Temporally resolved proteomics identifies nidogen-2 as a cotarget in pancreatic cancer that modulates fibrosis and therapy response.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by increasing fibrosis, which can enhance tumor progression and spread. Here, we undertook an unbiased temporal assessment of the matrisome of the highly metastatic KPC (Pdx1-Cre, LSL-KrasG12D/+, LSL-Trp53R172H/+) and poorly metastatic KPflC (Pdx1-Cre, LSL-KrasG12D/+, Trp53fl/+) genetically engineered mouse models of pancreatic cancer using mass spectrometry proteomics. Our assessment at early-, mid-, and late-stage disease reveals an increased abundance of nidogen-2 (NID2) in the KPC model compared to KPflC, with further validation showing that NID2 is primarily expressed by cancer-associated fibroblasts (CAFs). Using biomechanical assessments, second harmonic generation imaging, and birefringence analysis, we show that NID2 reduction by CRISPR interference (CRISPRi) in CAFs reduces stiffness and matrix remodeling in three-dimensional models, leading to impaired cancer cell invasion. Intravital imaging revealed improved vascular patency in live NID2-depleted tumors, with enhanced response to gemcitabine/Abraxane. In orthotopic models, NID2 CRISPRi tumors had less liver metastasis and increased survival, highlighting NID2 as a potential PDAC cotarget.

Human Organoids for Rapid Validation of Gene Variants Linked to Cochlear Malformations.

Developmental anomalies of the hearing organ, the cochlea, are diagnosed in approximately one-fourth of individuals with congenital deafness. Most patients with cochlear malformations remain etiologically undiagnosed due to insufficient knowledge about underlying genes or the inability to make conclusive interpretations of identified genetic variants. We used exome sequencing for genetic evaluation of hearing loss associated with cochlear malformations in three probands from unrelated families. We subsequently generated monoclonal induced pluripotent stem cell (iPSC) lines, bearing patient-specific knockins and knockouts using CRISPR/Cas9 to assess pathogenicity of candidate variants. We detected FGF3 (p.Arg165Gly) and GREB1L (p.Cys186Arg), variants of uncertain significance in two recognized genes for deafness, and PBXIP1(p.Trp574*) in a candidate gene. Upon differentiation of iPSCs towards inner ear organoids, we observed significant developmental aberrations in knockout lines compared to their isogenic controls. Patient-specific single nucleotide variants (SNVs) showed similar abnormalities as the knockout lines, functionally supporting their causality in the observed phenotype. Therefore, we present human inner ear organoids as a tool to rapidly validate the pathogenicity of DNA variants associated with cochlear malformations.

Examining missingness at the momentary level in clinical research using ecological momentary assessment: Implications for suicide research.

The use of intensive time sampling methods, such as ecological momentary assessment (EMA), has increased in clinical, and specifically suicide, research during the past decade. While EMA can capture dynamic intraindividual processes, repeated assessments increase participant burden, potentially resulting in low compliance. This study aimed to shed light on study-level and psychological variables, including suicidal ideation (SI), that may predict momentary prompt (i.e., prompt-to-prompt) completion. We combined data from three EMA studies examining mental health difficulties (N = 103; 10,656 prompts; 7144 completed), using multilevel models and machine learning to determine how well we can predict prompt-to-prompt completion and which variables are most important. The two most important variables in prompt-to-prompt completion were hours since the last prompt and time in study. Psychological variables added little predictive validity; similarly, trait-level SI demonstrated a small effect on prompt-to-prompt completion. Our study showed how study-level characteristics can be used to explain prompt-to-prompt compliance rates in EMA research, highlighting the potential for developing adaptive assessment schedules to improve compliance.

Coxiella burnetii-containing vacuoles interact with host recycling endosomal proteins Rab11a and Rab35 for vacuolar expansion and bacterial growth.

Introduction: Coxiella burnetii is a gram-negative obligate intracellular bacterium and a zoonotic pathogen that causes human Q fever. The lack of effective antibiotics and a licensed vaccine for Coxiella in the U.S. warrants further research into Coxiella pathogenesis. Within the host cells, Coxiella replicates in an acidic phagolysosome-like vacuole termed Coxiella-containing vacuole (CCV). Previously, we have shown that the CCV pH is critical for Coxiella survival and that the Coxiella Type 4B secretion system regulates CCV pH by inhibiting the host endosomal maturation pathway. However, the trafficking pattern of the 'immature' endosomes in Coxiella- infected cells remained unclear. Methods: We transfected HeLa cells with GFP-tagged Rab proteins and subsequently infected them with mCherry-Coxiella to visualize Rab protein localization. Infected cells were immunostained with anti-Rab antibodies to confirm the Rab localization to the CCV, to quantitate Rab11a and Rab35- positive CCVs, and to quantitate total recycling endosome content of infected cells. A dual-hit siRNA mediated knockdown combined with either immunofluorescent assay or an agarose-based colony-forming unit assay were used to measure the effects of Rab11a and Rab35 knockdown on CCV area and Coxiella intracellular growth. Results: The CCV localization screen with host Rab proteins revealed that recycling endosome-associated proteins Rab11a and Rab35 localize to the CCV during infection, suggesting that CCV interacts with host recycling endosomes during maturation. Interestingly, only a subset of CCVs were Rab11a or Rab35-positive at any given time point. Quantitation of Rab11a/Rab35-positive CCVs revealed that while Rab11a interacts with the CCV more at 3 dpi, Rab35 is significantly more prevalent at CCVs at 6 dpi, suggesting that the CCV preferentially interacts with Rab11a and Rab35 depending on the stage of infection. Furthermore, we observed a significant increase in Rab11a and Rab35 fluorescent intensity in Coxiella-infected cells compared to mock, suggesting that Coxiella increases the recycling endosome content in infected cells. Finally, siRNA-mediated knockdown of Rab11a and Rab35 resulted in significantly smaller CCVs and reduced Coxiella intracellular growth, suggesting that recycling endosomal Rab proteins are essential for CCV expansion and bacterial multiplication. Discussion: Our data, for the first time, show that the CCV dynamically interacts with host recycling endosomes for Coxiella intracellular survival and potentially uncovers novel host cell factors essential for Coxiella pathogenesis.

Lower extremity inter-joint coupling angles and variability during gait in pediatric hypermobility spectrum disorder.

Complex musculoskeletal complications in children with hypermobility spectrum disorder (HSD) include pain, proprioception deficits, and joint instability, which may result in movement dysfunction during walking. However, no studies have explored the inter-joint coordination deficits in children with HSD. The purpose of this study was to determine the lower extremity inter-joint coupling angles, patterns, and variability during walking in children with HSD compared to children without HSD (non-HSD). Ankle, knee, and hip kinematics during the stance phase of walking in 18 children with HSD and 18 children without HSD were measured using three-dimensional motion analysis. Coupling angles, patterns, and variability of hip-knee, hip-ankle, and knee-ankle were quantified in the sagittal, frontal, and transverse planes using vector coding techniques. Statistical modeling of coupling angles on sine and cosine scales and bootstrapped standard errors were used to compare coupling angles between HSD and non-HSD groups. Permutational multivariate analysis of variance and statistical non-parametric mapping two-sample t-tests were used to compare the coupling patterns and variability between HSD and non-HSD groups, respectively. Our results indicated that coupling angles, patterns, and variability were not significantly different between the groups. These findings suggest that lower extremity inter-joint coordination and its variability during walking might not be a promising area for further research or intervention in children with HSD. Further research could use other biomechanical methods to investigate coordination deficits in pediatric patients with HSD, and how aging and disease progression are associated with coordination deficits in individuals with HSD.

TransKinect: a computer vision and machine learning clinical decision support system for automatic independent wheelchair transfer technique assessment.

BACKGROUND: Physical and occupational therapists provide routine care for manual wheelchair users and are responsible for training and assessing the quality of transfers. These transfers can produce large loads on the upper extremity joints if improper sitting-pivot-technique is used. Methods to assess quality of transfers include the Transfer Assessment Instrument, a clinically validated tool derived from quantitative biomechanical features; however, adoption of this tool is low due to the complex usage requirements and speed of typical transfers. OBJECTIVE: The objective of this study is to develop and validate a computer vison and machine learning solution to better implement the Transfer Assessment Instrument in clinical settings. METHODS: The prototype system, TransKinect, consists of an infrared depth sensor and a custom software application; usability testing was carried out with fifteen therapists who performed two transfer assessments with the TransKinect. Proficiency in using features, usability, acceptability and satisfaction were analysed with validated surveys and themes were extracted from the qualitative feedback. RESULTS: The therapists were able to successfully complete the transfer quality assessments with 86.7 ± 5.4% proficiency. Total scores for System Usability Scale (77.6 ± 14.7%) and Questionnaire for User Interface Satisfaction (83.5 ± 8.7%) indicated that the system was usable and satisfactory. Qualitative feedback indicated that TransKinect was user-friendly, easy to learn, and had high potential. DISCUSSION: The results support TransKinect as a potential clinical decision support system for therapists for the comprehensive assessment of independent transfer technique. Future research is needed to investigate the utility and acceptance of TransKinect in real clinical environments. Implications for RehabilitationMachine learning and computer vision can be used to analyze transfer techniqueTransKinect is a usable and user-friendly means for therapists to automate analysisSummary reports and videos of transfers show high potential for clinical useAdoption of TransKinect can increase quality of care for manual wheelchair users.

Influence of Polygenic Background on the Clinical Presentation of Familial Hypercholesterolemia.

BACKGROUND: Heterozygous familial hypercholesterolemia (FH) is among the most common genetic conditions worldwide that affects ≈ 1 in 300 individuals. FH is characterized by increased levels of low-density lipoprotein cholesterol (LDL-C) and increased risk of coronary artery disease (CAD), but there is a wide spectrum of severity within the FH population. This variability in expression is incompletely explained by known risk factors. We hypothesized that genome-wide genetic influences, as represented by polygenic risk scores (PRSs) for cardiometabolic traits, would influence the phenotypic severity of FH. METHODS: We studied individuals with clinically diagnosed FH (n=1123) from the FH Canada National Registry, as well as individuals with genetically identified FH from the UK Biobank (n=723). For all individuals, we used genome-wide gene array data to calculate PRSs for CAD, LDL-C, lipoprotein(a), and other cardiometabolic traits. We compared the distribution of PRSs in individuals with clinically diagnosed FH, genetically diagnosed FH, and non-FH controls and examined the association of the PRSs with the risk of atherosclerotic cardiovascular disease. RESULTS: Individuals with clinically diagnosed FH had higher levels of LDL-C, and the incidence of atherosclerotic cardiovascular disease was higher in individuals with clinically diagnosed compared with genetically identified FH. Individuals with clinically diagnosed FH displayed enrichment for higher PRSs for CAD, LDL-C, and lipoprotein(a) but not for other cardiometabolic risk factors. The CAD PRS was associated with a risk of atherosclerotic cardiovascular disease among individuals with an FH-causing genetic variant. CONCLUSIONS: Genetic background, as expressed by genome-wide PRSs for CAD, LDL-C, and lipoprotein(a), influences the phenotypic severity of FH, expanding our understanding of the determinants that contribute to the variable expressivity of FH. A PRS for CAD may aid in risk prediction among individuals with FH.

Bridging the Evidence and Practice Gap in Chronic Kidney Disease: A System Thinking Approach to Population Health.

Chronic kidney disease (CKD) is common, costly, and life-limiting, requiring dialysis and transplantation in advanced stages. Although effective guideline-based therapy exists, the asymptomatic nature of CKD together with low health literacy, adverse social determinants of health, unmet behavioral health needs, and primary care providers' (PCP) limited understanding of CKD result in defects in screening and diagnosis. Care is fragmented between PCPs and specialty nephrologists, with limited time, expertise, and resources to address systemic gaps. In this article, the authors define how they classified defects in care and report the current numbers of patients exposed to these defects, both nationally and in their health system Accountable Care Organization. They describe use of the health system's three-pillar leadership model (believing, belonging, and building) to empower providers to transform CKD care. Believing entailed engaging individuals to believe defects in CKD care could be eliminated and were a collective responsibility. Belonging fostered the creation of learning communities that broke down silos and encouraged open communication and collaboration between PCPs and nephrologists. Building involved constructing a fractal management infrastructure with transparent reporting and shared accountability, which would enable success in innovation and transformation. The result is proactive and relational CKD care organized around the patient's needs in University Hospitals Systems of Excellence. Systems of excellence combine multiple domains of expertise to promote best practice guidelines and integrate care throughout the system. The authors further describe a preliminary pilot of the CKD System of Excellence in primary care.

Ketogenic diet modifies ribosomal protein dysregulation in KMT2D Kabuki syndrome.

BACKGROUND: Kabuki syndrome (KS) is a genetic disorder caused by DNA mutations in KMT2D, a lysine methyltransferase that methylates histones and other proteins, and therefore modifies chromatin structure and subsequent gene expression. Ketones, derived from the ketogenic diet, are histone deacetylase inhibitors that can 'open' chromatin and encourage gene expression. Preclinical studies have shown that the ketogenic diet rescues hippocampal memory neurogenesis in mice with KS via the epigenetic effects of ketones. METHODS: Single-cell RNA sequencing and mass spectrometry-based proteomics were used to explore molecular mechanisms of disease in individuals with KS (n = 4) versus controls (n = 4). FINDINGS: Pathway enrichment analysis indicated that loss of function mutations in KMT2D are associated with ribosomal protein dysregulation at an RNA and protein level in individuals with KS (FDR <0.05). Cellular proteomics also identified immune dysregulation and increased abundance of other lysine modification and histone binding proteins, representing a potential compensatory mechanism. A 12-year-old boy with KS, suffering from recurrent episodes of cognitive decline, exhibited improved cognitive function and neuropsychological assessment performance after 12 months on the ketogenic diet, with concomitant improvement in transcriptomic ribosomal protein dysregulation. INTERPRETATION: Our data reveals that lysine methyltransferase deficiency is associated with ribosomal protein dysfunction, with secondary immune dysregulation. Diet and the production of bioactive molecules such as ketone bodies serve as a significant environmental factor that can induce epigenetic changes and improve clinical outcomes. Integrating transcriptomic, proteomic, and clinical data can define mechanisms of disease and treatment effects in individuals with neurodevelopmental disorders. FUNDING: This study was supported by the Dale NHMRC Investigator Grant (APP1193648) (R.D), Petre Foundation (R.D), and The Sydney Children's Hospital Foundation/Kids Research Early and Mid-Career Researcher Grant (E.T).

Extruded alginate tubes with myogenic potential.

BACKGROUND: There are currently no proven methods to reverse muscle loss in humans, which is caused by trauma (e.g., volumetric muscle loss, VML), genetic neuromuscular diseases (e.g., muscular dystrophies, MDs), and accelerated senescence (e.g., sarcopenia). Since muscle tissue is capable of regeneration through muscle satellite cells (MuSCs), the implantation of autologous (or other) donor MuSCs and MuSC-derived myoblasts into host muscles can promote donor-cell-derived myogenesis. Direct injection or implantation of MuSCs or MuSC-derived myoblasts into host muscles only promotes minimal donor-cell-derived myogenesis, whereas implantation of MuSCs/myoblasts along with associated muscle tissue (muscle fibers, extracellular matrix, neurovascular pathways, etc.) gives better results. METHODS: We aim to leverage the benefits of constraining donor myogenic cells within a template that resembles muscle tissue. In this paper, we present a workflow for basic and translational studies aimed at promoting donor-cell-derived myogenesis to increase functional muscle mass in mice. Our workflow involves preparing a slurry of 10% sodium alginate mixed with myogenic cells in cell culture media, extruding the cell-containing slurry into 10% calcium lactate to form tubes, and implanting the cellularized alginate tubes into host muscle. RESULTS: Our data suggest that, the extruded alginate tubes can tolerate a peak stress of 1892 ± 527 mN, that the elastic range is at ~75-125% strain beyond initial length, and that the Young's modulus (stiffness) is 14.17 ± 1.68 %/mm2. Importantly, these mechanical properties render the alginate tubes suitable for a published technique known as minimally-invasive muscle embedding (MIME) that was developed by us to implant myogenic material into host muscle. MIME involves threading donor myogenic tissue into a needle track created within a host muscle. Cellularized alginate tubes implanted into the tibialis anterior muscle of previously euthanized mice had numerous hematoxylin-stained structures similar to nuclear staining, supporting the idea that our alginate tubes can support cell seeding. Alginate tubes that were seeded with MuSCs, incubated in MuSC/myoblast growth (i.e., proliferation) media for two days, incubated in myotube differentiation media for six days, and then minced and reseeded in new dishes, were able to promote in vitro myoblast outgrowth over several days. DISCUSSION: This pilot study is limited in its translational scope because it was performed in vitro and with previously euthanized mice. Additional studies are needed to confirm that cellularized alginate tubes can promote the de novo development of donor-cell-derived muscle fibers, which can contribute to contractile force production. CONCLUSION: Alginate tubes with MuSC/myoblasts can be generated by a simple extrusion method. The alginate tubes have sufficient mechanical strength to tolerate insertion into a host muscle, in a minimally-invasive manner, through a needle track. The cellularized alginate tubes demonstrate myogenic potential since they are capable of being maintained in culture conditions for several days, after which they can still facilitate myoblast outgrowth in a dish.

Quantitative Musculoskeletal Imaging of the Pediatric Shoulder.

ABSTRACT: Pediatric acquired and congenital conditions leading to shoulder pain and dysfunction are common. Objective, quantitative musculoskeletal imaging-based measures of shoulder health in children lag recent developments in adults. We review promising applications of quantitative imaging that tend to be available for common pediatric shoulder pathologies, especially brachial plexus birth palsy and recurrent shoulder instability, and imaging-related considerations of musculoskeletal growth and development of the shoulder. We highlight the status of quantitative imaging practices for the pediatric shoulder and highlight gaps where better care may be provided with advances in imaging technique and/or technology.

Carotid intima-medial thickness in patients with severe hypertriglyceridemia.

Background and aims: Severe hypertriglyceridemia (HTG), defined as plasma triglyceride (TG) concentration > 10 mmol/L, is relatively uncommon, and its implications for atherosclerotic cardiovascular disease (ASCVD) risk remain somewhat unclear. We evaluated the association between severe HTG and carotid intima-media thickness (IMT), a marker for ASCVD. Methods: We studied three clinical cohorts: 88 patients with severe HTG (mean TG level 20.6 mmol/L), 271 patients with familial hypercholesterolemia (FH) as a contrast group, and 70 normolipidemic controls. Carotid IMT was measured using standardized ultrasound imaging. Statistical analysis was conducted using one-way analysis of variance (ANOVA) to compare mean IMT values, analysis of covariance (ANCOVA) to adjust for confounding variables, specifically age and sex, as well as Spearman pairwise correlation analysis between variables. Results: Unadjusted mean carotid IMT was greater in severe HTG and FH groups compared to controls, however, this was no longer significant for severe HTG after adjustment for age and sex. In contrast, adjusted carotid IMT remained significantly different between the FH and control groups. Conclusions: Our findings suggest that extreme TG elevations in severe HTG patients are not significantly associated with carotid IMT, in contrast to the increased IMT seen in FH patients. These findings add perspective to the complex relationship between severe HTG and ASCVD risk.

Psychiatric mental health nurse practitioners experiencing therapeutic alliance while using tele-mental health: A phenomenological study.

BACKGROUND: Psychiatric mental health nurse practitioners have rapidly adopted and implemented tele-mental health in their practice; however it is unclear how this modality of care affects the experiential quality of therapeutic alliance, simply defined as the interpersonal working bond between provider and patient. OBJECTIVE: This study is the first to explore how psychiatric mental health nurse practitioners experience therapeutic alliance while using tele-mental health. DESIGN: Husserlian phenomenological qualitative study. PARTICIPANTS: A purposive, convenience sample of 17 American psychiatric mental health nurse practitioners who engaged in tele-mental health care were recruited online and interviewed. METHODS: Phenomenological interview transcripts recorded and later thematically coded in the qualitative software MaxQDA. RESULTS: From 1426 individual codes, five major themes and 16 subthemes were discovered. Overall, themes illuminated that psychiatric mental health nurse practitioners could build therapeutic alliance over tele-mental health using inherent interpersonal skills that had to be adapted to the technology. Adaptions included working with patient environmental factors, individual patient considerations, provider ambivalence, and technological observation shifting awareness and communication patterns. CONCLUSIONS: When adapting for the tele-mental health environment, psychiatric mental health nurse practitioners experienced building and sustaining therapeutic alliance with most patients. Unparalleled aspects of tele-mental health allowed for a fuller clinical picture and logistical convenience to see patients more often with ease for both the provider and patient. However, experiential aspects of therapeutic alliance created during in-person care could not be replaced with tele-mental health. In conclusion, participants concluded that a hybrid care model would enhance therapeutic alliance for most patients.

Maternal nano-titanium dioxide inhalation exposure alters placental cyclooxygenase and oxidant balance in a sexually dimorphic manner.

The placenta plays a critical role in nutrient-waste exchange between the maternal and fetal circulation, and thus impacts fetal growth and development. We have previously shown that nano-titanium dioxide (nano-TiO2) inhalation exposure during gestation decreased fetal female pup and placenta mass [1], which persists in the following generation [2]. In utero exposed females, once mated, their offspring's placentas had increased capacity for H2O2 production. Generation of oxidants such as hydrogen peroxide (H2O2), have been shown to impact cyclooxygenase activity, specifically metabolites such as prostacyclin (PGI2) or thromboxane (TXA2). Therefore, we hypothesized that maternal nano-TiO2 inhalation exposure during gestation results in alterations in placental production of prostacyclin and thromboxane mediated by enhanced H2O2 production in a sexually dimorphic manner. Pregnant Sprague-Dawley rats were exposed to nano-TiO2 aerosols or filtered air (sham--control) from gestational day (GD) 10-19. Dams were euthanized on GD 20, and fetal serum and placental tissue were collected based on fetal sex. Fetal placental zones (junctional zone (JZ) and labyrinth zone (LZ)) were assessed for xanthine oxidoreductase (XOR) activity, H2O2, and catalase activity, as well as 6-keto-PGF1α and TXB2 levels. Nano-TiO2 exposed fetal female LZ demonstrated significantly greater XOR activity compared to exposed males. Exposed fetal female LZ also demonstrated significantly diminished catalase activity compared to sham-control females. Exposed fetal female LZ had significantly increased abundance of 6-keto-PGF1α compared to sham-control females and increased TXB2 compared to exposed males. In the aggregate these data indicate that maternal nano-TiO2 inhalation exposure has a greater impact on redox homeostasis and PGI2/TXA2 balance in the fetal female LZ. Future studies need to address if treatment with an XO inhibitor during gestation can prevent diminished fetal female growth during maternal nano-TiO2 inhalation exposure.

A Histone Methylation-MAPK Signaling Axis Drives Durable Epithelial-Mesenchymal Transition in Hypoxic Pancreatic Cancer.

The tumor microenvironment in pancreatic ductal adenocarcinoma (PDAC) plays a key role in tumor progression and response to therapy. The dense PDAC stroma causes hypovascularity, which leads to hypoxia. Here, we showed that hypoxia drives long-lasting epithelial-mesenchymal transition (EMT) in PDAC primarily through a positive-feedback histone methylation-MAPK signaling axis. Transformed cells preferentially underwent EMT in hypoxic tumor regions in multiple model systems. Hypoxia drove a cell autonomous EMT in PDAC cells, which, unlike EMT in response to growth factors, could last for weeks. Furthermore, hypoxia reduced histone demethylase KDM2A activity, suppressed PP2 family phosphatase expression, and activated MAPKs to post-translationally stabilize histone methyltransferase NSD2, leading to an H3K36me2-dependent EMT in which hypoxia-inducible factors played only a supporting role. Hypoxia-driven EMT could be antagonized in vivo by combinations of MAPK inhibitors. Collectively, these results suggest that hypoxia promotes durable EMT in PDAC by inducing a histone methylation-MAPK axis that can be effectively targeted with multidrug therapies, providing a potential strategy for overcoming chemoresistance. SIGNIFICANCE: Integrated regulation of histone methylation and MAPK signaling by the low-oxygen environment of pancreatic cancer drives long-lasting EMT that promotes chemoresistance and shortens patient survival and that can be pharmacologically inhibited. See related commentary by Wirth and Schneider, p. 1739.

Development and validation of the self-injury stigma scale.

Non-suicidal self-injury is a prevalent and concerning behavior. Negative beliefs and stereotypes about NSSI are associated with negative outcomes, and negative, self-referential beliefs (e.g., self-stigmatizing beliefs) related to engagement in NSSI may be particularly harmful. Despite this, there is no validated measure specifically designed to assess for NSSI self-stigma. As this significantly hinders the ability to understand and quantify the effect of NSSI self-stigma, this study sought to validate the newly developed Self-Injury Stigma Scale (SISS). It was hypothesized the SISS would follow a four-factor structure that parallels a widely cited theoretical model of stigma. It was also hypothesized measures of shame and help-seeking self-stigma would be moderately, negatively, correlated with the SISS subscales, supporting the measure's validity. Participants from Study 1 were college students (n = 264, 65.8% female) with at least one lifetime NSSI act. A series of factor analytic models revealed a one-factor structure for the Application of Stigma subscale (i.e., third step of the four-step model). As this was the only SISS subscale to achieve an appropriate model fit, this scale alone was retained as the final SISS. The factor structure was tested via confirmatory factor analysis on a second sample (i.e., community participants with at least one past month of NSSI act; n = 240, 41.3% female). An acceptable fit on most, but not all, indices was reached. Convergent and discriminant validity were supported. The SISS retrospectively predicted past 3 month NSSI frequency and method versatility, and lifetime NSSI versatility, but not lifetime NSSI frequency or disclosure. Thus, the clinical utility of the SISS was partially supported. Findings offering evidence in favor of the SISS's appropriateness and utility as a measure of self-stigma of NSSI. Future work using this measure has the potential to clarify the risk associated with NSSI self-stigma and inform behavioral interventions.

The influence of women's age and fall history on foot and lower limb kinematics during transition step descent.

Falls are a major public health concern, with older women being at the greatest risk to experience a fall. Step descent increases the likelihood of a fall injury, yet the influence of age and fall history on lower extremity kinematics have not been extensively studied. The purpose of this study was to examine lower extremity and foot kinematics of women with and without a fall history during single step descent. Hip, knee, and foot kinematics of young women (n = 15, age = 22.6 ± 3.2 years), older women with no recent falls (n = 15, age = 71.6 ± 4.4 years), and older women with a fall history (n = 15, age = 71.5 ± 5.0 years) as they descended a 17 cm step were examined. Differences in initial contact angles and ROM during landing were examined with between group MANOVA tests. Distal foot initial contact angles were not significant between groups. For range of motion, both older groups went through greater hip extension (p = 0.003, partial η2 = 0.25), but less hip adduction (p = 0.002, partial η2 = 0.27) and less lateral midfoot dorsiflexion (p = 0.001, partial η2 = 0.28) than the younger women. The older fall group had reduced knee flexion (p = 0.004, partial η2 = 0.23) than the younger group, and the older non-fallers slightly plantarflexed at the medial midfoot (p = 0.005, partial η2 = 0.23) while the young women dorsiflexed. Thelanding phase ROMdifferences exhibited by the older adult groupsmayincrease the likelihood of a misstep, which may result in a fall.

Nontoxic Initiator Alternatives to TEMED for Redox Hydrogel Polymerization.

Polymeric hydrogels are versatile biomaterials, offering unique advantages in tunability and biocompatibility that make them well-suited to a range of applications. Cross-linking, a fundamental step in hydrogel fabrication, is often initiated using a toxic redox system, ammonium persulfate (APS), and tetramethylethylenediamine (TEMED), which hinders hydrogel utility in direct contact with cells (e.g., wound dressings). To overcome this limitation, we developed alternative redox gelation systems that serve as nontoxic replacements for TEMED. The alternate initiators were either synthetic or bioinspired amine-containing polymers, Glycofect and polyethylenimine (PEI). Used with APS, these initiator candidates produced hydrogels with short gelation time and comparable moduli to TEMED-based gels and underwent further mechanical testing and biocompatibility characterization. While achieving mechanical properties similar to those of the control, the gels based on Glycofect and PEI outperformed TEMED-based gels in two cell viability studies, with Glycofect-initiated gels displaying significantly higher cytocompatibility. Taken together, these results indicate that Glycofect may serve as a drop-in replacement for TEMED to fabricate hydrogels with improved biocompatibility.

Rhinoplasty Outcomes in Patients with Symptoms of Body Dysmorphia.

BACKGROUND: Patients with symptoms of body dysmorphia often seek consultation for aesthetic rhinoplasty. While body dysmorphic disorder is a formal psychiatric diagnosis, recent evidence indicates that patients with symptoms of this condition who seek rhinoplasty may experience increased satisfaction with their appearance following surgery. OBJECTIVES: To determine the psychological impact of rhinoplasty in patients screened pre-/postoperatively with a body dysmorphia screening questionnaire. METHODS: Retrospective chart review of patients who underwent aesthetic and/or functional rhinoplasty by a single surgeon (S.P.M.) from 6/2021- 4/2023. Adult patients with a complete pre- and postoperative body dysmorphic disorder-aesthetic surgery questionnaire (BDDQ-AS), Standardized Cosmesis and Health Nasal Outcomes Survey-Obstruction and Cosmesis (SCHNOS), and Visual Analog Scale (VAS) were included. Patient characteristics and outcomes were analyzed stratifying by BDDQ-AS screen. RESULTS: One-hundred fifteen patients (88% female) met criteria for inclusion. There was an 83% resolution rate in BDDQ-AS positive screening following rhinoplasty. Positive BDDQ-AS screening status pre- and postoperatively correlated with worse aesthetic satisfaction (all p<0.002). No patient reported outcome measures were indicative of which patients with a BDDQ-AS positive screen preoperatively would experience 'resolution' postoperatively. CONCLUSIONS: Body dysmorphia screening resolution following surgical intervention correlated with improved patient aesthetic satisfaction, pointing to a potential positive psychological impact of undergoing rhinoplasty.

RTF2 controls replication repriming and ribonucleotide excision at the replisome.

DNA replication through a challenging genomic landscape is coordinated by the replisome, which must adjust to local conditions to provide appropriate replication speed and respond to lesions that hinder its progression. We have previously shown that proteasome shuttle proteins, DNA Damage Inducible 1 and 2 (DDI1/2), regulate Replication Termination Factor 2 (RTF2) levels at stalled replisomes, allowing fork stabilization and restart. Here, we show that during unperturbed replication, RTF2 regulates replisome localization of RNase H2, a heterotrimeric enzyme that removes RNA from RNA-DNA heteroduplexes. RTF2, like RNase H2, is essential for mammalian development and maintains normal replication speed. However, persistent RTF2 and RNase H2 at stalled replication forks prevent efficient replication restart, which is dependent on PRIM1, the primase component of DNA polymerase α-primase. Our data show a fundamental need for RTF2-dependent regulation of replication-coupled ribonucleotide removal and reveal the existence of PRIM1-mediated direct replication restart in mammalian cells.