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Silica encapsulation under ambient conditions is commonly used to shield protein-based nanosystems from chemical stress. However, encapsulation-induced photo- and structural instabilities at elevated temperatures have been overlooked. Using bovine serum albumin-capped fluorescent gold nanoclusters (BSA-AuNCs) as a model, we demonstrated that chaperone/polymer layer-by-layer complexation can stabilize the template to resist encapsulation-induced fragmentation/reorganization and emission increases at 37 °C or higher temperatures. We first wrapped BSA-AuNCs with α-crystallin chaperones (α-Crys) to gain the highest thermal stability at a 1:50 molar ratio and then enfolded BSA-AuNC/α-Crys with thermoresponsive poly- N -isopropylacrylamide (PNIPAM) at 60 °C to shield silica interaction and increase the chaperone-client protein accessibility. The resulting BSA-AuNC/α-Crys/PNIPAM (BαP) was encapsulated by a sol-gel process to yield BαP-Si (∼80 ± 4.5 nm), which exhibited excellent structural integrity and photostability against chemical and thermal stresses. Moreover, targeted BαP-Si demonstrated prolonged fluorescence stability for cancer cell imaging. This template stabilization strategy for silica encapsulation is biocompatible and applicable to other protein-based nanosystems.

Background: Globally, women 15-24 years are at heightened risk of sexual violence victimization, a risk factor for adverse mental, physical, and behavioral health outcomes. Sexual violence is common at universities and most often perpetrated by men, yet few evidence-based prevention strategies targeting men have been tested in low- and middle-income countries. GlobalConsent is a six-module, web-based educational program adapted from an efficacious U.S.-based program. Nine months post-treatment in a randomized trial in Vietnam, GlobalConsent reduced men's sexually violent behavior (Odds Ratio [OR] = 0.71, 95%CI 0.50-1.00) and increased prosocial intervening behavior (OR = 1.51, 1.00-2.28) relative to an attention-control. Evidence regarding optimal implementation strategies for scale up is needed., Methods: We will randomize six medical universities in North, Central, and South Vietnam to deliver GlobalConsent using two different packages of implementation strategies that vary in intensity. Higher-intensity strategies will include greater 1) pre- and post-implementation engagement with university leaders and faculty, and 2) greater pre-implementation outreach, follow-up, and incentives for students to promote engagement and completion of GlobalConsent. Higher intensity universities will receive additional training and support for their added activities. We will compare implementation drivers and outcomes, intervention effectiveness, and cost-effectiveness across the two implementation bundles. Our mixed-methods comparative interrupted time-series design includes 1) qualitative interviews and quantitative surveys with university leaders and implementation teams to assess implementation barriers and facilitators; 2) repeated surveys with leaders and faculty, implementation teams, and male students to assess multilevel implementation drivers and outcomes, 3) repeated surveys with male students to assess behavioral outcomes (sexual violence and intervening behavior) and mediating variables (knowledge, attitudes, affect, and capacities), and 4) time diaries and cost tracking to assess cost effectiveness of the two implementation-strategies bundles., Discussion: This project is the first to assess packages of implementation strategies to deliver an efficacious web-based sexual-violence-prevention program for undergraduate men across all regions of Vietnam and synergizes with a violence-prevention training initiative (D43TW012188). This approach will produce rigorous evidence about how to disseminate GlobalConsent nationally, which holds promise to reduce gender-based health inequities linked to sexual violence as GlobalConsent is brought to scale., Trial Registration: NCT06443541. Retrospectively registered with clinicaltrials.gov., Competing Interests: Competing interests {28} KMY has submitted a “discovery statement” to the Office of Technology Transfer at Emory University for the development of GlobalConsent. Otherwise, the authors declare that they have no competing interests.

Background: Globally, women 15-24 years are at heightened risk of sexual violence victimization, a risk factor for adverse mental, physical, and behavioral health outcomes. Sexual violence is common at universities and most often perpetrated by men, yet few evidence-based prevention strategies targeting men have been tested in low- and middle-income countries. GlobalConsent is a six-module, web-based educational program adapted from an efficacious U.S.-based program. Nine months post-treatment in a randomized trial in Vietnam, GlobalConsent reduced men's sexually violent behavior (odds ratio [OR] = 0.71, 95%CI 0.50-1.00) and increased prosocial intervening behavior (OR = 1.51, 1.00-2.28) relative to an attention-control. Evidence regarding optimal implementation strategies for scale up is needed., Methods: We will randomize six medical universities in North, Central, and South Vietnam to deliver GlobalConsent using two different packages of implementation strategies that vary in intensity. Higher-intensity strategies will include greater (1) pre- and post-implementation engagement with university leaders and faculty and (2) greater pre-implementation outreach, follow-up, and incentives for students to promote engagement and completion of GlobalConsent. Higher intensity universities will receive additional training and support for their added activities. We will compare implementation drivers and outcomes, intervention effectiveness, and cost-effectiveness across the two implementation bundles. Our mixed-methods comparative interrupted time series design includes (1) qualitative interviews and quantitative surveys with university leaders and implementation teams to assess implementation barriers and facilitators; (2) repeated surveys with leaders and faculty, implementation teams, and male students to assess multilevel implementation drivers and outcomes; (3) repeated surveys with male students to assess behavioral outcomes (sexual violence and intervening behavior) and mediating variables (knowledge, attitudes, affect, and capacities); and (4) time diaries and cost tracking to assess cost-effectiveness of the two implementation-strategies bundles., Discussion: This project is the first to assess packages of implementation strategies to deliver an efficacious web-based sexual violence prevention program for undergraduate men across all regions of Vietnam and synergizes with a violence-prevention training initiative (D43TW012188). This approach will produce rigorous evidence about how to disseminate GlobalConsent nationally, which holds promise to reduce gender-based health inequities linked to sexual violence as GlobalConsent is brought to scale., Trial Registration: NCT06443541. Retrospectively registered with ClinicalTrials.gov. Registered on June 05, 2024., (© 2024. The Author(s).)

Rape myths-false but widely held beliefs that serve to deny and justify sexual aggression-present a major barrier to reporting and prevention of sexual violence in Vietnam and globally. Based on a parent study aimed at reducing sexual violence at two universities in Hanoi, we developed and assessed a contextualized measure of rape myths among young people in Vietnam. Items from previously validated rape myth acceptance (RMA) scales and data from qualitative research informed the development of 50 items, which were administered to Vietnamese 18-24-year-olds (n = 2,756 total, n = 1,798 cisgender women) via an anonymous link in February 2021. We used factor analysis to explore and test factor structure and multi-group factor analysis to assess measurement equivalence across gender. We calculated item-level discrimination and difficulty parameters and visualized information curves using item response theory analysis, informing the development of a short form. Four hypothesized subconstructs identified in the qualitative data emerged as factors: (1) "He didn't mean to"; (2) "She asked for it"; (3) "It wasn't really rape"; and (4) "Rape is a deviant event." A fifth factor, "She didn't protect herself," included four items from formative data. Confirming formative findings and prior literature, cisgender women had lower RMA than cisgender men, particularly on items related to victim-blaming. The Vietnamese Rape Myths Acceptance Scales were internally consistent and equivalent between cisgender men and women, capturing elements specific to the Vietnamese context and providing a tool for campus climate surveys and evaluations of sexual violence prevention programs., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Purpose: To assess the impact of ocular confounding factors on aqueous humor (AH) proteomic and metabolomic analyses for retinal disease characterization., Methods: This study recruited 138 subjects (eyes): 102 with neovascular age-related macular degeneration (nAMD), 18 with diabetic macular edema (DME), and 18 with cataract (control group). AH samples underwent analysis using Olink Target 96 proteomics and Metabolon's metabolomics platform Data analysis included correlation, differential abundance, and gene-set analysis., Results: In total, 756 proteins and 408 metabolites were quantified in AH. Total AH protein concentration was notably higher in nAMD (3.2-fold) and DME (4.1-fold) compared to controls. Pseudophakic eyes showed higher total AH protein concentrations than phakic eyes (e.g., 1.6-fold in nAMD) and a specific protein signature indicative of matrix remodeling. Unexpectedly, pupil-dilating drugs containing phenylephrine/tropicamide increased several AH proteins, notably interleukin-6 (5.4-fold in nAMD). Correcting for these factors revealed functionally relevant protein correlation clusters and disease-relevant, differentially abundant proteins across the groups. Metabolomics analysis, for which the relevance of confounder adjustment was less apparent, suggested insufficiently controlled diabetes and chronic hyperglycemia in the DME group., Conclusions: AH protein concentration, pseudophakia, and pupil dilation with phenylephrine/tropicamide are important confounding factors for AH protein analyses. When these factors are considered, AH analyses can more clearly reveal disease-relevant factors., Translational Relevance: Considering AH protein concentration, lens status, and phenylephrine/tropicamide administration as confounders is crucial for accurate interpretation of AH protein data.

Growing access to technology and media has presented new avenues of influence on youth attitudes and norms regarding sexuality and sexual violence, as well as new technological pathways through which to perpetrate sexual violence. The aim of this research was to understand contextual influences on and needs for scale-up of sexual violence prevention programming in the media-violence context of Vietnam. We conducted 45 interviews with high school teachers (n = 15), university lecturers (n = 15), and affiliates from youth-focused community service organizations (n = 15) from across Vietnam. Additionally, we conducted four sector-specific focus groups with a sub-sample of interview participants (k = 4, n = 22). Media and technology were brought up consistently in relation to sexual violence prevention and sexual health information. Key informants noted that, in Vietnam, generational differences in acceptability of sex and lack of comprehensive sexuality education intersect with new technological opportunities for exposure to sexual information and media. This creates a complex landscape that can promote sexual violence through priming processes, instigate mimicry of violent media, and presents new opportunities for the perpetration of sexual violence though technology. Development of comprehensive sexual education, including violence prevention education, is imperative, with consideration of age-specific needs for Vietnamese youth., (© 2024. The Author(s).)

Background: Sexual violence by young men against women is common, but efficacious primary prevention interventions tailored to men are limited in low- and middle-income settings like Vietnam. GlobalConsent, a web-based sexual violence prevention intervention tailored to university men in Hanoi, is efficacious. Implementation research is needed to understand facilitators and barriers to scaling GlobalConsent and prevention programs generally. We conducted qualitative research with key informants from three youth-focused organizational settings to understand the context of implementation in Vietnam., Methods: Interviews with university (n = 15), high-school (n = 15) and non-governmental (n = 15) key informants focused on perceptions about sexual violence among young people and prevention programming. Four focus group discussions with 22 interviewed informants, following the Consolidated Framework for Implementation Research, asked about facilitators and barriers to implementing GlobalConsent. Narratives were transcribed, translated, and coded inductively and deductively to identify salient themes., Results: Outer-setting influences included greater expectations for sex among young people alongside norms favoring men's sexual privilege, ostensibly ambiguous and lax laws on sexual violence, government ministries as bureaucratic but potential allies, external subject-matter experts, and the media. Inner-setting influences included variable cultures regarding openness to discuss sexual violence and equitable gender norms, variable departmental coordination, limited funding and 'red tape' especially in public institutions, inconsistent student access to technologies, and limited time and competing priorities among students and teachers. Several actors were considered influential, including institutional leaders, human-resource staff, the Youth Union, and student-facing staff. Important characteristics of individuals for implementation included subject-matter expertise, science or social science training, younger age, engagement in social justice related activities, and more open attitudes about sex. Regarding characteristics of sexual violence prevention programming, some participants preferred online formats for busy students while others suggested hybrid or in-person formats, peer education, and incentives. Participants generally accepted the content of GlobalConsent and suggested adding more content for women, ancillary support services, and adapted content for high-school students., Conclusions: Implementation of sexual violence prevention programs in youth-focused organizations in Vietnam requires multilevel strategies that connect outer-setting subject-matter experts with supportive inner-setting leaders and student-facing staff to overcome normative and organizational constraints, and thereby, to deliver institution-wide programming., (© 2023. The Author(s).)

Background: Sexual violence against women is prevalent worldwide. Prevention programs that treat men as allies and integrate a bystander framework are emerging in lower income settings, but evidence of their effectiveness is conflicting., Objective: This study aimed to test the impact of GlobalConsent on sexually violent behavior and prosocial bystander behavior among university men in Vietnam., Methods: We used a double-blind, parallel intervention versus control group design with 1:1 randomization at 2 universities. A total of 793 consenting heterosexual or bisexual men aged 18-24 years who matriculated in September 2019 were enrolled and assigned randomly to GlobalConsent or an attention-control adolescent health education (AHEAD) program. GlobalConsent is an adapted, theory-based, 6-module web-based intervention with diverse behavior change techniques and a locally produced serial drama. AHEAD is a customized, 6-module attention-control program on adolescent health. Both the programs were delivered to computers and smartphones over 12 weeks. Self-reported sexually violent behaviors toward women in the prior 6 months and prosocial bystander behaviors in the prior year were measured at 0, 6, and 12 months., Results: More than 92.7% (735/793) of men in both study arms completed at least 1 program module, and >90.2% (715/793) of men completed all 6 modules. At baseline, a notable percentage of men reported any sexually violent behavior (GlobalConsent: 123/396, 31.1%; AHEAD: 103/397, 25.9%) in the prior 6 months. Among men receiving GlobalConsent, the odds of reporting a high level (at least 2 acts) of sexually violent behavior at the endline were 1.3 times the odds at baseline. Among men receiving AHEAD, the corresponding odds ratio was higher at 2.7. The odds of reporting any bystander behavior at endline were 0.7 times the odds at baseline for GlobalConsent, and the corresponding odds ratio for AHEAD was lower at 0.5., Conclusions: Compared with a health attention-control condition, GlobalConsent has sustained favorable impacts on sexually violent behavior and prosocial bystander behavior among matriculating university men in Vietnam, who would otherwise face increasing risks of sexually violent behavior. GlobalConsent shows promise for national scale-up and regional adaptations., Trial Registration: ClinicalTrials.gov NCT04147455; https://clinicaltrials.gov/ct2/show/NCT04147455., International Registered Report Identifier (irrid): RR2-10.1186/s12889-020-09454-2., (©Kathryn M Yount, Yuk Fai Cheong, Irina Bergenfeld, Quach Thu Trang, Jessica M Sales, Yiman Li, Tran Hung Minh. Originally published in JMIR Public Health and Surveillance (https://publichealth.jmir.org), 27.01.2023.)

Sexual violence remains a global problem that disproportionately affects women. Though sexual violence interventions exist, few have been implemented in low- or middle-income countries, and none in Vietnam for young men. We adapted a sexual violence prevention intervention (RealConsent) developed for college men in the U.S. and conducted a randomized controlled trial of the adapted intervention (GlobalConsent) with college men in Vietnam. We assessed the effects of GlobalConsent on sexually violent behavior and prosocial bystander behavior, directly and through theoretically targeted mediators. The study design entailed a double-blind, parallel intervention-control-group design in two universities. Consenting heterosexual or bisexual men 18-24 years starting university in September 2019 (n = 793) completed a baseline survey and were assigned with 1:1 randomization to GlobalConsent or attention control. Both programs were web-based and lasted 12 weeks. Path analysis was performed to study the mediating effects of cognition/knowledge, beliefs/attitudes, affect, and efficacy/intention variables measured at six months on sexually violent behavior and prosocial bystander behavior measured at 12 months. In parallel multiple-mediator models, initiating GlobalConsent lowered the odds of sexually violent behavior mainly indirectly, via knowledge of sexual violence legality and harm and victim empathy and increased the odds of prosocial bystander behavior directly and indirectly, through knowledge of sexual violence legality and harm and bystander capacities. The efficacious direct and indirect effects of GlobalConsent support the cross-cultural applicability of its underlying theory of change and findings from mediation analyses of its sister program RealConsent, suggesting GlobalConsent's national scalability and adaptability across Southeast Asia., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Adolescence and emerging adulthood represent a period of heightened vulnerability to sexual violence (SV). While some research suggests that exposure to sexually explicit material (SEM) among adolescents and college students is associated with sexually violent behavior, our understanding of this relationship is limited. This study aimed to assess the relationship between prior exposure to several types of SEM and sexually violent behavior in a sample of first-year university men in Vietnam., Methods and Findings: A cohort of 739 first-year male university students completed three survey waves over 14 months, providing information on contact and non-contact sexually violent behavior, exposure to SEM, and other theorized confounders of the SEM-SV relationship. Controlling for these covariates, we estimated the average treatment effect of SEM on contact and non-contact SV using the propensity score method. We also conducted a dose-response analysis for the effect of violent SEM on SV based on frequency-of-exposure classes derived from latent class analysis. The majority of the sample reported exposure to SEM in the prior six months, with 41% of the sample reporting exposure to violent SEM. In propensity-adjusted models, exposure to violent SEM, but not other types, had a small but significant positive effect on contact and non-contact SV. These effects increased for frequent viewers of violent SEM. Models of contact SV showed endogeneity, warranting caution., Conclusions: Exposure to violent SEM is prevalent among university men in Vietnam and may be contributing to sexually violent behavior. Incorporating media literacy into SV prevention programs to mitigate these potential effects may be warranted., Competing Interests: The authors have declared that no competing interests exist.

Sexual violence is a persistent, underreported threat to the well-being of women and girls worldwide. In Vietnam, as elsewhere, myths and misconceptions around rape and other forms of sexual violence present a major barrier to reporting and prevention. Based on qualitative research from a parent study aimed at reducing sexual violence perpetration at universities in Vietnam, we sought to characterize commonly held myths among university students that may contribute to the perpetration and justification of such behaviors. Our analysis drew on focus group discussions with men ( n  = 69) and semi-structured interviews with men ( n  = 12) and women ( n  = 9) recruited from two universities in Hanoi. Conducted in April and May of 2018, interviews covered topics including gender norms, dating relationships, consent, and sexual violence, whereas focus groups conducted in October 2018 were designed to assess reactions to an existing sexual violence prevention program. Thematic analysis of interview and focus group transcripts revealed that myths and misconceptions about sexual violence persist among university-aged men and women. Chief among these were that rape results mainly from men's uncontrollable desire, that victims provoke rape by their "reckless" behavior, such as drinking or dressing provocatively, that "real" rape is characterized by physical force from the perpetrator and "fierce resistance" on the part of the victim, and that rape occurs only under a narrow set of circumstances. Although men and women endorsed most myths equally, justification of rape through victim blaming featured much less in women's narratives than in men's. Myths concerning false rape allegations, observed in Western populations, were not considered relevant to Vietnamese students. Findings informed the development of a contextualized rape myths acceptance scale for testing and use in Vietnam. A nuanced understanding of salient rape myths among male and female students may also inform university-based efforts to prevent sexual violence.

The emergent occurrence of sulfonamide species involving sulfadiazine (SDZ) and sulfamethazine (SMZ) in aquatic systems can cause a wide range of potential risks; hence, remediation strategies need to be necessary. Here, we develop the novel metal-organic framework-derived nanocomposite, and apply for the adsorption of SDZ and SMZ antibiotics. To assess the best-fitting kinetic (pseudo first-order, pseudo second-order) and isotherm (Langmuir, Freundlich, Temkin, Dubinin-Radushkevich, Redlich-Peterson, Sips, Toth, and Khan) models, a series of numerous statistical analysis was performed. Numerous error functions including squares of the errors (SSE), hybrid fractional error function (HYBRID), Marquardt's percent standard deviation (MPSD), and mean relative error (MRE) were also analyzed to assess the linear and nonlinear models. The results indicated that both linear and nonlinear kinetic models were mostly fitted well with pseudo second-order models (R adj ) 2  > 0.97. Although linear kinetics gave better (R adj ) 2 , error functions (MRE, SSE, HYBRID, and MPSD) were mostly higher than those of nonlinear kinetics. For adsorption isotherm, nonlinear Redlich-Peterson was the most compatible model with extremely high adjusted coefficients of determination (R adj ) 2  ~ 1.0000. While nonlinear Langmuir model gave relatively high (R adj ) 2 (0.9898-0.9960) and acceptable error functions, we found the considerable difference of error functions and parameters among four types of linear Langmuir (Types I, II, III, IV). The findings indicate potential errors as selecting one of linearized Langmuir types in equilibrium study. It is suggested that nonlinear models should be applied for better fitness., (© 2020. Springer-Verlag GmbH Germany, part of Springer Nature.)

Among crop fruit trees, the apricot (Prunus armeniaca) provides an excellent model to study divergence and adaptation processes. Here, we obtain nearly 600 Armeniaca apricot genomes and four high-quality assemblies anchored on genetic maps. Chinese and European apricots form two differentiated gene pools with high genetic diversity, resulting from independent domestication events from distinct wild Central Asian populations, and with subsequent gene flow. A relatively low proportion of the genome is affected by selection. Different genomic regions show footprints of selection in European and Chinese cultivated apricots, despite convergent phenotypic traits, with predicted functions in both groups involved in the perennial life cycle, fruit quality and disease resistance. Selection footprints appear more abundant in European apricots, with a hotspot on chromosome 4, while admixture is more pervasive in Chinese cultivated apricots. Our study provides clues to the biology of selected traits and targets for fruit tree research and breeding.

Resistance to Fusarium head blight (FHB) of spelt wheat was investigated in field trials carried out at three European locations between 2016 and 2018. Resistance was assessed after artificial inoculation by visual scoring of symptoms and the determination of the contamination of grains and glumes with the mycotoxin deoxynivalenol (DON). It was found that typical spelt traits such as tall plant height, lax spikes, and tough glumes play a role as passive resistance factors. Across all test environments, modern spelt varieties with a significantly reduced plant height showed a significantly higher susceptibility to FHB and a higher contamination of the grains with DON compared to old landraces/varieties and plant genetic resources. Similarly, the lowest mycotoxin levels in grains were found only in old landraces and varieties, while the highest DON concentration was observed mainly in modern varieties. The results obtained can be used for the selection of suitable parental material for breeding spelt with improved FHB resistance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Chrpová, Grausgruber, Weyermann, Buerstmayr, Palicová, Kozová, Trávníčková, Nguyen, Moreno Amores, Buerstmayr and Janovská.)

Catechol estrogens (CEs) are known to be toxic metabolites and the initiators of the oncogenesis of breast cancers via forming covalent adducts with DNAs. CEs shall also react with proteins, but their cellular protein targets remain unexplored. Here, we reported the identification of protein targets of CEs in the soluble cytosol of estrogen-sensitive breast cancer cells by multiple comparative proteomics using liquid chromatography-tandem mass spectrometry (LC-MS/MS) coupled with an improved click chemistry-based workflow. Multiple comparative proteomics composed of an experimental pair (probe versus solvent) and two control pairs (solvent versus solvent and probe versus solvent without enrichment) were studied using stable isotope dimethyl labeling. The use of 4-hydroxyethynylestradiol (4OHEE2) probe with an amide-free linker coupled with on-bead digestion and redigestion of the proteins cleaved from the beads was shown to greatly improve the recovery and identification of CE-adducted peptides. A total of 310 protein targets and 40 adduction sites were repeatedly ( n ≥ 2) identified with D/H (probe/solvent) ratio >4 versus only one identified with D/H >4 from the two control pairs, suggesting that our workflow imposes only a very low background. Meanwhile, multiple comparative D/H ratios revealed that CEs may downregulate many target proteins involved in the metabolism or detoxification, suggesting a negative correlation between CE-induced adduction and expression of proteins acting on the alleviation of stress-induced cellular damages. The reported method and data will provide opportunities to study the progression of estrogen metabolism-derived diseases and biomarkers.

Background: Sexual violence-any sexual act committed against a person without freely given consent-disproportionately affects women. Women's first experiences of sexual violence often occur in adolescence. In Asia and the Pacific, 14% of sexually experienced adolescent girls report forced sexual debut. Early prevention with men that integrates a bystander framework is one way to address attitudes and behavior while reducing potential resistance to participation., Methods: This paper describes a study protocol to adapt RealConsent for use in Vietnam and to test the impact of the adapted program-GlobalConsent-on cognitive/attitudinal/affective mediators, and in turn, on sexual violence perpetration and prosocial bystander behavior. RealConsent is a six-session, web-based educational entertainment program designed to prevent sexual violence perpetration and to enhance prosocial bystander behavior in young men. The program has reduced the incidence of sexual violence among men attending an urban, public university in the Southeastern United States. We used formative qualitative research and the Centers for Disease Control and Prevention's Map of the Adaptation Process to adapt RealConsent. We conducted semi-structured interviews with college men (n = 12) and women (n = 9) to understand the social context of sexual violence. We conducted focus group discussions with university men and stakeholders (n = 14) to elicit feedback on the original program. From these data, we created scripts in storyboard format of the adapted program. We worked closely with a small group of university men to elicit feedback on the storyboards and to refine them for acceptability and production. We are testing the final program-GlobalConsent-in a randomized controlled trial in heterosexual or bisexual freshmen men 18-24 years attending two universities in Hanoi. We are testing the impact of GlobalConsent (n = 400 planned), relative to a health-education attention control condition we developed (n = 400 planned), on cognitive/attitudinal/affective mediators, prosocial bystander behavior, and sexual violence perpetration., Discussion: This project is the first to test the impact of an adapted, theoretically grounded, web-based educational entertainment program to prevent sexual violence perpetration and to promote prosocial bystander behavior among young men in a middle-income country. If effective, GlobalConsent will have exceptional potential to prevent men's sexual violence against women globally., Trial Registration: U.S. National Library of Medicine Clinicaltrials.gov NCT04147455 on November 1, 2019 (Version 1). Retrospectively registered. Protocol amendments will be submitted to clinicaltrials.gov .

Abundant blood proteins adducted by active electrophiles are excellent markers to predict the risk of electrophile-induced toxicity. However, detecting endogenously adducted proteins by bottom-up selective (or parallel) reaction monitoring (SRM/PRM) is challenging because of the high variability in sample preparation and detection as well as low adduction levels. Here, we reported a new approach in developing PRM methods by combining intact protein measurement with standard additions to target optimal conditions for detecting catechol estrogens (CEs)-adducted human serum albumin (HSA). Blood serum was added with multiple amounts of CEs to obtain serum standards. Intact protein measurement revealed two linear ranges of adduction levels (adducted-CE/HSA): 0.34-0.42 ( R 2 > 0.94) and 0.81-8.54 ( R 2 > 0.96) against the amount of added CEs, respectively. Six adduction sites were identified by trypsin (K20, C34, K73, K281, H338, K378) or chymotrypsin (K20, C34, K378) digestion. PRM methods targeting all adducted/nonadducted peptide pairs based on chymotrypsin or trypsin digestion were developed, and the data were compared with those obtained by intact protein measurement. Correlation plots indicated that chymotrypsin-PRM leads to poor sensitivity and largely underestimated protein adduction levels. Trypsin-PRM leads to sensitive and highly correlated ( R 2 > 0.91) protein adduction levels with a detection limit below the endogenous level and relative standard deviation <25%. As a proof of concept, clinical serum samples were examined by trypsin-PRM, and a slightly higher adduction level was observed for the obesity group when compared with the healthy group. This is the first report on determining adduction levels of blood proteins for long-term exposure to CEs. The standard addition approach can be generally applied to protein adductomics with resolvable mass increments by intact protein measurement to accelerate the development of bottom-up methods close to the inherent limit.

Catechol estrogens (CEs) are metabolic electrophiles that actively undergo covalent interaction with cellular proteins, influencing molecular function. There is no feasible method to identify their binders in a living system. Herein, we developed a click chemistry-based approach using ethinylestradiol (EE2) as the precursor probe coupled with quantitative proteomics to identify protein targets of CEs and classify their binding strengths. Using in situ metabolic conversion and click reaction in liver microsomes, CEs-protein complex was captured by the probe, digested by trypsin, stable isotope labeled via reductive amination, and analyzed by liquid chromatography-mass spectrometry (LC-MS). A total of 334 liver proteins were repeatedly identified ( n ≥ 2); 274 identified proteins were classified as strong binders based on precursor mass mapping. The binding strength was further scaled by D/H ratio (activity probe/solvent): 259 strong binders had D/H > 5.25; 46 weak binders had 5.25 > D/H > 1; 5 nonspecific binders (keratins) had D/H < 1. These results were confirmed using spiked covalent control (strong binder) and noncovalent control (weak binder), as well as in vitro testing of cytochrome c (D/H = 5.9), which showed covalent conjugation with CEs. Many identified strong binders, such as glutathione transferase, catechol-O-methyl transferase, superoxide dismutase, catalase, glutathione peroxidase, and cytochrome c, are involved in cellular redox processes or detoxification activities. CE conjugation was shown to suppress the superoxide oxidase activity of cytochrome c, suggesting that CEs modification may alter the redox action of cellular proteins. Due to structural similarity and inert alkyne group, EE2 probe is very likely to capture protein targets of CEs in general. Thus, this strategy can be adopted to explore the biological impact of CEs modification in living systems.

Interactions of plant retrotransposons with different steps of biotic and abiotic stress-associated signaling cascades are still poorly understood. We perform here a finely tuned comparison of four tobacco retrotransposons (Tnt1, Tnt2, Queenti, and Tto1) responses to the plant elicitor cryptogein. We demonstrate that basal transcript levels in cell suspensions and plant leaves as well as the activation during the steps of defense signaling events are specific to each retrotransposon. Using antisense NtrbohD lines, we show that NtrbohD-dependent reactive oxygen species (ROS) production might act as negative regulator of retrotransposon activation., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

This article describes an action research project designed to engage women, health providers, and communities to respond to gender-based violence (GBV) in Vietnam. Based on results from in-depth interviews and group discussions, it considers the extent to which the project approaches were empowering for abused women. The results underscore the problems entailed in introducing systematic screening for gender-based violence into government health facilities in the low-resource setting of Vietnam, the importance of combining ideational change and rights components with support for abused women, and the difficulty of engaging male perpetrators.

The Arf1 exchange factor GBF1 (Golgi Brefeldin A resistance factor 1) and its effector COPI are required for delivery of ATGL (adipose triglyceride lipase) to lipid droplets (LDs). Using yeast two hybrid, co-immunoprecipitation in mammalian cells and direct protein binding approaches, we report here that GBF1 and ATGL interact directly and in cells, through multiple contact sites on each protein. The C-terminal region of ATGL interacts with N-terminal domains of GBF1, including the catalytic Sec7 domain, but not with full-length GBF1 or its entire N-terminus. The N-terminal lipase domain of ATGL (called the patatin domain) interacts with two C-terminal domains of GBF1, HDS (Homology downstream of Sec7) 1 and HDS2. These two domains of GBF1 localize to lipid droplets when expressed alone in cells, but not to the Golgi, unlike the full-length GBF1 protein, which localizes to both. We suggest that interaction of GBF1 with ATGL may be involved in the membrane trafficking pathway mediated by GBF1, Arf1 and COPI that contributes to the localization of ATGL to lipid droplets.

The Sec7 domain ADP-ribosylation factor (Arf) guanine nucleotide exchange factors (GEFs) are found in all eukaryotes, and are involved in membrane remodeling processes throughout the cell. This review is focused on members of the GBF/Gea and BIG/Sec7 subfamilies of Arf GEFs, all of which use the class I Arf proteins (Arf1-3) as substrates, and play a fundamental role in trafficking in the endoplasmic reticulum (ER)-Golgi and endosomal membrane systems. Members of the GBF/Gea and BIG/Sec7 subfamilies are large proteins on the order of 200 kDa, and they possess multiple homology domains. Phylogenetic analyses indicate that both of these subfamilies of Arf GEFs have members in at least five out of the six eukaryotic supergroups, and hence were likely present very early in eukaryotic evolution. The homology domains of the large Arf1 GEFs play important functional roles, and are involved in interactions with numerous protein partners. The large Arf1 GEFs have been implicated in several human diseases. They are crucial host factors for the replication of several viral pathogens, including poliovirus, coxsackievirus, mouse hepatitis coronavirus, and hepatitis C virus. Mutations in the BIG2 Arf1 GEF have been linked to autosomal recessive periventricular heterotopia, a disorder of neuronal migration that leads to severe malformation of the cerebral cortex. Understanding the roles of the Arf1 GEFs in membrane dynamics is crucial to a full understanding of trafficking in the secretory and endosomal pathways, which in turn will provide essential insights into human diseases that arise from misregulation of these pathways.

Mariner-like elements (MLEs) are class-II transposable elements that move within the genome of their hosts by means of a DNA-mediated "cut and paste" mechanism. MLEs have been identified in several organisms, from most of the phyla. Nevertheless, only a few of the sequences characterized contain an intact open reading frame. Investigation of the genome of a coastal crab, Pachygrapsus marmoratus, has identified nine Pacmmar elements, two of which have an open reading frame encoding a putatively functional transposase. Nucleic acid analyses and comparison with the previous data showed that the GC contents of MLEs derived from coastal organisms such as P. marmoratus are significantly higher than those of terrestrial MLEs and significantly lower than those of hydrothermal ones. Furthermore, molecular phylogeny analyses have shown that Pacmmar elements constitute a new lineage of the irritans subfamily within the mariner family.

The fungal pathogen Candida albicans is frequently associated with catheter-based infections because of its ability to form resilient biofilms. Prior studies have shown that the transcription factor Bcr1 governs biofilm formation in an in vitro catheter model. However, the mechanistic role of the Bcr1 pathway and its relationship to biofilm formation in vivo are unknown. Our studies of biofilm formation in vitro indicate that the surface protein Als3, a known adhesin, is a key target under Bcr1 control. We show that an als3/als3 mutant is biofilm-defective in vitro, and that ALS3 overexpression rescues the biofilm defect of the bcr1/bcr1 mutant. We extend these findings with an in vivo venous catheter model. The bcr1/bcr1 mutant is unable to populate the catheter surface, though its virulence suggests that it has no growth defect in vivo. ALS3 overexpression rescues the bcr1/bcr1 biofilm defect in vivo, thus arguing that Als3 is a pivotal Bcr1 target in this setting. Surprisingly, the als3/als3 mutant forms a biofilm in vivo, and we suggest that additional Bcr1 targets compensate for the Als3 defect in vivo. Indeed, overexpression of Bcr1 targets ALS1, ECE1, and HWP1 partially restores biofilm formation in a bcr1/bcr1 mutant background in vitro, though these genes are not required for biofilm formation in vitro. Our findings demonstrate that the Bcr1 pathway functions in vivo to promote biofilm formation, and that Als3-mediated adherence is a fundamental property under Bcr1 control. Known adhesins Als1 and Hwp1 also contribute to biofilm formation, as does the novel protein Ece1.

We examined the relationship between host survival and renal and splenic immune responses in a murine model of hematogenously disseminated candidiasis. Male BALB/c mice were infected via tail vein injection with wild-type C. albicans or with an isogenic, Deltaefg1/Deltaefg1 hypha-deficient mutant. Host survival, organ fungal burden, intracellular cytokine content of splenic and kidney lymphocytes, and whole-organ cytokine profiles were determined. Wild-type C. albicans induced type 2 splenocyte responses with both nonfatal and fatal inocula. In the kidney, conversely, wild-type inocula causing no or low mortality induced type 1 responses and 100% fatal inocula induced type 2 or interleukin-10 (IL-10)-dominant responses. Hypha-deficient mutant C. albicans caused no or low mortality while inducing type 1 responses in both the spleen and kidney. To our knowledge, this is the first demonstration that host survival during systemic infection correlates with the type of immune response engendered in a nonlymphoid, parenchymal organ and not with the response in the spleen. Furthermore, the results provide in vivo confirmation that hyphal formation by C. albicans induces type 2 or IL-10-dominant host responses in tissues.

The detection of the single base pair mutations at codon 6 of the beta-globin gene is important for the prenatal diagnosis of sickle-cell anaemia and SC disease. A novel procedure has been designed to create a restriction site at both the beta A and beta C alleles to facilitate the discrimination of haemoglobins A, S and C. The general principle of this procedure is to enzymatically amplify genomic DNA using a modified primer containing an altered 3'-terminal nucleotide to create these restriction sites. After this modified primer has been efficiently incorporated into amplified DNA, the PCR products are digested with the restriction enzymes Ava I and Sty I. Ava I recognizes a site in amplified DNA containing a beta A allele, and Sty I recognizes a site in DNA containing a beta C allele. Since the beta A and beta C alleles can be distinguished directly by the presence of a restriction site, the beta S allele can be identified indirectly. All three beta-globin alleles are easily distinguished by size and pattern of electrophoresed fragments on agarose gels. This procedure is specific and sensitive, thus permitting rapid, economical diagnosis of sickle-cell anaemia and SC disease.