Gain Loss Research Articles

Genome wide association analysis reveals the genetic sites of heat resistance in largemouth bass (Micropterus salmoides)

Largemouth bass (Micropterus salmoides) is an important commercially cultured freshwater fish. High temperature is a common challenge encountered in the summer cultivation of largemouth bass. In this study, we conducted a genome-wide association study using general linear models and linear mixed models to identify significant single nucleotide polymorphisms (SNPs) and candidate genes associated with loss of equilibrium (LOE) and weight gain (WG) during high temperature challenge. We collected fin tissue from 353 individuals of largemouth bass exposed to high water temperature (37.5 °C) and performed whole-genome resequencing. Through association analysis, we successfully identified 10 SNPs and 38 candidate genes that were significantly associated with LOE, and four SNPs and 19 candidate genes that were significantly associated with WG. Using quantitative PCR and Western blot, we tested the expression of eight genes related to LOE in heat-sensitive and heat-tolerant individuals, and identified significant differences in the expression of the following five genes: Tmub2, Ngfr, Hgf, Hdac6, and Sypl. For WG, we assessed the expression of three candidate genes, among which Mrc2l displayed significant differences in expression between the high WG group and low WG group. We discuss the functions and potential biological processes of candidate genes for LOE and WG. Our results provide basic genetic information for breeding largemouth bass with high temperature tolerance.

Abstract PO5-22-04: Genomic analysis of local recurrences following risk adapted breast radiotherapy in the IMPORT trials defines 'true recurrences' and 'new primaries'

Abstract Background: Most ipsilateral breast tumor recurrences (IBTR) following breast conserving surgery occur near the tumor bed (Veronisi et al, Ann Oncol 2001) and are considered to represent ‘true recurrence’ (derived from residual malignant cells of the index cancer). A smaller number occur elsewhere in the breast and are thought to be ‘new primaries’ (independently occurring cancer). The IMPORT LOW (Coles et al, The Lancet 2017) and IMPORT HIGH (Coles et al, The Lancet 2023) trials investigated adaptation of radiotherapy dose-volume to this spatially varying risk, testing partial breast irradiation and simultaneous integrated boost in patients with tumors at low and high risk of IBTR respectively. We analysed genomic relationships between index cancers and recurrences in these trials, to ascertain the frequency of true recurrences and unrelated new primaries, and their spatial distribution. Methodology: FFPE blocks were obtained from 137 patients who developed subsequent ipsi- or contralateral breast cancer: 66 from IMPORT HIGH and 71 from IMPORT LOW (4 with bilateral second cancers). DNA extracted from index and subsequent cancers underwent shallow whole genome sequencing (sWGS) using the Illumina NovaSeq 6000 system. Copy number profiles were derived from sWGS data using the R package QDNAseq(Scheinin et al, Genome Res 2014); those that failed QC criteria were removed. Relatedness of tumor pairs was determined using the R package Breakclone (Lips et al, Nat Genet 2022). This approach uses individual copy number aberration breakpoint position rather than events at chromosomal arm level. The p value cutoff for relatedness was 0.01; values between 0.01 and 0.05 were considered ambiguous. Researchers were blinded to clinical information prior to results. Results: In 80/137 tumor pairs both copy number profiles met QC criteria and clonal relationship could be ascertained. In total 26/80 were considered related, 20/80 ambiguous and 34/40 non-related (Table 1). 16/26 ipsilateral subsequent cancers in IMPORT HIGH were clonally related to the primary tumor and could be considered true recurrences. In 5/26 it was impossible to accurately call clonality as pairs shared only common copy number changes such as 1q or 8q gain and 16q loss. 5/26 were considered new primaries. In the lower risk IMPORT LOW cohort, subsequent cancers appear to have similar chances of being related or independent. As expected, most contralateral tumors in IMPORT LOW showed no evidence of a clonal relationship to the index tumors. In both trials an ambiguous subgroup of pairs shared some common copy number changes suggesting similar phenotype.5/14 contralateral tumors in IMPORT HIGH, in which some women were also at high risk of systemic relapse, shared very similar copy number profiles to the index tumor and may be metastases rather than new primaries. Discussion: A considerable number of tumors classed as IBTR appear genomically unrelated to the index cancer particularly in low risk cancer; this biology is relevant to clinical management both initially and at ‘recurrence’. The finding that 5/14 contralateral cancers in the high risk group appear to be clonally related suggests that some may be metastases. Results are being confirmed by targeted sequencing and will be correlated with recorded spatial and clinicopathological data and patient outcome. Future work will analyse spatial relationships more precisely using deformable image registration. *From the 77 patients with baseline circulating tumor DNA samples, 2 patients were excluded because their samples failed during assay quality control. Table. PIK3CA and ESR1 alterations detected in circulating tumor DNA at baseline and post abemaciclib treatment Citation Format: Sara Lightowlers, Maria Roman-Escorza, Elena Provenzano, Judith Bliss, Jason Carroll, H Y Charlie Chan, Clare Griffin, Joanne Haviland, Monica Jefford, Anna Kirby, Navita Somaiah, Mark Sydenham, Jenny Titley, John Yarnold, Charlotte Coles, Elinor Sawyer. Genomic analysis of local recurrences following risk adapted breast radiotherapy in the IMPORT trials defines 'true recurrences' and 'new primaries' [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-22-04.

Does ovariectomy exacerbate the cardiorespiratory and metabolic consequences of moderate intermittent hypoxia in female rats?

Intermittent hypoxia (IH) is a major consequence of sleep apnea (SA). IH causes respiratory dysfunction and contributes to numerous co-morbidities of SA including hypertension and obesity. The fact that menopause increases the prevalence of SA and related diseases in women indicates the importance of loss of ovarian function in the pathophysiology. Here, we determined whether experimental loss of ovarian function (ovariectomy; OVX) exacerbates the cardiorespiratory and metabolic consequences of IH. We explored neuroendocrine changes by assessing hormones known to be involved in energy balance, respiratory control, and stress responses (leptin, ACTH and corticosterone). We subjected adult (8 weeks) Sprague Dawley female rats to bilateral OVX (n = 13) or sham operation (n = 14). Rats were housed in pairs, and after two weeks of recovery, animals were exposed to room air (control; n = 16) or to a 7-day moderate IH protocol (n = 11). Within their cage, rats intermittently received a nitrogen-enriched hypoxic mixture, followed by room air for reoxygenation. Each bout of hypoxia lasted 30 seconds (nadir FIO2 = 0.10) and was followed by 5-minutes of reoxygenation (10 cycles/h, for 8hrs between 10h-18h). Apnea frequency, acute responses to hypoxia (FIO2 = 0.12; 120 sec) and hypercapnia (FICO2 = 0.05; 10 min) were measured by whole-body plethysmography. Blood pressure was assessed by the tail cuff method, and body weight was followed throughout the protocol. Plasma corticosterone and leptin were measured by multiplex assays. OVX increased apneic events, whereas IH reduced them; with the highest frequency observed in OVX females maintained in normoxia. Ventilatory responses (hypoxia and CO2) and blood pressure were not affected by IH or OVX. Over the course of the protocol, OVX increased weight gain whereas IH induced weight loss. OVX (but not IH) increased ACTH and corticosterone. IH (but not OVX) reduced leptin. Correlation matrix analysis showed that weight gain was the best predictor of apnea frequency (r = 0.759; p = 0.03). We conclude that loss of ovarian function and related weight gain have the greatest impact on respiratory function during sleep. Additionally, OVX-related activation of the stress pathways may contribute to this process. These results fail to demonstrate a role for OVX in exacerbating the cardiorespiratory and metabolic consequences of IH. Lack of weight gain following IH likely contributes to this finding. This study is supported by operating grants from the Canadian Institutes of Health Research (RK and VJ). NJM is supported by a Sentinel North Partnered Research Chair in Sleep Pharmacometabolism and a Canada Research Chair in Metabolism and Sleep Neurobiology. MG received a doctoral scholarship from the Fonds de Recherche du Québec — Santé (FRQS). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Abstract 2149: Smooth Muscle-derived Resident Vascular Adventitial Progenitor Cell Contribution To Cardiac Fibrosis And Inflammation Is Dependent On KLF4

Cardiac fibrosis is defined by the excessive accumulation of extracellular matrix (ECM) material resulting in cardiac tissue scarring and dysfunction. While it is commonly accepted that myofibroblasts are the major contributors to ECM deposition in cardiac fibrosis, their origin remains debated. By combining lineage tracing and RNA sequencing, our group made the paradigm-shifting discovery that a subpopulation of resident vascular stem cells residing within the vascular adventitia (termed AdvSca1-SM cells) originate from mature vascular smooth muscle cells (SMCs) through an in situ reprogramming process. SMC-to-AdvSca1-SM reprogramming and AdvSca1-SM cell maintenance is dependent on induction and activity of the transcription factor, KLF4. In contrast, loss of Klf4 expression promotes the transition of AdvSca1-SM cells to a myofibroblast phenotype in vascular injury and remodeling. However, the molecular mechanism whereby KLF4 regulates AdvSca1-SM phenotype remains unclear. Our recent preliminary data confirm the existence of perivascular cardiac AdvSca1-SM cells. Leveraging a highly specific AdvSca1-SM cell reporter system, single-cell RNA-sequencing (scRNA-seq), and spatial transcriptomic approaches, we demonstrate the profibrotic differentiation trajectory of AdvSca1-SM cells in the setting of Angiotensin II (AngII)-induced cardiac fibrosis. Differentiation is characterized by loss of stemness-related genes, including Klf4 , but gain of expression of a profibrotic phenotype. Importantly, these changes are recapitulated in human cardiac hypertrophic tissue, supporting the translational significance of profibrotic transition of AdvSca1-SM-like cells in human cardiomyopathy. Very surprisingly and paradoxically, AdvSca1-SM-specific genetic knockout of Klf4 prior to AngII treatment protects against cardiac inflammation and fibrosis, indicating that Klf4 is essential for the profibrotic response of AdvSca1-SM cells. Overall, our data reveal the contribution of AdvSca1-SM cells to myofibroblasts in the setting of cardiac fibrosis. KLF4 not only maintains the stemness of AdvSca1-SM cells, but also orchestrates their response to profibrotic stimuli, and may serve as a therapeutic target in cardiac fibrosis.

DFMO inhibition of neuroblastoma tumorigenesis.

Most high-risk neuroblastoma patients who relapse succumb to disease despite the existing therapy. We recently reported increased event-free and overall survival in neuroblastoma patients receiving difluoromethylornithine (DFMO) during maintenance therapy. The effect of DFMO on cellular processes associated with neuroblastoma tumorigenesis needs further elucidation. Previous studies have shown cytotoxicity with IC50 values >5-15 mM, these doses are physiologically unattainable in patients, prompting further mechanistic studies at therapeutic doses. We characterized the effect of DFMO on cell viability, cell cycle, apoptosis, neurosphere formation, and protein expression invitro using five established neuroblastoma cell lines (BE2C, CHLA-90, SHSY5Y, SMS-KCNR, and NGP) at clinically relevant doses of 0, 50, 100, 500, 1000, and 2500 μM. Limiting Dilution studies of tumor formation in murine models were performed. Statistical analysis was done using GraphPad and the level of significance set at p = 0.05. There was not a significant loss of cell viability or gain of apoptotic activity in the invitro assays (p > 0.05). DFMO treatment initiated G1 to S phase cell cycle arrest. There was a dose-dependent decrease in frequency and size of neurospheres and a dose-dependent increase in beta-galactosidase activity in all cell lines. Tumor formation was decreased in xenografts both with DFMO-pretreated cells and in mice treated with DFMO. DFMO treatment is cytostatic at physiologically relevant doses and inhibits tumor initiation and progression in mice. This study suggests that DFMO, inhibits neuroblastoma by targeting cellular processes integral to neuroblastoma tumorigenesis at clinically relevant doses.

Now or Later? Examining Social and Financial Decision-Making in Middle-to-Older Aged Adults.

Contextually driven decision-making is multidimensional, as individuals need to contend with prioritizing both competing and complementary demands. However, data is limited as to whether temporal discounting rates vary as a function of framing (gains vs. loss) and domain (monetary vs. social) in middle-to-older aged adults. It is also unclear whether socioaffective characteristics like social isolation and loneliness are associated with temporal discounting. Temporal discounting rates were examined across monetary gain, monetary loss, social gain, and social loss conditions in 140 adults aged 50-90 during the Omicron stage of the pandemic. Self-report measures assessed loneliness and social isolation levels. Results found evidence of steeper temporal discounting rates for gains as compared to losses in both domains. Social outcomes were also more steeply discounted than monetary outcomes, without evidence of an interaction with the framing condition. Socioeconomic and socioaffective factors were unexpectedly not associated with temporal discounting rates. Community-dwelling middle-to-older aged adults showed a preference for immediate rewards and devalued social outcomes more than monetary outcomes. These findings have implications for tailoring social and financial incentive programs for middle to later adulthood.

Dynamics of carbon and water vapor fluxes in three typical ecosystems of Heihe River Basin, Northwestern China

Understanding the dynamics of carbon and water vapor fluxes in arid inland river basin ecosystems is essential for predicting and assessing the regional carbon-water budget amid climate change. However, studies aiming to unravel the mechanisms driving the variations and coupling process of regional carbon-water budget in a changing environment in arid regions are limited. Here, we used the eddy covariance technique to analyze the relationship between CO2 and H2O fluxes in three typical ecosystems across the upper, middle, and lower reaches of an arid inland river basin in Northwestern China. Our results showed that all ecosystems acted as carbon sinks, with the alpine swamp meadow, cropland, and desert shrubland sequestrating −300.2 ± 0.01, −644.8 ± 2.9, and − 203.7 ± 22.5 g C m−2 yr−1, respectively. Air temperature (Ta) primarily controlled daily gross primary productivity (GPP) and net ecosystem CO2 exchange (NEE) in the irrigated cropland during the growing season, while soil temperature (Ts) and vapor pressure deficit (VPD) regulated these parameters in the alpine swamp meadow and desert shrubland. Additionally, Ta and net radiation (Rn) controlled daily evapotranspiration (ET) in cropland, while Ts and Rn regulated ET at other sites. Consequently, carbon and water vapor fluxes of all three ecosystems tended to be energy-limited during the growing season. The differential responses of carbon and water vapor fluxes in the upper, middle, and lower reaches of these ecosystems to biophysical factors determined their distinct coupling and variations in water use efficiency. Notably, the desert shrub ecosystem in the lower reach of the basin maintained a stable balance between carbon gain and water loss, indicating adaptation to aridity. This study provides valuable insights into the underlying mechanisms behind the changes in carbon and water vapor fluxes and water-use efficiency in arid river basin ecosystems.

Collapse Dynamics of Vector Vortex Beams in Kerr Medium with Parity–Time-Symmetric Lattice Modulation

Based on the two-dimensional (2D) nonlinear Schrödinger equation, we investigate the collapse dynamics of a vector vortex optical field (VVOF) in nonlinear Kerr media with parity–time (PT)-symmetric modulation. The critical power for the collapse of a VVOF in a Kerr-ROLP medium (Kerr medium with a real optical lattice potential) is derived. Numerical simulations indicate that the number, position, propagation distance, and collapse profile of the collapse of a VVOF in sine and cosine parity–time-symmetric potential (SCPT) Kerr media are closely related to the modulation depth, initial powers, and the topological charge number of a VVOF. The VVOF collapses into symmetric shapes during propagation in a Kerr-ROLP medium, and collapse shapes are sensitively related to the density of the PT-symmetric optical lattice potential. In addition, due to gain–loss, the VVOF will be distorted during propagation in the Kerr-SCPT medium, forming an asymmetric shape of collapse. The power evolution of the VVOF in a Kerr-SCPT medium as a function of the transmission distance with different modulating parameters and topological numbers is analyzed in detail. The introduction of PT-symmetric optical lattice potentials into nonlinear Kerr materials may provide a new approach to manipulate the collapse of the VVOF.

The Molecular Evolution of Melanoma Distant Metastases

The evolution of primary melanoma to lymph node and distant metastasis is incompletely understood. We examined the genomic diversity in melanoma progression in matched primary melanomas, lymph node and distant metastases from 17 patients. Fluorescence in situ hybridization (FISH) analysis revealed cancer cell fractions with monotonic copy number alterations (CNAs), including PHIP gain and PTEN loss, in the metastatic cascade. By contrast, the cancer cell fraction with CNAs for BPTF and MITF was reduced in lymph node metastases but increased in distant metastases. Separately, the cancer cell fraction with NCOA3 CNA was comparable in primary tumors and lymph nodes yet increased in distant metastases. These results suggest enrichment of the PI3K and MITF pathways in the transition through the metastatic cascade. By contrast, next-generation sequencing analysis did not identify a consistent pattern of changes in variant allele frequency (VAF) while revealing several intriguing findings, including decreased VAF in distant metastases and distinct drivers in lymph node versus distant metastases. These results provide evidence that distant melanoma metastasis does not always emanate from lymph node metastasis. These results enhance our understanding of clonal patterns of melanoma metastasis, with possible implications for targeted therapy and metastasis competency.

Agronomic adaptations to heat stress: sowing summer crops early in late winter.

Abstract CONTEXT: Summer crops are exposed to heat and drought stresses at critical stages during and after flowering, and their intensity and frequency are likely to increase with climate change. Agronomic stress avoidance offers the opportunity to temporally separate critical crop stages from heat and drought events, though it might require sowing cold sensitive summer crops early in late winter into colder than recommended soil temperatures. There is a need to understand how cold is too cold to sow summer crops early in late winter as well as what are the yield benefits and risks. OBJECTIVE: Here we quantify the likely benefits and trade-offs of sowing sorghum, a summer cereal, early in late winter as an adaptation to the increased frequency and intensity of heat and water stresses during flowering and grain filling. METHODS: Two seasons of multi-environment ( n =32) genotype by management trials were conducted across sub-tropical and semi-arid regions of eastern Australia. Environments (E) consisted of the combination of seasons, sites, three times of sowing (early, spring, and summer), and the use of supplementary irrigation. At each E a factorial combination of four plant populations (M) and eight commercial sorghum hybrids (G) were sown with three replications. Crop growth and yield components were measured, and the APSIM model was used to simulate all trials and treatments to quantify risks, and derive insights into functional relationships between simulated and measured environmental co-variates, and measured crop traits. RESULTS: The tested hybrids showed small differences in cold tolerance during crop establishment. Across the tested environments the G×M combinations produced up to 60% variation in treatment yields across environment yields, that varied between <0.5 to about 10t ha -1 , this translated into a ~8-fold variation in water use efficiency. Significant G×E and M×E interactions were observed for grain yield components. No G×M or G×E×M interactions were observed on yield or yield components. Early sowing was associated to a reduced risk of heat stress, and water use transfer from vegetative to reproductive stages. Early sowing in late winter or early spring resulted in no significant yield gain or yield loss when all sites and seasons are included in the analysis, and between 1-2 t ha -1 yield gains when the hottest sites and seasons are considered separately. The likelihood and severity of terminal drought stresses was also reduced with late winter and early spring sowings, improving yield quality parameters. CONCLUSIONS: Early sowing of sorghum can reduce the likelihood of heat stresses around flowering as well as the likelihood of terminal drought stresses. Advantages include reduced yield losses in the hottest seasons, and a transfer of water use to grain filling stages that resulted in increased grain yield and improved grain quality parameters. IMPLICATIONS: Agronomic adaptations offer opportunity to quickly adapt to the increase in the frequency and intensity of extreme hot events during critical crop stages. However, for the practice to be de-risked there is need to increase cold and chilling tolerance in sorghum, and or identify interventions that enhance seed germination and seedling vigour when the crop is sown early into cold soils.

Does the Understanding of Managing Variables among Pre-Service Early Childhood Teachers Correspond to Distinct Teaching Methods in Their Future Careers?

This study aims to examine the views of pre-service early childhood student teachers (referred to as student teachers) regarding the potential impact of an inquiry-based science course on their intention to utilize inquiry-based learning approaches, such as the Control of Variables Strategy (CVS) method. The study comprised a preliminary phase (N = 17) and a primary phase (N = 81). The participants’ level of understanding of the inquiry-based method was assessed using an open- and semi-open questionnaire. A 34-item Likert-type questionnaire was created using the Theory of Planned Behavior in the preliminary phase of the study and utilized in the primary phase to examine student teachers’ intentions to include the CVS technique into their lessons. The results showed student teachers’ strong intention to implement inquiry-based learning strategies, being supported by their positive estimations of managing the CVS method engagement in the classroom. However, their estimations of the significant others’ opinions and the personal gain–loss balance expected from engaging with the CVS method significantly hindered their intention. Most interestingly, the results of the study suggest that the various levels of comprehension of the CVS method attained by student teachers are connected to the adoption of distinct approaches in their future teaching endeavors. In light of these results, we discuss several teaching implications.

Safety of low weight gain or weight loss in pregnancies with class 1, 2, and 3 obesity: a population-based cohort study

There are concerns that current gestational weight gain recommendations for women with obesity are too high and that guidelines should differ on the basis of severity of obesity. In this study we investigated the safety of gestational weight gain below current recommendations or weight loss in pregnancies with obesity, and evaluated whether separate guidelines are needed for different obesity classes. In this population-based cohort study, we used electronic medical records from the Stockholm-Gotland Perinatal Cohort study to identify pregnancies with obesity (early pregnancy BMI before 14 weeks' gestation ≥30 kg/m2) among singleton pregnancies that delivered between Jan 1, 2008, and Dec 31, 2015. The pregnancy records were linked with Swedish national health-care register data up to Dec 31, 2019. Gestational weight gain was calculated as the last measured weight before or at delivery minus early pregnancy weight (at <14 weeks' gestation), and standardised for gestational age into z-scores. We used Poisson regression to assess the association of gestational weight gain z-score with a composite outcome of: stillbirth, infant death, large for gestational age and small for gestational age at birth, preterm birth, unplanned caesarean delivery, gestational diabetes, pre-eclampsia, excess postpartum weight retention, and new-onset longer-term maternal cardiometabolic disease after pregnancy, weighted to account for event severity. We calculated rate ratios (RRs) for our composite adverse outcome along the weight gain z-score continuum, compared with a reference of the current lower limit for gestational weight gain recommended by the US Institute of Medicine (IOM; 5 kg at term). RRs were adjusted for confounding factors (maternal age, height, parity, early pregnancy BMI, early pregnancy smoking status, prepregnancy cardiovascular disease or diabetes, education, cohabitation status, and Nordic country of birth). Our cohort comprised 15 760 pregnancies with obesity, followed up for a median of 7·9 years (IQR 5·8-9·4). 11 667 (74·0%) pregnancies had class 1 obesity, 3160 (20·1%) had class 2 obesity, and 933 (5·9%) had class 3 obesity. Among these pregnancies, 1623 (13·9%), 786 (24·9%), and 310 (33·2%), respectively, had weight gain during pregnancy below the lower limit of the IOM recommendation (5 kg). In pregnancies with class 1 or 2 obesity, gestational weight gain values below the lower limit of the IOM recommendation or weight loss did not increase risk of the adverse composite outcome (eg, at weight gain z-score -2·4, corresponding to 0 kg at 40 weeks: adjusted RR 0·97 [95% CI 0·89-1·06] in obesity class 1 and 0·96 [0·86-1·08] in obesity class 2). In pregnancies with class 3 obesity, weight gain values below the IOM limit or weight loss were associated with reduced risk of the adverse composite outcome (eg, adjusted RR 0·81 [0·71-0·89] at weight gain z-score -2·4, or 0 kg). Our findings support calls to lower or remove the lower limit of current IOM recommendations for pregnant women with obesity, and suggest that separate guidelines for class 3 obesity might be warranted. Karolinska Institutet and the Eunice Kennedy Shriver National Institute of Child Health and Human Development.

Supplementation with Lactiplantibacillus brevis GKEX Combined with Resistance Exercise Training Improves Muscle Mass, Strength Performance, and Body Fat Condition in Healthy Humans.

In addition to maintaining good exercise and dietary habits, recent studies have shown that probiotics may have potential benefits for muscle mass and strength. It is worth noting that the effects may vary depending on the specific strains used. To date, no studies have analyzed the effects of Lactiplantibacillus brevis in this context. Here, we combine the L. brevis strain GKEX with resistance training to further understand its effects on muscle mass, thickness, performance, and fat loss. In a six-week intervention for a double-blind randomized trial, 52 healthy subjects were divided into two groups (10 male and 16 female participants in each group): a placebo group (two capsules/day, containing 0 CFU of GKEX per capsule) and a GKEX group (two capsules/day, containing 1 × 1010 CFU of GKEX per capsule). Before the intervention, no differences were observed between the two groups in any of the tests (body composition, muscle thickness, exercise performance, and blood parameters). However, supplementation with GKEX significantly improved muscle mass and thickness, as well as grip strength, muscle strength, and explosive performance, when compared to the associated parameters before the intervention. Additionally, GKEX supplementation promoted a reduction in the body fat percentage (p < 0.05). Through analysis of the change amount, we observed that GKEX supplementation yielded significantly improved benefits when compared to the placebo group (p < 0.05). In summary, our findings support the notion that a six-week resistance exercise training program combined with L. brevis GKEX supplementation has superior additive effects that enhance muscle mass and strength performance, while also reducing body fat percentage. This intervention can promote muscle gain and fat loss.

Abstract 4575: FGFR3 altered bladder cancer exhibits a lineage-dependent vulnerability

Abstract Fibroblast Growth Factor 3 (FGFR3) genomic alterations are oncogenic drivers in ~15% of bladder cancer patients. A novel FGFR inhibitor, Erdafitinib, was FDA approved in 2019 based off a Phase II trial with an overall response rate of 40%. However, the majority of patients rapidly relapse at a median of 6 months, requiring combination therapies to prevent drug resistance. We hypothesized that a genome-wide CRISPR screen would identify Erdafitinib-sensitizing co-targets. We screened UM-UC-14, an S249C FGFR3 mutated bladder cancer cell line, and identified Cyclin C as a top hit in addition to known MAPK effectors such as ERK2 and SHP2. Cyclin C is a non-cell cycle related cyclin and part of the CDK8/19 kinase module of Mediator. Pharmacological validation in 4 FGFR3 altered cell lines confirmed a high degree of synergy between Erdafitinib and the CDK8/19 inhibitor Sel120, while no effect was seen in 2 wildtype cell lines, suggesting strong specificity. Moreover, the synergy in FGFR3-mutant cells was superior to that of SHP2 inhibitor SHP099 or the ERK inhibitor BVD-523. Similarly, GATA3 and PPARG also scored highly in our CRISPR screen. These transcription factors have been shown to be sufficient to enforce a luminal identity but not necessarily to maintain it. We validated these 3 genes as sensitizers and show in luminal cells that loss of either GATA3 or PPARG leads to loss of luminal markers and gain of basal markers. In contrast, cells treated with Sel120 showed, on RNAseq, an increase in basal markers without loss of luminal markers. Interestingly, our data suggests that the gain of basal markers, commonly associated with a worse prognosis, is associated with a strong sensitization to FGFR inhibition in FGFR3 alteration driven bladder cancer. We conducted a xenograft experiment confirming that mice responded well to single agent Erdafitinib, but without frank tumor regression. By contrast, Sel120+Erdafitinib induced complete responses in 8/10 mice compared to 6/10 mice for SHP099+Erdafitinib. Remarkably, after 60 days of daily drugging and a 20-day washout, the tumors did not grow back in the complete responders. Importantly, no loss of body weight or other signs of morbidity were found. Our results suggest an immediate and promising benefit in combining either CDK8/19 or SHP2 inhibitors with Erdafitinib in treating patients with FGFR3 alteration driven bladder cancers. Furthermore, we identified a strategy in targeting lineage modulators as sensitizers to existing therapies. Citation Format: Mohamad Karim Koleilat, Yan Zhu, Lawrence Kwong. FGFR3 altered bladder cancer exhibits a lineage-dependent vulnerability [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4575.

Abstract 1434: Modeling the 3q26 OncCassette using genetically engineered mice

Abstract Lung squamous cell carcinoma (LUSC) is a devastating disease, accounting for ~30% of all lung cancer diagnoses and ~40,000 deaths annually in the United States alone. LUSC patients suffer from high prevalence of relapse and a dismal 5-year survival of only ~24%. The lack of animal models that reflect the salient features of human disease to assess the safety and efficacy of drugs, and to explore the underlying molecular mechanisms of LUSC tumor biology, is a major impediment for LUSC patients. We have identified concomitant 3q26 copy number gain (CNG) and TP53 loss as a defining genetic alteration present in ~90% of LUSC tumors. Furthermore, we have identified and characterized three 3q26 oncogenes (PRKCI, ECT2 and SOX2) that genetically and biochemically collaborate to drive LUSC tumor formation. Using syngeneic mouse modeling, we have demonstrated that coordinate overexpression of Prkci, Ect2 and Sox2, and loss of Trp53 in ex vivo cultures of lung basal stem cells (LBSCs; a cell of origin for LUSC) drives transformation of these cells into tumors of exclusively LUSC histopathology. These findings lead to the hypothesis that PRKCI, ECT2 and SOX2 represent a multigenic driver of 3q26 CNG-driven LUSC, which we term the 3q26 OncCassette. Based in these findings, we have developed and begun characterizing the first autochthonous genetically engineered mouse model (GEMM) to study 3q26 OncCassette-driven tumorigenesis. Our novel immune competent GEMM will provide critical insight into tumor initiation and progression of preneoplastic lesions to malignant LUSC and allow characterization of the immune landscape in these tumors. Citation Format: Kayleah M. Meneses, Yi Liu, Capella R. Weems, Lee Jamieson, Duy T. Nguyen, Verline Justilien, Nicole R. Murray, Alan P. Fields. Modeling the 3q26 OncCassette using genetically engineered mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1434.

Abstract 6985: The oncogenic functions and action mechanism of UTP14A, a subunit required for 18S rRNA synthesis, in head and neck cancer

Abstract Head and neck cancer (HNC) is the sixth common malignancy in the world and &amp;gt; 90% of HNC cases are squamous cell carcinoma in origin. Oral cancer is one major HNC subtype. Despite the recent advancements in cancer diagnostics and therapeutics, the 5-yr survival rate of advanced HNC patients remains &amp;lt;50%. Ribosomes are the cellular factories responsible for making proteins. The hyper-activation of ribosome biogenesis initiated by oncogene gain or tumor suppressor gene loss plays a critical role in cancer initiation and progression. Ribosome biogenesis is a complex and energy intensive process that involves hundreds of factors. UTP14A gene encodes a member of the uridine triphosphate 14 family and is an essential component of a large ribonucleoprotein complex bound to the U3 small nucleolar RNA. Previous studies have demonstrated that UTP14A plays a key role in the synthesis of ribosomes and 18S rRNA, and is overexpressed in several malignancies. However, the biological functions and underlying mechanisms of UTP14A in HNC remain elusive. We found not only the increase of UTP14A mRNA expression in TCGA-HNC database but also its negative impact on patient clinical outcome. UTP14A depletion significantly impaired HNC cell proliferation, migration and invasion in vitro. Ectopic overexpression had the opposite effect, suggesting an oncogenic role for UTP14A in HNC. Through gene set enrichment analysis of pairwise RNA sequencing results, several genes involved in the process of epithelial mesenchymal transition (EMT) were potentially associated with manipulated UTP14A expression. We also detected a concordant change of lipid droplet numbers and its surface marker expression in UTP14A-manipulated cells, respectively, by BODIPY staining and immunhistochemical staining. Western blot analysis validated a positive relationship of EMT markers and COX-2, a potential mediator of lipid metabolism and EMT, with altered UTP14A expression. The involvement of COX2 in UTP14A mediation was confirmed by using both pharmacological inhibition and genetic manipulation. We also identified p38 as a central downstream effector but acting upstream from COX-2 in UTP14A mediation in oral cancer cells. We believe that our study of UTP14A-mediated oncogenic signaling axis can potentially be harnessed as a novel therapeutic target against HNC although more mechanistic studies may be needed. Citation Format: Li-Wha Wu, Chen-Ni Chang, Pulak K. Biswas, Sen-Tien Tsai. The oncogenic functions and action mechanism of UTP14A, a subunit required for 18S rRNA synthesis, in head and neck cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6985.

Abstract P351: The Associations of Short-Term, Chronic Localized and Chronic Widespread Pain With Life Expectancy Free of Major Chronic Diseases

Introduction: Pain is among the top underlying causes of years lived with disability. Hypothesis: Pain with higher chronicity and widespreadness is associated with shorter life expectancy free of major chronic diseases (cardiovascular disease, cancer, type 2 diabetes, and dementia), and longer lifespan lived with chronic diseases. Methods: The study included a total of 427,464 women and men from the UK Biobank study, free of major chronic diseases at baseline and with available information of pain (categorized as short-term pain, chronic localized pain, and chronic widespread pain), followed up until May 31, 2021. Population-based multistate life tables were used to calculate life expectancy with and without major chronic diseases. Results: Chronic localized pain was associated with significant but modest loss in healthy life expectancy and gain in lifespan lived with chronic diseases. Chronic widespread pain was associated with the largest loss in healthy life expectancy. Men and women with chronic widespread pain at age 50 years had 3.17 (95% confidence interval, 3.07 - 3.30) and 2.74 (2.66 - 2.83) shorter life expectancy free of major chronic diseases, 1.86 (1.73 - 1.97) and 1.84 (1.75 - 1.92) longer life expectancy with major chronic diseases as compared to individuals without pain ( Figure ). Chronic widespread pain was associated with ~15% and ~10% shorter life expectancy free of major chronic diseases in men and women at age 50 years, respectively, which increased to ~40% and ~30% after age 80 years. In terms of life expectancy in the presence of major chronic diseases, chronic widespread pain was constantly associated with ~25% and &gt;30% longer life expectancy as compared to those without pain in men and women, respectively. Conclusions: Chronic pain, especially chronic widespread pain, was associated with larger loss in healthy life expectancy and more lifespan lived with chronic diseases. Our study suggests chronic pain as a potential intervention target to improve quality of life.

The Impact of Quarantine on Physical Activity, Body Weight, and Eating Behaviors During the COVID-19 Pandemic in Saudi Arabia.

Background Since February 2020, the number of coronavirus disease 2019 (COVID-19) cases in Saudi Arabia has been rising. The Saudi Arabian government implemented strict lockdown measures in March 2020 in an effort to stop the spread of the virus. Globally, changes in socioeconomic levels and health during quarantine have been reported. Additional detrimental effects of the stay-at-home mandate include less physical activity among adult populations, anxiety, stress, and emotional eating. This study aimed to assess the impact of quarantine during the COVID‑19 pandemic on physical activity practice and weight. Methodology This cross-sectional study was conducted using a newly developed validated online questionnaire survey. The data were collected during quarantine in June 2020, including each adult who lived in Saudi Arabia during the quarantine. The data were reviewed and analyzed using SPSS version 21 (IBM Corp., Armonk, NY, USA). Results A total of 4,970 eligible participants completed the study survey. About 78.1% of the study participants performed physical activity inside the house during the pandemic. Overall, 30.9% reported increased body weight during the lockdown, and the most reported cause of increased body weight was feeling bored and empty. Regarding factors associated with physical activity and weight changes, 82.2% of participants aged 25-30 years performed physical activity during the pandemic. Increased weight was significantly higher among participants aged more than 30 years. Conclusions This study shows an increased level of physical activity among the study participants during the COVID-19 pandemic, especially among younger, female, and unmarried participants. More than half of the participants reported weight change, either weight gain or weight loss. Weight gain was higher in older participants. Regarding eating habits, there was an increased tendency toward eating a healthy diet.

Glucagon-Like Peptide-1 Receptor Agonists in Post-bariatric Surgery Patients: A Systematic Review and Meta-analysis.

A significant number of patients face the issue of weight gain (WG) or inadequate weight loss (IWL) post-bariatric surgery for obesity. Several studies have been published evaluating the role of glucagon-like peptide-1 receptor agonists (GLP1RA) for weight loss post-bariatric surgery. However, no systematic review and meta-analysis (SRM) till date has evaluated the efficacy, safety and tolerability of GLP1RA in this clinical scenario. Hence, this SRM aimed to address this knowledge gap. Databases were searched for randomized controlled trials (RCTs), case-control, cohort and observational studies involving use of GLP1RA in the intervention arm post-bariatric surgery. Primary outcome was weight loss post at least 3months of therapy. Secondary outcomes were evaluation of body composition parameters, total adverse events (TAEs) and severe adverse events (SAEs). From initially screened 1759 articles, 8 studies (557 individuals) were analysed. Compared to placebo, patients receiving liraglutide had significantly greater weight loss after 6-month therapy [MD - 6.0kg (95% CI, - 8.66 to - 3.33); P < 0.001; I2 = 79%]. Compared to liraglutide, semaglutide had significantly greater percent reduction in body weight after 6-month [MD - 2.57% (95% CI, - 3.91 to - 1.23); P < 0.001; I2 = 0%] and 12-month [MD - 4.15% (95% CI, - 6.96 to - 1.34); P = 0.004] therapy. In study by Murvelashvili et al. (2023), after 12-month therapy, semaglutide had significantly higher rates of achieving > 15% [OR 2.15 (95% CI, 1.07-4.33); P = 0.03; n = 207] and > 10% [OR 2.10 (95% CI, 1.19-3.71); P = 0.01; n = 207] weight loss. A significant decrease in fat mass [MD - 4.78kg (95% CI, - 7.11 to - 2.45); P < 0.001], lean mass [MD - 3.01kg (95% CI, - 4.80 to - 1.22); P = 0.001] and whole-body bone mineral density [MD - 0.02kg/m2 (95% CI, - 0.04 to - 0.00); P = 0.03] was noted with liraglutide. Current data is encouraging regarding use of GLP1RAs for managing WG or IWL post-bariatric surgery. Deterioration of bone health and muscle mass remains a concern needing further evaluation. The predefined protocol has been registered in PROSPERO having registration number of CRD42023473991.

Lower revascularization rates after high-speed rotational atherectomy compared to modified balloons in calcified coronary lesions: 5-year outcomes of the randomized PREPARE-CALC trial.

In the PREPARE-CALC trial, severely calcified lesion preparation with rotational atherectomy (RA) before biodegradable polymer sirolimus-eluting stent (SES) implantation demonstrated higher procedural success and comparable rates of acute lumen gain and late lumen loss compared to modified balloons (MB) (scoring/cutting). We aimed to analyze the 5-year outcomes of both lesion preparation strategies. PREPARE-CALC randomly assigned 200 patients 1:1 to MB or RA, followed by SES implantation. The principal endpoint of the current analysis was target vessel failure (TVF) at 5years. At 5years, MB had comparable rates of TVF to RA (19% vs. 21%, HR 1.14, 95% CI 0.60-2.16, p = 0.687). Subgroup analysis showed a lesion length treatment interaction, favoring MB for short lesions and RA for long ones (p for interaction = 0.042). Target lesion revascularization (TLR) was significantly less common with RA (12 vs. 3%, HR 0.28, 95% CI 0.08-0.98, p = 0.048). In a multivariate analysis, RA was independently protective against TLR (adj. HR 0.17, 95% CI 0.04-0.78, p = 0.022), while ostial lesions were associated with higher TLR independent of treatment strategy (adj. HR 11.3, 95% CI 2.98-42.6, p < 0.001). In patients with severely calcified coronary lesions, using MB or RA for lesion preparation followed by biodegradable polymer SES implantation was associated with comparable rates of TVF at 5years. However, a significant reduction of TLR was observed after RA. PREPARE-CALC is the first randomized trial showing potential clinical advantages of RA over MB during long-term follow-up. URL: https://www. gov . Unique identifier: NCT02502851.