Understanding the Intersection of Identity and Cancer Experience Among Racially, Ethnically, Gender and Sexual Minoritized Adolescents and Young Adults With Cancer.

OBJECTIVES: Adolescents and young adults (AYAs, 18-39 years) with cancer identifying as racially/ethnically minoritized or 2SLGBTQIA+ (Two-Spirit, lesbian, gay, bisexual, transgender, queer, intersex, asexual and "+" referring to other queer identities) have been underrepresented in cancer research. This study explores the aspects of identity that hold significance for these minoritized AYAs and how these facets impact their healthcare experiences. METHODS: Eligible participants comprised English-speaking AYAs who self-identified as racially/ethnically minoritized and/or 2SLGBTQIA+, were diagnosed with cancer between the ages of 15-39, currently aged > 18, and had received or were receiving cancer care within Canadian healthcare system. Additionally, four patient partners meeting the same criteria were recruited as research collaborators. Semi-structured one-on-one virtual interviews guided by an interview script were conducted, and qualitative analysis employed a framework approach. RESULTS: We recruited 23 participants from 4 Canadian provinces (mean age: 28, Range: 20-44); 17 identified as racially/ethnically minoritized, one as sexual/gender minoritized, and five as racially/ethnically and sexually/gender minoritized. Participants emphasized that their culture/ethnicity, religion/spirituality, sexuality, gender, family, career, and being an immigrant are important aspects of their identity, with only one participant recognizing their identity as a "person with cancer". A cancer diagnosis altered the aspects of identity deemed most significant by participants. Both visible and invisible aspects of identity shaped participants' experiences and influenced their level of trust in the healthcare system. CONCLUSION: Racially, ethnically, gender, or sexually minoritized AYAs with cancer place considerable importance on aspects of their identity that are shaped by their respective communities. Recognizing and respecting these identities are paramount for healthcare professionals to deliver safe and inclusive care.

SIRT5 stabilizes mitochondrial glutaminase and supports breast cancer tumorigenesis.

The mitochondrial enzyme glutaminase (GLS) is frequently up-regulated during tumorigenesis and is being evaluated as a target for cancer therapy. GLS catalyzes the hydrolysis of glutamine to glutamate, which then supplies diverse metabolic pathways with carbon and/or nitrogen. Here, we report that SIRT5, a mitochondrial NAD+-dependent lysine deacylase, plays a key role in stabilizing GLS. In transformed cells, SIRT5 regulates glutamine metabolism by desuccinylating GLS and thereby protecting it from ubiquitin-mediated degradation. Moreover, we show that SIRT5 is up-regulated during cellular transformation and supports proliferation and tumorigenesis. Elevated SIRT5 expression in human breast tumors correlates with poor patient prognosis. These findings reveal a mechanism for increasing GLS expression in cancer cells and establish a role for SIRT5 in metabolic reprogramming and mammary tumorigenesis.

Creating a Vision for a Healthier Workforce Using a Systems-Based Approach.

CONTEXT: The public health system faces unprecedented challenges due to the pandemic, racism, health inequity, and the politicization of public health. At all levels of the system, the workforce is experiencing distress, burnout, safety issues, and attrition. Public health is being challenged to demonstrate and justify its impact and value, while also leveraging opportunities for learning and system strengthening. PROGRAM: To explore the current state and identify opportunities to strengthen the public health system, the Region 7 Midwestern Public Health Training Center (MPHTC), with support from Engaging Inquiry, embarked on a distinctive type of systems analysis, called "dynamic systems mapping." IMPLEMENTATION: This approach brought together diverse sectors of public health partners in the region to develop a rich contextual narrative and system-level understanding to highlight and align existing and emergent strengths, areas for growth, and tangible goals for the immediate- and long-term sustainability of local and regional health. EVALUATION: Focus groups and workshops were conducted with diverse practitioners to identify upstream causes and downstream effects of 11 key forces driving system behavior. These focus groups resulted in the development of a visual map that MPHTC is utilizing to identify opportunities for leverage, develop strategies to maximize the potential impact of these leverage points, as well as facilitate continuous learning. DISCUSSION: Public health utilization of systems mapping is a valuable approach to strengthening local and national system responses to current and future public health needs. Outcomes and lessons learned from the systems mapping process are discussed.

Virtual Skills Training Format for Teaching Intrauterine Contraception Insertion During Coronavirus Disease 2019.

The coronavirus disease 2019 pandemic resulted in the curtailment of face-to-face clinical skills training in 2020. To meet Title X workforce needs, the National Clinical Training Center for Family Planning transitioned onsite intrauterine contraception training to a virtual format using online didactic material and livestreamed training and telementoring. Videos demonstrated the placement and removal of intrauterine contraceptives, and all necessary supplies were shipped directly to participants. Attendees reported a high level of skill uptake and impact on their practice. This pilot study suggests that virtual skills training is suitable for providers unable to travel to in-person events.

Animal Models of Peripheral Pain: Biology Review and Application for Drug Discovery.

Pain is a complex constellation of cognitive, unpleasant sensory, and emotional experiences that primarily serves as a survival mechanism. Pain arises in the peripheral nervous system and pain signals synapse with nerve tracts extending into the central nervous system. Several different schemes are used to classify pain, including the underlying mechanism, tissues primarily affected, and time-course. Numerous animal models of pain, which should be employed with appropriate Institutional Animal Care and Use approvals, have been developed to elucidate pathophysiology mechanisms and aid in identification of novel therapeutic targets. The variety of available models underscores the observations that pain phenotypes are driven by several distinct mechanisms. Pain outcome measurement encompasses both reflexive (responses to heat, cold, mechanical and electrical stimuli) and nonreflexive (spontaneous pain responses to stimuli) behaviors. However, the question of translatability to human pain conditions and potential treatment outcomes remains a topic of continued scrutiny. In this review we discuss the different types of pain and their mechanisms and pathways, available rodent pain models with an emphasis on type of pain stimulations and pain outcome measures and discuss the role of pathologists in assessing and validating pain models.

Volitional media multitasking: awareness of performance costs and modulation of media multitasking as a function of task demand.

In two experiments, we sought to determine whether (a) people are aware of the frequently observed performance costs associated with engaging in media multitasking (Experiment 1), and (b) if so, whether they modulate the extent to which they engage in multitasking as a function of task demand (Experiment 2). In Experiment 1, participants completed a high-demand task (2-back) both independently and while a video was simultaneously presented. To determine whether people were sensitive to the impact that the concurrent video had on primary-task performance, subjective estimates of performance were collected following both trial types (No-Video vs. Video trials), as were explicit beliefs about the influence of the video on performance. In Experiment 2, we modified our paradigm by allowing participants to turn the video on and off at their discretion, and had them complete either a high-demand task (2-back) or a low-demand task (0-back). Findings from Experiment 1 indicated that people are sensitive to the magnitude of the decrement that media multitasking has on primary-task performance. In addition, findings from Experiment 2 indicated that people modulate the extent to which they engage in media multitasking in accordance with the demands of their primary task. In particular, participants completing the high-demand task were more likely to turn off the optional video stream compared to those completing the low-demand task. The results suggest that people media multitask in a strategic manner by balancing considerations of task performance with other potential concerns.

Duchenne and Becker Muscular Dystrophies: A Review of Animal Models, Clinical End Points, and Biomarker Quantification.

Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are neuromuscular disorders that primarily affect boys due to an X-linked mutation in the DMD gene, resulting in reduced to near absence of dystrophin or expression of truncated forms of dystrophin. Some newer therapeutic interventions aim to increase sarcolemmal dystrophin expression, and accurate dystrophin quantification is critical for demonstrating pharmacodynamic relationships in preclinical studies and clinical trials. Current challenges with measuring dystrophin include the variation in protein expression within individual muscle fibers and across whole muscle samples, the presence of preexisting dystrophin-positive revertant fibers, and trace amounts of residual dystrophin. Immunofluorescence quantification of dystrophin can overcome many of these challenges, but manual quantification of protein expression may be complicated by variations in the collection of images, reproducible scoring of fluorescent intensity, and bias introduced by manual scoring of typically only a few high-power fields. This review highlights the pathology of DMD and BMD, discusses animal models of DMD and BMD, and describes dystrophin biomarker quantitation in DMD and BMD, with several image analysis approaches, including a new automated method that evaluates protein expression of individual muscle fibers.

Viewing Telehealth Through the Lens of a Community Benefit Mission.

INTRODUCTION: This article reviews the intersection of common objectives between telehealth initiatives launched by health systems and community health (specifically community benefit). Telehealth initiatives can benefit communities and play a potential role in improving population health. There is great opportunity to better integrate telehealth activities into community health strategies and community benefit reporting (for nonprofit hospitals) while building the case for telehealth service expansion. METHODS: Through descriptive, hypothetical cases, we demonstrate the intersection of the triple aim, specifically improving population-level health, and three common objectives between telehealth and community benefit: (1) improving access to care, (2) enhancing community health, and (3) advancing medical or healthcare knowledge. CONCLUSIONS: This article is intended to spur inspiration for possible telehealth and community health/community benefit connections.

Commentary: Roles for Pathologists in a High-throughput Image Analysis Team.

Historically, pathologists perform manual evaluation of H&E- or immunohistochemically-stained slides, which can be subjective, inconsistent, and, at best, semiquantitative. As the complexity of staining and demand for increased precision of manual evaluation increase, the pathologist's assessment will include automated analyses (i.e., "digital pathology") to increase the accuracy, efficiency, and speed of diagnosis and hypothesis testing and as an important biomedical research and diagnostic tool. This commentary introduces the many roles for pathologists in designing and conducting high-throughput digital image analysis. Pathology review is central to the entire course of a digital pathology study, including experimental design, sample quality verification, specimen annotation, analytical algorithm development, and report preparation. The pathologist performs these roles by reviewing work undertaken by technicians and scientists with training and expertise in image analysis instruments and software. These roles require regular, face-to-face interactions between team members and the lead pathologist. Traditional pathology training is suitable preparation for entry-level participation on image analysis teams. The future of pathology is very exciting, with the expanding utilization of digital image analysis set to expand pathology roles in research and drug development with increasing and new career opportunities for pathologists.

Collaboration among Missouri nonprofit hospitals and local health departments: content analysis of community health needs assessments.

OBJECTIVES: We identified the levels of joint action that led to collaboration between hospitals and local health departments (LHDs) using the hospital's community health needs assessments (CHNAs). METHODS: In 2014, we conducted a content analysis of Missouri nonprofit hospitals (n = 34) CHNAs, and identified hospitals based on previously reported collaboration with LHDs. We coded the content according to the level of joint action. A comparison sample (n = 50) of Missouri nonprofit hospitals provided the basic comparative information on hospital characteristics. RESULTS: Among the hospitals identified by LHDs, 20.6% were "networking," 20.6% were "coordinating," 38.2% were "cooperating," and 2.9% were "collaborating." Almost 18% of study hospitals had no identifiable level of joint action with LHDs based on their CHNAs. In addition, comparison hospitals were more often part of a larger system (74%) compared with study hospitals (52.9%). CONCLUSIONS: The results of our study helped develop a better understanding of levels of joint action from a hospital perspective. Our results might assist hospitals and LHDs in making more informed decisions about efficient deployment of resources for assessment processes and implementation plans.

Myofilament dysfunction as an emerging mechanism of volume overload heart failure.

Two main hemodynamic overload mechanisms [i.e., volume and pressure overload (VO and PO, respectively] result in heart failure (HF), and these two mechanisms have divergent pathologic alterations and different pathophysiological mechanisms. Extensive evidence from animal models and human studies of PO demonstrate a clear association with alterations in Ca(2+) homeostasis. By contrast, emerging evidence from animal models and patients with regurgitant valve disease and dilated cardiomyopathy point toward a more prominent role of myofilament dysfunction. With respect to VO HF, key features of excitation-contraction coupling defects, myofilament dysfunction, and extracellular matrix composition will be discussed.

Effects of a myofilament calcium sensitizer on left ventricular systolic and diastolic function in rats with volume overload heart failure.

Aortocaval fistula (ACF)-induced volume overload (VO) heart failure (HF) results in progressive left ventricular (LV) dysfunction. Hemodynamic load reversal during pre-HF (4 wk post-ACF; REV) results in rapid structural but delayed functional recovery. This study investigated myocyte and myofilament function in ACF and REV and tested the hypothesis that a myofilament Ca(2+) sensitizer would improve VO-induced myofilament dysfunction in ACF and REV. Following the initial sham or ACF surgery in male Sprague-Dawley rats (200-240 g) at week 0, REV surgery and experiments were performed at weeks 4 and 8, respectively. In ACF, decreased LV function is accompanied by impaired sarcomeric shortening and force generation and decreased Ca(2+) sensitivity, whereas, in REV, impaired LV function is accompanied by decreased Ca(2+) sensitivity. Intravenous levosimendan (Levo) elicited the best inotropic and lusitropic responses and was selected for chronic oral studies. Subsets of ACF and REV rats were given vehicle (water) or Levo (1 mg/kg) in drinking water from weeks 4-8. Levo improved systolic (% fractional shortening, end-systolic elastance, and preload-recruitable stroke work) and diastolic (τ, dP/dtmin) function in ACF and REV. Levo improved Ca(2+) sensitivity without altering the amplitude and kinetics of the intracellular Ca(2+) transient. In ACF-Levo, increased cMyBP-C Ser-273 and Ser-302 and cardiac troponin I Ser-23/24 phosphorylation correlated with improved diastolic relaxation, whereas, in REV-Levo, increased cMyBP-C Ser-273 phosphorylation and increased α-to-ß-myosin heavy chain correlated with improved diastolic relaxation. We concluded that Levo improves LV function, and myofilament composition and regulatory protein phosphorylation likely play a key role in improving function.

Describing the continuum of collaboration among local health departments with hospitals around the community health assessments.

BACKGROUND: Hospitals and local health departments (LHDs) are under policy requirements from the Affordable Care Act and accreditation standards through the Public Health Accreditation Board. Tax exempt hospitals must perform a community health needs assessment (CHNA), similar to the community health assessment (CHA) required for LHDs. These efforts have led to a renewed interest in hospitals and LHDs working together to achieve common goals. PURPOSE: The purpose of this study is to gain a better understanding of levels of joint action leading toward collaboration between LHDs and hospitals and describe collaboration around CHAs. METHODS: Local health departments were selected on the basis of reporting collaboration (n = 26) or unsure about collaboration (n = 29) with local hospitals. Local health departments were surveyed regarding their relationship with local hospitals. For LHDs currently collaborating with a hospital, a collaboration continuum scale was calculated. Appropriate nonparametric tests, chi-squares, and Spearman's rank correlations were conducted to determine differences between groups. RESULTS: A total of 44 LHDs responded to the survey (80.0%). Currently collaborating LHDs were more likely to be interested in accreditation and to refer to their CHA 5 or more times a year compared to the unsure LHDs. In the analysis, a collaboration continuum was created and is positively correlated with aspects of the CHA and CHA process. CONCLUSIONS: This study is the first attempt to quantify the level of collaboration between LHDs and hospitals around CHAs. Better understanding of the levels of joint action required may assist LHDs in making informed decisions regarding deployment of resources on the path to accreditation.

PTEN loss in glioma cell lines leads to increased extracellular vesicles biogenesis and PD-L1 cargo in a PI3K-dependent manner.

Phosphatase and Tensin Homologue (PTEN) is one of the most frequently lost tumor suppressors in cancer and the predominant negative regulator of the PI3K/AKT signaling axis. A growing body of evidence has highlighted the loss of PTEN with immuno-modulatory functions including the upregulation of the programmed death ligand-1 (PD-L1), an altered tumor derived secretome that drives an immunosuppressive tumor immune microenvironment (TIME), and resistance to certain immunotherapies. Given their roles in immunosuppression and tumor growth, we examined whether the loss of PTEN would impact the biogenesis, cargo, and function of extracellular vesicles (EVs) in the context of the anti-tumor associated cytokine interferon-γ (IFN-γ). Through genetic and pharmacological approaches, we show that PD-L1 expression is regulated by JAK/STAT signaling, not PI3K signaling. Instead, we observe that PTEN loss positively upregulates cell surface levels of PD-L1 and enhances the biogenesis of EVs enriched with PD-L1 in a PI3K-dependent manner. We demonstrate that because of these changes, EVs derived from glioma cells lacking PTEN have a greater ability to suppress T cell receptor (TCR) signaling. Taken together, these findings provide important new insights into how the loss of PTEN can contribute to an immunosuppressive TIME, facilitate immune evasion, and highlight a novel role for PI3K signaling in the regulation of EV biogenesis and the cargo they contain.

A multiprotein signaling complex sustains AKT and mTOR/S6K activity necessary for the survival of cancer cells undergoing stress.

Cancer cells encounter stresses during tumor progression and metastatic spread, however, how they survive these challenges is not fully understood. We now identify a mechanism for cancer cell survival through the discovery of a multiprotein signaling complex that includes the GTPase Cdc42, the Cdc42 GEF/effector protein Dock7, AKT, mTOR and the mTORC1 regulatory partners TSC1, TSC2, and Rheb. This pro-survival signaling complex sustains the activated state of AKT by preventing its dephosphorylation at Ser473 during serum starvation, resulting in a low but critical activation of a Raptor-independent mTOR/S6K activity. We demonstrate that the Dock7 DHR1 domain, previously of unknown function, is responsible for preserving AKT phosphorylation through an interaction requiring its C2-like motif. Collectively, these findings help address long-standing questions of how Cdc42 signals mTOR activation by elucidating the unique functions of its signaling partner Dock7 as an AKT regulator necessary for resistance to anoikis and apoptosis in cancer cells.

The case for the green banana: Using fruit to improve tenaculum placement simulation.

OBJECTIVES: The purpose of this study was to identify a fruit that would resemble the 'feel' of a uterine cervix when compared to plastic pelvic models. STUDY DESIGN: A group of experienced clinicians tested six different fruits and evaluated their suitability as a cervical simulation for tenaculum placement. RESULTS: An unripe (green) banana was rated the highest in seven out of eight characteristics and was the only fruit to achieve a consensus for overall simulation above 70%. CONCLUSIONS: A green banana is an affordable and accessible option to simulate a cervix for procedures requiring tenaculum placement on a cervix.

Inhibiting Glutamine Metabolism Blocks Coronavirus Replication in Mammalian Cells.

Developing therapeutic strategies against COVID-19 has gained widespread interest given the likelihood that new viral variants will continue to emerge. Here we describe one potential therapeutic strategy which involves targeting members of the glutaminase family of mitochondrial metabolic enzymes (GLS and GLS2), which catalyze the first step in glutamine metabolism, the hydrolysis of glutamine to glutamate. We show three examples where GLS expression increases during coronavirus infection of host cells, and another in which GLS2 is upregulated. The viruses hijack the metabolic machinery responsible for glutamine metabolism to generate the building blocks for biosynthetic processes and satisfy the bioenergetic requirements demanded by the 'glutamine addiction' of virus-infected host cells. We demonstrate how genetic silencing of glutaminase enzymes reduces coronavirus infection and that newer members of two classes of small molecule allosteric inhibitors targeting these enzymes, designated as SU1, a pan-GLS/GLS2 inhibitor, and UP4, which is specific for GLS, block viral replication in mammalian epithelial cells. Overall, these findings highlight the importance of glutamine metabolism for coronavirus replication in human cells and show that glutaminase inhibitors can block coronavirus infection and thereby may represent a novel class of anti-viral drug candidates. Teaser: Inhibitors targeting glutaminase enzymes block coronavirus replication and may represent a new class of anti-viral drugs.

To bind or not to bind: addressing the question of object representation in visual short-term memory.

Visual short-term memory (VSTM) is a capacity limited resource, which is consistently estimated to hold about four visual items at a time. There is, however, debate in the literature about what constitutes an "item" and how resources are allocated within VSTM. Some research suggests information is stored in VSTM as discrete objects; however, there is also evidence suggesting that within-object features alter VSTM performance. The present study addresses the question of whether VSTM load effects reflect the number of discrete objects and/or the number of within-object features. An electrophysiological correlate of VSTM--the contralateral delay activity (CDA)--was measured while participants performed a lateralized change-detection task, in which to-be-remembered items varied in the number of features and locations. Each trial contained either a solitary simple feature (shape, color, or orientation) or one of two multifeature arrays: three features presented at three separate locations or three features bound at one location. While presenting multiple features--regardless of whether they are at discrete locations or bound within a single object--resulted in greater CDA amplitude relative to a solitary feature, there was a dissociation in the distribution of activity between the two multifeature conditions, such that the CDA at site P1/P2 was sensitive to the number of discrete objects, while activity at P7/P8 was most enhanced when multiple features were bound in one object. The findings demonstrate the inhomogeneity of the CDA and suggest this electrophysiological marker may reflect both discrete object individuation/separation and flexible feature-feature binding in VSTM.

Elucidation of an mTORC2-PKC-NRF2 pathway that sustains the ATF4 stress response and identification of Sirt5 as a key ATF4 effector.

Proliferating cancer cells are dependent on glutamine metabolism for survival when challenged with oxidative stresses caused by reactive oxygen species, hypoxia, nutrient deprivation and matrix detachment. ATF4, a key stress responsive transcription factor, is essential for cancer cells to sustain glutamine metabolism when challenged with these various types of stress. While it is well documented how the ATF4 transcript is translated into protein as a stress response, an important question concerns how the ATF4 message levels are sustained to enable cancer cells to survive the challenges of nutrient deprivation and damaging reactive oxygen species. Here, we now identify the pathway in triple negative breast cancer cells that provides a sustained ATF4 response and enables their survival when encountering these challenges. This signaling pathway starts with mTORC2, which upon sensing cellular stresses arising from glutamine deprivation or an acute inhibition of glutamine metabolism, initiates a cascade of events that triggers an increase in ATF4 transcription. Surprisingly, this signaling pathway is not dependent on AKT activation, but rather requires the mTORC2 target, PKC, which activates the transcription factor Nrf2 that then induces ATF4 expression. Additionally, we identify a sirtuin family member, the NAD+-dependent de-succinylase Sirt5, as a key transcriptional target for ATF4 that promotes cancer cell survival during metabolic stress. Sirt5 plays fundamental roles in supporting cancer cell metabolism by regulating various enzymatic activities and by protecting an enzyme essential for glutaminolysis, glutaminase C (GAC), from degradation. We demonstrate that ectopic expression of Sirt5 compensates for knockdowns of ATF4 in cells exposed to glutamine deprivation-induced stress. These findings provide important new insights into the signaling cues that lead to sustained ATF4 expression as a general stress-induced regulator of glutamine metabolism, as well as highlight Sirt5 an essential effector of the ATF4 response to metabolic stress.

A T-cell engaging bispecific antibody with a tumor-selective bivalent folate receptor alpha binding arm for the treatment of ovarian cancer.

The use of T-cell engagers (TCEs) to treat solid tumors is challenging, and several have been limited by narrow therapeutic windows due to substantial on-target, off-tumor toxicities due to the expression of low levels of target antigens on healthy tissues. Here, we describe TNB-928B, a fully human TCE that has a bivalent binding arm for folate receptor alpha (FRα) to selectively target FRα overexpressing tumor cells while avoiding the lysis of cells with low levels of FRα expression. The bivalent design of the FRα binding arm confers tumor selectivity due to low-affinity but high-avidity binding to high FRα antigen density cells. TNB-928B induces preferential effector T-cell activation, proliferation, and selective cytotoxic activity on high FRα expressing cells while sparing low FRα expressing cells. In addition, TNB-928B induces minimal cytokine release compared to a positive control TCE containing OKT3. Moreover, TNB-928B exhibits substantial ex vivo tumor cell lysis using endogenous T-cells and robust tumor clearance in vivo, promoting T-cell infiltration and antitumor activity in mouse models of ovarian cancer. TNB-928B exhibits pharmacokinetics similar to conventional antibodies, which are projected to enable favorable administration in humans. TNB-928B is a novel TCE with enhanced safety and specificity for the treatment of ovarian cancer.