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1.
Trauma Violence Abuse ; : 15248380241253045, 2024 May 28.
Article in English | MEDLINE | ID: mdl-38804688

ABSTRACT

This scoping review aims to summarize current research on help-seeking behaviors and patterns among children who have experienced commercial sexual exploitation of children (CSEC) victimization and/or are at high risk of exploitation. Because the literature on help-seeking specific to CSEC victimization is limited, the current review was expanded to summarize findings from help-seeking research for children who have experienced harms that often co-occur with CSEC, such as maltreatment and homelessness. The authors searched three large article databases (PsycInfo, Ovid MEDLINE, and Web of Science) for articles that were (a) empirical; (b) measured or used the term "help-seeking" in their framing and/or results; (c) included children between the ages of 12 and 18; (d) primarily focused on children from the United States; (e) published in English. Ultimately, 22 articles met all inclusion criteria and were included in the final review. Results suggest that cisgender white females are the most likely to seek help, followed by sexual and gender minority children. Cisgender male children were the least likely to engage in help-seeking behaviors. Universally, children were more likely to engage in informal help-seeking rather than formal help-seeking, with younger children being more likely to reach out to parents and older children more likely to reach out to friends. Importantly, ethnically diverse children were under-represented, leading to limited information about how and why these children seek help. Implications for help-seeking by children impacted by commercial sexual exploitation are discussed, providing guidance for programming and research related to CSEC prevention.

2.
BMC Psychiatry ; 23(1): 940, 2023 12 13.
Article in English | MEDLINE | ID: mdl-38093256

ABSTRACT

BACKGROUND: The current study aims to better understand the mental health and subjective well-being of investigators and forensic examiners exposed to child sexual abuse material (CSAM) by examining which components of this work are associated with elevated mental health conditions and decreased well-being, as well as the intra-personal and organizational variables that may mitigate harm and improve well-being. METHODS: Police investigators, forensic examiners, and others connected with the criminal justice system from across the United States who were exposed to CSAM as part of their professions (N = 500) completed an anonymous online survey. Participants were recruited through connections with the National Criminal Justice Training Center. RESULTS: Duration, frequency, amount, and content of CSAM exposure was not related to poorer mental health with the exception of exposure to violent CSAM which was related to elevated post-traumatic stress symptoms. Several agency-level practices and policies, such as the availability of an Officer Wellness Program and more frequently knowing the final case resolution, were related to better mental health and well-being. Harm mitigation strategies, such as talking to other officers investigating the case and taking breaks from the material being viewed, were also related to better mental health. CONCLUSIONS: Findings indicate that police agencies have options for implementing agency-level procedures and practices that have the potential to reduce the negative impact of CSAM investigations. Additionally, many investigators use strategies that are correlated with greater well-being, suggesting opportunities for improving training programs.


Subject(s)
Child Abuse, Sexual , Child Abuse , Child , Humans , United States , Mental Health , Police/psychology , Criminal Law
3.
J Am Soc Mass Spectrom ; 34(12): 2864-2867, 2023 Dec 06.
Article in English | MEDLINE | ID: mdl-37971787

ABSTRACT

Hydroxyl radical protein footprinting (HRPF) is a mass-spectrometry-based method for studying protein structures, interactions, conformations, and folding. This method is based on the irreversible labeling of solvent-exposed amino acid side chains by hydroxyl radicals. While catalase is commonly used as a quencher after the labeling of a protein by the hydroxyl radicals to efficiently remove the remaining hydrogen peroxide, it has some disadvantages. Catalase quenching adds a relatively high amount of protein to the sample, limiting the sensitivity of the method due to dynamic range issues and causing significant issues when dealing with more complex samples. We evaluated dimethylthiourea (DMTU) as a replacement for catalase in the quenching HRPF reactions. We observed that DMTU is highly effective at quenching HRPF oxidation. DMTU does not cause the background protein issues that catalase does, resulting in an increased number of protein identifications from complex mixtures. We recommend the replacement of catalase quenching with DMTU for all HRPF experiments.


Subject(s)
Hydroxyl Radical , Protein Footprinting , Hydroxyl Radical/chemistry , Catalase , Protein Footprinting/methods , Proteins/chemistry , Oxidation-Reduction
4.
Anal Chem ; 95(44): 16131-16137, 2023 11 07.
Article in English | MEDLINE | ID: mdl-37878603

ABSTRACT

Monitoring protein structure before and after environmental alterations (e.g., different cell states) can give insights into the role and function of proteins. Fast photochemical oxidation of proteins (FPOP) coupled with mass spectrometry (MS) allows for monitoring of structural rearrangements by exposing proteins to OH radicals that oxidize solvent-accessible residues, indicating protein regions undergoing movement. Some of the benefits of FPOP include high throughput and a lack of scrambling due to label irreversibility. However, the challenges of processing FPOP data have thus far limited its proteome-scale uses. Here, we present a computational workflow for fast and sensitive analysis of FPOP data sets. Our workflow, implemented as part of the FragPipe computational platform, combines the speed of the MSFragger search with a unique hybrid search method to restrict the large search space of FPOP modifications. Together, these features enable more than 10-fold faster FPOP searches that identify 150% more modified peptide spectra than previous methods. We hope this new workflow will increase the accessibility of FPOP to enable more protein structure and function relationships to be explored.


Subject(s)
Peptides , Proteome , Mass Spectrometry/methods , Solvents , Oxidation-Reduction
5.
Neurourol Urodyn ; 42(8): 1769-1776, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37614056

ABSTRACT

BACKGROUND: Despite effective treatments for fecal incontinence (FI), FI remains underdiagnosed and undertreated. This study sought to characterize and compare rates of, delays in, experiences with, and barriers to care seeking for FI among men and women. METHODS: This study was a secondary analysis of electronic survey data collected from adults with FI. The survey included the use of a validated instrument to assess FI severity and questions that now comprise a validated instrument for assessing barriers to care seeking for FI in women. Descriptive analyses characterized differences between men and women. χ2 testing for categorical variables and t-testing for normally distributed continuous variables compared statistical differences. RESULTS: The sample (N = 548) was predominately female (84%), non-Hispanic White (90%), and insured (96%), with a mean Vaizey score (13.4 ± 5.3) consistent with moderate or severe FI. Care seeking rates (p = 0.81) and symptom duration before care seeking (p = 0.23) did not differ between women and men, but women were more likely than men to be told that effective treatments exist; most male and female respondents who sought care were offered treatment. Very few respondents had been asked about FI by a healthcare provider. Whereas normative thinking, limited life impact, and believing that a healthcare provider could not help were more common barriers to care seeking among men, avoidance, fear, and discouragement were more common in women. CONCLUSIONS: Men and women with FI seek care at similar rates and after experiencing symptoms for a similar duration of time. Very few patients with FI have been screened for it by a healthcare provider. Barriers to FI care seeking are different for women and men, and men are less likely than women to be informed about effective treatments by a healthcare provider.


Subject(s)
Fecal Incontinence , Adult , Humans , Male , Female , Fecal Incontinence/diagnosis , Fecal Incontinence/therapy , Surveys and Questionnaires , Patient Acceptance of Health Care , Health Services Accessibility , Quality of Life
6.
bioRxiv ; 2023 Jun 05.
Article in English | MEDLINE | ID: mdl-37333157

ABSTRACT

Monitoring protein structure before and after perturbations can give insights into the role and function of proteins. Fast photochemical oxidation of proteins (FPOP) coupled with mass spectrometry (MS) allows monitoring of structural rearrangements by exposing proteins to OH radicals that oxidize solvent accessible residues, indicating protein regions undergoing movement. Some of the benefits of FPOP include high throughput and lack of scrambling due to label irreversibility. However, the challenges of processing FPOP data have thus far limited its proteome-scale uses. Here, we present a computational workflow for fast and sensitive analysis of FPOP datasets. Our workflow combines the speed of MSFragger search with a unique hybrid search method to restrict the large search space of FPOP modifications. Together, these features enable more than 10-fold faster FPOP searches that identify 50% more modified peptide spectra than previous methods. We hope this new workflow will increase the accessibility of FPOP to enable more protein structure and function relationships to be explored.

7.
Lancet Psychiatry ; 10(3): 209-219, 2023 03.
Article in English | MEDLINE | ID: mdl-36804072

ABSTRACT

BACKGROUND: The antipsychotic, clozapine, is the only licensed drug against the treatment-resistant symptoms that affect 20-30% of people with schizophrenia. Clozapine is markedly underprescribed, partly because of concerns about its narrow therapeutic range and adverse drug reaction profile. Both concerns are linked to drug metabolism, which varies across populations globally and is partly genetically determined. Our study aimed to use a cross-ancestry genome-wide association study (GWAS) design to investigate variations in clozapine metabolism within and between genetically inferred ancestral backgrounds, to discover genomic associations to clozapine plasma concentrations, and to assess the effects of pharmacogenomic predictors across different ancestries. METHODS: In this GWAS, we analysed data from the UK Zaponex Treatment Access System clozapine monitoring service as part of the CLOZUK study. We included all available individuals with clozapine pharmacokinetic assays requested by their clinicians. We excluded people younger than 18 years, or whose records contained clerical errors, or with blood drawn 6-24 h after dose, a clozapine or norclozapine concentration less than 50 ng/mL, a clozapine concentration of more than 2000 ng/mL, a clozapine-to-norclozapine ratio outside of the 0·5-3·0 interval, or a clozapine dose of more than 900 mg/day. Using genomic information, we identified five biogeographical ancestries: European, sub-Saharan African, north African, southwest Asian, and east Asian. We did pharmacokinetic modelling, a GWAS, and a polygenic risk score association analysis using longitudinal regression analysis with three primary outcome variables: two metabolite plasma concentrations (clozapine and norclozapine) and the clozapine-to-norclozapine ratio. FINDINGS: 19 096 pharmacokinetic assays were available for 4760 individuals in the CLOZUK study. After data quality control, 4495 individuals (3268 [72·7%] male and 1227 [27·3%] female; mean age 42·19 years [range 18-85]) linked to 16 068 assays were included in this study. We found a faster average clozapine metabolism in people of sub-Saharan African ancestry than in those of European ancestry. By contrast, individuals with east Asian or southwest Asian ancestry were more likely to be slow clozapine metabolisers than those with European ancestry. Eight pharmacogenomic loci were identified in the GWAS, seven with significant effects in non-European groups. Polygenic scores generated from these loci were associated with clozapine outcome variables in the whole sample and within individual ancestries; the maximum variance explained was 7·26% for the metabolic ratio. INTERPRETATION: Longitudinal cross-ancestry GWAS can discover pharmacogenomic markers of clozapine metabolism that, individually or as polygenic scores, have consistent effects across ancestries. Our findings suggest that ancestral differences in clozapine metabolism could be considered for optimising clozapine prescription protocols for diverse populations. FUNDING: UK Academy of Medical Sciences, UK Medical Research Council, and European Commission.


Subject(s)
Antipsychotic Agents , Clozapine , Humans , Male , Female , Adolescent , Young Adult , Adult , Middle Aged , Aged , Aged, 80 and over , Clozapine/therapeutic use , Genome-Wide Association Study , Pharmacogenetics , Antipsychotic Agents/therapeutic use , Antipsychotic Agents/pharmacokinetics , United Kingdom
8.
Am J Community Psychol ; 71(3-4): 344-354, 2023 06.
Article in English | MEDLINE | ID: mdl-36609746

ABSTRACT

The purpose of the current study was to examine the diffusion effects of a youth-led sexual violence prevention program (i.e., Youth Voices in Prevention [Youth VIP]). Specifically, social network analysis was used to measure the extent to which Youth VIP changed behaviors for 1172 middle and high school youth who did not attend program events but were friends with Youth VIP participants and completed the first and final survey (approximately 2 years apart). Findings suggest that there was considerable interpersonal communication about Youth VIP among the students generated by program participation. Specifically, youth with friends who participated in Youth VIP were more likely to report hearing their friends talk about Youth VIP and reported talking to their friends about Youth VIP compared with those not connected to Youth VIP participants. However, there were no diffusion effects found for behavioral outcomes (i.e., bystander intervention behavior, violence victimization, and perpetration). Given the mixed findings, further research is needed to determine the extent to which youth-led sexual violence prevention initiatives lead to changes in broader community-wide changes in youths' behaviors.


Subject(s)
Crime Victims , Sex Offenses , Humans , Adolescent , Adult , Sex Offenses/prevention & control , Violence/prevention & control , Sexual Behavior , Schools
9.
J Am Soc Mass Spectrom ; 34(3): 417-425, 2023 Mar 01.
Article in English | MEDLINE | ID: mdl-36700916

ABSTRACT

Many cancer drugs fail at treating solid epithelial tumors with hypoxia and insufficient drug penetration thought to be contributing factors to the observed chemoresistance. Owing to this, it is imperative to evaluate potential cancer drugs in conditions as close to in vivo as possible, which is not always done. To address this, we developed a mass spectrometry-based protein footprinting method for exploring the impact of hypoxia on protein in 3D colorectal cancer cells. Our group has previously extended the protein footprinting method fast photochemical oxidation of proteins (FPOP) for live cell analysis (IC-FPOP); however, this is the first application of IC-FPOP in a 3D cancer model. In this study, we perform IC-FPOP on intact spheroids (Spheroid-FPOP) using a modified version of the static platform incubator with an XY movable stage (PIXY) FPOP platform. We detected modification in each of three spheroid layers, even the hypoxic core. Pathway analysis revealed protein modifications in over 10 distinct protein pathways, including some involved in protein ubiquitination; a process modulated in cancer pathologies. These results demonstrate the feasibility of Spheroid-FPOP to be utilized as a tool to interrogate protein interactions within a native tumor microenvironment.


Subject(s)
Antineoplastic Agents , Neoplasms , Humans , Protein Footprinting/methods , Proteins/analysis , Mass Spectrometry/methods , Oxidation-Reduction , Spheroids, Cellular/chemistry , Tumor Microenvironment
11.
Prev Sci ; 23(7): 1230-1240, 2022 10.
Article in English | MEDLINE | ID: mdl-35230613

ABSTRACT

Self-directed violence (SDV) is a significant public health issue for adolescents and emerging adults, and yet youth exposure to prevention messaging and youth perspectives on SDV prevention needs are understudied. The current study sought to better understand the ways in which a national sample of youth and emerging adults were exposed to suicide prevention programs or conversations. A sample of 1031 young people ages 13-23 were recruited nationally through social media. Survey questions asked about SDV prevention exposure. Open-ended questions asked youth to suggest additional information they desired about SDV. A majority of participants (87%) reported that they had received prevention exposure from at least one source (i.e., family, online, attending a talk, or formal program) with few differences by demographic characteristics. However, sexual and gender minority (SGM) youth reported accessing more SDV prevention information online compared to other youth. Overall, youth had many ideas about what additional information they needed, including how to help someone at risk for SDV and how to access information about mental health. While the majority of youth are receiving some SDV prevention messages, there is variation in how they get this information, and survey participants still felt they were missing important information. Findings highlight the need to resource more comprehensive SDV prevention for youth and young adults.


Subject(s)
Adolescent Behavior , Suicide Prevention , Adolescent , Adolescent Behavior/psychology , Adult , Humans , Motivation , Sexual Behavior , Violence , Young Adult
12.
Prev Sci ; 23(8): 1379-1393, 2022 11.
Article in English | MEDLINE | ID: mdl-35303249

ABSTRACT

Involving youth in developing and implementing prevention programs to reduce sexual violence (SV) has the potential to improve prevention outcomes. However, there has been little focus on youth-led SV prevention programs, and limited evaluation research to help guide efforts. The current study examined the effectiveness of Youth Voices in Prevention (Youth VIP) leadership retreats on SV victimization and perpetration, forms of violence related to SV (e.g., bullying), SV bystander behaviors and readiness, and perceptions of norms related to SV prevention. Results identified mixed findings for program impact, with variations in outcomes that can help guide future youth-led prevention program initiatives. Youth attending a large "kick-off" leadership retreat (that was less youth-led that subsequent smaller retreats) later reported more bystander behaviors, but also reported increased perpetration and victimization, compared to non-attending youth. However, youth attending smaller, more focused leadership retreats held during the school year, reported reductions in sexual harassment perpetration and improved bystander behaviors and attitudes compared to non-attending youth. Evaluation of moderator variables suggests that program impact was generally stronger for younger participants, sexual minority youth, and non-White youth (which were largely Native American youth in this sample). Findings suggest promise for youth-led prevention work but also highlight the need for testing the impact of different training structures and modalities. Clinical trials number: NCT03207386.


Subject(s)
Crime Victims , Sex Offenses , Sexual Harassment , Adolescent , Humans , Leadership , Sex Offenses/prevention & control , Violence/prevention & control , Sexual Harassment/prevention & control
13.
Violence Vict ; 37(2): 222-243, 2022 04 01.
Article in English | MEDLINE | ID: mdl-35264454

ABSTRACT

A coordinated response by a trained multidisciplinary team (MDT) can help support child sex trafficking (CST) victims, but little is known about factors that influence the development and sustainability of MDTs in this work. An online survey was conducted with 171 professionals who attended a Multidisciplinary Team Child Sex Trafficking (MDT-CST) training to identify factors related to team growth. Increased MDT success was related to: (1) the presence of a CST-specific advocacy organization in the community; (2) other community agencies active in supporting CST victims (e.g., SANE nurses, faith-based organizations, and runaway shelters); (3) a greater breadth of professional representation on the MDT; and (4) agency leadership support for the CST action plan. Most of the MDTs sustained and increased their coordination with other community agencies over time, but the study identified that growth is improved when administrators support team efforts and there are resources and supports for CST victims elsewhere in the community.


Subject(s)
Human Trafficking , Child , Family , Humans , Patient Care Team , Surveys and Questionnaires
14.
Chem Rev ; 122(8): 7532-7561, 2022 04 27.
Article in English | MEDLINE | ID: mdl-34633178

ABSTRACT

Hydroxyl radical protein footprinting (HRPF) coupled to mass spectrometry has been successfully used to investigate a plethora of protein-related questions. The method, which utilizes hydroxyl radicals to oxidatively modify solvent-accessible amino acids, can inform on protein interaction sites and regions of conformational change. Hydroxyl radical-based footprinting was originally developed to study nucleic acids, but coupling the method with mass spectrometry has enabled the study of proteins. The method has undergone several advancements since its inception that have increased its utility for more varied applications such as protein folding and the study of biotherapeutics. In addition, recent innovations have led to the study of increasingly complex systems including cell lysates and intact cells. Technological advances have also increased throughput and allowed for better control of experimental conditions. In this review, we provide a brief history of the field of HRPF and detail recent innovations and applications in the field.


Subject(s)
Hydroxyl Radical , Protein Footprinting , Hydroxyl Radical/chemistry , Mass Spectrometry , Protein Folding , Protein Footprinting/methods , Proteins/chemistry
15.
J Interpers Violence ; 37(9-10): NP7984-NP8005, 2022 05.
Article in English | MEDLINE | ID: mdl-33246375

ABSTRACT

Statutory rape laws are intended to protect adolescents from harm as a result of sexual activity with older individuals. In this pursuit, many, but far from all, states' statutory rape laws differentiate younger and older offenders. In effect, many of these states differentiate offenders who are 21 and older from those who are under 21. It is unknown, however, whether and how the dynamics of statutory rape vary depending on the age of the offender. To explore the contribution of offender age to the dynamics of statutory rape, data were collected from the records of 105 statutory rape cases referred to a child abuse assessment center over a 63-month period. Records included detailed reports on case history and victims' family history, mental health and health-risking behaviors, medical examination results, and forensic interview summaries. Cases of offenders11.Because these cases are under investigation, "offenders" here refers to "suspects." For ease of reading, we use "offenders" to be consistent with prior research terminology. under 21 were compared to the cases of offenders 21 years and older. Compared to cases of younger offenders, cases of offenders 21 years and older more often involved slightly older adolescent victims. Independent of age, victims with offenders 21 and over compared with offenders under 21 were over six times as likely to have a prior history of a high, versus low, number of psychosocial problems and were seven times more likely to experience multiple forms of coercion versus no coercion by the offender. Findings suggest that statutory rape prevention, policy and response strategies should consider the critical ways that case characteristics and victims differ according to this key offender attribute.


Subject(s)
Child Abuse , Criminals , Rape , Adolescent , Child , Humans , Sexual Behavior , Social Problems
16.
LGBT Health ; 9(1): 34-42, 2022 01.
Article in English | MEDLINE | ID: mdl-34747660

ABSTRACT

Purpose: The study purpose was to examine how the COVID-19 pandemic may be differentially impacting the well-being of sexual and gender minority (SGM) youth compared with their non-SGM counterparts. Specifically, we looked at perceptions of mental and physical health impact and relevant aspects of coping with the pandemic such as engagement in prosocial activities that might ultimately promote resilience. Methods: Survey data were collected between November 27, 2020, and December 11, 2020, from 990 adolescents and emerging adults aged 13-23 years. Participants were recruited through advertisements on Facebook and Instagram with an effort to overrecruit SGM youth. Questions asked youth about their perception of the impact of the COVID-19 pandemic on their physical and mental health, the types of prosocial activities engaged in during the pandemic, and how they felt they were doing now versus before the pandemic. Results: SGM youth reported a significantly greater impact from the pandemic than non-SGM youth. Both SGM youth and cisgender heterosexual females reported greater impact on their mental health compared with cisgender heterosexual males, with cisgender sexual minority females and gender minority youth reporting the highest mental health impact. Gender minority youth also reported feeling less connected to their families and less safe at home as a result of the pandemic. Conclusion: Health professionals, educators, and advocates need to help assess SGM youth for negative impact from the pandemic and help them connect with other youth and support resources online as they move forward. Youth may also benefit from programs connecting them to volunteer work and neighbors.


Subject(s)
COVID-19 , Sexual and Gender Minorities , Adolescent , Adult , Female , Gender Identity , Humans , Male , Pandemics , SARS-CoV-2 , Sexual Behavior
17.
J Interpers Violence ; 37(17-18): NP17052-NP17072, 2022 09.
Article in English | MEDLINE | ID: mdl-34107811

ABSTRACT

Engaging vulnerable populations in research is a critical focus for researchers seeking to find ways to improve safety and well-being for broader populations. Vulnerabilities often co-occur, meaning that individuals may experience vulnerabilities in multiple facets of their life (e.g., victim of abuse and mental health diagnoses). Unfortunately, many vulnerable populations remain underinvestigated due to difficulty reaching, engaging, and safely including such populations in research. While most researchers assert the importance of including vulnerable populations in research, few actively and successfully recruit them. Many vulnerable populations are understandably weary of research participation. Victims and survivors of domestic minor sex trafficking (DMST) are one such vulnerable population, in that identification may incur substantial risk for DMST victim's physical safety and well-being. In addition, DMST victims and survivors often experience co-occurring vulnerabilities including substance use, histories of abuse, delinquency, and poor mental health. Accordingly, they are notoriously difficult to recruit for research participation. The current paper uses a detailed case example to describe the recruitment and retention strategies used by one researcher to promote DMST survivor research participation. A candid account of challenges, successes, and lessons learned is offered in service of building methodological techniques for recruitment that both honor participant experiences while championing methodological rigor. Findings detail the importance of trust and a trauma-informed approach to qualitative data collection and analysis. Authors include specific strategies to promote comfort and candor amid traumatic response in ways that may empower research participants. A detailed discussion of how such strategies may be adapted for vulnerable populations beyond DMST victims and survivors is provided.


Subject(s)
Human Trafficking , Substance-Related Disorders , Human Trafficking/psychology , Humans , Respect , Survivors , Vulnerable Populations
18.
Psychol Violence ; 12(6): 403-412, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36588656

ABSTRACT

Objective: Most studies of peer sexual violence (SV) prevention programs for adolescents focus on program outcomes or feasibility and acceptability; few examine how exposure levels or dosage affects impact. The present study examined the effects of attending multiple community-located youth-led prevention events, as compared to attending one or none, on peer violence (PV)-related attitudes and behaviors. Method: Middle and high school students (M age at first wave = 13.7; 53.2% female; 76.5% White; 21.0% Native American) responded to surveys across 3 years. Logistic regression analyses compared students who attended one community-based event, two or more events, and zero events on sexual violence victimization, any other violence perpetration/victimization, social norms, denial of the problem of sexual violence, and bystander behaviors. Results: After controlling for exposure to longer prevention leadership training as well as baseline outcome levels, youth who participated in two or more community prevention events showed lower perpetration over time, improved prevention attitudes, and more helpful bystander actions in response to peer sexual violence. No significant differences were noted for attendance at one community-based event. Conclusions: Impact of out of school prevention events on youth behavior depends on more vigorous engagement than one-time contacts. Community-based prevention programs can utilize youth-led engagement strategies to help increase youth participation and resulting benefits.

19.
Proteomics ; 21(21-22): e2000295, 2021 11.
Article in English | MEDLINE | ID: mdl-34463027

ABSTRACT

Hydroxyl radical protein footprinting (HRPF) utilizes hydroxyl radicals to covalently modify solvent exposed regions of proteins. When coupled with mass spectrometry, HRPF can provide insightful information on protein structural changes including inside cells. However, the complex mixture of proteins and modifications makes identification a complicated task. To search all of the HRPF-induced modification combinations across the full proteome, requires substantial computational power and still can take days to search. To drastically decrease processing time and improve identifications, a novel cloud-based search engine, Bolt, was used to search for HRPF modifications in comparison to a commonly used search engine, Sequest. A 35% increase in the identification of modified peptides was observed in Bolt compared to Sequest with a decrease in computation time.


Subject(s)
Hydroxyl Radical , Protein Footprinting , Peptides , Proteome , Search Engine , Workflow
20.
J Am Soc Mass Spectrom ; 32(7): 1644-1647, 2021 Jul 07.
Article in English | MEDLINE | ID: mdl-34170666

ABSTRACT

Fast photochemical oxidation of proteins (FPOP) has demonstrated the ability to inform on the higher order structure of proteins. Recent technological advances have extended FPOP to live cells (IC-FPOP) using multiple cell lines and in vivo (IV-FPOP) using C. elegans. These innovations allow proteins to be studied in their native cellular environment. Hydroxyl radicals are generated via the photoloysis of hydrogen peroxide. Hydrogen peroxide is a signaling molecule that can induce changes to some proteins in the cell limiting the proteins that can be studied by IC-FPOP. Here, we evaluate the sulfate radical anion as a footprinting label in IC-FPOP with sodium persulfate as the precursor. Our findings show a 1.5-fold increase in the number of modified proteins compared to IC-FPOP using hydroxyl radicals at the same precursor concentration demonstrating the amenability of this radical with IC-FPOP.


Subject(s)
Indicators and Reagents/chemistry , Proteins , Sulfates/chemistry , HEK293 Cells , Humans , Oxidants, Photochemical , Oxidation-Reduction , Protein Footprinting/methods , Proteins/analysis , Proteins/chemistry , Sodium Compounds/chemistry
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