Regulation of cargo selection in exosome biogenesis and its biomedical applications in cancer.

Extracellular vesicles (EVs), including exosomes, are increasingly recognized as potent mediators of intercellular communication due to their capacity to transport a diverse array of bioactive molecules. They assume vital roles in a wide range of physiological and pathological processes and hold significant promise as emerging disease biomarkers, therapeutic agents, and carriers for drug delivery. Exosomes encompass specific groups of membrane proteins, lipids, nucleic acids, cytosolic proteins, and other signaling molecules within their interior. These cargo molecules dictate targeting specificity and functional roles upon reaching recipient cells. Despite our growing understanding of the significance of exosomes in diverse biological processes, the molecular mechanisms governing the selective sorting and packaging of cargo within exosomes have not been fully elucidated. In this review, we summarize current insights into the molecular mechanisms that regulate the sorting of various molecules into exosomes, the resulting biological functions, and potential clinical applications, with a particular emphasis on their relevance in cancer and other diseases. A comprehensive understanding of the loading processes and mechanisms involved in exosome cargo sorting is essential for uncovering the physiological and pathological roles of exosomes, identifying therapeutic targets, and advancing the clinical development of exosome-based therapeutics.

Activated Leukocyte Cell Adhesion Molecule Regulates the Expression of Interleukin-33 in RSV Induced Airway Inflammation by Regulating MAPK Signaling Pathways.

PURPOSE: The respiratory syncytial virus (RSV) is a common respiratory virus that causes acute lower respiratory tract infectious diseases, particularly in young children and older individuals. Activated leukocyte cell adhesion molecule (ALCAM) is a membrane glycoprotein expressed in various cell types, including epithelial cells, and is associated with inflammatory responses and various cancers. However, the precise role of ALCAM in RSV-induced airway inflammation remains unclear, and our study aimed to explore this gap in the literature. METHODS: C57BL/6 wild-type, ALCAM knockout mice and airway epithelial cells were infected with RSV and the expression of ALCAM and inflammatory cytokines were measured. We also conducted further experiments using Anti-ALCAM antibody and recombinant ALCAM in airway epithelial cells. RESULTS: The expression levels of ALCAM and inflammatory cytokines increased in both RSV-infected mice and airway epithelial cells. Interestingly, IL-33 expression was significantly reduced in ALCAM-knockdown cells compared to control cells following RSV infection. Anti-ALCAM antibody treatment also reduced IL-33 expression following RSV infection. Furthermore, the phosphorylation of ERK1/2, p38, and JNK was diminished in ALCAM-knockdown cells compared to control cells following RSV infection. Notably, in the control cells, inhibition of these pathways significantly decreased the expression of IL-33. In vivo study also confirmed a reduction in inflammation induced by RSV infection in ALCAM deficient mice compared to wild-type mice. CONCLUSION: These findings demonstrate that ALCAM contributes to RSV-induced airway inflammation at least partly by influencing IL-33 expression through mitogen-activated protein kinase signaling pathways. These results suggest that targeting ALCAM could be a potential therapeutic strategy for alleviating IL-33-associated lung diseases.

Brain structural correlates of subjective sleepiness and insomnia symptoms in shift workers.

Background: Studies on the brain structures of shift workers are limited; thus, this cross-sectional study aimed to compare the brain structures and the brain structural correlates of subjective sleepiness and insomnia symptoms between shift workers and non-shift workers. Methods: Shift workers (n = 63) and non-shift workers (n = 58) completed questionnaires assessing subjective sleepiness and insomnia symptoms. Cortical thickness, cortical surface area, and subcortical volumes were measured by magnetic resonance imaging. The brain morphometric measures were compared between the groups, and interaction analyses using the brain morphometric measures as the dependent variable were performed to test the interactions between the study group and measures of sleep disturbance (i.e., subjective sleepiness and insomnia symptoms). Results: No differences in cortical thickness, cortical surface area, or subcortical volumes were detected between shift workers and non-shift workers. A single cluster in the left motor cortex showed a significant interaction between the study group and subjective sleepiness in the cortical surface area. The correlation between the left motor cortex surface area and the subjective sleepiness level was negative in shift workers and positive in non-shift workers. Significant interaction between the study group and insomnia symptoms was present for the left/right putamen volumes. The correlation between the left/right putamen volumes and insomnia symptom levels was positive in shift workers and negative in non-shift workers. Conclusion: Left motor cortex surface area and bilateral putamen volumes were unique structural correlates of subjective sleepiness and insomnia symptoms in shift workers, respectively.

Ileoanal pouch revision and excision surgery in a newly established pouch center: requirements and costs for service provision.

Complications of ileoanal pouch surgery affecting function and quality of life may require surgical correction or pouch excision. The management of patients with pouch dysfunction requires a multidisciplinary approach and demand for service provision include multiple healthcare professionals and resources. The aim of this study is to present the service requirements, and surgical outcomes for redo pouch surgery and pouch excision, with cost analysis of the required resources. All patients undergoing surgery for revision or excision of the ileoanal pouch from June 2021 to May 2023 were prospectively included. Patient undergoing only diagnostic procedures, or perineal procedures were excluded. Outcomes within 30 days of surgery were collected, including readmissions and re-operations. Cost analysis of all investigations, outpatient appointments and procedures prior to pouch revision or pouch excision was conducted. Twenty patients were included during the 24 months study period: 13 underwent abdominal revisional pouch surgery, 7 had ileoanal pouch excision. 15 patients (75%) were tertiary referrals from other hospitals in the UK. The median interval between index IPAA surgery and revision was 113 months. Three multidisciplinary clinical appointments, two imaging modalities, and at least one invasive day-surgery procedure were required for each patient prior to surgery. Expertise and infrastructure are needed for indication and peri-operative management of patients with pouch dysfunction requiring pouch revision or pouch excision. We estimated a starting cost of £22.605 ($29.589) for provision of pouch revision or excision surgery for investigations and treatments from referral to the pouch unit to surgery. This likely represents an underestimate as only accounts for procedures performed since referral with pouch dysfunction.

Development and validation of a self-management self-efficacy scale for premature birth prevention (SMSE-PBP) for women of childbearing age.

BACKGROUND: This study aimed to develop and evaluate the validity and reliability of a self-management self-efficacy for premature birth prevention (SMSE-PBP) in women of childbearing age (WCA). METHODS: Instrument development and validation were undertaken in three phases: conceptualization, item generation and evaluation of content validity, and evaluation of construct and concurrent validity and reliability. Data were analyzed using exploratory and second-order confirmatory factor analyses, and concurrent validity was examined using Pearson's correlation coefficients. The reliability was analyzed using omega hierarchical and Cronbach's ⍺. RESULTS: Content validity was assessed by experts and cognitive interviews of WCA. The SMSE-PBP consists of a second-order 3-dimension and 10-factor scale with 60 items; therefore, the construct and concurrent validity of the SMSE-PBP were supported. The omega values were 0.93 for pre-pregnancy SMSE-PBP, 0.92 for pregnancy SMSE-PBP, and 0.94 for hospital SMSE-PBP. Cronbach's ⍺ was 0.88 for pre-pregnancy SMSE-PBP, 0.96 for pregnancy SMSE-PBP, and 0.96 for hospital SMSE-PBP. CONCLUSIONS: The SMSE-PBP scale is valid and reliable for WCA; it is helpful for WCA and health professionals to assess women's SMSE-PBP and pre-pregnancy, pregnancy, or hospital SMSE-PBP. The next steps should include assessing the relationship with pregnancy health behaviors.

Crystallographic and Photophysical Analysis on Facet-Controlled Defect-Free Blue-Emitting Quantum Dots.

The burgeoning demand for commercializing self-luminescing quantum dot (QD) light-emitting diodes (LEDs) has stimulated extensive research into environmentally friendly and efficient QD materials. Hydrofluoric acid (HF) additive improves photoluminescence (PL) properties of blue-emitting ZnSeTe QDs, ultimately reaching a remarkable quantum yield (QY) of 97% with an ultranarrow peak width of 14 nm after sufficient HF addition. The improvement in optical properties of the QDs is accompanied by a morphology change of the particles, forming cubic-shaped defect-free ZnSeTe QDs characterized by a zinc blende (ZB) crystal structure. This treatment improves the QD-emitting properties by facilitating facet-specific growth, selectively exposing stabilized (100) facets, and reducing the lattice disorders. The facet-specific growth process gives rise to defect-free monodispersed cubic dots that exhibit remarkably narrow and homogeneous PL spectra. Meticulous time-resolved spectroscopic studies allow an understanding of the correlation between ZnSeTe QDs' particle shape and performance following HF addition. These investigations shed light on the intricacies of the growth mechanism and the factors influencing the PL efficiency of the resulting QDs. The findings significantly contribute to understanding the role of HF treatment in tailoring the optical properties of ZnSeTe QDs, thereby bringing it closer to the realization of highly efficient and bright QD-LEDs for various practical applications.

Evaluation of the anti-androgenic and cytotoxic effects of benzophenone-3 in male Sprague-Dawley rats.

Benzophenone-3 (BP-3, 2-hydroxy-4-methoxybenzophenone, oxybenzone) is one of the most widely used types of benzophenone organic sunscreen. However, this compound is a potentially harmful toxicant. The aim of this study was 2-fold to: (1) utilize a Hershberger bioassay in vivo in castrated male Sprague-Dawley rats to investigate the anti-androgenic activities of BP-3, and (2) use in vitro a methyl tetrazolium assay to compare the toxicity between Leydig cells (TM3 cells) and mouse fibroblast (NIH-3T3) cell lines. In the Hershberger assay, rats were divided into 6 groups (each of n = 7): a vehicle control, negative control, positive control, PB-3 low (40 mg/kg), BP-3 intermediate (200 mg/kg), and BP-3 high (1000 mg/kg)-dose. The weight of the ventral prostate was significantly decreased at BP-3 doses of 200 or 1,000 mg/kg/day. In addition, the levator anibulbocavernosus muscle weights were also significantly reduced at BP-3 doses of 40, 200, or 1,000 mg/kg/day. In the MTT assay, the viability of NIH-3T3 mouse fibroblast cells was within the normal range. However, the TM3 mouse testis Leydig cell viability was significantly lowered in a concentration-dependent manner. Therefore, data indicate that BP-3 might exert in vivo anti-androgenic and in vitro cytotoxic effects in cells associated with the male reproductive system compared to normal non-reproductive cells.Abbreviation: BP-3: benzophenone-3; CG: Cowper's gland; DMEM: Dulbecco's modified Eagle's medium; DMSO: dimethyl sulfoxide; GP: glans penis; LABC: levator anibulbocavernosus muscle; MTT: methyl tetrazolium; NC: negative control; PC: positive control; SV: seminal vesicle; TP: testosterone propionate; VC: vehicle control; VP: ventral prostate.

Suture Stenting After Sialendoscopy: A Novel Technique That Reduces Risk of Recurrent Parotitis.

OBJECTIVE: Chronic sialadenitis is associated with decreased quality of life and recurrent infections. While sialendoscopy with stenting is effective in relieving symptoms of sialadenitis, currently available stents are rigid and poorly tolerated by patients, leading to early removal and potential for adverse scarring. This study examines whether sutures can be used as a stenting material to improve patient comfort and reduce recurrence risk. METHODS: This is a retrospective cohort study of a consecutive series of adult patients with chronic sialadenitis undergoing sialendoscopy with or without suture stenting. Data were collected between 2014 and 2018 with a 3-year follow-up period ending in 2021. The primary outcome measure was recurrence of sialadenitis within 3 years of surgery. Secondary outcomes were stent dislodgement and patient-reported discomfort. RESULTS: We included 63 patients with parotid sialadenitis of whom 28 underwent suture stenting and 35 did not receive stenting after sialendoscopy. Stents were well tolerated, with a mean duration of 34.5 days, and only 2 of 28 stents (7.1%) accidentally dislodged within the first week. Suture stenting significantly reduced symptom recurrence after sialendoscopy (OR = 0.09, 95% CI 0.02-0.45, p = 0.003; 3-year sialadenitis recurrence rate: 7.1% vs. 45.7%, p = 0.005). Cox multivariate regression for clinicodemographic variables showed an HR of 0.04 (95% CI 0.01-0.19, p < 0.001) for the risk of symptom recurrence. CONCLUSIONS AND RELEVANCE: Suture stenting after sialendoscopy is low cost, available across all institutions, well-tolerated by patients, and highly efficacious in reducing risk of recurrent sialadenitis after sialendoscopy. LEVEL OF EVIDENCE: 3 Laryngoscope, 134:614-621, 2024.

A longitudinal molecular and cellular lung atlas of lethal SARS-CoV-2 infection in K18-hACE2 transgenic mice.

BACKGROUND: The global pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to approximately 500 million cases and 6 million deaths worldwide. Previous investigations into the pathophysiology of SARS-CoV-2 primarily focused on peripheral blood mononuclear cells from patients, lacking detailed mechanistic insights into the virus's impact on inflamed tissue. Existing animal models, such as hamster and ferret, do not faithfully replicate the severe SARS-CoV-2 infection seen in patients, underscoring the need for more relevant animal system-based research. METHODS: In this study, we employed single-cell RNA sequencing (scRNA-seq) with lung tissues from K18-hACE2 transgenic (TG) mice during SARS-CoV-2 infection. This approach allowed for a comprehensive examination of the molecular and cellular responses to the virus in lung tissue. FINDINGS: Upon SARS-CoV-2 infection, K18-hACE2 TG mice exhibited severe lung pathologies, including acute pneumonia, alveolar collapse, and immune cell infiltration. Through scRNA-seq, we identified 36 different types of cells dynamically orchestrating SARS-CoV-2-induced pathologies. Notably, SPP1+ macrophages in the myeloid compartment emerged as key drivers of severe lung inflammation and fibrosis in K18-hACE2 TG mice. Dynamic receptor-ligand interactions, involving various cell types such as immunological and bronchial cells, defined an enhanced TGFß signaling pathway linked to delayed tissue regeneration, severe lung injury, and fibrotic processes. INTERPRETATION: Our study provides a comprehensive understanding of SARS-CoV-2 pathogenesis in lung tissue, surpassing previous limitations in investigating inflamed tissues. The identified SPP1+ macrophages and the dysregulated TGFß signaling pathway offer potential targets for therapeutic intervention. Insights from this research may contribute to the development of innovative diagnostics and therapies for COVID-19. FUNDING: This research was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (2020M3A9I2109027, 2021R1A2C2004501).

Nanographene-Fused Expanded Carbaporphyrin Tweezers.

A nanographene-fused expanded carbaporphyrin (5) and its BF2 complex (6) were synthesized. Single-crystal X-ray structures revealed that 5 and 6 are connected by two hexa-peri-hexabenzocoronene (HBC) units and two dipyrromethene or BODIPY units, respectively. As prepared, 5 and 6 both show nonaromatic character with figure-of-eight carbaoctaphyrin (1.1.1.0.1.1.1.0) cores and adopt tweezers-like conformations characterized by a partially confined space between the two constituent HBC units. The distance between the HBC centers is >10 Å, while the dihedral angles between the two HBC planes are 30.5 and 35.2° for 5 and 6, respectively. The interactions between 5 and 6 and fullerene C60 were studied both in organic media and in the solid state. Proton NMR spectral titrations of 5 and 6 with C60 revealed a 1:1 binding mode for both macrocycles. In toluene-d8, the corresponding binding constants were determined to be 1141 ± 17 and 994 ± 10 M-1 for 5 and 6, respectively. Single-crystal X-ray diffraction structural analyses confirmed the formation of 1:1 fullerene inclusion complexes in the solid state. The C60 guests in both complexes are found within triangular pockets composed of two HBC units from the tweezers-like receptor most closely associated with the bound fullerene, as well as an HBC unit from an adjacent host. Femtosecond transient absorption measurements revealed subpicosecond ultrafast charge separation between 5 (and 6) and C60 in the complexes. To the best of our knowledge, the present report provides the first example wherein a nanographene building block is incorporated into the core of a porphyrinic framework.

Factors affecting patients who attempted suicide in the emergency department due to the prolonged pandemic of COVID-19.

OBJECTIVE: This study examined the characteristics of patients who attempted suicide in the emergency department before and during the COVID-19 pandemic. METHODS: We compared data from patients in the emergency department following suicide attempts between January 2018 and December 2021. The patients were categorized into two groups: "pre-COVID-19" and "during COVID-19" pandemic. RESULTS: The findings revealed an increasing trend of suicide attempts during the study period. Suicide attempts were reported at 1,107 before the COVID-19 pandemic and 1,356 during the COVID-19 pandemic. Patients who attempted suicide during the COVID-19 pandemic were younger (38.0±18.5 years vs. 40.7±18.4 years, P<0.01), had a smaller proportion of men (36% vs. 44%, P<0.01), and had fewer medical comorbidities (20.2% vs. 23.6%, P<0.05). The group during the COVID-19 pandemic reported better hygiene conditions (50.5% vs. 40.8%, P<0.01) and lower alcohol consumption (27.7% vs. 37.6%, P<0.01). Patients who attempted suicide during the COVID-19 pandemic had higher rates of use of psychiatric medications and previous suicide attempts. The most common reasons for the suicide attempt were unstable psychiatric disorders (38.8%), poor interpersonal relationships (20.5%), and economic difficulties (14.0%). Drug poisoning (44.1%) was the most common method of suicide attempts. Subgroup analysis with patients who attributed their suicide attempts to COVID-19 revealed a higher level of education (30.8%) and employment status (69.2%), with economic difficulties (61.6%) being the primary cause of suicide attempts. CONCLUSION: These findings suggest that the prolonged duration of the COVID-19 pandemic and its effects on social and economic factors have influenced suicide attempts.

Development of generalizable automatic sleep staging using heart rate and movement based on large databases.

Purpose: With the advancement of deep neural networks in biosignals processing, the performance of automatic sleep staging algorithms has improved significantly. However, sleep staging using only non-electroencephalogram features has not been as successful, especially following the current American Association of Sleep Medicine (AASM) standards. This study presents a fine-tuning based approach to widely generalizable automatic sleep staging using heart rate and movement features trained and validated on large databases of polysomnography. Methods: A deep neural network is used to predict sleep stages using heart rate and movement features. The model is optimized on a dataset of 8731 nights of polysomnography recordings labeled using the Rechtschaffen & Kales scoring system, and fine-tuned to a smaller dataset of 1641 AASM-labeled recordings. The model prior to and after fine-tuning is validated on two AASM-labeled external datasets totaling 1183 recordings. In order to measure the performance of the model, the output of the optimized model is compared to reference expert-labeled sleep stages using accuracy and Cohen's κ as key metrics. Results: The fine-tuned model showed accuracy of 76.6% with Cohen's κ of 0.606 in one of the external validation datasets, outperforming a previously reported result, and showed accuracy of 81.0% with Cohen's κ of 0.673 in another external validation dataset. Conclusion: These results indicate that the proposed model is generalizable and effective in predicting sleep stages using features which can be extracted from non-contact sleep monitors. This holds valuable implications for future development of home sleep evaluation systems.

Association between social networks and symptoms of post-traumatic stress during the pandemic: Cohort study in South Korea.

BACKGROUND: This study estimated the association between pre-pandemic social network properties and symptoms of posttraumatic stress during the coronavirus disease 2019 (COVID-19) pandemic in South Korea. METHODS: We conducted four online mental health surveys during COVID-19 (from March 14, 2020, to December 11, 2021) among individuals enrolled in a community-based cohort study (previously recruited from 2013 to 2018). Among 4060 people interviewed at the pre-pandemic baseline, 2652 individuals (men = 951, women = 1701) who responded to at least one of the four surveys conducted were included. At baseline, each individual's social network, including size and average closeness, was measured in an egocentric way. Post Traumatic Stress Disorder Checklist for DSM-5 (PCL-5) was measured at each survey point. A generalized linear model and a generalized linear mixed model were used after stratified by sex. RESULTS: Among men, network size was negatively associated with total PCL-5 score (b = -0.42, SE = 0.16, p = 0.002). The magnitude of the association increased over time and was maximal by the 4th survey (b = -0.65, SE = 0.31, p = 0.037). Among women, average closeness was negatively associated with the total PCL-5 score (b = -1.16, SE = 0.37, p = 0.002). In analyses disaggregated by symptom clusters, social networks were associated explicitly with symptoms of intrusion and mood but not with avoidance and arousal symptoms. CONCLUSION: In this cohort of adults followed for more than a year during the outbreak of COVID-19 in South Korea, social networks established before the onset of the COVID-19 pandemic had a protective association against developing symptoms of PTSD during the first two years of the pandemic.

Current status of ß-lactam antibiotic use and characterization of ß-lactam-resistant Escherichia coli from commercial farms by integrated broiler chicken operations in Korea.

ß-Lactam antibiotics are one of the most clinical importance in human and veterinary medicine because they are used for both preventive and therapeutic purposes against several gram-positive, gram-negative, and anaerobic organisms. In this study, it was confirmed that ß-lactams (81.1%) were found to be significantly prescribed the most among 74 farms in 5 integrated broiler operations, and single prescription (84.6%), 2-day (41.5%) or 3-day (40.0%) administration, and 15 to 22 d of age (67.7%) administration was significantly higher in the farms (P < 0.05). Among the E. coli isolated from 74 farms in 5 integrated broiler operations, ß-lactam-resistant E. coli isolates were detected more frequently in fecal sample (94.6%) than in dust sample (60.8%) (P < 0.05). The prevalence of MDR in ß-lactam-resistant isolates, ranging from 88.1 to 96.5%, was significantly higher than that in non-ß-lactam-resistant isolates (P < 0.05), without significant differences among operations. Of 466 ß-lactam-resistant isolates, 432 (92.7%) isolates harbored ß-lactamase genes. The non-extended-spectrum ß-lactamase (ESBL) gene blaTEM-1 (81.8%) showed the highest prevalence among isolates, followed by the non-ESBL gene blaTEM-135 (6.4%) (P < 0.05). Five ESBL genes, SHV-12, OXA-1, CTX-M-27, CTX-M-55, and CTX-M-65, were found in 0.9 to 6.0% of the isolates. The pAmpC gene blaCMY-2 was detected in 17 isolates (3.6%). These results suggest that feces and dust are important reservoirs of antimicrobial-resistant bacteria, highlighting the need to strengthen farm management regulations, such as cleaning, disinfection, and litter disposal and to reduce the use of antibiotics in broiler operations.

Circadian misalignment alters resting-state functional connectivity of the salience network in rotating shift workers.

STUDY OBJECTIVES: This study compared resting-state functional connectivity (rsFC) of the salience network (SN) between rotating shift workers (RSWs) and controls. Furthermore, we examined whether rsFC of the SN was correlated with sleep, emotion, cognition, and attention. METHODS: The 60 RSWs and 57 controls enrolled in this study completed self-report questionnaires and sleep diaries to assess subjective sleep quality, and polysomnography and actigraphy to evaluate objective sleep and 24-hour rest-activity rhythm parameters. The participants also underwent resting-state functional magnetic resonance imaging and structural T1 scans. We performed a seed-based rsFC analysis of the SN using the anterior cingulate cortex (ACC) and anterior insula (AI) as seed regions. Furthermore, AI and ACC rsFC were compared in RSWs and controls, and we analyzed correlations between rsFC and variables of interest showing significant group differences. RESULTS: Compared with controls, RSWs showed reduced rsFC between the ACC and right insula, and increased rsFC of the ACC with the left occipital lobe and right superior frontal gyrus extending to the supplementary motor area (SFG/SMA). Moreover, RSWs showed reduced rsFC between the right AI and right superior parietal lobule (SPL). Finally, rsFC between the ACC and right AI was correlated with 24-hour rest-activity rhythmicity. CONCLUSIONS: Although RSWs did not show sleep disturbance, emotional distress, cognitive impairment, or attention deficits, alterations of right insula, left occipital lobe, right SFG/SMA, and right SPL rsFC in the SN indicate that impairments in salience detection and top-down attentional control may emerge in shift workers over time.

Combined inhibition of Bcl-2 family members and YAP induces synthetic lethality in metastatic gastric cancer with RASA1 and NF2 deficiency.

BACKGROUND: Targetable molecular drivers of gastric cancer (GC) metastasis remain largely unidentified, leading to limited targeted therapy options for advanced GC. We aimed to identify molecular drivers for metastasis and devise corresponding therapeutic strategies. METHODS: We performed an unbiased in vivo genome-wide CRISPR/Cas9 knockout (KO) screening in peritoneal dissemination using genetically engineered GC mouse models. Candidate genes were validated through in vivo transplantation assays using KO cells. We analyzed target expression patterns in GC clinical samples using immunohistochemistry. The functional contributions of target genes were studied through knockdown, KO, and overexpression approaches in tumorsphere and organoid assays. Small chemical inhibitors against Bcl-2 members and YAP were tested in vitro and in vivo. RESULTS: We identified Nf2 and Rasa1 as metastasis-suppressing genes through the screening. Clinically, RASA1 mutations along with low NF2 expression define a distinct molecular subtype of metastatic GC exhibiting aggressive traits. NF2 and RASA1 deficiency increased in vivo metastasis and in vitro tumorsphere formation by synergistically amplifying Wnt and YAP signaling in cancer stem cells (CSCs). NF2 deficiency enhanced Bcl-2-mediated Wnt signaling, conferring resistance to YAP inhibition in CSCs. This resistance was counteracted via synthetic lethality achieved by simultaneous inhibition of YAP and Bcl-2. RASA1 deficiency amplified the Wnt pathway via Bcl-xL, contributing to cancer stemness. RASA1 mutation created vulnerability to Bcl-xL inhibition, but the additional NF2 deletion conferred resistance to Bcl-xL inhibition due to YAP activation. The combined inhibition of Bcl-xL and YAP synergistically suppressed cancer stemness and in vivo metastasis in RASA1 and NF2 co-deficiency. CONCLUSION: Our research unveils the intricate interplay between YAP and Bcl-2 family members, which can lead to synthetic lethality, offering a potential strategy to overcome drug resistance. Importantly, our findings support a personalized medicine approach where combined therapy targeting YAP and Bcl-2, tailored to NF2 and RASA1 status, could effectively manage metastatic GC.

π-Extended Hexapyrrolylbenzenes: Exploring Charge-Transfer Phenomena in Donor-Acceptor Propellers.

A family of propeller-shaped donor-acceptor hexapyrrolylbenzenes (HPBs) were designed and synthesized by sequential nucleophilic substitution of hexafluorobenzene with π-extended pyrroles. In particular, four hybrids were obtained, containing various combinations of electron-rich and electron-poor acenaphthylene-fused pyrroles. Additionally, to probe the efficiency of ortho transfer interactions, a system was designed containing unique donor and acceptor subunits spatially separated with four unfunctionalized pyrroles. DFT calculations showed propeller-shaped geometries of all HPB molecules and separation of frontier molecular orbitals between donor and acceptor subunits. Steady-state and time-resolved photophysical measurements revealed charge-transfer (CT) character of the emission with strong positive dependence on solvent polarity. The principal CT pathway involves ortho-positioned pairs of donors and acceptors and requires bending of the acceptor in the excited state.