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Although the United States Food & Drug Administration (FDA) has provided guidance on the control of drug degradants for prescription drugs, there is less guidance on how to set degradant specifications for FDA OTC monograph drugs. Given that extensive impurity testing was not part of the safety paradigm in original OTC monographs, a weight of evidence (WOE) approach to qualify OTC degradants is proposed. This approach relies on in silico tools and read-across approaches alongside standard toxicity testing to determine safety. Using several drugs marketed under 21 CFR 341 as case studies, this research demonstrates the utility of a WOE approach across data-rich and data-poor degradants. Based on degradant levels ranging from 1 to 4% of the maximum daily doses of each case study drug and 10th percentile body weight data for each patient group, children were recognized as having the highest potential exposure relative to adults per body mass. Depending on data availability and relationship to the parent API, margins of safety (MOS) or exposure margins were calculated for each degradant. The findings supported safe use, and indicated that this contemporary WOE approach could be utilized to assess OTC degradants. This approach is valuable to establish specifications for degradants in OTCs. • There is a need to establish safety profiles of common degradants in OTC drugs. • In silico tools were integrated with traditional safety data to evaluate degradants. • Approaches used applied to data-rich and data-poor degradants. • A weight-of-evidence approach was used to establish safety of common OTC cough/cold degradants. [ABSTRACT FROM AUTHOR]

In zebrafish, sensory epithelia and neuroblasts of the inner ear form simultaneously in abutting medial and lateral domains, respectively, in the floor of the otic vesicle. Previous studies support regulatory roles for Fgf and Wnt, but how signaling is coordinated is poorly understood. We investigated this problem using pharmacological and transgenic methods to alter Fgf or Wnt signaling from early placodal stages to evaluate later changes in growth and patterning. Blocking Fgf at any stage reduces proliferation of otic tissue and terminates both sensory and neural specification. Wnt promotes proliferation in the otic vesicle but is not required for sensory or neural development. However, sustained overactivation of Wnt laterally expands sensory epithelia and blocks neurogenesis. pax2a , sp5a and sp5l are coregulated by Fgf and Wnt and show overlapping expression in the otic placode and vesicle. Gain- and loss-of-function studies show that these genes are together required for Wnt's suppression of neurogenesis, as well as some aspects of sensory development. Thus, pax2a , sp5a and sp5l are critical for mediating Fgf and Wnt signaling to promote spatially localized sensory and neural development. [Display omitted] • Sensory & neural fates arise simultaneously in abutting regions of the otic vesicle. • Fgf is required for both fates while Wnt promotes sensory & blocks neural fates. • Downstream factors pax2a , sp5a and sp5l help facilitate precise spatial patterning. [ABSTRACT FROM AUTHOR]

Epstein-Barr virus (EBV) is known to manipulate its cellular environment to enhance viral replication, which can lead to dysregulation of cellular machinery, setting the stage for potential future disease. Previous research showed that under rapamycin-mediated inhibition of mTORC1, EBV lytic protein production was altered in a cell-type specific manner, suggesting that EBV differentially activates or utilizes signaling pathways in B versus epithelial cells. Here we correlated activation of the mTORC1, ERK1/2, and p38 pathways in relation to EBV lytic replication and discovered that activation of MAPK-interacting kinase 1/2 (Mnk1/2) was strongly associated with EBV lytic replication. Inhibition studies revealed that Mnk1/2 activated lytic replication in epithelial cells yet acted as an inhibitor of lytic replication in B cells. The ability of lytic epithelial cells to migrate, a potentially important aspect of metastasis, was also demonstrated to be dependent upon Mnk1/2 activity. • EBV lytic replication triggers activation of cellular Mnk, further promoting EBV lytic replication within epithelial cells. • Mnk stimulates the cell migration capability of lytic Epstein-Barr virus-positive epithelial cells. • Treatment of EBV-positive cells with rapamycin induces Mnk, in turn elevating lytic replication and migration of these cells. [ABSTRACT FROM AUTHOR]

[Display omitted] • Transcriptomic data is used to reveal a new putative DNA virus. • The DNA virus may belong to the Mininucleoviridae. • This is the fourth virus to be identified in this new viral family. • The viral sequence data was derived from an amphipod host. The Mininucleoviridae are crustacean-infecting viruses thought to drive mortality across aquatic biomes. Three have been characterised from Carcinus maenas, Panulirus argus, and Dikerogammarus haemobaphes. We screened 202 SRA datasets (NCBI) for novel mininucleoviruses from 44 amphipod species. Three metatranscriptome datasets from Gammarus lacustris contained sequences with similarity to Dikerogammarus haemobaphes mininucleovirus. Assembly resulted in 19 transcripts, 16 were putatively polycistronic. The putative Gammarus lacustris mininucleovirus shares 46 homologues with other mininucleoviruses (similarity range: 24.07 – 78.2 %). The transcripts from this putative virus highlight its likely association with the Mininucleoviridae. [ABSTRACT FROM AUTHOR]

To determine whether the Memorial Sloan Kettering Frailty Index (MSK-FI) is associated with decision-making in older women surgically treated for advanced-stage ovarian cancer. We retrospectively applied the MSK-FI to women ≥70 years with newly diagnosed advanced-stage ovarian cancer surgically treated at our institution from 01/2001–05/2017. MSK-FI components, including 10 comorbidities and functional assessment, were extracted from medical records. The MSK-FI ranges from 0 to 11, with higher scores indicating greater frailty. The primary outcome was the association between frailty and rate of primary debulking surgery (PDS), for which a multivariable logistic regression was used, adjusted for stage and histology. We identified 430 women treated with PDS (n = 231, 54%) or neoadjuvant chemotherapy/interval debulking (n = 199, 46%) with complete data. MSK-FI score distribution was: "0", 95 patients (22%); "1", 172 (40%); "2", 89 (21%); and "3+", 74 (17%). More-frail patients were less likely to have undergone PDS (OR for a unit increase of MSK-FI: 0.64; 95%CI, 0.53–0.77; p < 0.0001). Grade 3+ complications and unintended intensive care admission occurred in 40 (9%) and 38 (9%) women, respectively, but were not associated with frailty (OR 1.21; 95%CI, 0.96–1.52; p = 0.11). More-frail patients were more likely to delay postoperative chemotherapy (non-linear association p = 0.009) and less likely to enroll in research (OR 0.84; 95%CI, 0.70–1.00; p = 0.049). Greater frailty was associated with poorer overall survival (HR 1.16; 95%CI, 1.05–1.30; p = 0.005). Frailty, as calculated by the MSK-FI, is strongly associated with treatment approach in older women with advanced ovarian cancer, suggesting objective or subjective correlates of the MSK-FI influence decision-making. • Higher MSK-FI score was associated with forgoing primary debulking surgery. • Higher MSK-FI score was associated with delays in chemotherapy and worse overall survival. • Major complications or unintended intensive care unit admission were not associated with frailty. [ABSTRACT FROM AUTHOR]

To compare survival after nodal assessment using a sentinel lymph node (SLN) algorithm versus comprehensive pelvic and paraaortic lymphadenectomy (LND) in serous or clear cell endometrial carcinoma, and to compare survival in node-negative cases. Three-year recurrence-free survival (RFS) and overall survival were compared between one institution that used comprehensive LND to the renal veins and a second institution that used an SLN algorithm with ultra-staging with inverse-probability of treatment weighting (IPTW) derived from propensity scores to adjust for covariate imbalance between cohorts. 214 patients were identified (118 SLN cohort, 96 LND cohort). Adjuvant therapy differed between the cohorts; 84% and 40% in the SLN and LND cohorts, respectively, received chemotherapy ± radiation therapy. The IPTW-adjusted 3-year RFS rates were 69% and 80%, respectively. The IPTW-adjusted 3-year OS rates were 88% and 77%, respectively. The IPTW-adjusted hazard ratio (HR) for the association of surgical approach (SLN vs LND) with progression and death was 1.46 (95% CI: 0.70–3.04) and 0.44 (95% CI: 0.19–1.02), respectively. In the 168 node-negative cases, the IPTW-adjusted 3-year RFS rates were 73% and 91%, respectively. The IPTW-adjusted 3-year OS rates were 88% and 86%, respectively. In this subgroup, IPTW-adjusted HR for the association of surgical approach (SLN vs LND) with progression and death was 3.12 (95% CI: 1.02–9.57) and 0.69 (95% CI: 0.24–1.95), respectively. OS was not compromised with the SLN algorithm. SLN may be associated with a decreased RFS but similar OS in node-negative cases despite the majority receiving chemotherapy. This may be due to differences in surveillance. • Overall survival in non-endometrioid endometrial carcinoma appears uncompromised using a sentinel lymph node algorithm. • With negative nodes, recurrence-free survival is shorter with a sentinel node algorithm, but overall survival is similar. • Lymphatic recurrences do not appear increased with sentinel lymph node assessment in non-endometrioid endometrial carcinoma. [ABSTRACT FROM AUTHOR]

Objective: The objective of this study was to explore barriers and facilitators to healthy eating among low-income Latino adolescents using an intervention development framework.Methods: Semi-structured interviews (n = 30) were conducted with Latino youth ages 13-17 who had overweight or obesity at a safety-net clinic in San Francisco, CA. Adolescent beliefs and attitudes regarding healthy eating and individual, family, and community level barriers and facilitators were elicited. Interviews were analyzed using an inductive approach and the Capability-Opportunity-Motivation (COM-B) model.Results: Participants had capability gaps; while they demonstrated basic nutrition knowledge, they also held significant misconceptions about healthy eating, equating "organic" with healthy and failing to recognize sugar in a number of beverages and foods. Families were a source of support through role modeling and purchasing fresh produce, yet in many cases also undermined adolescents' healthy eating goals through purchases of high calorie low nutrient food, an opportunity facilitator and challenge. By contrast, peers were mostly a negative influence due to frequent consumption of high calorie low nutrient food. The school environment posed opportunity challenges as participants found school lunch unpalatable and had ready access to unhealthy options nearby. Participants were motivated to improve their eating habits but often not resilient in the face of obstacles.Conclusions: Interventions to promote healthy eating among low-income Latino adolescents should address common nutritional misconceptions, target families as well as teens, consider peer influences, and advocate for policy approaches that improve the school food environment. [ABSTRACT FROM AUTHOR]

[Display omitted] • Pontogammarus roubustoides is an invader with a large complement of symbionts. • Symbiotic diversity spans 13 groups, including viruses. • Histopathology is used to explore potentially pathogenic symbionts. • With this baseline screen, we are better equipped to assess parasite introduction risk to other areas. Biological invasions may act as conduits for pathogen introduction. To determine which invasive non-native species pose the biggest threat, we must first determine the symbionts (pathogens, parasites, commensals, mutualists) they carry, via pathological surveys that can be conducted in multiple ways (i.e., molecular, pathological, and histological). Whole animal histopathology allows for the observation of pathogenic agents (virus to Metazoa), based on their pathological effect upon host tissue. Where the technique cannot accurately predict pathogen taxonomy, it does highlight pathogen groups of importance. This study provides a histopathological survey of Pontogammarus robustoides (invasive amphipod in Europe) as a baseline for symbiont groups that may translocate to other areas/hosts in future invasions. Pontogammarus robustoides (n = 1,141) collected throughout Poland (seven sites), were noted to include a total of 13 symbiotic groups: a putative gut epithelia virus (overall prevalence = 0.6%), a putative hepatopancreatic cytoplasmic virus (1.4%), a hepatopancreatic bacilliform virus (15.7%), systemic bacteria (0.7%), fouling ciliates (62.0%), gut gregarines (39.5%), hepatopancreatic gregarines (0.4%), haplosporidians (0.4%), muscle infecting microsporidians (6.4%), digeneans (3.5%), external rotifers (3.0%), an endoparasitic arthropod (putatively: Isopoda) (0.1%), and Gregarines with putative microsporidian infections (1.4%). Parasite assemblages partially differed across collection sites. Co-infection patterns revealed strong positive and negative associations between five parasites. Microsporidians were common across sites and could easily spread to other areas following the invasion of P. robustoides. By providing this initial histopathological survey, we hope to provide a concise list of symbiont groups for risk-assessment in the case of a novel invasion by this highly invasive amphipod. [ABSTRACT FROM AUTHOR]

Development of neuronal and glial populations in the dorsal root ganglia (DRG) is required for detection of touch, body position, temperature, and noxious stimuli. While female-male differences in somatosensory perception have been previously reported, no study has examined global sex differences in the abundance of DRG cell types, and the developmental origin of these differences has not been characterized. To investigate whether sex-specific differences in neuronal and glial cell types arise in the DRG during development, we performed single-cell mass cytometry analysis on sex-separated DRGs from 4 separate litter replicates of postnatal day 0 (P0) C57/BL6 mouse pups. In this analysis, we observed that females had a higher abundance of total neurons (p = 0.0266), as well as an increased abundance of TrkB+ (p = 0.031) and TrkC+ (p = 0.04) neurons for mechanoreception and proprioception, while males had a higher abundance of TrkA+ (p = 0.025) neurons for thermoreception and nociception. Pseudotime comparison of the female and male datasets indicates that male neurons are more mature and differentiated than female neurons at P0. These findings warrant further studies to determine whether these differences are maintained across development, and their impact on somatosensory perception. [Display omitted] • Sexual dimorphism exists within broad and specific subpopulations of neurons. • No significant sexual dimorphism is observed in glial cell types or leukocytes. • Females have a higher relative abundance of immature neurons. [ABSTRACT FROM AUTHOR]

Abstract Cognitive flexibility is often examined using task-switch paradigms, whereby individuals either switch between tasks or repeat the same task on successive trials. The behavioral costs of switching in terms of accuracy and reaction time are well-known, but the oscillatory dynamics underlying such costs are poorly understood. Herein, we examined 25 healthy adults who performed a task-switching paradigm during magnetoencephalography (MEG). All MEG data were transformed into the time-frequency domain and significant oscillatory responses were imaged separately per condition (i.e., switch, repeat) using a beamformer. To determine the impact of task-switching on the neural dynamics, the resulting images were examined using paired-samples t -tests. Whole-brain correlations were also computed using the switch-related difference images (switch – repeat) and the switch-related behavioral data (i.e., switch costs). Our key results indicated stronger decreases in alpha and beta activity, and greater increases in gamma activity in nodes of the cingulo-opercular and fronto-parietal networks during switch relative to repeat trials. In addition, behavioral switch costs were positively correlated with switch-related differences in right frontal and inferior parietal alpha activity, and negatively correlated with switch effects in anterior cingulate and right temporoparietal gamma activity. In other words, participants who had a greater decrease in alpha or increase in gamma in these respective regions had smaller behavioral switch costs, which suggests that these oscillations are critical to supporting cognitive flexibility. In sum, we provide novel data linking switch effects and gamma oscillations, and employed a whole-brain approach to directly link switch-related oscillatory differences with switch-related performance differences. Highlights • Task-switching costs are well known, but their oscillatory signature is unclear. • Adults performed a task-switching paradigm during magnetoencephalography (MEG). • MEG data were subjected to a beamformer and advanced oscillatory analysis methods. • Task-switching distinctly modulated behavior, alpha, beta, and gamma oscillations. • Switch effects on alpha and gamma oscillations were tied to behavioral switch costs. [ABSTRACT FROM AUTHOR]

Abstract A network of predominantly left-lateralized brain regions has been linked to verbal working memory (VWM) performance. However, the impact of memory load on the oscillatory dynamics serving VWM is far less understood. To further investigate this, we had 26 healthy adults perform a high-load (6 letter) and low-load (4 letter) variant of a VWM task while undergoing magnetoencephalography (MEG). MEG data were evaluated in the time-frequency domain and significant oscillatory responses spanning the encoding and maintenance phases were reconstructed using a beamformer. To determine the impact of load on the neural dynamics, the resulting images were examined using paired-samples t -tests and virtual sensor analyses. Our results indicated stronger increases in frontal theta activity in the high- relative to low-load condition during early encoding. Stronger decreases in alpha/beta activity were also observed during encoding in bilateral posterior cortices during the high-load condition, and the strength of these load effects increased as encoding progressed. During maintenance, stronger decreases in alpha activity in the left inferior frontal gyrus, middle temporal gyrus, supramarginal gyrus, and inferior parietal cortices were detected during high- relative to low-load performance, with the strength of these load effects remaining largely static throughout maintenance. Finally, stronger increases in occipital alpha activity were observed during maintenance in the high-load condition, and the strength of these effects grew stronger with time during the first half of maintenance, before dissipating during the latter half of maintenance. Notably, this was the first study to utilize a whole-brain approach to statistically evaluate the temporal dynamics of load-related oscillatory differences during encoding and maintenance processes, and our results highlight the importance of spatial, temporal, and spectral specificity in this regard. Highlights • The impact of load on working memory-related oscillations has been inconsistent. • Adults performed a load-varying verbal working memory task during MEG. • MEG data were subjected to a beamformer and advanced oscillatory analysis methods. • Load distinctly modulated behavior, encoding- and maintenance-related oscillations. • Load-related effects were dynamic and involved a network of left cortical regions. [ABSTRACT FROM AUTHOR]

Highlights • Multi-cohort, longitudinal investigation delineated trajectories of youth psychopathy. • Findings revealed substantial malleability in developmental course of psychopathy. • Chronically high trajectory exhibited most deleterious outcomes in later adulthood. • Unique pathways exert considerable influence on future antisocial and criminal behaviors. Abstract This multi-cohort study delineates developmental trajectories of psychopathic features across childhood and adolescence (ages 7–16) and investigates associations with adult outcomes (ages ∼ 23–34). Although most youth demonstrated consistently low levels of psychopathic features, approximately 10–15% followed a chronically high trajectory. A similar number (∼14%) displayed initially high levels that decreased over time, while others (∼10–20%) followed an increasing pattern. Boys in the chronically high trajectory exhibited the most deleterious adult outcomes and some evidence suggested that youth in the decreasing subgroup experienced fewer maladaptive outcomes than those in the increasing and high groups. Findings revealed substantial malleability in the developmental course of psychopathic features and suggest that unique pathways may exert considerable influence on future engagement in antisocial and criminal behaviors. [ABSTRACT FROM AUTHOR]

Abstract Objectives To compare oncologic outcomes in the staging of deeply invasive endometrioid endometrial carcinoma (EEC) using a sentinel lymph node algorithm (SLN) versus pelvic and paraaortic lymphadenectomy to the renal veins (LND); to compare outcomes in node-negative cases. Methods At two institutions, patients with deeply invasive (≥50% myometrial invasion) EEC were identified. One institution used LND (2004–2008), the other SLN (2005–2013). FIGO stage IV cases were excluded. Clinical characteristics and follow-up data were recorded. Results 176 patients were identified (LND, 94; SLN, 82). SLN patients were younger (p = 0.003) and had more LVSI (p < 0.001). 9.8% in the SLN and 29.8% in the LND cohorts, respectively, received no adjuvant therapy (p < 0.001). There was no association between type of assessment and recurrence; adjusted hazard ratio (aHR; LND vs. SLN) 0.87 (95%CI 0.40, 1.89) PFS. After controlling for age and adjuvant therapy, there was no association between assessment method and OS (aHR 2.54; 95%CI 0.81, 7.91). The node-negative cohort demonstrated no association between survival and assessment method: aHR 0.69 (95%CI 0.23, 2.03) PFS, 0.81 (95%CI 0.16, 4.22) OS. In the node-negative cohort, neither adjuvant EBRT+/-IVRT (HR 1.63; 95%CI 0.18, 14.97) nor adjuvant chemotherapy+/-EBRT+/-IVRT (HR 0.49; 95%CI 0.11, 2.22) were associated with OS, compared to no adjuvant therapy or IVRT-only. Conclusion Use of an SLN algorithm in deeply invasive EEC does not impair oncologic outcomes. Survival is excellent in node-negative cases, irrespective of assessment method. Adjuvant chemotherapy in node-negative patients does not appear to impact outcome. Highlights • SLN assessment does not adversely affect outcomes in deeply invasive endometrioid endometrial cancer. • Survival is excellent in node-negative cases regardless of nodal assessment method. • Nodal recurrence rates are similar between SLN and full LND. [ABSTRACT FROM AUTHOR]

Abstract The Friars Formation of San Diego County, California, has yielded a middle Eocene mammalian fauna from the early part of the Uintan North American Land Mammal Age. Prior research on the primate fauna from the Friars Formation provides evidence of one notharctine and multiple omomyine species, but many specimens collected since the early 1980s remain unstudied. Here we describe three new omomyine genera from the Friars Formation. These new taxa range in estimated body mass from about 119 g to 757 g, and substantially expand the diversity of middle Eocene omomyoids known from Southern California. Resolution of the phylogenetic relationships of the new Friars Formation omomyines is complicated by the fact that different character-taxon matrices and tree building methods produce different results. Nevertheless, all preliminary phylogenetic analyses are congruent in recovering a close relationship between the three new genera and the omomyines Macrotarsius , Omomys , Ourayia , and Utahia. Prior research has documented a shift in omomyoid diversity in North America from the anantomophine-rich Bridgerian to the omomyine-rich Uintan. Our description of three new Uintan omomyine taxa from the Friars Formation further emphasizes these opposite trends in anaptomorphine and omomyine species richness during the middle Eocene. All three of the new taxa are currently known from only the Friars Formation in San Diego County, California. Four of the previously known omomyoid genera from Southern California (Dyseolemur, Chumashius, Yaquius, and Stockia) are also endemic to the region, further highlighting the provincial character of primate faunas in Utah, Southern California, and West Texas during the Uintan. [ABSTRACT FROM AUTHOR]

It is suggested that joint architecture of the extant cercopithecid forelimb differentiates terrestrial from arboreal quadrupedal species. Linear dimensions of forelimb joint morphology have also been used to assign fossil species to locomotor categories. However, many primates use a mix of terrestrial and arboreal behaviors, which can be problematic when developing models of behavior reconstruction using morphological variation. The current study uses multivariate analyses to identify morphology related to substrate use in primates, including determination of semiterrestriality. Measurements collected from distal humeri and proximal ulnae of 49 extant cercopithecid primate species were selected based on studies indicating that they could individually predict substrate use. Analyses including one-way analysis of variance, principal components, and discriminant functions were conducted to assess their ability to differentiate between arboreal and terrestrial substrate use. The functions created in these analyses are then applied to data from fossil specimens from the Hadar sequence, Ethiopia, sampling both the Hadar and overlying Busidima Formations, to retrodict possible substrate behavior of fossil monkeys at Hadar through time. As this study is designed to identify function and behavior rather than phylogeny, the taxonomic assignment of the fossil specimens is sometimes uncertain, but substrate behavior can still be inferred. Results suggest that substrate use, including semiterrestrial behavior, in extant and extinct primates can be inferred successfully from multivariate analyses based on joint morphology of the monkey elbow. This study reveals that the ecological distribution of primarily terrestrial fossil primate species of the Hadar sequence is comparable to modern-day communities in habitats similar to those reconstructed for the Hadar members. [ABSTRACT FROM AUTHOR]

Synthetic cannabinoids (SCBs), synonymous with ‘K2’, ‘Spice’ or ‘synthetic marijuana’, are psychoactive drugs of abuse that frequently result in clinical effects and toxicity more severe than those classically associated with Δ 9 -tetrahydrocannabinol such as extreme agitation, hallucinations, supraventricular tachycardia, syncope, and seizures. JWH-018 is one of the earliest compounds identified in various SCB products, and our laboratory previously demonstrated that JWH-018 undergoes extensive metabolism by cytochromes P450 (P450), binds to, and activates cannabinoid receptors (CBRs). The major enzyme involved in the metabolism of JWH-018 is CYP2C9, a highly polymorphic enzyme found largely in the intestines and liver, with *1 being designated as the wild type, and *2 and *3 as the two most common variants. Three different major products have been identified in human urine and plasma: JWH-018 (ω)-OH, JWH-018 (ω-1)-OH(R), and JWH-018 (ω-1)-OH(S). The (ω-1)-OH metabolite of JWH-018 is a chiral molecule, and is thus designated as either (ω-1)-OH(R) or (ω-1)-OH(S). Here, in vitro enzyme kinetic assays performed with human recombinant CYP2C9 variants (*1, *2, and *3) revealed that oxidative metabolism by CYP2C9*3 resulted in significantly less formation of (ω)-OH and (ω-1)-OH metabolites. Surprisingly, CYP2C9*2 was roughly 3.6-fold more efficient as the CYP2C9*1 enzyme based on V max /K m , increasing the rate of JWH-018 metabolism and allowed for a much more rapid elimination. These results suggest that genetic polymorphisms of P450 enzymes result in the production of varying levels of biologically active JWH-018 metabolites in some individuals, offering a mechanistic explanation for the diverse clinical toxicity often observed following JWH-018 abuse. [ABSTRACT FROM AUTHOR]

Objectives Psychopathy is associated with severe forms of antisocial and violent behavior in adults. There is also a rapidly growing body of research focused on extending features of adult psychopathy downward to youth. To date however, the degree to which these features can be consistently and comparatively assessed at these younger ages, remains unclear. This study addresses this issue by investigating measurement invariance of underlying features of psychopathy across childhood and adolescence in a racially diverse sample of youth. Methods Three cohorts of youth (n = 1517) were assessed annually from childhood to adolescence (ages  ∼7–16). Underlying features of psychopathy commonly assessed in youth (e.g. lack of guilt, impulsivity) were examined within a longitudinal bi-factor framework using multi-dimensional item-response theory (IRT) techniques. Differential item functioning was used to assess invariance across development and participant’s race (African-American and Caucasian), using two distinct approaches: (1) traditional item-response theory (IRT) methods; and (2) a recently developed Bayesian structural equation modeling (BSEM) approach. Results Psychopathy features assessed in this study exhibited measurement consistency across development (∼ages 7–16) and were found to tap into the same underlying construct as intended across measurement occasions, and equivalently for African-American and Caucasian youth. Results were similar when assessed using traditional IRT procedures for longitudinal invariance testing and when implementing the more recent BSEM methodology. Conclusions Findings provide the first evidence that features of psychopathy can be assessed consistently in youth and improve our understanding of important developmental and sociocultural factors associated these features during earlier periods of development. [ABSTRACT FROM AUTHOR]

With the release and use of the Becton Dickenson FACS Diva Software, the use of Area as the default parameter came into play. As such, the use of area as a calculated parameter, methods were needed to be employed to ensure doublet discrimination and proper display on standard FSC/SSC. Improper setting of forward area scaling can alter the display cell populations. This combined with improper area gating strategy can lead to doublet inclusion which in sorting rare events can compromise sort purity. In extreme cases where area scaling with the individual lasers is ignored, differences can exist between Area and Height where compensation will likely not be optimal, particularly if one parameter – usually height is saturated. In addition, area scaling can impact population grouping. As FSC and individual laser area scaling is a function of event size, the most common error is to accept the setting determined by CS&T, which are 3.2 µm particles and proceed with the sample(s) without regard to the sample’s actual size. With cellular events smaller or more likely larger than the CS&T beads, this will make the area scaling settings less than optimal. Analysis and sorting rare events with populations larger than the CS&T beads can be compromised if adjustments in FSC area scaling are not addressed. Proper FSC and laser area scaling must be determined empirically for each sample. Examples of the effects of sample size on area scaling will be presented in addition to gating and templates for determining area scaling. [ABSTRACT FROM AUTHOR]

This study examined the influence of younger siblings on children’s understanding of second-order false belief. In a representative community sample of firstborn children ( N = 229) with a mean age of 7 years ( SD = 4.58), false belief was assessed during a home visit using an adaptation of a well-established second-order false belief narrative enacted with Playmobil figures. Children’s responses were coded to establish performance on second-order false belief questions. When controlling for verbal IQ and age, the existence of a younger sibling predicted a twofold advantage in children’s second-order false belief performance, yet this was the case only for firstborns who experienced the arrival of a sibling after their second birthday. These findings provide a foundation for future research on family influences on social cognition. [ABSTRACT FROM AUTHOR]

Latino children are more likely to be obese than non-Hispanic white children, and feeding patterns that begin in infancy may contribute to this disparity. The objective of this study was to elucidate beliefs and practices related to the introduction of solids and solid food feeding in the first year of life among low-income Latino parents residing in Northern California. We conducted 26 semi-structured interviews that explored the timing of introduction of solids, selection of foods to serve to infants, feeding strategies, sources of information on solid food feeding and concerns about infant weight. We found that most parents relied on traditional practices in selecting first foods for infants and had a strong preference for homemade food, which was often chicken soup with vegetables. Parents generally described responsive feeding practices; however a minority used pressuring practices to encourage infants to eat more. Very few parents practiced repeated gentle introduction of unfamiliar food to increase acceptance. High calorie low nutrient foods were typically introduced at around 12 months of age and parents struggled to limit such foods once children were old enough to ask for them. Parents were concerned about the possibility of infants becoming overweight and considered health care providers to be an important source of information on infant weight status. The results of this study can be used to inform the development of interventions to prevent obesity in Latino children with similar demographics to our study population. [ABSTRACT FROM AUTHOR]

Following the Reasoned Action Approach, the aim of this study was to explore consumers' top-of-mind food safety beliefs about local beef. Beef consumers recruited from farmers' markets (N = 101) and grocery stores (N = 174) across the state of Alabama participated in face-to-face intercept surveys. The survey included closed- and open-ended questions designed to elicit consumers' food safety beliefs about local beef. Results indicate that beef safety was not a top-of-mind concern for a majority of participants, however of the total number of participants familiar with the term "local beef" (n = 168, 61%), a majority (n = 105, 63%) associated local beef with improved food safety. Content analysis of verbatim text revealed that consumers believed local beef was safer because they possess greater knowledge about the product and less shipping was involved. Respondents also believe that locally processed meat is derived from small-scale operations which provided the assurance that local beef is more likely to meet U.S. regulatory standards and therefore be safer. Consumers believe they have more oversight of local beef due to both their relationships with supply chain actors and proximity which also provided food safety assurances. [ABSTRACT FROM AUTHOR]

Objective We assessed the association between reproductive and hormonal factors and ovarian cancer incidence characterized by estrogen receptor-α (ERα) and progesterone receptor (PR) status. Methods Tissue microarrays were used to assess ERα and PR expression among 197 Nurses' Health Study (NHS), 42 NHSII and 76 New England Case-Control Study (NECC) ovarian cancer cases. NHS/NHSII cases were matched to up to 4 controls (n = 954) on diagnosis date and birth year. NECC controls (n = 725) were frequency matched on age. Cases were considered receptor positive if ≥ 1% of tumor cells stained positive. Associations by ERα and PR status were assessed using polytomous logistic regression. p-Value for heterogeneity was calculated using a likelihood ratio test. Results 45% of ovarian tumors were PR (+), 78% were ERα (+) and 45% were ERα (+)/PR (+), while 22% were ERα(–)/PR(–). Postmenopausal status was associated with an increased risk of PR(–) tumors (OR: 2.07; 95%CI: 1.15–3.75; p-heterogeneity = 0.01) and age at natural menopause was inversely associated with PR(–) tumors (OR, per 5 years: 0.77; 95%CI: 0.61–0.96; p-het = 0.01). Increasing duration of postmenopause was differentially associated by PR status (p-het = 0.0009). Number of children and tubal ligation were more strongly associated with ERα(–) versus ERα (+) tumors (p-het = 0.002 and 0.05, respectively). No differential associations were observed for oral contraceptive or hormone therapy use. Conclusions Postmenopausal women have an increased risk of developing PR(–) ovarian tumors compared to premenopausal women. The associations observed for ovarian cancer differ from those seen for breast cancer suggesting that the biology for tumor development through ERα and PR pathways may differ. [ABSTRACT FROM AUTHOR]

Microcystins (MCs) are potent toxins released during cyanobacterial blooms. Clay minerals are implicated in trapping MCs within soil particles in surface waters and sediments. In the absence of molecular characterization, the relevance of previously proposed adsorption mechanisms is lacking. Towards obtaining this characterization, we conducted Monte Carlo simulations combined with molecular dynamics relaxation of two MC variants, MC-leucine-arginine (MC-LR) and MC-leucine-alanine (MC-LA), adsorbed on hydrated montmorillonite with different electrolytes. The resulting adsorbate structures revealed how MC conformations and aqueous conditions dictate binding interactions at the mineral surface. Electrostatic coupling between the arginine residue and a carboxylate in MC-LR excluded the participation of arginine in mediating adsorption on montmorillonite in a NaCl solution. However, in a CaCl 2 solution, the complexation of Ca by two carboxylate moieties in MC-LR changed the MC conformation, which allowed arginine to mediate electrostatic interaction with the mineral. By contrast, due to the lack of arginine in MC-LA, complexation of Ca by only one carboxylate in MC-LA was required to favor Ca-bridging interaction with the mineral. Multiple water-bridged H-bonding interactions were also important in anchoring MCs at the mineral surface. Our modeling results offer molecular insights into the structural and chemical factors that can control the fate of MCs at water-mineral interfaces. [ABSTRACT FROM AUTHOR]

Reactive oxygen species (ROS) are by-products of photosynthesis and respiration in plant tissues. Abiotic and biotic stressors also induce the production and temporary accumulation of ROS in plants, including hydrogen peroxide (H 2 O 2 ), whereby they can act as secondary messengers/chemical mediators in plant defense signaling and lead to programmed cell death. H 2 O 2 acts as a hub for critical information flow in plants. Despite such key roles in fundamental cellular processes, reliable determination of H 2 O 2 levels in plant tissues is hard to achieve. We optimized an Amplex Red-based quantitation method for H 2 O 2 estimation from plant tissue lysate. The standard limit of detection and quantitation was determined as 6 and 18 picomol respectively. In this study we also quantified constitutive and/or induced levels of H 2 O 2 in three model plants, Pinus nigra (Austrian pine), Oryza sativa (rice), and Arabidopsis thaliana . Overall, assay sensitivity was in the nmol g −1 FW range. Commonly used additives for H 2 O 2 extraction such as activated charcoal, ammonium sulfate, perchloric acid, polyvinylpolypyrrolidone, and trichloroacetic acid either degraded H 2 O 2 directly or interfered with the Amplex Red assay. Finally, We measured stability of Amplex Red working solution over one month of storage at −80 °C and found it to be significantly stable over time. With appropriate modifications, this optimized method should be applicable to any plant tissue. [ABSTRACT FROM AUTHOR]

A history of safe use is a backbone of safety assessments for many current probiotic species, however, there is no global harmonization regarding requirements for establishing probiotic safety for use in foods and supplements. As probiotic manufacturers are increasingly seeking to use new strains, novel species, and next-generation probiotics, justification based on a significant history of use may be challenged. There are efforts underway by a variety of stakeholders, including the United States Pharmacopeia (USP), to develop best practices guidelines for assessing the quality and safety of probiotics. A current initiative of the USP seeks to provide expert advice specific to safety considerations for probiotics. Toward this goal, this review provides a helpful summary guide to global regulatory guidelines. We question the suitability of traditional animal toxicology studies designed for testing chemicals for relevance in assessing probiotic safety. This includes discussion of the use of excessive dose levels, the length of repeated dose toxicity studies needed, and the most suitable animal species used in toxicology studies. In addition, the importance of proper manufacturing practices with regard to final product safety are also included. Thus, an outline of essential parameters of a comprehensive safety assessment for a probiotic are provided. • United States Pharmacopeia (USP) expert advice specific for safety considerations for probiotics. • Summary guide of global regulatory guidelines for assessing probiotic safety. • Suitability of traditional animal toxicology studies for use in studying the safety of probiotics. • Essential parameters of a comprehensive safety assessment for a probiotic dietary supplement product. [ABSTRACT FROM AUTHOR]

The adoption of comprehensive tobacco policies by colleges and universities may help reduce student tobacco use. To this end, The American Cancer Society's Tobacco-Free Generation Campus Initiative (TFGCI) awarded grants to 106 higher learning institutions to adopt 100% tobacco-free campus policies. This study measured changes in student tobacco use, reported exposure to secondhand smoke, and support for types of tobacco policies among five TFGCI grantee institutions who implemented 100% tobacco-free policies. Students at five U.S. TFGCI grantee institutions completed two independent cross-sectional online surveys regarding tobacco use, exposure to secondhand smoke, and policy attitudes, once before (n = 2499) and once after (n = 1667) their campuses adopted a tobacco-free policy. Students were less likely to report current cigarette smoking (aOR: 0.73, 95% C.I.: 0.63, 0.85) and exposure to secondhand smoke on campus (aOR: 0.42, 95% C.I.: 0.23, 0.76) following the policy change. In contrast, students were more likely to report past 30-day use of electronic nicotine delivery systems (ENDS) (aOR: 2.16, C.I.: 1.77, 2.63) following the policy change, despite the policy's inclusion of all tobacco and nicotine products. Tobacco-free campus policies can be associated with decreases in tobacco product use and environmental smoke exposure. The extent of their effectiveness may vary by product and the inclusion of tailored messaging, cessation support, and enforcement approaches. To discourage use of these products among students, colleges and universities should adopt 100% tobacco-free policies, monitor product use trends, offer cessation support and messaging customized for specific groups and products, and utilize a comprehensive enforcement strategy. [ABSTRACT FROM AUTHOR]

Cellular therapies utilizing T cells expressing chimeric antigen receptors (CARs) have garnered significant interest due to their clinical success in hematological malignancies. Unfortunately, this success has not been replicated in solid tumors, with only a small fraction of patients achieving complete responses. A number of obstacles to effective CAR-T cell therapy in solid tumors have been identified including tumor antigen heterogeneity, poor T cell fitness and persistence, inefficient trafficking and inability to penetrate into the tumor, immune-related adverse events due to on-target/off-tumor toxicity, and the immunosuppressive tumor microenvironment. Many preclinical studies have focused on improvements to CAR design to try to overcome some of these hurdles. However, a growing body of work has also focused on the use of local and/or regional delivery of CAR-T cells as a means to overcome poor T cell trafficking and inefficient T cell penetration into tumors. Most trials that incorporate locoregional delivery of CAR-T cells have targeted tumors of the central nervous system - repurposing an Ommaya/Rickham reservoir for repeated delivery of cells directly to the tumor cavity or ventricles. Hepatic artery infusion is another technique used for locoregional delivery to hepatic tumors. Locoregional delivery theoretically permits increased numbers of CAR-T cells within the tumor while reducing the risk of immune-related systemic toxicity. Studies to date have been almost exclusively phase I. The growing body of evidence indicates that locoregional delivery of CAR-T cells is both safe and feasible. This review focuses specifically on the use of locoregional delivery of CAR-T cells in clinical trials. [Display omitted] [ABSTRACT FROM AUTHOR]

The use of natural products (NPs), including herbal medicines and other dietary supplements, by North Americans continues to increase across all age groups. This population has access to conventional medications, with significant polypharmacy observed in older adults. Thus, the safety of the interactions between multi-ingredient NPs and drugs is a topic of paramount importance. Considerations such as history of safe use, literature data from animal toxicity and human clinical studies, and NP constituent characterization would provide guidance on whether to assess NP-drug interactions experimentally. The literature is replete with reports of various NP extracts and constituents as potent inhibitors of drug metabolizing enzymes, and transporters. However, without standard methods for NP characterization or in vitro testing, extrapolating these reports to clinically-relevant NP-drug interactions is difficult. This lack of a clear definition of risk precludes clinicians and consumers from making informed decisions about the safety of taking NPs with conventional medications. A framework is needed that describes an integrated robust approach for assessing NP-drug interactions; and, translation of the data into formulation alterations, dose adjustment, labelling, and/or post-marketing surveillance strategies. A session was held at the 41st Annual Summer Meeting of the Toxicology Forum in Colorado Springs, CO, to highlight the challenges and critical components that should be included in a framework approach. [ABSTRACT FROM AUTHOR]

Impairments in executive skills broadly span across multiple childhood epilepsy syndromes and can adversely affect quality of life. Bilingualism has been previously shown to correlate with enhanced executive functioning in healthy individuals. This study sought to determine whether the bilingual advantage in executive functioning exists in the context of pediatric epilepsy. We retrospectively analyzed neuropsychological data in 52 children with epilepsy and compared executive function scores in monolingual versus bilingual children with epilepsy while controlling for socioeconomic status and ethnicity. Bilingual children performed significantly better on the Working Memory Index than did monolingual children. There were no significant differences on the remaining executive function variables. The bilingual advantage appears to persist for working memory in children with epilepsy. These findings suggest that bilingualism is potentially a protective variable in the face of epilepsy-related working memory dysfunction. [ABSTRACT FROM AUTHOR]

Previous studies in rat hepatocytes demonstrated that insulin-dependent apolipoprotein (apo) B degradation (IDAD) is lost when cells are maintained for 3 d under enriched culture conditions. Loss of IDAD correlates with increased expression of protein tyrosine phosphatase 1B (PTP1B) known to be associated with resistance to insulin signaling in the liver. McArdle RH7777 hepatoma (McA) cells cultured in serum containing medium are resistant to IDAD; demonstrate a 30% increase in apo B secretion, and express increased levels of PTP1B protein and mRNA. In addition, insulin-stimulated Class I phosphatidylinositide 3-kinase (PI3K) activity of anti-pY immunoprecipitates is severely blunted. IDAD resistance in McA cells correlates with diminished translocation of insulin-stimulated pY-IRS1 to intracellular membranes. Incubation of McA cells with RK682, a protein tyrosine phosphatase inhibitor, is sufficient to restore IDAD in resistant McA cells. Overall, results further support the importance of Class I PI3K activity in IDAD, and suggest that loss of this activity is sufficient to cause resistance. Although other factors are involved in downstream events including sortilin binding to apo B, autophagy, and lysosomal degradation, loss of signal generation and reduced localization of Class I PI3K to intracellular membranes plays a significant role in IDAD resistance. [ABSTRACT FROM AUTHOR]

The selfish herd hypothesis, as proposed by Hamilton (1971, Journal of Theoretical Biology , 31 , 295–311), is a powerful hypothesis to explain emergent grouping behaviour by individuals acting in their own self-interest. However, immediately after prey detect a predator, the prey group may undergo a rapid disassembly, called a flash expansion, which might be considered a ‘repulsive herd’. Although flash expansion occurs in bird flocks, fish schools and insect swarms, few empirical or simulation studies have directly examined it or tested whether there are differences among its members. In addition, although flash expansion is typically thought of as a near-simultaneous movement of individuals away from the group centre, little data has been collected to verify this. We performed an empirical study to test whether the overall movement of individuals within a flash expansion is away from (1) the first individual to startle, (2) the geometric centre or (3) the point of highest density. We videotaped replicate swarms of marked whirligig beetles (Gyrinidae: Dineutes ) during flash expansion and determined their trajectories. Overall, individuals moved away from the geometric centre more strongly than from the density maximum or the first to respond (starter). The geometric centre hypothesis was also supported by the lack of polarization of the group and that the bearing angle was away from the geometric centre. The starter was more likely to be a female at the edge of a group, and she moved more quickly than others and favoured the centre of the group. This is one of the first detailed examinations of flash expansion and the individual differences within it. Future empirical and simulation studies of the movement rules and emergent properties of flash expansion are needed to better understand the collective motion of other animals. [ABSTRACT FROM AUTHOR]

Objective: In the context of a food purchasing environment filled with advertising and promotions, and an increased desire from policy makers to guide individuals toward choosing healthier foods, this study tests whether priming methods that use healthy food adverts to increase preference for healthier food generalize to a representative population.Methods: In two studies (Study 1 n = 143; Study 2 n = 764), participants were randomly allocated to a prime condition, where they viewed fruit and vegetable advertisements, or a control condition, with no advertisements. A subsequent forced choice task assessed preference between fruits and other sweet snacks. Additional measures included current hunger and thirst, dietary restraint, age, gender, education and self-reported weight and height.Results: In Study 1, hunger reduced preferences for fruits (OR (95% CI) = 0.38 (0.26-0.56), p <0.0001), an effect countered by the prime (OR (95% CI) = 2.29 (1.33-3.96), p = 0.003). In Study 2, the effect of the prime did not generalize to a representative population. More educated participants, as used in Study 1, chose more fruit when hungry and primed (OR (95% CI) = 1.42 (1.13-1.79), p = 0.003), while less educated participants' fruit choice was unaffected by hunger or the prime.Conclusion: This study provides preliminary evidence that the effects of adverts on healthy eating choices depend on key individual traits (education level) and states (hunger), do not generalize to a broader population and have the potential to increase health inequalities arising from food choice. [ABSTRACT FROM AUTHOR]

Objective In the context of a food purchasing environment filled with advertising and promotions, and an increased desire from policy makers to guide individuals toward choosing healthier foods, this study tests whether priming methods that use healthy food adverts to increase preference for healthier food generalize to a representative population. Methods In two studies (Study 1 n = 143; Study 2 n = 764), participants were randomly allocated to a prime condition, where they viewed fruit and vegetable advertisements, or a control condition, with no advertisements. A subsequent forced choice task assessed preference between fruits and other sweet snacks. Additional measures included current hunger and thirst, dietary restraint, age, gender, education and self-reported weight and height. Results In Study 1, hunger reduced preferences for fruits (OR (95% CI) = 0.38 (0.26–0.56), p  <   0.0001), an effect countered by the prime (OR (95% CI) = 2.29 (1.33–3.96), p  = 0.003). In Study 2, the effect of the prime did not generalize to a representative population. More educated participants, as used in Study 1, chose more fruit when hungry and primed (OR (95% CI) = 1.42 (1.13–1.79), p  = 0.003), while less educated participants' fruit choice was unaffected by hunger or the prime. Conclusion This study provides preliminary evidence that the effects of adverts on healthy eating choices depend on key individual traits (education level) and states (hunger), do not generalize to a broader population and have the potential to increase health inequalities arising from food choice. [ABSTRACT FROM AUTHOR]

Both positive affect and positive close relationships contribute significantly to overall well-being. This review examines the literature assessing associations between positive affect and positive indices within close relationships across the life span. Specifically, the reviewed research includes parent–child relationships, friendships, and romantic relationships in relation to a variety of positive emotions and happiness more generally. This review also highlights several processes that may serve as partial mechanisms linking positive close relationships and positive affect including the interpersonal regulation and coregulation of positive emotion and the biological processes involved in experiences of positive affect and close relationships. Throughout the review, evidence of bidirectional, reciprocal associations between positive affect and positive close relationships is emphasized. Based on the current state of the literature, future directions for research in this area are considered. [ABSTRACT FROM AUTHOR]

The human brain is considerably larger and more energetically costly than that of other primate species. As such, discovering how human ancestors were able to provide sufficient energy to their brains is a central theme in the study of hominin evolution. However, many discussions of metabolism frequently omit the different ways in which energy, primarily glucose, is used once made available to the brain. In this review, we discuss two glucose metabolic pathways, oxidative phosphorylation and aerobic glycolysis, and their respective contributions to the energetic and anabolic budgets of the brain. While oxidative phosphorylation is a more efficient producer of energy, aerobic glycolysis contributes essential molecules for the growth of the brain and maintaining the structure of its cells. Although both pathways occur in the brain throughout the lifetime, aerobic glycolysis is a critical pathway during development, and oxidative phosphorylation is highest during adulthood. We outline how elevated levels of aerobic glycolysis may support the protracted neurodevelopmental sequence of humans compared with other primates. Finally, we review the genetic evidence for differences in metabolic function in the brains of primates and explore genes that may provide insight into how glucose metabolism may differ across species. [ABSTRACT FROM AUTHOR]

Access to cancer care, especially surgery, is limited in rural areas. However, the specific reasons rural patient populations do not receive surgery for non–small cell lung cancer (NSCLC) is unknown. We investigated geographic disparities in reasons for failure to receive guideline-indicated surgical treatment for patients with potentially resectable NSCLC. The National Cancer Database was used to identify patients with clinical stage I-IIIA (N 0-N1) NSCLC between 2004 and 2018. Patients from rural areas were compared to urban areas, and the reason for nonreceipt of surgery was evaluated. Adjusted odds of (1) primary nonsurgical management, (2) surgery being deemed contraindicated due to risk, (3) surgery being recommended but not performed, and (4) overall failure to receive surgery were determined. The study included 324,785 patients with NSCLC with 42,361 (13.0%) from rural areas. Overall, 62.4% of patients from urban areas and 58.8% of patients from rural areas underwent surgery (P < 0.001). Patients from rural areas had increased odds of (1) being recommended primary nonsurgical management (adjusted odds ratio [aOR]: 1.14, 95% confidence interval [CI]: 1.05-1.23), (2) surgery being deemed contraindicated due to risk (aOR: 1.19, 95% CI: 1.07-1.33), (3) surgery being recommended but not performed (aOR: 1.13, 95% CI: 1.01-1.26), and (4) overall failure to receive surgery (aOR: 1.21, 95% CI: 1.13-1.29; all P < 0.001). There are geographic disparities in the management of NSCLC. Rural patient populations are more likely to fail to undergo surgery for potentially resectable disease for every reason examined. [ABSTRACT FROM AUTHOR]

Abstract: Background: Hprt-Cre doubles the prevalence of homozygous null embryos per litter versus heterozygous breedings without decreasing litter size. Resulting mutant embryos are genotypically and phenotypically equivalent between strategies. We set out to confirm the effectiveness of this approach with other alleles and hypothesized that it would increase efficiency in generating compound mutants. Materials and methods: Null mutants for Cyp26b1, Pitx2, and Shh were generated with Hprt-Cre from conditional alleles as were double and triple allelic combinations of Fgfr2IIIb, Raldh2, and Cyp26b1. Embryos were genotyped and phenotyped by whole mount photography, histology, and immunohistochemistry. Results: Fifty percent of Hprt-Cre litters were homozygous null for Cyp26b1 (15/29) and Pitx2 (75/143), with phenotypic and genotypic equivalence to mutants from standard heterozygous breedings. In multi-allele breedings, mutant embryos constituted half of litters without significant embryo loss. In contrast, Shh breedings yielded a smaller ratio of embryos carrying two recombined alleles (6 of 16), with a significant litter size reduction because of early embryonic lethality (16 live embryos from 38 deciduae). Conclusions: Hprt-Cre can be used to efficiently generate large numbers of mutant embryos with a number of alleles. Compound mutant generation was equally efficient. However, efficiency is reduced for genes whose protein product potentially interacts with the Hprt pathway (e.g., Shh). [Copyright &y& Elsevier]

Abstract: Background: The mechanism of intestinal atresia formation remains undefined. Atresia in fibroblast growth factor receptor 2IIIb (Fgfr2IIIb −/− ) mutant mouse embryos is preceded by endodermal apoptosis and involution of the surrounding mesoderm. We have observed that involution of the atretic segment is preceded by the downregulation of Sonic hedgehog (SHH) in the endoderm, which is a critical organizer of the intestinal mesoderm. We hypothesized that supplementation of Fgfr2IIIb −/− intestinal tracts with exogenous SHH protein before atresia formation would prevent involution of the mesoderm and rescue normal intestinal development. Methods: In situ hybridization was performed on control and Fgfr2IIIb −/− intestinal tracts for Shh or forkhead box protein F1 (FoxF1) between embryonic (E) day 11.5 and E12.0. Control and Fgfr2IIIb −/− intestinal tracts were harvested at E10.5 and cultured in media supplemented with fibroblast growth factor (FGF) 10 + SHH, or FGF10 with a SHH-coated bead. In situ hybridization was performed at E12.5 for Foxf1. Results: SHH and Foxf1 expression were downregulated during intestinal atresia formation. Media containing exogenous FGF10 + SHH did not prevent colonic atresia formation (involution). A SHH protein point source bead did induce Foxf1 expression in controls and mutants. Conclusions: Shh and Foxf1 expression are disrupted in atresia formation of distal colon, thereby serving as potential markers of atretic events. Application of exogenous SHH (in media supplement or as a point source bead) is sufficient to induce Foxf1 expression, but insufficient to rescue development of distal colonic mesoderm in Fgfr2IIIb −/− mutant embryos. Shh signal disruption is not the critical mechanism by which loss of Fgfr2IIIb function results in atresia formation. [Copyright &y& Elsevier]

Abstract: Objective: To evaluate the effects of a theory-based physical activity and healthy eating intervention aimed at reducing stroke risk factors among overweight/obese and sedentary older adults. Methods: Between 12/2009–1/2011, participants (n=69) were randomly assigned to an 8-week group motivational intervention or biweekly newsletters by mail. Physical activity (blinded pedometer, 7-day recall) body composition, theoretical mediator, and dietary (24-hour recall) variables were measured at pre-test and post-test. The physical activity and dietary outcomes are reported. Results: For outcome measures, the follow-up was 90% for the intervention group (n=29) and 91% for the control group (n=34) for this sample. Statistically significant differences in the 7-day physical activity self-report were noted at post-test in the intervention group. The dietary measures were not statistically significant at post-test; however, the intervention group increased the quantity of vegetable servings. Conclusion: Limited efficacy testing was supported for a combined physical activity-dietary intervention, framed by a wellness-motivation theory, and designed to reduce stroke risk factors in older adults who are sedentary and overweight/obese. Limitations were identified and recommendations for additional research provided. [Copyright &y& Elsevier]

Abstract: Background: Pancreatic cancer cells exist in a hypoxic microenvironment containing numerous factors that impact tumor survival, proliferation, and metastasis. MicroRNAs (miRs) are differentially expressed in cancer but also altered by hypoxia. We hypothesized that hypoxia could induce expression of miR-21, an oncomir in pancreatic cancer cells. Materials and methods: We examined how hypoxia regulates miR-21 expression in pancreatic cancer cell lines (BxPC-3, AsPC-1) by stem-loop RT-PCR. Chromatin immunoprecipitation assays were used to study how hypoxia alters hypoxia-inducible factor (HIF)-1α binding to the hypoxia response element of miR-21. BxPC-3 and AsPC-1 cells were transfected with a constitutively stable HIF-1α subunit or vector control (pcDNA3.1) to determine the influence of miR-21 in normoxia. The effect of mature miR-21 sense and antisense oligonucleotides on proliferation and apoptosis in hypoxic and normoxic conditions was assessed via WST-1 assay and flow cytometry. Results: MiR-21 levels increased in all cell lines grown in hypoxic conditions versus normoxia, whereas siRNA targeting HIF-1α reduced miR-21 expression. Hypoxic conditions resulted in direct binding of HIF-1α to the predicted binding site in miR-21. Transfection with a constitutively stable HIF-1α expression plasmid in normoxia resulted in upregulated miR-21, similar to that seen in hypoxia. Cells transfected with antisense constructs targeting miR-21 had reduced proliferation and increased apoptosis in normoxia, whereas miR-21 overexpression abrogated hypoxia-associated reductions in proliferation. Conclusions: MiR-21 is induced by hypoxia in pancreatic cancer cells via HIF-1α upregulation. MiR-21 overexpression allows cells to avoid apoptosis in a hypoxic microenvironment. Inhibition of miR-21 expression may increase cellular susceptibility to hypoxia in pancreatic cancer. [Copyright &y& Elsevier]

Abstract: As new influenza virus strains emerge, finding new mechanisms to control infection is imperative. In this study, we found that we could control influenza infection of mammalian cells by altering the level of glucose given to cells. Higher glucose concentrations induced a dose-specific increase in influenza infection. Linking influenza virus infection with glycolysis, we found that viral replication was significantly reduced after cells were treated with glycolytic inhibitors. Addition of extracellular ATP after glycolytic inhibition restored influenza infection. We also determined that higher levels of glucose promoted the assembly of the vacuolar-type ATPase within cells, and increased vacuolar-type ATPase proton-transport activity. The increase of viral infection via high glucose levels could be reversed by inhibition of the proton pump, linking glucose metabolism, vacuolar-type ATPase activity, and influenza viral infection. Taken together, we propose that altering glucose metabolism may be a potential new approach to inhibit influenza viral infection. [Copyright &y& Elsevier]

• Chronic stress increases fecal corticosterone metabolites (FCMs) in lactating rats. • FCMs did not change across lactation in control rats. • FCMs in chronically-stressed rats increased in late lactation. • This change may be due to lactation stage and/or repeated exposure to the stressor. • This study highlights the need for repeated temporal sampling across lactation. Maternal exposure to stressors during lactation has previously been demonstrated to impact various aspects of milk synthesis and to have long-term physiological effects on offspring. Much of the current literature investigating the effects of stress during lactation has used acute stressors, and the studies investigating the effects of chronic stressors largely focus on neurological changes. Further, temporal variation in glucocorticoids across lactation in response to stressors has rarely been assessed. The present work uses a novel male intruder paradigm to model the effects of chronic stress on maternal fecal corticosterone metabolites (FCMs) in Sprague-Dawley rats across lactation. FCM levels were elevated in chronically-stressed mothers relative to the control group. Further, FCMs in the stress group were time-dependent either due to repeated exposure to the stressor or lactation stage. Together, this work demonstrates the efficacy of this established paradigm in increasing circulating glucocorticoids in lactating rats. These results highlight the need for repeated temporal sampling, as glucocorticoid levels in response to a chronic stressor may change across lactation. [ABSTRACT FROM AUTHOR]

Conflicting guidelines regarding universal pediatric cholesterol screening were released between 2011 and 2019, but the impact on screening rates remains understudied. The purpose of this study was to examine trends in pediatric cholesterol screening rates within a single institution in the United States and their association with release of national guidelines, local educational tools, and electronic health record (EHR) modifications. Order placement was defined as ordering a high-density lipoprotein cholesterol level in a patient aged 9-21 years with ≥1 well visit in prior 3 years. Order placement rate (OPR) was calculated per month using 3 months' moving average smoothing and analyzed based on date, patient age, and specialty of ordering clinician. Timing of educational tools, EHR modifications, and national guideline release were analyzed for changes in OPR. Prior to release of 2011 guidelines recommending universal pediatric cholesterol screening, pediatrician OPR was 35% (95% CI: 29-43%) compared to 8% (7-11%) for family physicians. For both specialties, OPR increased after 2011 guidelines, educational initiatives, and EHR changes, but decreased after 2016, with a larger decrease for family physicians (p < 0.001 for all). OPR was consistently higher for pediatricians than for family physicians during the study period, with largest OPR changes correlating with release of guidelines. The findings from the study suggest that conflicting guidelines may contribute to lower overall OPR, and to different screening rates for children cared for by pediatricians compared to family physicians. [ABSTRACT FROM AUTHOR]

[Display omitted] • We know little about the parasites that co-invade with invasive hosts. • There are a predicted 323 invasive crustaceans. • Only 31.2% of invasive crustaceans have one or more known symbionts. • Invasive symbionts/parasites pose variable threats to local ecosystems. • Some invasive parasites cause harm to wildlife, farming, human health, and the economy. • Other invasive parasites protect the local ecology by controlling the invasive populations. • Parasites need to be accounted for during invasion risk assessment. Invasive non-native species (INNS) pose a risk as vectors of parasitic organisms (Invasive Parasites). Introducing invasive parasites can result in ecological disturbances, leading to biodiversity loss and native species illness/mortality, but occasionally can control INNS limiting their impact. Risks to human health and the economy are also associated with INNS and invasive parasites; however, we understand little about the diversity of symbiotic organisms co-invading alongside INNS. This lack of clarity is an important aspect of the 'One Health' prerogative, which aims to bridge the gap between human, wildlife, and ecosystem health. To explore symbiont diversity associated with the invasive crustacean group (including: crab, lobster, crayfish, shrimp, amphipod, isopod, copepod, barnacle, other) (n = 323) derived from 1054 aquatic invertebrates classed as INNS across databases, we compile literature (year range 1800–2017) from the native and invasive range to provide a cumulative symbiont profile for each species. Our search indicated that 31.2% of INN crustaceans were known to hold at least one symbiont, whereby the remaining 68.8% had no documented symbionts. The symbiont list mostly consisted of helminths (27% of the known diversity) and protists (23% of the known diversity), followed by bacteria (12%) and microsporidians (12%). Carcinus maenas , the globally invasive and extremely well-studied green crab, harboured the greatest number of symbionts (n = 72). Additional screening is imperative to become more informed on invasive symbiont threats. We reveal that few studies provide truly empirical data that connect biodiversity loss with invasive parasites and suggest that dedicated studies on available systems will help to provide vital case studies. Despite the lack of empirical data, co-invasive parasites of invasive invertebrates appear capable of lowering local biodiversity, especially by causing behavioural change and mortality in native species. Alternatively, several invasive parasites appear to protect ecosystems by controlling the impact and population size of their invasive host. We provide a protocol that could be followed to explore symbiont diversity in invasive groups as part of our case studies. The consequence of limited parasite screening of INNS, in addition to the impacts invasive parasites impart on local ecologies, are explored throughout the review. We conclude in strong support of the 'One Health' prerogative and further identify a need to better explore disease in invasion systems, many of which are accountable for economic, human health and ecological diversity impacts. [ABSTRACT FROM AUTHOR]

Abstract: The neuronal composition of the insula in primates displays a gradient, transitioning from granular neocortex in the posterior-dorsal insula to agranular neocortex in the anterior-ventral insula with an intermediate zone of dysgranularity. Additionally, apes and humans exhibit a distinctive subdomain in the agranular insula, the frontoinsular cortex (FI), defined by the presence of clusters of von Economo neurons (VENs). Studies in humans indicate that the ventral anterior insula, including agranular insular cortex and FI, is involved in social awareness, and that the posterodorsal insula, including granular and dysgranular cortices, produces an internal representation of the body''s homeostatic state. We examined the volumes of these cytoarchitectural areas of insular cortex in 30 primate species, including the volume of FI in apes and humans. Results indicate that the whole insula scales hyperallometrically (exponent = 1.13) relative to total brain mass, and the agranular insula (including FI) scales against total brain mass with even greater positive allometry (exponent = 1.23), providing a potential neural basis for enhancement of social cognition in association with increased brain size. The relative volumes of the subdivisions of the insular cortex, after controlling for total brain volume, are not correlated with species typical social group size. Although its size is predicted by primate-wide allometric scaling patterns, we found that the absolute volume of the left and right agranular insula and left FI are among the most differentially expanded of the human cerebral cortex compared to our closest living relative, the chimpanzee. [Copyright &y& Elsevier]

Abstract: A transposon insertional mutagenesis spore library of the pathogen Bacillus anthracis was screened to identify mutants altered in germination kinetics. One mutant exhibited an accelerated rate of germination in association with disruption of benK. This gene encodes a putative protein with high homology to membrane transporters that facilitate benzoate transport. We hypothesized that BenK may not be only spore associated, but also have a vegetative cell role. A reporter strain with a translational fusion of benK to green fluorescent protein demonstrated that full-length BenK was present in vegetative cells and that a BenK degradation product was present in spores by detecting the reporter using fluorescence and Western blot analysis. A minimum inhibitory concentration assay indicated that vegetative cells of a benK::Kan mutant were more susceptible to the antimicrobial effects of Na-benzoate. The mutant spores germinated to a greater extent within 1 h than the wild type in an in vitro fluorescence assay. The disruption of benK also resulted in spores that were less readily phagocytosed in a macrophage assay. Despite these altered in vitro phenotypes, no apparent effect of the BenK protein on virulence in the intranasal mouse model or the guinea pig competitive assay was observed. This work shows that, although the BenK protein does not impact fitness or virulence in an infection model, it is involved in other aspects of both the spore and vegetative forms of the organism. [Copyright &y& Elsevier]

Abstract: 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-3β) 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. [Copyright &y& Elsevier]

Abstract: The study aim was to examine changes in food cue-elicited cravings and the macronutrient content of craved foods across menstrual cycle phases in a non-eating disordered sample. Thirty-five college females attended laboratory sessions in the late follicular and late luteal phases. In each session they completed a measure of state food craving before and after exposure to preferred, high fat/high sugar chocolate candy. Candy consumption following cue exposure was measured during an ad libitum “taste test.” Additionally, participants rated their desire to eat foods differing systematically and significantly in macronutrient content. Ovulation was confirmed with luteinizing hormone detection kits. Results show that whereas the food cue increased cravings, this effect did not differ between cycle phases examined. The macronutrient content of foods desired also did not differ significantly between cycle phases, however, a non-significant trend suggested that high fat/high complex carbohydrate and low fat/high protein foods were more strongly desired in the late luteal phase. Amount of chocolate candy eaten did not differ between cycle phases. These results suggest that cravings for high fat/high sugar foods do not differ between menstrual cycle phases examined, whereas cravings for other foods may fluctuate across cycle phases in non-eating disordered women. [Copyright &y& Elsevier]

Abstract: Purpose: The etiology of intestinal atresia remains elusive but has been ascribed to a number of possible events including in utero vascular accidents, failure of recanalization of the intestinal lumen, and mechanical compression. Another such event that has been postulated to be a cause in atresia formation is disruption in notochord development. This hypothesis arose from clinical observations of notochord abnormalities in patients with intestinal atresias as well as abnormal notochord development observed in a pharmacologic animal model of intestinal atresia. Atresias in this model result from in utero exposure to Adriamycin, wherein notochord defects were noted in up to 80% of embryos that manifested intestinal atresias. Embryos with notochord abnormalities were observed to have ectopic expression of Sonic Hedgehog (Shh), which in turn was postulated to be causative in atresia formation. We were interested in determining whether disruptions in notochord development or Shh expression occurred in an established genetic model of intestinal atresia and used the fibroblast growth factor receptor 2IIIb homozygous mutant (Fgfr2IIIb−/−) mouse model. These embryos develop colonic atresias (100% penetrance) and duodenal atresias (42% penetrance). Methods: Wild-type and Fgfr2IIIb−/− mouse embryos were harvested at embryonic day (E) 10.5, E11.5, E12.5, and E13.5. Whole-mount in situ hybridization was performed on E10.5 embryos for Shh. Embryos at each time point were harvested and sectioned for hematoxylin-eosin staining. Sections were photographed specifically for the notochord and resulting images reconstructed in 3-D using Amira software. Colons were isolated from wild-type and Fgfr2IIIb−/− embryos at E10.5, then cultured for 48 hours in Matrigel with FGF10 in the presence or absence of exogenous Shh protein. Explants were harvested, fixed in formalin, and photographed. Results: Fgfr2IIIb−/− mouse embryos exhibit no disruptions in Shh expression at E10.5, when the first events in atresia formation are known to occur. Three-dimensional reconstructions failed to demonstrate any anatomical disruptions in the notochord by discontinuity or excessive branching. Culture of wild-type intestines in the presence of Shh failed to induce atresia formation in either the duodenum or colon. Cultured Fgfr2IIIb−/− intestines developed atresias of the colon in either the presence or absence of Shh protein. Conclusions: Although disruptions in notochord development can be associated with intestinal atresia formation, in the Fgfr2IIIb−/− genetic animal model neither disruptions in notochord development nor the presence of exogenous Shh protein are causative in the formation of these defects. [Copyright &y& Elsevier]

Abstract: Activation induced deaminase (AID) is globally targeted to immunoglobulin loci, preferentially focused to switch (S) regions and variable (V) regions, and prone to attack hotspot motifs. Nevertheless, AID deamination is not exclusive to Ig loci and the rules regulating AID targeting remain unclear. Transcription is critically required for class switch recombination and somatic hypermutation. Here, I consider the unique features associated with S region transcription leading to RNA polymerase II pausing, that in turn promote the introduction of activating chromatin remodeling, histone modifications and recruitment of AID to targeted S regions. These findings allow for a better understanding of the interplay between transcription, AID targeting and mistargeting to Ig and non-Ig loci. [Copyright &y& Elsevier]