Two-tailed Unpaired T-test Research Articles

Diesel exhaust particles activate macrophages and IRF5 expression in atherosclerosis

Abstract Introduction Over 20% of global deaths related to cardiovascular disease have been attributed in part to air pollution, particularly fine particulate matter (PM2.5) (1). In highly trafficked urban areas, smaller ultrafine diesel exhaust particles (DEP) represent a significant proportion of PM2.5. DEP is associated with the progression of atherosclerosis and increased plaque susceptibility to rupture (2). We have previously shown that interferon regulatory factor 5 (IRF5) has a key role in necrotic core formation, a distinguishing feature of vulnerable atherosclerotic plaques, by rendering macrophages defective at efferocytosis (3). Pronounced elevation of IRF5 is also seen in symptomatic carotid plaques, particularly in the rupture-prone shoulder regions (4). Purpose We examined the effect of DEP on macrophage phenotype and IRF5 expression in atherosclerosis. Methods Bone marrow-derived macrophages (BMDMs) from C57BL/6 mice, differentiated with GM-CSF (to mimic inflammatory macrophages) or M-CSF (mimicking homeostatic resident macrophages) were incubated with PBS or DEP (10 µg/mL) in vitro for 18 hours. Expression of inflammatory cytokines (IL-6, IL-12α), macrophage polarisation markers CD11c (itgax, inflammatory) and CD206 (mrc1, resident anti-inflammatory) and IRF5 was quantified by real-time PCR. In addition, high fat-fed ApoE-/- mice were exposed to saline or DEP (35 µL at 1 mg/mL) twice weekly for 5 weeks by pulmonary administration. The pan-macrophage marker CD64, CD206 and IRF5 were quantified in paraffin-embedded sections of brachiocephalic artery lesions by immunohistochemistry. Results In M-CSF-differentiated murine macrophages incubated with DEP, significant fold increases in gene expression (relative to PBS control) were observed: IRF5 (1.4-fold), IL-6 (31.5-fold) and IL-12α (5.8-fold) while CD206 (0.8-fold) decreased (Figure 1a, n = 5, p<0.05, two-tailed paired t-tests). CD11c expression was unchanged. No statistically significant differences were detected in GM-CSF-differentiated macrophages. In the brachiocephalic arteries of DEP-exposed ApoE-/- mice we observed increases in CD64+ macrophages (28.2±7.4 vs 15.5±5.6) and IRF5 (16.6±3.6 vs 10.1±3.4) expression, while CD206 expression decreased (4.1±0.9 vs 11.0±1.2) in comparison to saline-exposed mice (Figure 1b mean±SD, n = 5, p<0.05, two-tailed unpaired t-tests, data expressed as % of intima area). Conclusion DEP promotes CD64+ macrophages and IRF5 expression in murine atherosclerosis, while inflammation and IRF5 is induced in vitro, but only in M-CSF-differentiated murine BMDMs. This suggests that DEP has no impact on inflammatory macrophages but could switch homeostatic resident macrophages to a more inflammatory phenotype via the activation of IRF5. Therefore, IRF5 is a promising candidate for further investigation into the initial macrophage response to inhaled particles that leads to the exacerbation of vascular disease.

7143 Near Term Cardiometabolic Outcomes in Transgender Boys and Men in the First Year of Gender-Affirming Testosterone Therapy

Abstract Disclosure: J. Dey: None. M. Widlansky: None. S. Cabrera: None. Background: Gender affirming testosterone (T) therapy is used to masculinize transgender adolescent men (TMs), resulting in elevated T and suppressed estradiol (E2). In cis-females, hypoestrogenism reduces vaso-protective E2-mediated estrogen receptor expression, decreasing nitric oxide vasodilation and increasing susceptibility to future vascular complications. Whether T promotes similar changes in TMs remains poorly understood. Methods: A longitudinal observational study is conducted in TMs aged 12-30yrs over the 1st yr of T, with visits at baseline (BL; pre-T), 6mo, and 1yr, compared to cis-males (CMs) with endogenous T. Primary outcome is change in brachial artery flow-mediated dilation (FMD%), an in vivo measure of vascular endothelial health, after 1yr of T. To detect a significant relative change in FMD%, 60 participants are needed (40 TMs, 20 CMs). Metabolic profile is assessed by fasting lipids, glucose, and insulin. Body composition is assessed by BMI and DEXA. Differences in mean values between and within a cohort were analyzed using the unpaired two-tailed T-test assuming unequal and equal variances, respectively. Results: To date, 51 participants are enrolled; the 21 participants who fully completed the study are included here (13 TMs, 8 CMs). At BL, TMs were post-menarchal with a mean age of 15.6yrs, ht 162.6cm, and BMI z-score 0.47. At BL, 46% were on progestin-only menstrual suppression. CMs had a mean age of 18.4yrs, ht 170.7cm, and BMI z-score 0.13. Cohorts were similar for age, height, and BMI.At BL, FMD% was higher in TMs than CMs (8.2 vs 4.6, p=0.01) but not at 1yr (6.9 vs 6.2, p=0.6). FMD% did not change in TMs receiving T over 1yr (8.2 vs 6.9, p=0.3). At BL, the brachial artery resting diameter (mm) was lower in TMs than CMs (2.6 vs 3.5, p<0.01) and remained such at 1-yr. Resting diameter increased in TMs over 1yr of T (2.6 vs 3.1, p<0.01) but not in CMs. There were no differences in resting or hyperemic flow velocity, or FMD (mm) between or within groups. At BL, % lean body mass (%LBM) was lower in TMs than CMs (60 vs 72, p<0.01) and estimated percent visceral adiposity tissue (%VAT) was higher (26 vs 14, p<0.01). In TMs, %LBM increased (60 vs 66, p <0.01) and %VAT decreased (26 vs 21, p<0.01) over 1yr of T, such that TMs and CMs were similar at 1yr. At BL, TMs had lower T but similar E2 (46 vs 27 pg/mL, p=0.2) to CMs. At 1-yr, TMs and CMs had similar T (556 vs 574 ng/dL, p=0.9) and E2 (52 vs 29, p=0.1). In TMs, T did not change E2. Lipid and glycemic indices did not change over 1yr in TMs. Conclusion: Initial analyses suggest that T does not affect FMD; however, FMD% interpretation is confounded by the small number of participants, progestins at BL in TMs, and the lower resting brachial artery diameter in TMs. The stable E2 in TMs on T may explain the stable FMD%. Overall, our findings suggest T results in relative stability of vascular and metabolic function and a beneficial shift in body composition in TMs. Presentation: 6/2/2024

Musculoskeletal imaging fellowship program directors: A study of educational paths, and scholarly engagement in the United States

PurposeThe overall aim of the study is to analyze and provide a comprehensive understanding of the demographics, educational backgrounds, and scholarly activities of musculoskeletal imaging fellowship program directors across the United States. MethodsA list of all members of the Society of Skeletal Radiology was obtained and musculoskeletal imaging fellowship program directors across the US were included. Publicly available online sources were used to gather demographic and educational information about each musculoskeletal imaging fellowship program director, which included the online curriculum vitae from the program websites, Health Grades, Doximity, and Elsevier's Scopus database. Demographic and educational data including age, gender, educational background (medical school, residency, fellowship), additional degrees, academic rank, prior leadership positions, and metrics of scholarly activity were recorded. Fellowships in diagnostic musculoskeletal radiology along with additional degrees were recorded. A two-tailed unpaired t-test was used to calculate the difference between means of scholarly activity between male and female PDs. ResultsIn this study encompassing 92 programs across the United States, the majority (88) were dedicated to pure Musculoskeletal (MSK) Imaging Fellowship, while one each offered combined training in MSK and Body Imaging, MSK, and Emergency Imaging, MSK Sports, and Interventional Spine, and Pediatric MSK Imaging. Program directors were identified for 90 out of 92 programs, revealing a regional distribution of 29 in the South (31.5 %), 24 in the North East (26.1 %), 20 in the Midwest (21.74 %) and 19 in the West (20.65 %). Gender analysis unveiled a predominantly male representation, with 71 male directors compared to 17 female directors, while age ranged from 30 to 70 years, with a mean age of 47.17 ± 7.4 years. Medical school backgrounds predominantly featured MD degrees (80), followed by IMG (7) and DO (4) degrees, with the most common IMG source being India. Faculty positions were mainly distributed among Assistant Professors (35), Associate Professors (32), and Professors (11). Research output metrics showcased a mean of 41.943 publications and an h-index of 10.625. Extra degrees were obtained by 31 directors, with common additions being other fellowships, MBAs, MS degrees, and PhDs. Notably, some directors held previous or current leadership positions, while a few had completed residencies outside of Radiology or pursued fellowships beyond MSK Imaging. ConclusionMusculoskeletal imaging fellowship program directors across the United States are predominantly male, with approximately 8 % having graduated from international medical schools. The most common training pathway for these directors is completing a diagnostic radiology residency followed by a musculoskeletal radiology fellowship. This study highlighted the gender disparity within this leadership group and emphasized the diverse educational backgrounds that contribute to the field of musculoskeletal imaging.

Characteristics of a Spray-Dried Porcine Blood Meal for Aedes aegypti Mosquitoes.

Research into mosquito-borne illnesses faces hurdles because feeding fresh animal blood to rear female mosquitoes presents logistical, economic, and safety challenges. In this study, a shelf-stable additive (spray-dried porcine blood; SDPB) hypothesized to supply accessible hemoglobin was evaluated within an alternative meal (AM) containing whey powder and PBS for rearing the yellow fever mosquito Aedes aegypti. LC-MS/MS proteomics, microbial assays, and particle reduction techniques confirmed and characterized the functionality of hemoglobin in SDPB, while engorgement, fecundity, egg viability, and meal stability bioassays assessed AM performance. Chemical assays supported hemoglobin as the phagostimulant in SDPB with aggregates partially solubilized in the AM that can be more accessible via particle reduction. Unpaired two-tailed t-tests indicate that the AM stimulates oogenesis (t11 = 13.6, p = 0.003) and is stable under ambient (1+ y; t12 = 0.576, p = 0.575) and aqueous (14 d; t12 = 0.515, p = 0.639) conditions without decreasing fecundity. Egg hatch rates for the ninth generation of AM-reared Ae. aegypti were 50-70+%. With further development, this meal may serve as a platform for mass rearing or studying effects of nutritional additives on mosquito fitness due to its low cost and stability. Future work may examine tuning spray drying parameters and resulting impacts on hemoglobin agglomeration and feeding.

Abstract P346: Expansion of meningeal lymphatic vessels in DOCA-salt hypertension

The meningeal lymphatic system is critical for proper brain waste clearance and maintenance of the local meningeal neuroimmune niche. Meningeal lymphatic function is impaired during aging and neurodegenerative diseases and is associated with the development of cognitive impairment. There is a bidirectional communication between meningeal lymphatic function and dural T cells, whereby the drainage of T cells into the deep cervical lymph nodes depends on functional lymphatic vessels, and meningeal lymphatic function is itself modulated by T cell derived cytokines. We have previously shown that IL-17A producing T cells accumulate in the dura during deoxycorticosterone acetate (DOCA)-salt hypertension and contribute to the impairment of neurovascular coupling and cognitive function. However, the mechanisms by which these T cells accumulate in the dura remain to be determined. Here, we tested the hypothesis that the meningeal lymphatic vessels are impaired during DOCA-salt hypertension, thus contributing to dural T cell accumulation. 12-week old C57BL6 male mice were implanted with a s.c. DOCA pellet and received 0.9% NaCl in the drinking water for 21 days (n=4). Control mice underwent a sham surgery (n=4). Blood pressure was monitored by tail-cuff plethysmography. Meningeal lymphatic vessels were assessed by immunohistochemistry using the lymphatic vessel marker LYVE1. In a separate group of mice, we assessed meningeal lymphatic function by delivering Alexa647-Ovalbumin or IgG-RPE into the cisterna magna and measuring fluorescence in the deep cervical lymph nodes one hour after injection. DOCA-salt mice developed increased systolic blood pressure (control: 117.7 ± 4.9 vs DOCA: 141.5 ± 2.9mmHg). We observed no difference in the CD31+ dural sinus area between control and DOCA-salt (control: 11.482 ± 0.718 vs DOCA: 11.816 ± 0.662 mm 2 ). We found a significant increase in LYVE1+ area surrounding the dural sinuses in DOCA-salt (control: 0.508 ± 0.039 vs DOCA: 0.816 ± 0.105 mm 2 ; p=0.0329 by unpaired two-tailed t-test) and an increase in LYVE1+/CD31+ % area (control: 4.476 ± 0.462 vs DOCA: 6.840 ± 0.609 %; p=0.0213 by unpaired two-tailed t-test) which was not different between the sagittal sinus or the transverse sinus. Contrary to our hypothesis, our data suggests that the meningeal lymphatic vessels are expanded, not retracted, during DOCA-salt hypertension. The interaction between meningeal lymphatics and dura immunity in hypertension remains to be determined.

Z-Spectral MRI Quantifies the Mass and Metabolic Activity of Adipose Tissues With Fat-Water-Fraction and Amide-Proton-Transfer Contrasts.

Brown adipose tissue (BAT) is metabolically activatable and plays an important role in obesity and metabolic diseases. With reduced fat-water-fraction (FWF) compared with white adipose tissue (WAT), BAT mass and its functional activation may be quantified with Z-spectra MRI, with built-in FWF and the metabolic amide proton transfer (APT) contrasts. To investigate if Z-spectral MRI can quantify the mass and metabolic activity of adipose tissues. Prospective. Seven groups of 8-week-old male rats, including two groups (n = 7 per group) for in vivo MRI study and five groups (n = 5 per group) for ex vivo validation; 12 young and healthy volunteers with 6 male and 6 female. The 7 T small animal and 3 T clinical systems, T2-weighted imaging, Rapid Acquisition with Relaxation Enhancement (RARE) readout based chemical exchange saturation transfer (CEST) Z-spectral MRI sequence. Quantified FWF and APT from Z-spectra in rats before and after norepinephrine (NE) stimulation and in healthy human subjects; ex vivo measurements of total proteins in BAT from rats. Two-tailed unpaired Student's t-tests and repeated measures ANOVA. P-value <0.05 was considered significant. Decreased FWF (from 39.6% ± 7.2% before NE injection to 16.4% ± 7.2% 120 minutes after NE injection, P < 0.0001) and elevated APT (from 1.1% ± 0.5% before NE injection to 2.9% ± 0.5% 120 minutes after NE injection, P < 0.0001) signals in BAT were observed with in vivo Z-spectral MRI in rats injected with NE at 7 T MRI. At clinical 3 T, Z-spectral MRI was used to quantify the FWF (58.5% ± 7.2% in BAT and 73.7% ± 6.5% in WAT with P < 0.0001) and APT (2.6% ± 0.8% in BAT and 0.9% ± 0.3% in WAT with P < 0.0001) signals in healthy volunteers. APT signals of BAT were negatively correlated with the BMI in humans (r = 0.71). Endogenous Z-spectral MRI was demonstrated to simultaneously quantify BAT mass and function based on its FWF and APT contrasts. 1.

Abstract Tu007: The Impact of Shear-Thinning Hydrogel Delivery on Extracellular Vesicle Cardiac Retention

Background: Extracellular vesicles (EVs) are essential mediators of intercellular communication. EVs secreted by cardiosphere-derived cells (CDC-EVs) hold promise as vectors in the context of cardiac repair and remodeling through modulation of cardiomyocyte apoptosis and the immune response following injury. Cardiac retention of CDC-EVs is a critical factor influencing their therapeutic efficacy as first-pass uptake and washout from the myocardium remains a variable. We hypothesized that loading CDC-EVs in a shear-thinning, hybrid hydrogel would provide sustained myocardial delivery and reduced EV extraction by the lungs. Methods: The hydrogel system was constructed via a two-step approach utilizing Michael-addition reaction and host-guest complexation. This system incorporates four components: (1) an adamantane-functionalized multi-arm polyethylene glycol, (2) monosubstituted β-cyclodextrin-maleimide, (3) a thiolated hyaluronic acid, and (4) a heparin molecule containing thiol moieties. Following LAD ligation, intramyocardial injections of DiR-labeled EVs or hydrogel-encapsulated DiR-labeled EVs were performed on mice. Animals were euthanized after 24 hours and organs harvested. Whole organ biofluorescence was performed using IVIS Spectrum imaging. Results: Hydrogel-encapsulated EVs exhibited enhanced cardiac retention 24 hours post-MI when compared with EVs without hydrogel (17 ± 4.4 vs. 3.4 ± 0.9, mean radiant efficiency as a % of dose ± SEM, p&lt;0.01 using unpaired two-tailed t-test). Mice treated with hydrogel encapsulated EVs showed reduced EV retention in the lungs compared to controls treated with EVs alone (5.7 ± 0.9 vs. 10.6 ± 1.4, mean radiant efficiency as a % of dose ± SEM, p&lt;0.05 using unpaired two-tailed t-test). Conclusion: Usage of shear-thinning hydrogel as an EV delivery vehicle enhances EV retention in the heart and decreases off-target uptake of EVs following acute MI. This finding suggests that hydrogel delivery may be a promising strategy to improve the therapeutic efficacy of CDC-EVs for cardiac repair.

Abstract Tu133: Cardiac Adaptations and Mitochondrial Protection Through Long Noncoding RNAs Regulation in Mice on a Ketogenic Diet

Background: Heart failure, characterized by a metabolic imbalance, leads to impaired mitochondrial ATP production. Ketone bodies not only serve as an alternate energy source but also mitigate heart failure by reducing cardiac remodeling and inflammation. The role of long noncoding RNAs (lncRNAs) in cardiac remodeling and mitochondrial metabolism is increasingly appreciated, and this study examines the effect of ketogenic diet on cardiac lncRNA regulation and mitochondrial protection. Methods and Results: In a six-month study, 10-week-old male and female C57BL/6J mice were assigned to either a control diet (10% fat, 20% protein, 70% carbohydrate) or a ketogenic diet (Keto; 80% fat, 15% protein, 5% carbohydrate). Data were analyzed using unpaired two-tailed t-tests (GraphPad Prism) and reported as mean ± SE. The keto diet significantly increased blood ketone bodies (n=36/group), indicating ketosis, and led to weight gain as well as a significant decrease in heart mass relative to body weight, suggesting cardiac metabolic adaptation (Fig 1A-B) . The cardiac expression analysis of lncRNAs Anril, Caren, Carmn, Gm15441, Malat1, Miat, and Oip5-as1 via qPCR (n=13/group) revealed significant upregulation of Anril and Malat1 in the Keto hearts ( Fig 1C-D ), indicating their potential regulatory roles under ketogenic conditions. Sex-specific differences were not observed in the lncRNA expressions. Key proteins involved in mitochondrial biogenesis, protection, and metabolic adaptation, including Nrf2 and PGC1α, were significantly increased at both mRNA and protein levels (n=6-10) in the hearts of mice on keto diet ( Fig 1E-H ). Conclusion: A six-month ketogenic diet in mice promotes cardiometabolic adaptations characterized by a reduction in relative heart mass and activation of key proteins involved in mitochondrial biogenesis and protection possibly through lncRNAs Anril and Malat1. These findings shed light on novel molecular targets and the potential of ketogenic diet/ketone bodies in treating heart failure.

Activation of dendritic cells by a synthesized small molecule for breast cancer vaccination.

e13149 Background: Dendritic cell-based vaccination is a promising immunotherapy for treating various types of cancer. However, tumor-loaded dendritic cells alone have limited treatment efficacy. We have designed a small molecule mimicking the action of damage-associated molecular pattern molecules as an adjuvant for activating dendritic cells in vitro, which demonstrated augmented treatment efficacy. Methods: We aimed at exploring the roles of the synthesized small molecule in augmenting the efficacy of dendritic cell-based cancer vaccination to target triple negative breast cancer. Breast cancer mouse model is constructed using a 4T1 tumor cell line. Data were analyzed using a two-tailed unpaired t-test. Results: Mouse dendritic cells could be activated in vitro by the small molecule as an adjuvant evidenced by increased expression of MHC-I, MHC-II and CD40. Augmented motility of dendritic cells was also observed after treatment with the adjuvant. The signaling of NFkB is remarkably activated in dendritic cells by the adjuvant, which facilitates the maturation of dendritic cells. Engraftment of adjuvant-primed dendritic cells loaded with 4T1 tumor antigen into 4T1 tumor-bearing mice efficiently reduced the tumor size and improved survival, compared with the group treated with dendritic cells only. After injection of adjuvant-activated dendritic cells, both CD4 T cell and CD8 T cell populations were expanded in the spleen and lymph nodes draining the tumor sites. Furthermore, the adjuvant could promote the production of IFN-γ and IL-2, cytokines required for T cell activation and expansion. Next, splenocytes were obtained from individual mice after dendritic cell-based treatment. Upon re-encounter with the tumor antigen in vitro, remarkable expansion of T cell populations with a memory cell feature was confirmed in splenocytes derived from mice that received adjuvant-treated dendritic cells. Consistently, production of TNF-α, IFN-γ and granzyme B, was profoundly increased in these T cells associated with the adjuvant treatment. Conclusions: Our work suggests a novel dendritic cell-based cancer immunotherapy by using a small molecule mimicking the action of damage-associated molecular pattern molecules, which have documented immune potentiation activities.

Revolutionizing global hematology-oncology education: Assessing the two-year impact of a digital platform on socioeconomic equity and inclusivity against gender disparities.

e21000 Background: Social media has transformed the global exchange of knowledge, with 77.4% of cancer patients relying on it as a primary source of information especially in resource-limited areas. Given that a third of social media posts contain misinformation that can have negative ramifications, MedNews Week (MNW) was developed as a digital platform featuring presentations by global authorities to enhance global oncology education through its freely accessible programming. In light of the gender disparity in speakers at scientific symposia, where females represent &lt;30%, MNW strived to combat gender inequity by featuring them in equal proportion to males. The goal of this study was to evaluate the ability of MNW’s virtual oncology education platform to combat socioeconomic (SE) education inequity while promoting gender inclusivity. Methods: From January 2022 to 2024, MNW hosted 36 distinguished global oncology leaders as Keynote Speakers (Mean H-index=50.09). Data on viewership and outreach were collected from MNW's social media accounts. A mixed-method approach was employed to determine audience and speaker demographics, and engagement. Results: MNW achieved a Healthcare Social Score amongst the top 0.07 percentile of global healthcare influencers on Twitter. Over the past year, MNWoncology programming experienced a 10.5-fold increase in mean attendance, reaching 743,991 people in 95 countries globally, with 23 classified as low-to-middle SE status. In 2023, MNW followers exhibited a female predominance (48.8%), mainly from non-healthcare fields (60.0%). MNW showcased gender parity among oncology keynote speakers, with female lecturers drawing similar attendance as males (p=0.89, two-tailed unpaired t-test). Conclusions: MNW has established a growing digital platform, while removing financial barriers to effectively engage lay audiences, particularly in underserved regions. MNW's commitment to empowering female speakers exemplifies its dedication to inclusivity in a historically imbalanced field. This underscores MNW's success in fostering inclusivity, by driving engagement of those traditionally underserved, and positively impacting oncology education globally. [Table: see text]

Association of chr11q13 amplification with survival and recurrence in patients with surgically treated hepatocellular carcinoma.

e16253 Background: Hepatocellular carcinoma (HCC) is a highly lethal and aggressive malignant tumor characterized by high rates of recurrence and metastasis, and low survival outcomes. The current treatments available for HCC have shown limited success, and the prevalence of genomic characterization and their association with survival and recurrence in HCC patients are largely unknown. Methods: The investigation enrolled 1,221 Chinese individuals diagnosed with primary HCC. This study collected samples including tumor and paired tumor-free tissues and clinical characteristics. Among them, 348 patients' clinical outcome data, including overall survival (OS) and recurrence-free survival (RFS) were gathered. Genomic analysis was conducted through panel sequencing by OrigiMed. Hazard ratios were calculated employing both univariate and multivariate Cox proportional hazards regression models. The log-rank test and Kaplan-Meier plots were used to assess comparison of clinical outcomes by various prognostic factors. For other statistical analyses, a two-tailed unpaired t-test and Fisher's exact test were applied. Results: With a median age of 54, 87% of the cases were male. Pertaining to risk factors, 74% cases had HBV infection and 62% cases were cirrhosis. Meanwhile, serum AFP levels greater (or less) than 400 ng/mL were present in 299 cases (or 485 cases). Panel sequencing identified the most prevalent mutations ( &gt; 10%, excluding copy number variations) in TP53 (60%), TERT (40%), CTNNB1 (21%), AXIN1 (14%), LRP1B (14%), ARID1A (13%), RB1 (12%) and TSC2 (11%). The most frequently amplified genes were located at chr11q13, including CCND1 (9%), FGF19 (9%), FGF4 (7%) and FGF3 (7%). Survival analysis indicated that patients with Chr11q13 amp had significantly shorter OS and RFS. Further studies revealed a higher incidence of Chr11q13 amp in patients at advanced stages III/IV (64/564, 11%) compared to early-stage I/II (38/523, 7%), and in patients with vascular invasion (31/220, 14%) versus those without (44/599, 7%). No significant differences were observed in tumor mutational burden and HLA-I evolutionary divergence between patients with various Chr11q13 amp statuses. Furthermore, multivariate Cox proportional hazards regression models were utilized to examine the relationship between Chr11q13 amp status, characteristics and outcomes. Among the 248 patients with comprehensive clinical characteristics and available OS and RFS data, analyses indicated significant negative correlations between Chr11q13 amp and embolus with both OS and RFS. Conclusions: Our study elucidates that the presence of Chr11q13 amp was significantly associated with worse survival and increased risk of recurrence among a large cohort of Chinese patients with HCC. This discovery yields important insights into the understanding of HCC and has the potential to benefit personalized therapy.

Changes in Compression Pressure of Elastic Stockings for the Lower Limbs During Cesarean Section: A Prospective Observational Study.

Background Postpartum peripheral nerve injuries can impact recovery. Elastic stockings are recommended for thromboembolism prevention, although concerns about entrapment neuropathy exist. In this prospective observational study, we investigated the differential compressions caused by wearing elastic stockings before and after anesthesia, as well as changes in the diameters of the lower leg and ankle in parturient women undergoing spinal anesthesia for elective cesarean section (CS). Methods Eighteen pregnant women, classified by the American Society of Anesthesiologists as having physical status 2, underwent lower leg measurements taken before a CS. Elastic stockings were applied, and compression pressure was measured at pre-anesthesia, post-surgery, and six hours post-return to a hospital room. Fluid, blood loss, urine output, and neuropathy presence were recorded. For all parameters, changes at the three time points were compared for the primary analysis. For secondary analysis, participants were categorized as having intraoperative blood loss greater than (group P) or less than 1,000 g (group N), and factors were compared with pre-anesthesia and six hours post-return to a room. Data were analyzed and presented using a one-way analysis of variance with Bonferroni correction for multiple comparisons or unpaired two-tailed t-tests for pairwise comparison. Results None of the women had postoperative entrapment neuropathy. Six patients had >1,000 g of blood loss. Compression significantly increased from pre-anesthesia (left 13.6 ± 2.4, 95% CI: 12.18 to 14.52; right 13.4 ± 2.4, 95% CI: 12.41 to 14.69) to post-surgery (left, 17.4 ± 2.6, 95% CI: 15.68 to 18.12; right, 16.9 ± 2.6, 95% CI: 16.20 to 18.70) (p < 0.01). Compression pressure at post-surgery differed significantly between group P (left, 15.3 ± 1.3; right, 14.7 ± 1.8; 95% CI: -4.98 to -0.32) and group N (left, 18.1 ± 2.9; right, 17.8 ± 2.4; 95% CI: -5.38 to -0.26) (p < 0.05). The results are expressed as mean ± standard deviation, with P-values <0.05 indicating statistical significance. Conclusions In this study, no neuropathy occurred; however, over-compression risk with elastic stockings, especially when exceeding recommended pressure levels, was highlighted. Balancing thromboembolism prevention and over-compression risks is crucial for patients undergoing CSs with spinal anesthesia.

Real-World Efficacy of Ensitrelvir in Hospitalized Patients With COVID-19 in Japan: A Retrospective Observational Study.

The coronavirus disease 2019 (COVID-19) pandemic necessitates continuously evaluating antiviral treatments, especially for high-risk groups, including older individuals. This study aimed to compare the efficacy of three antiviral drugs, including remdesivir, molnupiravir, and ensitrelvir, in hospitalized patients as measured by our own institution's antigen test, focusing on outcomes, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen levels, hospitalization duration, and fever resolution. This retrospective observational study was conducted at Yoshida Hospital, Asahikawa City, Japan, enrolling 154 patients who received antiviral treatment upon COVID-19 diagnosis from July 1, 2022, to September 15, 2023. The diagnosis was confirmed by proprietary antigen tests or loop-mediated isothermal amplification assays. Patients who received treatment outside the hospital or with consistently negative antigen results were excluded. Drug administration was determined by attending physicians, considering oral administration challenges and renal dysfunction. The data were statistically analyzed using an unpaired two-tailed Student's t-test and one-way analysis of variance complemented by the Tukey post-hoc test for detailed group comparisons. No significant differences were observed in the initial antigen levels among the treatment groups. By day 10, the ensitrelvir group showed lower antigen levels than the other groups, but not significantly. The ensitrelvir group had a higher antigen-negative conversion rate and a significantly shorter hospital stay than the molnupiravir group. However, no significant differences were noted in the fever resolution time among the groups. This study suggests the potential benefits of ensitrelvir in reducing antigen levels and hospitalization duration. However, the overall efficacy of the antiviral agents for symptomatic relief appears similar. These findings underscore the need for further research to optimize COVID-19 management by considering personalized treatment approaches and long-term outcomes.

Role of hydroxymethylglutharyl-coenzyme A reductase in the induction of stem-like states in breast cancer

PurposeDe novo synthesis of cholesterol and its rate-limiting enzyme, 3-hydroxy-3-methylglutharyl-coenzyme A reductase (HMGCR), is deregulated in tumors and critical for tumor cell survival and proliferation. However, the role of HMGCR in the induction and maintenance of stem-like states in tumors remains unclear.MethodsA compiled public database from breast cancer (BC) patients was analyzed with the web application SurvExpress. Cell Miner was used for the analysis of HMGCR expression and statin sensitivity of the NCI-60 cell lines panel. A CRISPRon system was used to induce HMGCR overexpression in the luminal BC cell line MCF-7 and a lentiviral pLM-OSKM system for the reprogramming of MCF-7 cells. Comparisons were performed by two-tailed unpaired t-test for two groups and one- or two-way ANOVA.ResultsData from BC patients showed that high expression of several members of the cholesterol synthesis pathway were associated with lower recurrence-free survival, particularly in hormone-receptor-positive BC. In silico and in vitro analysis showed that HMGCR is expressed in several BC cancer cell lines, which exhibit a subtype-dependent response to statins in silico and in vitro. A stem-like phenotype was demonstrated upon HMGCR expression in MCF-7 cells, characterized by expression of the pluripotency markers NANOG, SOX2, increased CD44 +/CD24low/ −, CD133 + populations, and increased mammosphere formation ability. Pluripotent and cancer stem cell lines showed high expression of HMGCR, whereas cell reprogramming of MCF-7 cells did not increase HMGCR expression.ConclusionHMGCR induces a stem-like phenotype in BC cells of epithelial nature, thus affecting tumor initiation, progression and statin sensitivity.

Antibacterial efficacy and mechanism of Cyprinus carpio chemokine-derived L-10 against multidrug-resistant Escherichia coli infections

ObjectivesAntimicrobial resistance has raised concerns regarding untreatable infections and poses a growing threat to public health. Rational design of new AMPs is an ideal solution to this threat. MethodsIn this study, we designed, modified, and synthesised an excellent AMP, L-10, based on the original sequence of the Cyprinus carpio chemokine. All experimental data were presented as the mean ± standard deviation (SD), and the two-tailed unpaired T-test method was used to analyze all data. ResultsL-10 exhibited excellent antibacterial activity with negligible toxicity and improved the efficacy of a broad class of antibiotics against MDR Gram-negative pathogens, including tetracycline, meropenem, levofloxacin, and rifampin. Mechanistic studies have suggested that L-10 targets the bacterial membrane components, LPS and PG, to disrupt bacterial membrane integrity, thereby exerting antibacterial effects and enhancing the efficacy of antibiotics. Moreover, in animal infection models, L-10 significantly increased the survival rate of infected animals and effectively reduced the tissue bacterial load and inflammatory factor levels. In addition to its direct antibacterial activity, L-10 dramatically reduced pulmonary pathological alterations in a mouse model of endotoxemia and suppressed LPS-induced proinflammatory cytokines in vitro and in vivo. Lastly, L-10 was successfully expressed in Pichia pastoris and maintained antimicrobial activity against MDR Gram-negative pathogens in vivo and in vitro. ConclusionCollectively, these results reveal the potential of L-10 as an ideal candidate against MDR bacterial infections and provide new insights into the design, development, and clinical application of AMPs.

Progression of an Artificial Intelligence Chatbot (ChatGPT) for Pediatric Cardiology Educational Knowledge Assessment.

Artificial intelligence chatbots, like ChatGPT, have become powerful tools that are disrupting how humans interact with technology. The potential uses within medicine are vast. In medical education, these chatbots have shown improvements, in a short time span, in generalized medical examinations. We evaluated the overall performance and improvement between ChatGPT 3.5 and 4.0 in a test of pediatric cardiology knowledge. ChatGPT 3.5 and ChatGPT 4.0 were used to answer text-based multiple-choice questions derived from a Pediatric Cardiology Board Review textbook. Each chatbot was given an 88 question test, subcategorized into 11 topics. We excluded questions with modalities other than text (sound clips or images). Statistical analysis was done using an unpaired two-tailed t-test. Of the same 88 questions, ChatGPT 4.0 answered 66% of the questions correctly (n = 58/88) which was significantly greater (p < 0.0001) than ChatGPT 3.5, which only answered 38% (33/88). The ChatGPT 4.0 version also did better on each subspeciality topic as compared to ChatGPT 3.5. While acknowledging that ChatGPT does not yet offer subspecialty level knowledge in pediatric cardiology, the performance in pediatric cardiology educational assessments showed a considerable improvement in a short period of time between ChatGPT 3.5 and 4.0.

Brain morphometric changes in children born as small for gestational age without catch up growth.

Most infants born as small for gestational age (SGA) demonstrate catch up growth by 2-4 years, but some fail to do so. This failure is associated with several health risks, including neuropsychological development issues. However, data on the morphological characteristics of the brains of infants born as SGA without achieving catch up growth are lacking. This study aims to determine the structural aspects of the brains of children born as SGA without catch up growth. We conducted voxel- and surface-based morphometric analyses of 1.5-T T1-weighted brain images scanned from eight infants born as SGA who could not achieve catch up growth by 3 years and sixteen individuals with idiopathic short stature (ISS) to exclude body size effects. Growth hormone (GH) secretion stimulation tests were used to rule out GH deficiency in all SGA and ISS cases. The magnetic resonance imaging data were assessed using Levene's test for equality of variances and a two-tailed unpaired t-test for equality of means. The Benjamini-Hochberg procedure was used to apply discovery rate correction for multiple comparisons. Morphometric analyses of both t-statical map and surface-based analyses using general linear multiple analysis determined decreased left insula thickness and volume in SGA without catch up growth compared with ISS. The brain scans of patients with SGA who lack catch up growth indicated distinct morphological disparities when compared to those with ISS. The discernible features of brain morphology observed in patients born as SGA without catch up growth may improve understanding of the association of SGA without catch up growth with both intellectual and psychological outcomes.

Direct observation of NMR transverse relaxation in nanopatterned clusters of iron oxide particles.

We aim to verify predictions showing T2 relaxation rate of nanoparticle clusters and its dependence on spacing, size, geometry, and pulse sequence. We performed a laboratory validation study using nanopatterned arrays of iron oxide nanoparticles to precisely control cluster geometry and image diverse samples using a 4.7T MRI scanner with a T2 -weighted fast spin-echo multislice sequence. We applied denoising and normalization to regions of interest and estimated relative R2 for each relevant nanoparticle array or nanocluster array. We determined significance using an unpaired two-tailed t-test or one-way analysis of variance and performed curve fitting. We measured a density-dependent T2 effect (p = 8.9976 × 10-20 , one-way analysis of variance) and insignificant effect of cluster anisotropy (p = 0.5924, unpaired t-test) on T2 relaxation. We found negative quadratic relationships (-0.0045[log τD ]2 -0.0655[log τD ]-2.7800) for single nanoparticles of varying sizes and for clusters (-0.0045[log τD ]2 -0.0827[log τD ]-2.3249) for diffusional correlation time τD = rp 2 /D. Clusters show positive quadratic relationships for large (3.8615 × 10-6 [dpp /rp ]2 -9.3853 × 10-5 [dpp /rp ]-2.0393) and exponential relationships for small (-2.0050[dpp /rp ]0.0010 ) clusters. Calculated R2 peak values also align well with in silico predictions (7.85 × 10-4 ms compared with 1.47 × 10-4 , 4.23 × 10-4 , and 5.02 × 10-4 ms for single iron oxide nanoparticles, 7.88 × 10-4 ms compared with 5.24 × 10-4 ms for nanoparticle clusters). Our verification affirms longstanding in silico predictions and demonstrates aggregation-dependent behavior in agreement with previous Monte Carlo simulation studies.

SAT106 Use Of Data Sharing Feature Of Dexcom G6 Sensor Is Associated With Improved Glycemic Control: A Comparative Analysis

Abstract Disclosure: B. Sadek: None. Introduction: The use of Continuous Glucose Monitoring (CGM) sensors has significantly increased in part due to the seamless integration with smartphones. The Dexcom G6 CGM automatically sends real-time glucose readings to a compatible smart device. Moreover, the Dexcom Clarity app provides analytic tools and allows users to continuously share glucose data with their healthcare professionals, enabling remote monitoring of patients' glycemic control. Although multiple studies have shown better glucose control in patients who use CGM systems, what remains unexplored is if glucose metrics are further improved in those who share CGM data with healthcare providers. In this study, we compared glycemic outcomes among patients who use the sharing feature of the Dexcom G6 sensor to share their data with their healthcare providers to those who do not. Method: We conducted a single-center cross-sectional study in a community hospital with patients who use the Dexcom G6 CGM (n=201). Data was obtained from Dexcom Clarity for Healthcare Professionals, an online platform for clinicians to review patients' glucose patterns and statistics through interactive reports. The following data was collected for each patient: date of the last upload, data sharing status, average glucose, standard deviation, glucose management indicator (GMI) (%), glucose ranges (categorized as very high, high, in range, low, very low), sensor usage, and coefficient of variation (%). Two-tailed unpaired Student's t-tests were used to analyze the CGM metrics based on data sharing status. Significance was defined as a p-value less than 0.05. Results: 123 (61.19%) of the study participants shared data with healthcare providers. Participants with sharing data 'on' were significantly younger (49±15 years, p-value = 0.0007) and had increased sensor usage (90±21%, p-value = 0.0012), compared to those with sharing off. Participants who shared data spent more time in range (57±24 %, p-value = 0.0435), indicating a higher percentage of readings with glucose levels in a healthy range (70-180 mg/dL). Lastly, average glucose was lower in participants who shared data, although not statistically significant (179 ±44 mg/dL, p=0.08). Conclusion: The data-sharing feature on Dexcom G6 was utilized more frequently by younger people and was associated with a higher percentage of sensor usage. In addition, using the data-sharing feature of the Dexcom G6 could lead to a clinically meaningful improvement in glycemic control. Presentation: Saturday, June 17, 2023

Phosphatase and Tensin Homolog (PTEN) Expression as a Surrogate Biomarker Correlated With the Depth of Invasion in Cutaneous Malignant Melanoma.

Objective The aim of this study is to evaluate the expression of the phosphatase and tensin homolog (PTEN), which is a tumor suppressor gene that is implicated in the pathogenesis of cutaneous malignant melanoma, in normal skin and melanoma tissue samples. The study also aimed to correlate PTEN expression levels with various clinicopathological parameters of melanoma lesions, thus highlighting the utility of PTEN expression as a prognostic biomarker for melanoma. Study design Immunohistochemistry (IHC) staining was performed on tissue microarray samples representing normal skin and melanoma biopsies of different clinicopathological parameters. Tissue photomicrographs were evaluated with Aperio ImageScope, which has a positive-pixel-counting algorithm built in. Subsequently, a histochemical score (H-score) was derived from the percentage of positive cells (%-staining) and their staining intensity. The H-scores were averaged in groups of tissue samples representing the different melanomas' tumor (T), node (N), and distant metastasis (M), also known as TNM parameters, as set forth by the American Joint Committee on Cancer (AJCC) classification. The mean H-scores were statistically compared using a two-tailed unpaired t-test. Results The PTEN protein expression was measured by IHC and found to be correlated with tumor thickness (T), which is a reliable indicator for survival rates. Specifically, PTEN was significantly downregulated in tumors with a thickness over 2 mm (T3+T4) compared to tumors with a thickness at or below 2 mm (T1+T2). Conclusions The PTEN protein expression, as measured by immunohistochemistry, helped differentiate between tumors with a thickness over 2 mm and tumors with a thickness at or below 2 mm, suggesting PTEN as a potential surrogate marker for the melanoma's invasion depth along with possible prognostic implications. Longitudinal studies evaluating risk stratification based on the expression of PTEN are needed to establish the utility of this promising biomarker in the clinic as an adjunct for pathological examination.