Stealth oxime ether lipid vesicles promote delivery of functional DsiRNA in human lung cancer A549 tumor bearing mouse xenografts.

We previously reported that hydroxylated oxime ether lipids (OELs) efficiently deliver functional Dicer substrate siRNAs (DsiRNAs) in cells. Here, we explored in vivo utility of these OELs, using OEL4 as a prototype and report that surface modification of the OEL4 formulations was essential for their in vivo applications. These surface-modified OEL4 formulations were developed by inclusion of various PEGylated lipids. The vesicle stability and gene knock-down were dependent on the PEG chain length. OEL4 containing DSPE-PEG350 and DSPE-PEG1000 (surprisingly not DSPE2000) promoted gene silencing in cells. In vivo studies demonstrated that OEL4 vesicles formulated using 3 mol% DSPE-PEG350 accumulate in human lung cancer (A549-luc2) xenografts in mice and exhibit a significant increase in tumor to liver ratios. These vesicles also showed a statistically significant reduction of luciferase signal in tumors compared to untreated mice. Taken together, the scalable OEL4:DSPE-PEG350 formulation serves as a novel candidate for delivery of RNAi therapeutics.

One-Pot, One-Step Synthesis of Drug-Loaded Magnetic Multimicelle Aggregates.

Lipid-based formulations provide a nanotechnology platform that is widely used in a variety of biomedical applications because it has several advantageous properties including biocompatibility, reduced toxicity, relative ease of surface modifications, and the possibility for efficient loading of drugs, biologics, and nanoparticles. A combination of lipid-based formulations with magnetic nanoparticles such as iron oxide was shown to be highly advantageous in a growing number of applications including magnet-mediated drug delivery and image-guided therapy. Currently, lipid-based formulations are prepared by multistep protocols. Simplification of the current multistep procedures can lead to a number of important technological advantages including significantly decreased processing time, higher reaction yield, better product reproducibility, and improved quality. Here, we introduce a one-pot, single-step synthesis of drug-loaded magnetic multimicelle aggregates (MaMAs), which is based on controlled flow infusion of an iron oxide nanoparticle/lipid mixture into an aqueous drug solution under ultrasonication. Furthermore, we prepared molecular-targeted MaMAs by directional antibody conjugation through an Fc moiety using Cu-free click chemistry. Fluorescence imaging and quantification confirmed that antibody-conjugated MaMAs showed high cell-specific targeting that was enhanced by magnetic delivery.

Positive Caregiving Styles Attenuating Effects of Cumulative Trauma Among Youth Who Commit Sexual Crimes.

Among youth who commit sexual crimes, childhood trauma experiences have been linked to a host of outcomes including trauma symptom expression. Furthermore, research has begun assessing differential rates of cumulative adverse childhood experiences (ACEs) between youth who commit sexual and non-sexual crimes. Yet, few studies have comparatively examined rates of cumulative traumatic events using more robust measures of adversity. There is scant research on outcomes associated cumulative trauma among youth who commit sexual crimes including linkages to trauma symptoms and how positive parenting styles (PCSs) can attenuate in this relationship. This study aims to compare cumulative trauma experiences between youth who commit sexual (n = 112) and non-sexual crimes (n = 224). Among youth who commit sexual crimes, we test how PCSs mitigate or moderate the effects of cumulative trauma on trauma symptoms. Results from independent samples t-tests revealed significant group differences on singular and cumulative traumatic experiences, and trauma symptomatology, where youth who commit sexual crimes had higher rates of all. Stepwise linear regressions revealed mitigating effects of maternal PCSs and tests of moderation revealed main and interaction effects, where intermediate positive maternal caregiving styles for youth with higher cumulative trauma were associated with greater trauma symptomatology. Implications and limitations are discussed.

From Tube to Structure: SPA Cryo-EM Workflow Using Apoferritin as an Example.

In this chapter, we present an overview of a standard protocol to achieve structure determination at high resolution by Single Particle Analysis cryogenic Electron Microscopy using apoferritin as a standard sample. The purified apoferritin is applied to a glow-discharged support and then flash frozen in liquid ethane. The prepared grids are loaded into the electron microscope and checked for particle spreading and ice thickness. The microscope alignments are performed and the data collection session is setup for an overnight data collection. The collected movies containing two-dimensional images of the apoferritin sample are then processed to obtain a high-resolution three-dimensional reconstruction.

Resilience Following Child Maltreatment: Definitional Considerations and Developmental Variations.

Resilience following childhood maltreatment has received substantial empirical attention, with the number of studies on this construct growing exponentially in the past decade. While there is ample interest, inconsistencies remain about how to conceptualize and assess resilience. Further, there is a lack of consensus on how developmental stage influences resilience and how protective factors affect its expression. The current systematic review uses a developmental lens to synthesize findings on resilience following child maltreatment. Specifically, this article consolidates the body of empirical literature in a developmentally oriented review, with the intention of inclusively assessing three key areas-the conceptualization of resilience, assessment of resilience, and factors associated with resilience in maltreatment research. A total of 67 peer-reviewed, quantitative empirical articles that examined child maltreatment and resilience were included in this review. Results indicate that some inconsistencies in the literature may be addressed by utilizing a developmental lens and considering the individual's life stage when selecting a definition of resilience and associated measurement tool. The findings also support developmental variations in factors associated with resilience, with different individual, relational, and community protective factors emerging based on life stage. Implications for practice, policy, and research are incorporated throughout this review.

"Disbelief and Sadness": First-Year Health Profession Students' Perspectives on Medical Errors.

OBJECTIVES: The aims of the study were to examine the reactions of first-year health profession students to medical errors and determine whether differences exist between disciplines. METHODS: After viewing the Team STEPPS Program's Susan Sheridan video that describes two separate medical errors, students from anesthesia assistant, medical imaging, medicine, nursing, physical therapy, and physician assistant programs provided unstructured open responses reflecting on initial impressions of medical errors depicted in the film. Student responses were assessed via inductive coding techniques and thematic analysis and stratified by discipline. Frequencies of key themes were calculated. Descriptive analyses characterized respondents and χ2 tests compared responses between disciplines. RESULTS: In a review of 373 student responses (80% response rate), 255 students expressed an emotion-based reply, of which 93.75 were negative with such comments as they felt horrified, appalled, and disappointed by the patient's experience. Of the 227 students who commented on blame or fault, 70% felt that a system error was at fault, 45% felt that it was the provider and only 1.3% stated that it was the patient's fault. Of the students who mentioned the context or situation, just more than half felt that the episode was preventable and there was a causal relationship between clinic workers' actions and the medical error. Finally, a high percentage of students had a solutions-oriented response, noting the importance of teamwork and communication in the avoidance of medical errors. CONCLUSIONS: First-year health profession students responded to medical error with strong emotion and distress. Their responses largely represent an individualistic view of healthcare in both the causes and solutions to medical errors. No differences in response themes were observed by discipline. This study represents our students' emotional responses to a medical error scenario. The qualitative responses and reactions of students to the Sheridan video offered opportunities to tease out nuances that would otherwise be unavailable in a typical attitudes survey. We noted a individualistic view of healthcare in both the causes and solutions to medical errors. We view these results as an opportunity for interprofessional education in systems-level approaches to improve patient safety. Curricular efforts in interprofessional education, collaborative practice, and patient safety should be driven by these results.

Flagellar Structures from the Bacterium Caulobacter crescentus and Implications for Phage ϕ CbK Predation of Multiflagellin Bacteria.

Caulobacter crescentus is a Gram-negative alphaproteobacterium that commonly lives in oligotrophic fresh- and saltwater environments. C. crescentus is a host to many bacteriophages, including ϕCbK and ϕCbK-like bacteriophages, which require interaction with the bacterial flagellum and pilus complexes during adsorption. It is commonly thought that the six paralogs of the flagellin gene present in C. crescentus are important for bacteriophage evasion. Here, we show that deletion of specific flagellins in C. crescentus can indeed attenuate ϕCbK adsorption efficiency, although no single deletion completely ablates ϕCbK adsorption. Thus, the bacteriophage ϕCbK likely recognizes a common motif among the six known flagellins in C. crescentus with various degrees of efficiency. Interestingly, we observe that most deletion strains still generate flagellar filaments, with the exception of a strain that contains only the most divergent flagellin, FljJ, or a strain that contains only FljN and FljO. To visualize the surface residues that are likely recognized by ϕCbK, we determined two high-resolution structures of the FljK filament, with and without an amino acid substitution that induces straightening of the filament. We observe posttranslational modifications on conserved surface threonine residues of FljK that are likely O-linked glycans. The possibility of interplay between these modifications and ϕCbK adsorption is discussed. We also determined the structure of a filament composed of a heterogeneous mixture of FljK and FljL, the final resolution of which was limited to approximately 4.6 Å. Altogether, this work builds a platform for future investigations of how phage ϕCbK infects C. crescentus at the molecular level.IMPORTANCE Bacterial flagellar filaments serve as an initial attachment point for many bacteriophages to bacteria. Some bacteria harbor numerous flagellin genes and are therefore able to generate flagellar filaments with complex compositions, which is thought to be important for evasion from bacteriophages. This study characterizes the importance of the six flagellin genes in C. crescentus for infection by bacteriophage ϕCbK. We find that filaments containing the FljK flagellin are the preferred substrate for bacteriophage ϕCbK. We also present a high-resolution structure of a flagellar filament containing only the FljK flagellin, which provides a platform for future studies on determining how bacteriophage ϕCbK attaches to flagellar filaments at the molecular level.

Court-Affiliated Diversion Programs for Prostitution-Related Crimes: A Comprehensive Review of Program Components and Impact.

OBJECTIVE: Court diversion programs offer alternative treatment interventions in lieu of punitive sanctions. Programs have recently been developed for women arrested for prostitution, with a recognition that many of these individuals frequently experience multiple forms of violence and experience multiple barriers to exiting sex work. This review aims to (a) examine programmatic components used across programs, and (b) identify the diversionary programs' impact on participants. METHODS: Studies were identified by entering key search words into three electronic databases and by conducting a citation search. RESULTS: Nine articles were included in the review. Although programs varied in structure, services, and length of time, studies indicated a range of positive outcomes for participants. CONCLUSIONS: Results help to illuminate future directions for criminal justice practice, policy, and research.

Defining resilience in maltreated children from the practitioners' perspectives: A qualitative study.

BACKGROUND: Increasingly, studies have focused on understanding positive outcomes in children who have been maltreated and the factors that contribute to resilience. However, there is no universally accepted definition of resilience, thus hindering the ability to make comparisons across studies and to use such information to inform interventions to foster resilience. OBJECTIVE: The current study sought to address this gap by examining definitions of resilience in practitioners who work directly with maltreated children. PARTICIPANTS AND SETTING: 27 participants were recruited through two agencies that serve victims of child maltreatment in an urban Midwestern city. METHODS: Through a series of 27 qualitative interviews, the current study examined the following research question: "How is resilience defined and understood by practitioners working with children who have experienced child maltreatment?" Thematic coding and analysis were used to analyze the data. RESULTS: Findings suggest five unique themes described by practitioners as their definition of resilience: (a) surviving; (b) thriving; (c) perseverance; (d) reconciling and integrating traumatic experiences into healthy identity development; and (e) advocating for self. CONCLUSIONS: Our findings highlight the spectral and nuanced nature of resilience among maltreated children. Implications for theory, research and practice are discussed.

The Type and Timing of Child Maltreatment as Predictors of Adolescent Cigarette Smoking Trajectories.

Background: There is limited research examining the association between child maltreatment and cigarette smoking as a specific type of adolescent substance use, and research examining high-risk samples and variations based on maltreatment type and timing remain sparse. Objectives: The primary aim of the study was to examine the relationship between child maltreatment and cigarette smoking trajectories. Methods: Latent class growth analysis and multinomial logistic regression were performed on 903 youth drawn from the Longitudinal Studies of Child Abuse and Neglect (LONGSCAN). Results: Three distinct classes of cigarette smoking trajectories were identified: (1) Stable no/low cigarette use (61%); (2) Gradually increasing cigarette use (30%); and (3) Sharply increasing cigarette use (9%). Physical abuse during early childhood and adolescence predicted membership in the sharply increasing cigarette use class. Neglect during early childhood predicted membership in the gradually increasing cigarette use class. Conclusions: Findings suggest that interventions for adolescent cigarette smoking should integrate trauma-informed approaches. Further, the results highlight early childhood and adolescence as particularly vulnerable periods with respect to the influence of physical abuse and neglect on cigarette smoking, pointing to the need for additional maltreatment prevention efforts during these developmental stages.

Criminogenic Needs as Intervening Factors in the Relation Between Insecure Attachments and Youth Sexual Violence.

There is a strong theoretical and research base demonstrating the link between attachment styles and adolescent sexual offending. However, this relationship may be best explained by deficit-based mediational pathways including criminogenic needs such as emotional or affect regulation and callousness. Grady, Levenson, and Bolder propose a framework that details criminogenic needs as intermediary variables in the attachment-sexual offending relationship. Using data on adolescents adjudicated of sexual and nonsexual crimes in a Western state (N = 200), two structural equation models (SEM) tested direct and indirect relationships between ambivalent and avoidant attachment styles (in separate models), dysregulation including cognitive and behavioral transitions, emotional control, and inhibited/impulsive behaviors, callousness, delinquency, and offending type (sexual or nonsexual offending). Results revealed statistically significant direct pathways between variables of interest and a multimediational effect of dysregulation and callousness in the relationship between insecure attachments and sexual offending. Treatment, policy, and research implications are discussed.

Child protective services decision-making: The role of children's race and county factors.

The current study investigates the role of race and county characteristics in substantiation and out-of-home placement decisions in the United States. Using multilevel models, we analyzed data from counties in the United States available through the National Child Abuse and Neglect Data Systems and Adoption and Foster Care Analysis and Reporting System to investigate the interactions between children's race and the context in which they live. Our sample consisted exclusively of children whose cases had been investigated; therefore, we were able to focus on the role played by race and county characteristics in substantiation and out-of-home placement decisions made by Child Protective Services, net of the heightened risk factors (or potential biases) that lead to disparate rates of reporting. Adjusting for state and county of investigation, Black, American Indian/Alaskan Native, and multiracial children were more likely than White (non-Hispanic) children to be substantiated or placed out of home, whereas Asian children were less likely to be substantiated or placed out of home. Notably, differences across groups are far smaller in magnitude when demographic and geographic differences are taken into account. Higher county-level poverty, percentages of Black residents, and juvenile arrest rates were associated with lower odds of substantiation and out-of-home placement among investigated children, whereas an elevated percentage of single-headed households was associated with higher odds of both outcomes. We also found that living in a rural county was associated with greater odds of substantiation but lower odds of out-of-home placement. Important differences by race were found for these associations. (PsycINFO Database Record (c) 2020 APA, all rights reserved).

The cryo-EM structure of African swine fever virus unravels a unique architecture comprising two icosahedral protein capsids and two lipoprotein membranes.

African swine fever virus (ASFV) is a complex nucleocytoplasmic large DNA virus (NCLDV) that causes a devastating swine disease currently present in many countries of Africa, Europe, and Asia. Despite intense research efforts, relevant gaps in the architecture of the infectious virus particle remain. Here, we used single-particle cryo-EM to analyze the three-dimensional structure of the mature ASFV particle. Our results show that the ASFV virion, with a radial diameter of ∼2,080 Å, encloses a genome-containing nucleoid surrounded by two distinct icosahedral protein capsids and two lipoprotein membranes. The outer capsid forms a hexagonal lattice (triangulation number T = 277) composed of 8,280 copies of the double jelly-roll major capsid protein (MCP) p72, arranged in trimers displaying a pseudo-hexameric morphology, and of 60 copies of a penton protein at the vertices. The inner protein layer, organized as a T = 19 capsid, confines the core shell, and it is composed of the mature products derived from the ASFV polyproteins pp220 and pp62. Also, an icosahedral membrane lies between the two protein layers, whereas a pleomorphic envelope wraps the outer capsid. This high-level organization confers to ASFV a unique architecture among the NCLDVs that likely reflects the complexity of its infection process and may help explain current challenges in controlling it.

Trauma and Adolescents Who Engage in Sexually Abusive Behavior: A Review of the Literature.

While recent research has established the abuse and victimization histories of youth who engage in sexually abusive behavior, there is still debate regarding the significance of trauma and abuse as developmental antecedents to sexual offending in adolescence. Moreover, research has not conclusively identified the types, context, and timing of adverse experiences that act as catalysts for subsequent sexual perpetration. The present comprehensive literature review has one primary aim: Explore the adverse childhood experiences of youth who engage in sexually abusive behavior, considering whether trauma may be a consistent developmental antecedent to offending. Search method, inclusion criteria, and screening protocol are described, and the articles included in the review (N = 13) are qualitatively analyzed using thematic coding. Findings indicate that traumatic experiences and symptoms may be of developmental and etiological significance for subsequent sexually abusive behavior in adolescence. Implications for research, treatment, and policy are discussed, with the hope the findings will act as impetus to develop and deliver targeted treatment and prevention programs to sexually abusive youth who have histories of victimization.

A Formative Qualitative Evaluation to Inform Implementation of a Research Participation Enhancement and Advocacy Training Program for Diverse Seniors: The DREAMS Program.

This article reports on a formative qualitative evaluation conducted to identify potential barriers and facilitators prior to implementation of a two-part educational intervention designed to increase older adults' participation in clinical research. We conducted focus groups with 35 older adults from diverse racial, ethnic, and socioeconomic backgrounds (Mean age = 74.5 ± 7.4 years), and six clinical researchers involved in aging research. Two older adult patient stakeholder advisors participated as members of the research team. Findings from thematic analysis showed that common barriers to research participation included older adults' fear and mistrust of researchers and the medical community as well as misperceptions and negative attitudes about clinical research. A commonly held belief was that most clinical research is conducted by "greedy" pharmaceutical companies who are more interested in profits than finding cures for disease. Results provide useful insights regarding enrolling members of hard-to-reach older adult populations in clinical research.

Abuse disclosures of youth with problem sexualized behaviors and trauma symptomology.

BACKGROUND: The majority of youth with problem sexualized behaviors (PSB) have substantiated experiences of abuse or exposures to violence (Silovsky & Niec, 2002). Little is known about specific abuse experiences that may differentiate youth with PSB from those without. Few studies have examined the types of abuse associated with post-traumatic stress symptomology. OBJECTIVE: The current study explored two research questions: (1) Do children with PSB differ from children without PSB in terms of their abuse disclosures?; and (2) Are the types of abuse disclosed associated with the child's scores on a post-traumatic stress measure?. PARTICIPANTS & SETTING: Data were analyzed for youth (N = 950) ages 3-18 years who completed a clinical assessment at a child advocacy center in the Midwest during the 2015 calendar year. METHODS: Youth completed assessments that included a forensic interview and either the Trauma Symptom Checklist for Young Children (TSCYC) for children ages 3-10 years, or the Trauma Symptom Checklist for Children (TSCC) for children ages 11-16 years. Bivariate logistic regression was used to answer the research questions. RESULTS: Findings indicated that youths who disclosed offender to victim fondling were less likely to disclose PSB (OR = 0.460, p = .026), and children exposed to pornography were more likely to disclose PSB (OR = 3.252, p = .001). Additionally, youth who disclosed physical abuse (OR = 1.678, p = .001) or victim to offender sexual contact (OR = 2.242, p = .003) had higher odds of clinically significant trauma scores. CONCLUSIONS: Implications for practitioners and future research directions are discussed.

Maternal Caregiving Practices and Child Abuse Experiences as Developmental Antecedents to Insecure Attachments: Differential Pathways Between Adolescents Who Commit Sexual and Non-Sexual Crimes.

Grady, Levenson, and Bolder's etiological framework contextualizes sexual offending behaviors through a trauma and attachment lens to suggest child abuse experiences (CAE) are associated with attachment problems that contribute to sexual offending. To empirically test a portion of this theory, this cross-sectional study employed a structural equation mediated-moderated model on a sample of juvenile justice-involved youth (N = 505) who committed sexual (n = 355) and non-sexual (n = 150) offenses. Results revealed a good model fit and statistically significant direct effects between maternal harsh and indifferent caregiving styles and insecure attachment. However, CEAs did not mediate the relationship between caregiving styles and insecure attachment. Results also revealed a moderation effect whereby youth who commit sexual crimes who report harsher and more indifferent maternal caregiving styles were more likely to have higher CAEs relative to youth who commit non-sexual crimes. Implications are discussed using a public health perspective of prevention.

Biological Applications at the Cutting Edge of Cryo-Electron Microscopy.

Cryo-electron microscopy (cryo-EM) is a powerful tool for macromolecular to near-atomic resolution structure determination in the biological sciences. The specimen is maintained in a near-native environment within a thin film of vitreous ice and imaged in a transmission electron microscope. The images can then be processed by a number of computational methods to produce three-dimensional information. Recent advances in sample preparation, imaging, and data processing have led to tremendous growth in the field of cryo-EM by providing higher resolution structures and the ability to investigate macromolecules within the context of the cell. Here, we review developments in sample preparation methods and substrates, detectors, phase plates, and cryo-correlative light and electron microscopy that have contributed to this expansion. We also have included specific biological applications.

The Morphology and Assembly of Respiratory Syncytial Virus Revealed by Cryo-Electron Tomography.

Human respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract disease in young children. With repeat infections throughout life, it can also cause substantial disease in the elderly and in adults with compromised cardiac, pulmonary and immune systems. RSV is a pleomorphic enveloped RNA virus in the Pneumoviridae family. Recently, the three-dimensional (3D) structure of purified RSV particles has been elucidated, revealing three distinct morphological categories: spherical, asymmetric, and filamentous. However, the native 3D structure of RSV particles associated with or released from infected cells has yet to be investigated. In this study, we have established an optimized system for studying RSV structure by imaging RSV-infected cells on transmission electron microscopy (TEM) grids by cryo-electron tomography (cryo-ET). Our results demonstrate that RSV is filamentous across several virus strains and cell lines by cryo-ET, cryo-immuno EM, and thin section TEM techniques. The viral filament length varies from 0.5 to 12 µm and the average filament diameter is approximately 130 nm. Taking advantage of the whole cell tomography technique, we have resolved various stages of RSV assembly. Collectively, our results can facilitate the understanding of viral morphogenesis in RSV and other pleomorphic enveloped viruses.

Promotion of virus assembly and organization by the measles virus matrix protein.

Measles virus (MeV) remains a major human pathogen, but there are presently no licensed antivirals to treat MeV or other paramyxoviruses. Here, we use cryo-electron tomography (cryo-ET) to elucidate the principles governing paramyxovirus assembly in MeV-infected human cells. The three-dimensional (3D) arrangement of the MeV structural proteins including the surface glycoproteins (F and H), matrix protein (M), and the ribonucleoprotein complex (RNP) are characterized at stages of virus assembly and budding, and in released virus particles. The M protein is observed as an organized two-dimensional (2D) paracrystalline array associated with the membrane. A two-layered F-M lattice is revealed suggesting that interactions between F and M may coordinate processes essential for MeV assembly. The RNP complex remains associated with and in close proximity to the M lattice. In this model, the M lattice facilitates the well-ordered incorporation and concentration of the surface glycoproteins and the RNP at sites of virus assembly.