Search

Women with human immunodeficiency virus (HIV) infection have higher rates of concurrent human papillomavirus (HPV) infection and cervical dysplasia than do HIV-uninfected women. They are also more commonly infected with multiple HPV types simultaneously. To determine the prevalence of different HPV genotypes in a group of HIV-infected women and to correlate these findings with cervical cytological results, we studied a group of 229 women attending a university-based HIV clinic during a 7-year period. When cervicovaginal lavage specimens, the reverse line-blot assay, and DNA sequencing were used, the most commonly detected HPV types (in decreasing order of frequency) were 56, 53, 16, 58, 52, MM7, MM8, and 33. These results contrast sharply with similar studies of HIV-uninfected women, in whom HPV-16 and -18 generally predominate. In our study, the HPV types most commonly associated with low-grade squamous intraepithelial lesions (SILs) were 56 and 53. Types most commonly associated with high-grade SILs were 52 and 58. High-risk HPV types other than 16 and 18 are often found in HIV-infected women and are frequently associated with abnormal cervical cytological results in this setting. These observations have implications for the design of future HPV vaccines. Copyright © 2006 Infectious Diseases Society of America [ABSTRACT FROM AUTHOR]

Abstract Background Vitamin D deficiency is a re-emerging epidemic, especially in minority populations. Vitamin D is crucial not only for bone health but for proper brain development and functioning. Low levels of vitamin D are associated with depression, seasonal affective disorder, and schizophrenia in adults, but little is known about vitamin D and mental health in the pediatric population. Methods One hundred four adolescents presenting for acute mental health treatment over a 16-month period were assessed for vitamin D status and the relationship of 25-OH vitamin D levels to severity of illness, defined by presence of psychotic features. Results Vitamin D deficiency (25-OH D levels Conclusions Vitamin D deficiency and insufficiency are both highly prevalent in adolescents with severe mental illness. The preliminary associations between vitamin D deficiency and presence of psychotic features warrant further investigation as to whether vitamin D deficiency is a mediator of illness severity, result of illness severity, or both. Higher prevalence of vitamin D deficiency but no greater risk of psychosis in African Americans, if confirmed, may have special implications for health disparity and treatment outcome research.

CONTEXT: Quality of life (QOL) for patients is now a worldwide concern, but there are few specific instruments developed by modular approach to measure outcomes in chronic diseases. OBJECTIVES: This study aimed to develop and validate the System of Quality of Life Instruments for Chronic Diseases-General Module (QLICD-GM). METHODS: Based on the World Health Organization's definition of QOL and programmed decision procedures, the QLICD-GM was developed using focus group discussions, pilot tests of 201 cases, and field tests of 620 cases representing seven different chronic diseases. The number of items in the final version was reduced to 30 from a 73-item pool, and the psychometric properties of the scale were evaluated by indicators, such as validity and reliability coefficients, standardized response mean, and statistical methods of correlational analysis, t-tests, and structural equation modeling. RESULTS: Correlational and structural equation model analyses confirmed good construct validity with root mean square error of approximation of 0.0606, nonnormed fit index of 0.941, and comparative fit index of 0.947. Test-retest reliability coefficients for all domains and facets were higher than 0.70, with a range of 0.71-0.92, and the internal consistency alphas for most domains and facets were higher than 0.65 (ranging from 0.52 to 0.89). All domains and overall scores of the QLICD-GM had a statistically significant change after treatment, with moderate effect size. CONCLUSION: The QLICD-GM has good validity, reliability, and better responsiveness compared with the SF-36(R) Health Survey and can be used as the general module for chronic diseases. [ABSTRACT FROM AUTHOR]

SARS-CoV-2 infections led to a worldwide pandemic in 2020. As of 2024, therapeutics against SARS-CoV-2 have continued to be desirable. NSP14 is a dual-function methyltransferase (MTase) and exonuclease (ExoN) with key roles in SARS-CoV-2 genome propagation and host immune system evasion. In this work, we developed high-throughput screening (HTS) assays for NSP14 MTase and ExoN activities. We screened both activities against a collection of 40,664 compounds. A total of 1677 initial hit compounds were identified, cherrypicked, counterscreened for assay interference, and screened for off-target selectivity. We identified 396 and 174 high-quality hits against the MTase and ExoN activities, respectively. Along with inhibitors for individual activities, we identified dual-activity inhibitors, including a novel inhibitor that is not competitive with any substrate and interacts with a putative allosteric binding site. This study represents the largest published screen of SARS-CoV-2 NSP14 MTase and ExoN activities to date and culminates in a pipeline for the NSP14 drug discovery.

Mutations in RAS and PI3Kα are major drivers of human cancer. Their interaction plays a crucial role in activating PI3Kα and amplifying the PI3K-AKT-mTOR pathway. Disrupting RAS-PI3Kα interaction enhances survival in lung and skin cancer models and reduces tumor growth and angiogenesis, although the structural details of this interaction remain unclear. Here, we present structures of KRAS, RRAS2, and MRAS bound to the catalytic subunit (p110α) of PI3Kα, elucidating the interaction interfaces and local conformational changes upon complex formation. Structural and mutational analyses highlighted key residues in RAS and PI3Kα impacting binding affinity and revealed isoform-specific differences at the interaction interface in RAS and PI3K isoforms, providing a rationale for their differential affinities. Notably, in the RAS-p110α complex structures, RAS interaction with p110α is limited to the RAS-binding domain and does not involve the kinase domain. This study underscores the pivotal role of the RAS-PI3Kα interaction in PI3Kα activation and provides a blueprint for designing PI3Kα isoform-specific inhibitors to disrupt this interaction., Competing Interests: Competing interests: All authors, except F.M., have no competing interests. F.M. is a consultant for Ideaya Biosciences, Kura Oncology, Leidos Biomedical Research, Pfizer, Daiichi Sankyo, Amgen, PMV Pharma, OPNA-IO, and Quanta Therapeutics, has received research grants from Boehringer-Ingelheim, and is a consultant for and cofounder of BridgeBio Pharma., (© 2025. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Phosphatidylinositol 4,5-bisphosphate 3-kinases (PI3K) are a family of kinases whose activity affects pathways needed for basic cell functions. As a result, PI3K is one of the most mutated genes in all human cancers and serves as an ideal therapeutic target for cancer treatment. Expanding on work done by other groups we improved protein yield to produce stable and pure protein using a variety of modifications including improved solubility tag, novel expression modalities, and optimized purification protocol and buffer. By these means, we achieved a 40-fold increase in yield for p110α/p85α and a 3-fold increase in p110α. We also used these protocols to produce comparable constructs of the β and δ isoforms of PI3K. Increased yield enhanced the efficiency of our downstream high throughput drug discovery efforts on the PIK3 family of kinases., Competing Interests: Declaration of Competing interest The authors declare they have no conflicts of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Tobacco-etch-virus (TEV) protease is the workhorse of many laboratories in which protein expression is the linchpin of downstream experiments. TEV protease is remarkable in its sequence specificity as the cleavage sequence rarely appears in higher organisms and its ability to cleave fusion tag proteins from proteins of interest. Herein we report work done on large-scale production of TEV protease using different promotors, media, fusion tags, and expression platforms. During our work we detected post-translational modification (gluconoylation and phosphogluconoylation) of TEV protease and the subsequent effects this has on the purity of the protein. Subsequently we made our pgl plus bacteria that negates these modifications and their effects. We also introduce a GFP-based assay for measurement of activity and ultimately a new set of protocols for producing 400-500 mg/L TEV protease., Competing Interests: Conflicts of interest The authors declare they have no conflicts of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Humanity is facing a polycrisis, including climate change, loss of biodiversity, and economic inequalities. This commentary examines the role of physical activity (PA) research in the context of climate change, highlighting 3 perspectives: PA as a victim, as a perpetrator, and as part of the solution. The first perspective conceptualizes PA as a victim of climate change, as heat waves, extreme weather events, and high ozone levels pose a threat to health-enhancing PA and sports; adaptation strategies are needed and potentially lifesaving. The second perspective describes PA as a perpetrator to climate change due to the significant carbon footprint of some forms of PA and sport. The third perspective focuses on PA as a solution to climate change, as some forms of PA-such as active transport and urban gardening-have both individual and planetary health benefits. In addition, PA provides health benefits in extreme situations by reducing stress and maintaining health in times of crisis. Each of these 3 perspectives can guide future research in the field of PA and health.

The diversity of chemical and structural attributes of proteins makes it inherently difficult to produce a wide range of proteins in a single recombinant protein production system. The nature of the target proteins themselves, along with cost, ease of use, and speed, are typically cited as major factors to consider in production. Despite a wide variety of alternative expression systems, most recombinant proteins for research and therapeutics are produced in a limited number of systems: Escherichia coli, yeast, insect cells, and the mammalian cell lines HEK293 and CHO. Recent interest in Vibrio natriegens as a new bacterial recombinant protein expression host is due in part to its short doubling time of ≤ 10 min but also stems from the promise of compatibility with techniques and genetic systems developed for E. coli. We successfully incorporated V. natriegens as an additional bacterial expression system for recombinant protein production and report improvements to published protocols as well as new protocols that expand the versatility of the system. While not all proteins benefit from production in V. natriegens, we successfully produced several proteins that were difficult or impossible to produce in E. coli. We also show that in some cases, the increased yield is due to higher levels of properly folded protein. Additionally, we were able to adapt our enhanced isotope incorporation methods for use with V. natriegens. Taken together, these observations and improvements allowed production of proteins for structural biology, biochemistry, assay development, and structure-based drug design in V. natriegens that were impossible and/or unaffordable to produce in E. coli., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

The small GTPase Rat sarcoma virus proteins (RAS) are key regulators of cell growth and involved in 20-30% of cancers. RAS switches between its active state and inactive state via exchange of GTP (active) and GDP (inactive). Therefore, to study active protein, it needs to undergo nucleotide exchange to a non-hydrolysable GTP analog. Calf intestine alkaline phosphatase bound to agarose beads (CIP-agarose) is regularly used in a nucleotide exchange protocol to replace GDP with a non-hydrolysable analog. Due to pandemic supply problems and product shortages, we found the need for an alternative to this commercially available product. Here we describe how we generated a bacterial alkaline phosphatase (BAP) with an affinity tag bound to an agarose bead. This BAP completely exchanges the nucleotide in our samples, thereby demonstrating an alternative to the commercially available product using generally available laboratory equipment., Competing Interests: Declaration of Competing interest The authors declare they have no conflicts of interest with the contents of this article., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Development of new targeted inhibitors for oncogenic KRAS mutants may benefit from insight into how a given mutation influences the accessibility of protein residues and how compounds interact with mutant or wild-type KRAS proteins. Targeted proteomic analysis, a key validation step in the KRAS inhibitor development process, typically involves both intact mass- and peptide-based methods to confirm compound localization or quantify binding. However, these methods may not always provide a clear picture of the compound binding affinity for KRAS, how specific the compound is to the target KRAS residue, and how experimental conditions may impact these factors. To address this, we have developed a novel top-down proteomic assay to evaluate in vitro KRAS4B-compound engagement while assessing relative quantitation in parallel. We present two applications to demonstrate the capabilities of our assay: maleimide-biotin labeling of a KRAS4B G12D cysteine mutant panel and treatment of three KRAS4B proteins (WT, G12C, and G13C) with small molecule compounds. Our results show the time- or concentration-dependence of KRAS4B-compound engagement in context of the intact protein molecule while directly mapping the compound binding site.

Background: Accurate and fast measurement of physical activity is important for surveillance. Even though many physical activity questionnaires (PAQ) are currently used in research, it is unclear which of them is the most reliable, valid, and easy to use. This systematic review aimed to identify existing brief PAQs, describe and compare their measurement properties, and assess their level of readability., Methods: We performed a systematic review based on the PRISMA statement. Literature searches were conducted in six scientific databases. Articles were included if they evaluated validity and/or reliability of brief (i.e., with a maximum of three questions) physical activity or exercise questionnaires intended for healthy adults. Due to the heterogeneity of studies, data were summarized narratively. The level of readability was calculated according to the Flesch-Kincaid formula., Results: In total, 35 articles published in English or Spanish were included, evaluating 32 distinct brief PAQs. The studies indicated moderate to good levels of reliability for the PAQs. However, the majority of results showed weak validity when validated against device-based measurements and demonstrated weak to moderate validity when validated against other PAQs. Most of the assessed PAQs met the criterion of being "short," allowing respondents to complete them in less than one minute either by themselves or with an interviewer. However, only 17 questionnaires had a readability level that indicates that the PAQ is easy to understand for the majority of the population., Conclusions: This review identified a variety of brief PAQs, but most of them were evaluated in only a single study. Validity and reliability of short and long questionnaires are found to be at a comparable level, short PAQs can be recommended for use in surveillance systems. However, the methods used to assess measurement properties varied widely across studies, limiting the comparability between different PAQs and making it challenging to identify a single tool as the most suitable. None of the evaluated brief PAQs allowed for the measurement of whether a person fulfills current WHO physical activity guidelines. Future development or adaptation of PAQs should prioritize readability as an important factor to enhance their usability., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Tcymbal et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

KRAS , the most frequently mutated oncogene in human cancer, produces two isoforms, KRAS4a and KRAS4b, through alternative splicing. These isoforms differ in exon 4, which encodes the final 15 residues of the G-domain and hypervariable regions (HVRs), vital for trafficking and membrane localization. While KRAS4b has been extensively studied, KRAS4a has been largely overlooked. Our multidisciplinary study compared the structural and functional characteristics of KRAS4a and KRAS4b, revealing distinct structural properties and thermal stability. Position 151 influences KRAS4a's thermal stability, while position 153 affects binding to RAF1 CRD protein. Nuclear magnetic resonance analysis identified localized structural differences near sequence variations and provided a solution-state conformational ensemble. Notably, KRAS4a exhibits substantial transcript abundance in bile ducts, liver, and stomach, with transcript levels approaching KRAS4b in the colon and rectum. Functional disparities were observed in full-length KRAS variants, highlighting the impact of HVR variations on interaction with trafficking proteins and downstream effectors like RAF and PI3K within cells.

We present a novel method to determine engagement and specificity of KRAS4B-targeting compounds in vitro. By employing top-down mass spectrometry (MS), which analyzes intact and modified protein molecules (proteoforms), we can directly visualize and confidently characterize each KRAS4B species within compound-treated samples. Moreover, by employing targeted MS2 fragmentation, we can precisely localize each compound molecule to a specific residue on a given KRAS4B proteoform. This method allows us to comprehensively evaluate compound specificity, clearly detect nonspecific binding events, and determine the order and frequency with which they occur. We provide two proof-of-concept examples of our method employing publicly available compounds, along with detailed protocols for sample preparation, top-down MS data acquisition, targeted proteoform MS2 fragmentation, and analysis of the resulting data. Our results demonstrate the concentration dependence of KRAS4B-compound engagement and highlight the ability of top-down MS to directly map compound binding location(s) without disrupting the KRAS4B primary structure. Our hope is that this novel method may help accelerate the identification of new successful targeted inhibitors for KRAS4B and other RAS isoforms., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Even though the importance of physical activity policy monitoring has increased in the last decade, there is a lack of understanding what different approaches exist and which methodology they employ. In order to address this research gap, this review attempts to map existing approaches of physical activity policy monitoring and to analyse methodological aspects, especially with regards to the roles of governments and researchers., Methods: A systematic search was conducted in five scientific databases (PubMed, Scopus, SportDiscus, Psycinfo, Web of Knowledge) in July 2021, and the identified records were screened independently by two reviewers. Records were included if they (a) focused on the monitoring of public policies to promote PA, (b) allowed to compare policies across time, across nations/regions or across policy sectors, and (c) were written in English, German or Russian. During full text analysis, information on methodological aspects was extracted and studies were categorized based on the level of government involvement., Results: The search yielded in a total of 112 studies. 86 of these studies (76.8%) followed a research-driven approach (little or no government involvement) while only two studies (1.8%) were based on a government-driven approach (led by governments). The remaining 24 studies (21.4%) were based on a co-production approach (strong collaboration between researchers and governments). All in all, 18 different tools for physical activity policy monitoring were identified; key examples are the Report Cards on Physical Activity for Children and Youth (research-driven approach), the HEPA Monitoring Framework (government-driven approach) and the HEPA Policy Audit Tool (co-production approach)., Conclusions: The level of government involvement in policy monitoring differs significantly, and research-driven, government-driven and co-production approaches can be distinguished. These approaches have different strengths and weaknesses, and can be linked to distinct theories of change and models on research-policy relations. Increasing awareness on the implications of these approaches is key to improve the understanding and further development of physical activity policy monitoring., (© 2023. The Author(s).)

Introduction: While there are several approaches to collect basic information on physical activity (PA) promotion policies, some governments require more in-depth overviews on the situation in their country. In Germany, the Federal Ministry of Health expressed its interest in collecting detailed data on target group specific PA promotion, as relevant competences are distributed across a wide range of political levels and sectors. This study describes the development of a policy brief on physical activity promotion for children and adolescents in Germany. In particular, it addresses two major gaps in the current literature by systematically assessing good practice examples and "routine practices," i.e., PA promotion activities already taking place on large scale and regular basis., Materials and Methods: Based on relevant national and international guidelines, the TARGET:PA tool was co-produced by researchers and ministry officials. It includes (1) PA recommendations, (2) national prevalence rates, (3) recommendations for PA promotion, and data on national (4) routine practices, (5) good practice projects and (6) policies. Data were collected for children and adolescents in Germany using desk research, semi-structured interviews and secondary data analysis., Results: A policy brief and scientific background document were developed. Results showed that 46% of the 4-5-year-olds fulfil WHO recommendations but only 15% of the 11-17-year-olds, and that girls are less active than boys. Currently, in Germany no valid data are available on the PA behaviour of children under the age of three. An overview of routine practices for PA promotion for children and adolescents was compiled, and experts were asked to critically assess their effectiveness, reach and durability. Overall, 339 target group specific projects for PA promotion were found, with 22 classified as examples of good practice. National PA policies for children and adolescents were identified across different sectors and settings., Conclusion: The study provides a comprehensive overview of the current status of PA promotion for children and adolescents in Germany. The co-production of the policy brief was a strength of the study, as it allowed researchers to take the needs of ministry officials into account, and as it supported the immediate uptake of results in the policymaking process. Future studies should test the applicability of the TARGET:PA tool to different target groups and countries., 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 © 2023 Messing, Gelius, Abu-Omar, Marzi, Beck, Geidl, Grüne, Tcymbal, Reimers and Pfeifer.)

RIT1 is a RAS guanosine triphosphatase (GTPase) that regulates different aspects of signal transduction and is mutated in lung cancer, leukemia, and in the germline of individuals with Noonan syndrome. Pathogenic RIT1 proteins promote mitogen-activated protein kinase (MAPK) hyperactivation; however, this mechanism remains poorly understood. Here, we show that RAF kinases are direct effectors of membrane-bound mutant RIT1 necessary for MAPK activation. We identify critical residues in RIT1 that facilitate interaction with membrane lipids and show that these are necessary for association with RAF kinases and MAPK activation. Although mutant RIT1 binds to RAF kinases directly, it fails to activate MAPK signaling in the absence of classical RAS proteins. Consistent with aberrant RAF/MAPK activation as a driver of disease, we show that pathway inhibition alleviates cardiac hypertrophy in a mouse model of RIT1 mutant Noonan syndrome. These data shed light on the function of pathogenic RIT1 and identify avenues for therapeutic intervention.

Introduction: Regular physical activity provides many health benefits to older adults. As a well-known form of physical activity, cycling can be an appropriate means for older people to meet WHO recommendations and to improve their health. In addition, cycling can help to protect the environment and reduce greenhouse gas emissions. The primary aim of this scoping review is to identify the currently available scientific evidence and gaps of research in this field., Methods: A systematic search in seven databases resulted in 7,192 studies. After the exclusion of duplicates, studies were screened by two independent reviewers in a two-stage process. Based on previously defined inclusion criteria, 123 studies were included. Data extraction was based on a descriptive analytical method, and seven categories for the main topics of studies were developed. Data were extracted by three reviewers to analyze different characteristics of included articles such as age range, study design, data type, gender, type of bicycle, and country of origin., Results: The included studies covered the following topics: (1) traffic safety, (2) cycling as physical activity or for transport, (3) health benefits, (4) environmental factors, (5) facilitators and barriers, (6) application of technology and (7) promotion of cycling. Results show that the majority of studies were performed in both younger (60-79 years) and older (80+ years) adults. Most studies had an observational study design, used conventional bicycles, and were based on quantitative methods. Researchers from the United States, Netherlands, and Japan published the highest number of studies related to cycling., Discussion: Traffic safety was the most prevalent focus of the included studies. Gaps were identified with regard to studies focusing on the promotion of cycling, application of technology, as well as facilitators and barriers of cycling. While research on traffic safety should continue to be a high priority for public health, potentially more research should focus on how to get older people to bicycle more. This is warranted by the proven individual and planetary health benefits of cycling and the urgency of combating climate change., 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., (© 2023 Kardan, Akter, Iqbal, Tcymbal, Messing, Gelius and Abu-Omar.)

We present the results of a large, US$8.9 million campaign-wide field experiment, conducted among 2 million moderate- and low-information persuadable voters in five battleground states during the 2020 US presidential election. Treatment group participants were exposed to an 8-month-long advertising programme delivered via social media, designed to persuade people to vote against Donald Trump and for Joe Biden. We found no evidence that the programme increased or decreased turnout on average. We found evidence of differential turnout effects by modelled level of Trump support: the campaign increased voting among Biden leaners by 0.4 percentage points (s.e. = 0.2 pp) and decreased voting among Trump leaners by 0.3 percentage points (s.e. = 0.3 pp) for a difference in conditional average treatment effects of 0.7 points (t 1,035,571  = -2.09; P = 0.036; [Formula: see text] points; 95% confidence interval = -0.014 to 0). An important but exploratory finding is that the strongest differential effects appear in early voting data, which may inform future work on early campaigning in a post-COVID electoral environment. Our results indicate that differential mobilization effects of even large digital advertising campaigns in presidential elections are likely to be modest., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)