniques, functional imaging of the fetus has the potential to assess the earliest point of neural maturation and provide insight into the patterning and sequence of normal and abnormal brain development.

Cross-hemispheric functional connectivity in the human fetal brain.

Compelling evidence indicates that psychiatric and developmental disorders are generally caused by disruptions in the functional connectivity (FC) of brain networks. Events occurring during development, and in particular during fetal life, have been implicated in the genesis of such disorders. However, the developmental timetable for the emergence of neural FC during human fetal life is unknown. We present the results of resting-state functional magnetic resonance imaging performed in 25 healthy human fetuses in the second and third trimesters of pregnancy (24 to 38 weeks of gestation). We report the presence of bilateral fetal brain FC and regional and age-related variation in FC. Significant bilateral connectivity was evident in half of the 42 areas tested, and the strength of FC between homologous cortical brain regions increased with advancing gestational age. We also observed medial to lateral gradients in fetal functional brain connectivity. These findings improve understanding of human fetal central nervous system development and provide a basis for examining the role of insults during fetal life in the subsequent development of disorders in neural FC.

Indirubin-3'-(2,3 dihydroxypropyl)-oximether (E804) is a potent modulator of LPS-stimulated macrophage functions.

Indirubin is a deep-red bis-indole isomer of indigo blue, both of which are biologically active ingredients in Danggui Longhui Wan, an ancient Chinese herbal tea mixture used to treat neoplasia and chronic inflammation and to enhance detoxification of xenobiotics. Multiple indirubin derivatives have been synthesized and shown to inhibit cyclin-dependent kinases (CDKs) and glycogen-synthase kinase (GSK-3ß) with varying degrees of potency. Several indirubins are also aryl hydrocarbon receptor (AhR) agonists, with AhR-associated activities covering a wide range of potencies, depending on molecular structure. This study examined the effects of indirubin-3'-(2,3 dihydroxypropyl)-oximether (E804), a novel indirubin with potent STAT3 inhibitory properties, on basal and LPS-inducible activities in murine RAW264.7 macrophages. Using a focused commercial qRT-PCR array platform (SuperArray®), the effects of E804 on expression of a suite of genes associated with stress and toxicity were determined. Most genes up-regulated by LPS treatment were suppressed by E804; including LPS-induced expression of pro-inflammatory cytokines and receptors, apoptosis control genes, and oxidative stress response genes. Using qRT-PCR as a follow up to the commercial arrays, E804 treatment suppressed LPS-induced COX-2, iNOS, IL-6 and IL-10 gene expression, though the effects on iNOS and COX-2 protein expression were less dramatic. E804 also inhibited LPS-induced secretion of IL-6 and IL-10. Functional endpoints, including iNOS and lysozyme enzymatic activity, phagocytosis of fluorescent latex beads, and intracellular killing of bacteria, were also examined, and in each experimental condition E804 suppressed activities. Collectively, these results indicate that E804 is a potent modulator of pro-inflammatory profiles in LPS-treated macrophages.

The B-cell superantigen Finegoldia magna protein L causes pulmonary inflammation by a mechanism dependent on MyD88 but not B cells or immunoglobulins.

OBJECTIVE AND DESIGN: To determine whether Finegoldia magna protein L (PL) causes lung inflammation and, if so, whether the response is dependent on its immunoglobulin (Ig)-binding B-cell superantigenic property. MATERIAL: Pulmonary inflammatory reactions were analyzed at various time points after intratracheal administration of PL to various strains of mice. RESULTS: PL caused peribronchial and perivascular inflammation that peaked at 18-24 h. Polymorphonuclear cells (PMNs) began to accumulate in bronchoalveolar lavage fluid (BALF) of PL-challenged mice by 4 h and accounted for >90% of leukocytes by 18-24 h. Inflammation was marked by the appearance of MIP-2, KC, TNF-α, and IL-6 in the BALF with peak levels attained 4 h after PL administration. PL-induced pulmonary inflammation was associated with increased airway hyper-reactivity following inhalation of methacholine. The inflammatory reaction was unabated in mice lacking B cells and immunoglobulins. In contrast, PL-induced inflammation was abrogated in MyD88-deficient mice. PL-induced responses required alveolar macrophages. CONCLUSIONS: These results strongly suggest that PL-induced lung inflammation is dependent on an innate MyD88-dependent pathway rather than the Ig-binding properties of this microbial B cell superantigen. We propose that this pulmonary inflammatory reaction is caused by the interaction of PL with a Toll-like receptor expressed on alveolar macrophages.

Food safety perceptions and practices of older adults.

OBJECTIVES: Older adults are considered more vulnerable to foodborne illness due to lowered immune function. We compared the food safety perceptions and practices of older and younger adults and determined associations with demographic characteristics. METHODS: We focused on 1,317 participants > or = 60 years of age from the U.S. Food and Drug Administration's 2006 Food Safety Survey, a telephone survey of a nationally representative sample of American consumers. We used data on participants < 60 years of age to compare younger and older adults, and used Pearson's Chi-square tests to determine whether perceptions and practices differed by age, gender, level of education, living arrangement, and race/ethnicity. We conducted multiple logistic regression analysis to assess relationship of demographic characteristics and food safety perceptions with food safety practices of older adults. RESULTS: We found that adults > or = 60 years of age were more likely to follow recommended food safety practices than those < 60 years of age. Sixty-six percent of adults > or = 60 years of age reported eating potentially hazardous foods in the past year compared with 81% of adults < 60 years of age. Among people > or = 60 years of age, women, those with less education, and nonwhite individuals generally had better food safety practices and a greater awareness of food safety risk. CONCLUSIONS: These findings suggest that certain subsets of the older adult population are less likely to follow recommended food safety practices and, thus, are at greater risk of foodborne illness. Food safety education for older adults should target men and those with more education and higher incomes.

Dietary patterns and survival of older adults.

BACKGROUND: Recent research has linked overall dietary patterns to survival in older adults. OBJECTIVES: The objective of this study was to determine the dietary patterns of a cohort of older adults, and to explore associations of these dietary patterns with survival over a 10-year period. A secondary goal was to evaluate participants' quality of life and nutritional status according to their dietary patterns. DESIGN: The Health, Aging, and Body Composition Study is a prospective cohort study of 3,075 older adults. In this study, all-cause mortality was assessed from baseline through Year 10. Food intake was estimated with a modified Block food frequency questionnaire, and dietary patterns of 2,582 participants with complete data were derived by cluster analysis. RESULTS: Six dietary pattern clusters were identified, including a Healthy Foods cluster, characterized by higher intake of low-fat dairy products, fruit, whole grains, poultry, fish, and vegetables. Both the High-Fat Dairy Products and Sweets and Desserts clusters had a 1.4-fold higher risk of mortality than the Healthy Foods cluster after adjusting for potential confounders. The Healthy Foods cluster also had significantly more years of healthy life and more favorable levels of selected nutritional biomarkers than the other clusters. CONCLUSIONS: A dietary pattern consistent with current guidelines to consume relatively high amounts of vegetables, fruit, whole grains, poultry, fish, and low-fat dairy products may be associated with superior nutritional status, quality of life and survival in older adults.

Sleep and delinquency: does the amount of sleep matter?

Sleep, a key indicator of health, has been linked to a variety of indicators of well-being such that people who get an adequate amount generally experience greater well-being. Further, a lack of sleep has been linked to a wide range of negative developmental outcomes, yet sleep has been largely overlooked among researchers interested in adolescent delinquency. The purpose of this study was to explore the relationship between hours of sleep and delinquent behavior among adolescents by using data from Wave 1 of the National Longitudinal Study of Adolescent Health (n = 14,382; 50.2% female, 63.5% white). A series of negative binomial regressions showed that youth who typically sleep seven or fewer hours per night reported significantly more property delinquency than youth who sleep the recommended 8-10 h. Further, youth who reported sleeping 5 or fewer hours per night reported significantly more violent delinquency than youth who reported sleeping the recommended number of hours per night. The findings suggest that sleep is an important, and overlooked, dimension of delinquent behavior and studies that focus on adolescent health should further investigate the effects of insufficient sleep. Finally, the authors recommend that sleep and other relevant health behaviors be considered in the context of more comprehensive approaches to delinquency prevention and intervention.

Relationships of dietary patterns with body composition in older adults differ by gender and PPAR-γ Pro12Ala genotype.

PURPOSE: Dietary patterns may better capture the multifaceted effects of diet on body composition than individual nutrients or foods. The objective of this study was to investigate the dietary patterns of a cohort of older adults, and examine relationships of dietary patterns with body composition. The influence of a polymorphism in the peroxisome proliferator-activated receptor-γ (PPAR-γ) gene was considered. METHODS: The Health, Aging and Body Composition (Health ABC) Study is a prospective cohort study of 3,075 older adults. Participants' body composition and genetic variation were measured in detail. Food intake was assessed with a semi-quantitative food frequency questionnaire (Block Dietary Data Systems, Berkeley, CA), and dietary patterns of 1,809 participants with complete data were derived by cluster analysis. RESULTS: Six clusters were identified, including a 'Healthy foods' cluster characterized by higher intake of low-fat dairy products, fruit, whole grains, poultry, fish and vegetables. An interaction was found between dietary patterns and PPAR-γ Pro12Ala genotype in relation to body composition. While Pro/Pro homozygous men and women in the 'Healthy foods' cluster did not differ significantly in body composition from those in other clusters, men with the Ala allele in the 'Healthy foods' cluster had significantly lower levels of adiposity than those in other clusters. Women with the Ala allele in the 'Healthy foods' cluster differed only in right thigh intermuscular fat from those in other clusters. CONCLUSIONS: Relationships between diet and body composition in older adults may differ by gender and by genetic factors such as PPAR-γ Pro12Ala genotype.

Toll-like receptor 7-induced naive human B-cell differentiation and immunoglobulin production.

BACKGROUND: Toll-like receptors contribute to the establishment of adaptive immune responses. OBJECTIVE: The reported studies were conducted to examine the effects of the Toll-like receptor (TLR)-7 ligand, resiquimod, on human naive B-cell differentiation. METHODS: Naive human B cells were cultured with resiquimod in the presence or absence of IL-2 and IL-10. Secreted IgM and IgG were measured by ELISA, and IL-6, IL-10, and IFN-alpha were measured by a multiplex protein array. Cell proliferation was assessed by measuring [(3)H]thymidine uptake. mRNA for activation-induced cytidine deaminase and I(gamma 1)-C(mu) circle transcripts was measured by means of RT-PCR. RESULTS: Resiquimod induced the production of IgM and, to a lesser extent, IgG by naive human B cells in association with the secretion of IL-6 and IL-10, and a weak proliferative response. IL-2 and IL-10 synergized with resiquimod in markedly augmenting resiquimod-induced IgM and IgG production and proliferation. Resiquimod also stimulated production of IgG by B cells isolated from the blood of a patient with the X-linked hyper-IgM syndrome, with a greater response when these cells were costimulated with IL-2 and IL-10. The stimulated naive B cells from healthy volunteers displayed molecular evidence of immunoglobulin class-switch recombination-namely the appearance of activation-induced cytidine deaminase and I(gamma 1)-C(mu) circle transcripts. CONCLUSION: Perturbation of TLR-7 on naive human B cells can lead to the induction of immunoglobulin class switch and IgG production in the absence of B-cell receptor cross-linking and CD40-CD40L interaction. The results are relevant to vaccine development and mechanisms by which microbial infection may lead to autoimmunity.

A psychometric investigation of the Suicide Opinion Questionnaire.

The factor structure of the Suicide Opinion Questionnaire (SOQ) was investigated in 2 studies. In the first study, the SOQ was administered to 568 participants to investigate the relative strength of 3 competing latent structure models previously identified in the literature. Confirmatory factor analyses provided no support for any of the prior models. An exploratory factor analysis of these data resulted in a weak 2-factor structure accounting for only 15.33% of the common variance. In the second study, this 2-factor structure was evaluated using SOQ data from 288 college students. Confirmatory factor analysis results failed to support the 2-factor model. Based on these results, the authors recommend that future research efforts should focus on developing a more contemporary and psychometrically sound measure of attitudes toward suicide.

Dietary glycemic index and glycemic load and the risk of type 2 diabetes in older adults.

BACKGROUND: It is unclear whether immediate dietary effects on blood glucose influence the risk of developing type 2 diabetes. OBJECTIVE: The objective of this study was to examine whether the dietary glycemic index (GI) and glycemic load (GL) were associated with the risk of type 2 diabetes in older adults. DESIGN: The Health, Aging, and Body Composition Study is a prospective cohort study of 3075 adults who were 70-79 y old at baseline (n=1898 for this analysis). The intakes of specific nutrients and food groups and the risk of type 2 diabetes over a 4-y period were examined according to dietary GI and GL. RESULTS: Dietary GI was positively associated with dietary carbohydrate and negatively associated with the intakes of protein, total fat, saturated fat, alcohol, vegetables, and fruit. Dietary GL was positively associated with dietary carbohydrate, fruit, and fiber and negatively associated with the intakes of protein, total fat, saturated fat, and alcohol. Persons in the higher quintiles of dietary GI or GL did not have a significantly greater incidence of type 2 diabetes. CONCLUSIONS: These findings do not support a relation between dietary GI or GL and the risk of type 2 diabetes in older adults. Because dietary GI and GL show strong nutritional correlates, the overall dietary pattern should be considered.

Pathogenesis of B-cell superantigen-induced immune complex-mediated inflammation.

Staphylococcal protein A (SpA) is representative of a new class of antigens, the B-cell superantigens (SAgs). These antigens bind to the Fab regions of immunoglobulin molecules outside their complementarity-determining regions. SpA, the best-studied B-cell SAg, reacts with the Fabs of most VH3+ immunoglobulins, which are expressed on 30 to 60% of human peripheral B cells. Therefore, B-cell SAgs like SpA have great potential to elicit inflammatory responses in vivo. We previously reported that the interaction of SpA with VH3+ immunoglobulin molecules leads to activation of the complement cascade and produces a histologic pattern of inflammation in the skin of a rabbit indicative of immune complex injury. To elucidate the cellular and molecular events contributing to this type of unconventional immune complex-mediated inflammation, we established a mouse peritoneal Arthus reaction model. Mice treated intravenously with human polyclonal immunoglobulin G (IgG), followed by intraperitoneal injection of SpA, showed neutrophil influx into the peritoneal cavity with peak numbers appearing at 8 h. This inflammatory reaction was dependent on the interaction of SpA with VH3+ IgG. Mast cells, FcgammaRIII, complement components, and tumor necrosis factor alpha play obligatory roles, and the reaction is associated with the local release of the CXC chemokines macrophage inflammatory protein 2 and KC. The data provide further compelling evidence for the induction of immune complex-mediated injury by a B-cell SAg and highlight important factors contributing to the pathogenesis of this novel type of inflammatory reaction.

Dietary glycemic index and load, measures of glucose metabolism, and body fat distribution in older adults.

BACKGROUND: Recent evidence suggests that the rate of carbohydrate digestion and absorption may influence the development of type 2 diabetes. OBJECTIVE: The aim of this study was to examine associations of dietary glycemic index and glycemic load with predictors of type 2 diabetes in older adults. DESIGN: This study evaluated cross-sectional relations of dietary glycemic index and glycemic load with measures of glucose metabolism and body fat distribution in participants of the Health, Aging and Body Composition Study, a prospective cohort study of adults aged 70-80 y (n = 2248). RESULTS: In men, dietary glycemic index was positively associated with 2-h glucose (P for trend = 0.04) and fasting insulin (P for trend = 0.004), inversely associated with thigh intramuscular fat (P for trend = 0.02), and not significantly associated with fasting glucose, glycated hemoglobin, or visceral abdominal fat. Dietary glycemic load was inversely associated in men with visceral abdominal fat (P for trend = 0.02) and not significantly associated with fasting glucose, 2-h glucose, glycated hemoglobin, fasting insulin, or thigh intramuscular fat. In women, although dietary glycemic index and load were not significantly related to any measures of glucose metabolism or body fat distribution, the association between dietary glycemic index and 2-h glucose was nearly significant (P for trend = 0.06). CONCLUSION: The findings of this cross-sectional study indicate an association between dietary glycemic index and selected predictors of type 2 diabetes in older adults, particularly in men.

A further investigation of category learning by inference.

Categories are learned in many ways besides by classification, for example, by making inferences about classified items. One hypothesis is that classifications lead to the learning of features that distinguish categories, whereas inferences promote the learning of the internal structure of categories, such as the typical features. Experiment 1 included single-feature and full-feature classification tests following either classification or inference learning. Consistent with predictions, inference learners did better on the single tests but worse on the full tests. Experiment 2 further showed that inference learners, unlike classification learners, were no better at classifying items that they had seen at study compared with equally typical items they had not seen at study. Experiment 3 showed that features queried about during inference learning were classified better than ones not queried about, although even the latter features showed some learning on single-feature tests. The discussion focuses on how different types of category learning lead to different category representations.