Virginia Commonwealth University

The biology of GZ17-6.02 alone and more so in combination with either of the standard-of-care agents etoposide or carboplatin killed MYCN overexpressing neuroblastoma (NB) cells is unknown. The methods involved in this study are in-cell immunoblotting, trypan blue exclusion, plasmid and siRNA transfection, assessment of autophagy using a plasmid expressing LC3-GFP-RFP. GZ17-6.02 (602) comprises, by mass, a ratio of curcumin (1.0), harmine (1.3), and isovanillin (7.7). In tumors dosed with 602, the ratio becomes curcumin (1.0), harmine (16), and isovanillin (6.1) (602NR). GZ17-6.02 activated ATM, AMPK, ULK1, ATG13, and PERK and inactivated ERBB1, ERBB2, ERBB3, ERBB4, AKT, mTORC1, mTORC2, SRC, NFκB, YAP, and eIF2α. 602 enhanced autophagosome formation and autophagic flux that was amplified when it was combined with etoposide or carboplatin. Compared with 602, 602NR caused significantly greater autophagosome formation that was also amplified when in combination with chemotherapy and which was reduced ~40% by knockdown of ATM or AMPKα and abolished by knockdown of Beclin1 or ATG5. Knockdown of ATM or AMPKα significantly reduced tumor cell death caused by 602 of 602NR, whereas endoplasmic reticulum stress (eIF2α) and macroautophagy (Beclin1, ATG5) were more effective at maintaining tumor cell survival. Combined knockdown of Beclin1 and the death receptor CD95 almost abolished the antitumor actions of 602 and 602NR. 602, and more so 602NR, kills MYCN NB cells and interacts with standard-of-care chemotherapeutics to cause further killing via autophagy and death receptor signaling.

A 3D‐printed origami‐inspired magnetic scaffold has been developed to investigate the influence of physical cues on guided cellular proliferation in a 3D microenvironment. Microscale channels are first constructed and populated with NIH/3T3 fibroblast and/or A549 cancer cell clusters that are initially bioprinted within the channels. Once these channels are fully populated, a permanent magnet is applied to fold the scaffolds. By varying the channel width and incorporating an intermediate extracellular matrix hydrogel (IE) layer along with origami folding, the scaffold provides geometric and gravitational cues to influence cellular proliferation. In both monoculture and coculture, i) cells tend to proliferate more in a tapered manner, ii) scaffolds with enhanced media flow lead to a higher volume of cell growth, and iii) cells form homogeneous distributions under gravity after dispersion. In coculture, the expansion of fibroblast clusters within their seeded channels increased, facilitating the proliferation of cancer cell clusters into the non‐seeded channels. This origami scaffold offers valuable insights into tissue engineering and cancer research, serving as a versatile tool for examining cellular interactions and growth dynamics.

Air pollution, particularly fine particulate matter (PM2.5), has been associated with various health issues, but its effects on skin health, specifically skin redness, remain underexplored. This study aims to examine the relationship between PM2.5 exposure and skin redness, with a focus on the role of sebum production in different age groups. A total of 472 participants from two communities in Taiwan in two age groups (20–59 years, n=240; over 60 years, n=232) were included in the study. PM2.5 exposure levels were estimated using land use regression models based on participants’ residential addresses. Skin redness area was assessed using the VISIA Imaging System. Linear regression analyses were conducted to examine the association between PM2.5 and redness area, adjusting for demographic, lifestyle, and ultraviolet exposure. Results showed a significant positive association between PM2.5 levels and redness area in both age groups. In the 20-59 age group, each unit increase in PM2.5 corresponded to a 1.70-unit increase in redness area (95% CI: 0.32 – 3.07, p < 0.01), while in the over-60 group, the increase was 2.63 units (95% CI: 1.19 – 4.08, p < 0.001). Additionally, porphyrins showed a positive association with redness area among the 20-59 age group (p < 0.05), while no significant association was found in the over-60 group. This study suggests a linkage between PM2.5 exposure and skin redness area, indicating that air pollution may be a contributing factor to skin health issues. The findings suggest that the interaction between lipophilic and carcinogenic substances in PM2.5 and porphyrins could elevate redness area levels and potentially increase the risk of chronic skin conditions and skin cancer.

Sustainability of permanent magnets is an increasingly important topic as technologies such as electric vehicles or advanced wind turbines are further developed. A potential alternative lies in extraterrestrial sources, such as the asteroid Psyche, which is rich in a tetrataenite that has high magnetocrystalline anisotropy and high saturation magnetization, making it a potential permanent magnet. Using a robot to extract and collect tetrataenite provides an avenue for replacing rare-earth element-free magnets where possible, but the state-of-the-art end effectors for grasping magnetic materials are inappropriate for these conditions. Operating in space introduces a significant constraint in the form of energy utilization, and most magnetic grippers are constructed with electromagnets, which are quite energy-intensive. We propose a design that instead uses an actuate set of parallel linear Halbach arrays to toggle between the strong and weak magnetic fields to grasp and drop objects, respectively. As part of the design process, field simulations were performed to ensure proper interaction between the arrays in the grasping and dropping configurations. To verify this data, the field of a single Halbach array was measured and compared to the simulations. A gripper prototype was fabricated and tested with a collaborative UR5e robot using several objects (4140 alloy steel) in varying orientations to determine the payload limits of the gripper, which was approximated to a maximum of 1.50 kg. This is correlated to magnetic field simulations in all grasping trials showing field strength and variance to provide a rough estimate for the gripper’s ability to grasp varied sizes of objects.

Merkel cell carcinoma (MCC) is an aggressive malignant neuroendocrine tumor of the skin. While considerable advances in the management of MCC have been made, the clinical utility of potential biomarkers remains unclear. Biomarkers play a crucial role in assessing prognosis, diagnosis, and therapeutic interventions in many chronic diseases and may be similarly beneficial in MCC. This chapter focuses on examining potential biomarkers in MCC. Through conducting a literature search, 22 biomarkers were identified and classified based on the classification standards provided by the World Health Organization (WHO). These biomarkers have the potential to aid in screening, diagnosis, prognosis, monitoring, and pharmacodynamic properties of MCC.

Hidradenitis suppurativa (HS) is an inflammatory skin condition involving dysregulation of the innate and adaptive immune responses. Although the precise etiology is not yet elucidated, the pathophysiology of HS is multifactorial and associated with genetic and environment factors. There is currently no cure for HS. Biomarkers have been identified in the blood and tissue of patients with the disease, and the levels of which may be informative about diagnosis, prognosis, treatment efficacy, and monitoring comorbidities for this condition. These biomarkers encompass a wide range of molecules and include inflammatory cytokines, chemokine receptors, growth factors, markers of serum inflammation, and imaging features (Napolitano et al. in Clin Cosmet Investig Dermatol 10:105–115, 2017). Together, these biomarkers serve as important indicators of disease progression and response to treatment. The World Health Organization (WHO) categorizes biomarkers into screening, diagnostic, prognostic, monitoring, and pharmacodynamics. The chapter will discuss the role of biomarkers in HS, focusing on diagnosis, prognosis, monitoring, and therapeutic targets.

Melanoma is a skin cancer that originates from melanocytes, cells that sit in the basal layer of the epidermis and synthesize melanin. The gold standard for the diagnosis is histopathologic evaluation; however, melanoma can show a wide range of histopathologic features, including subtle histologic atypia, which can pose diagnostic challenges. Additionally, there are other melanocytic tumors that remain incompletely understood, designated as melanocytic tumors of uncertain malignant potential (MELTUMP). Biomarkers could potentially address limitations in disease screening, diagnosis, prognosis, and management of melanoma. This chapter focuses on analyzing the biomarkers most closely associated with clinical outcomes in melanoma patients. We conducted a literature review of 14 biomarkers and used classification standards provided by the World Health Organization (WHO). In this review, we discuss the measurable levels of antigens, proteins, and antibodies associated with melanoma diagnosis, prognosis, and response to treatment. The presence and extent of immunohistochemical staining, the serum levels of biomarkers, and the presence of inflammatory cell infiltrates can be analyzed to identify patients who qualify for further testing for melanoma, to distinguish malignant melanoma from benign nevi, to differentiate melanoma from other tumor types, to predict patient outcomes, to monitor the progression of melanoma, and to predict response to therapies.

The metazoan lifespan is determined in part by a complex signaling network that regulates energy metabolism and stress responses. Key signaling hubs in this network include insulin/IGF‐1, AMPK, mTOR, and sirtuins. The Hippo/Mammalian Ste20‐like Kinase1 (MST1) pathway has been reported to maintain lifespan in Caenorhabditis elegans, but its role has not been studied in higher metazoans. In this study, we report that overexpression of Hpo, the MST1 homolog in Drosophila melanogaster, decreased lifespan with concomitant changes in lipid metabolism and aging‐associated gene expression, while RNAi Hpo depletion increased lifespan. These effects were mediated primarily by Hpo‐induced transcriptional activation of the RNA‐binding protein maternal expression at 31B (Me31b)/RCK, resulting in stabilization of mRNA‐encoding a lipolytic hormone, Akh. In mouse adipocytes, Hpo/Mst1 mediated adipocyte differentiation, phosphorylation of RNA‐binding proteins such as Rck, decapping MRNA 2 (Dcp2), enhancer Of MRNA decapping 3 (Edc3), nucleolin (NCL), and glucagon mRNA stability by interacting with Rck. Decreased lifespan in Hpo‐overexpressing Drosophila lines required expression of Me31b, but not DCP2, which was potentially mediated by recovering expression of lipid metabolic genes and formation of lipid droplets. Taken together, our findings suggest that Hpo/Mst1 plays a conserved role in longevity by regulating adipogenesis and fatty acid metabolism.

Background Limited understanding of the biology predisposing certain human papillomavirus-related (HPV+) oropharyngeal squamous cell carcinomas (OPSCCs) to relapse impedes therapeutic personalization. We aimed to identify molecular traits that distinguish recurrence-prone tumors. Methods 50 HPV+ OPSCCs that later recurred (cases) and 50 non-recurrent controls matched for stage, therapy, and smoking history were RNA-sequenced. Groups were compared by gene set enrichment analysis, and select differences were validated by immunohistochemistry. Features discriminating groups were scored in each tumor using gene set variation analysis, and scores were evaluated for recurrence prediction ability. Results Cases downregulated pathways linked to anti-tumor immunity (FDR-adjusted p<.05) and contained fewer tumor-infiltrating lymphocytes (p<.001), including cytotoxic T-cells (p=.005). Cases also upregulated pathways related to cell division and other aspects of tumor progression. Upregulated and downregulated pathways were respectively used to define a tumor progression score (TPS) and immune suppression score (ISS) for each tumor. Correlation between TPS and ISS (r=.603, p<.001) was potentially explained by observed upregulation of DNA repair pathways in cases, which might enhance their progression directly and by limiting cytosolic DNA-induced inflammation. Accordingly, cases contained fewer double-strand breaks based on staining for phospho-RPA32 (p=.006) and γ-H2AX (p=.005) and downregulated the cytosolic DNA sensing pathway. A combined score derived from TPS and ISS optimized recurrence prediction and stratified survival in a manner generalizable to three external cohorts. Conclusions We describe a potential link in HPV+ OPSCCs between reduced DNA damage and other tumor-intrinsic and immune-related contributors to recurrence risk, opening opportunities to detect and target this high-risk biology.

Importance According to survey data, 12.8% of households experienced food insecurity in 2022, with 7.7% of households experiencing low food security and 5.1% experiencing very low food security. Nearly one-third of households with incomes below the federal poverty threshold are food insecure. Food insecurity is one among a multitude of medical, psychological, and social conditions common among economically disadvantaged households. Objective The US Preventive Services Task Force (USPSTF) commissioned a systematic review to evaluate the evidence on the benefits and harms of screening for food insecurity in the health care setting. Population Children, adolescents, and adults. Evidence Assessment The USPSTF concludes that the evidence is insufficient and the balance of benefits and harms for screening for food insecurity on health outcomes in the primary care setting cannot be determined. Recommendation The USPSTF concludes that the current evidence is insufficient to assess the balance of benefits and harms of screening for food insecurity on health outcomes in the primary care setting. (I statement)

Objective: In this review, we will examine the more common endpoints incorporated in randomized controlled trials (RCTs) and their strength of evidence, focusing on the definition of what constitutes a clinically meaningful change. We will also reflect on the perspective of patients and their families regarding the design of RCTs in amyotrophic lateral sclerosis (ALS). Methods: Authors performed a scoping review of the literature around clinical meaningfulness in the ALS field and the minimum clinically important difference to deem a treatment effective. Results: The use of survival as an RCT endpoint, as well as the ALS functional rating scale-revised slope, has been criticized, and their relevance for patients remains debated. Biomarkers are promising alternatives as surrogate endpoints, but currently, only cerebrospinal fluid and plasma neurofila-ments have emerged as reliable and sensitive biomarkers of disease progression. Incorporating patients' preferences and priorities for their care when treatments are selected is important to minimize the burden of care and limit the potential harms of overtreatment. Patients' interest in and acceptance of a new therapy is also determined by its impact on their quality of life. Discussion and conclusion: While scientifically sound trials must be conducted, this must be balanced with patient expectations of limiting trial burden, duration and placebo usage. An important approach in uniting these diverging needs is the inclusion of people with ALS and their organizations to advise in the design and execution of clinical trials, facilitating the design of RCTs more focused on patients' expectations while retaining a high scientific rigor.