Search Results (2,009)

Objective: The fetal-to-neonatal transition is marked by profound cardio-respiratory changes. Infections emerging within the first 48 h after birth may influence early cardiovascular adaptation. We aimed to evaluate the association between early infection/inflammation markers and vital parameters in neonates during the first 15 min after birth. Methods: This is a secondary outcome parameter post-hoc analysis of data derived from a prospective observation study. Preterm and term neonates with cerebral oxygen saturation (crSO2) monitoring (INVOS 5100C) during the first 15 min after birth and available inflammatory markers (C-reactive protein [CRP], leukocytes, immature-to-total neutrophils ratio [IT ratio]) within 48 h after birth were included. Heart rate (HR) and arterial oxygen saturation (SpO₂) were continuously recorded during the first 15 min. Inflammatory markers obtained at 16–24 and 24–48 h after birth were correlated with crSO₂, SpO₂, and HR at minute 5, 10 and 15. Results: Sixty-eight neonates were included (median (IQR) gestational age 34.0 (32.0; 35.9) weeks, birth weight 1900 (1488; 2542) grams). CRP within the first 24 h correlated negatively with crSO₂ (r = −0.314; p = 0.011) and with SpO₂ (r = −0.393; p = 0.001) at minute 15. IT ratio within 24 h correlated negatively with crSO₂ at minute 5 (r = −0.367; p = 0.005), 10 (r = −0.273; p = 0.035), and 15 (r = −0.306; p = 0.013), and with SpO₂ at minute 5 (r = −0.327; p = 0.008). IT ratio at 24–48 h correlated negatively with crSO₂ at minute 15 (r = −0.384, p = 0.012). No significant correlations were observed with HR. Leukocytes within the first 24 h after birth correlated negatively with crSO2 at minute 5 (r = −0.265; p = 0.046). Conclusions: Early inflammatory markers, particularly CRP and the IT ratio, are associated with cerebral and systemic oxygenation during immediate postnatal transition. These findings suggest a potential association between early inflammatory activation and oxygenation dynamics; however, given the observational design and modest correlation strength, the results should be interpreted cautiously and do not allow conclusions regarding causality or underlying mechanisms. Full article

Background and Objectives: Metaplastic breast carcinoma (MBC) is a rare, aggressive malignancy that is often resistant to conventional chemotherapy and characterized by a triple-negative phenotype. While immune checkpoint inhibition shows promise, the prognostic significance and distribution of programmed death-ligand 1 (PD-L1) expression within the heterogeneous architecture of MBC remain poorly understood. This study aimed to evaluate PD-L1 expression and the density of tumor-infiltrating lymphocytes (TILs) to clarify their roles in patient stratification and overall survival (OS). Materials and Methods: We retrospectively analyzed 48 MBC cases diagnosed between 2010 and 2025. PD-L1 expression was quantified using the Combined Positive Score (CPS) with the 22C3 antibody clone across diverse histological components. The density of stromal TIL density was assessed following internationally standardized guidelines. Clinical outcomes and clinicopathological parameters, including metastasis, lymphovascular invasion (LVI), and histological subtype, were correlated with biomarker status using Kaplan–Meier survival analysis and Cox proportional hazards regression models. Results: PD-L1 positivity (CPS ≥1) was identified in 72.9% of cases, one of the highest rates documented in literature. Notably, an inverse relationship was observed with PD-L1-negative tumors, which exhibited significantly higher rates of distant metastasis (46.2% vs. 17.1%; p = 0.039). Multivariate analysis confirmed that low density of TILs (HR = 9.66; p = 0.016), metastasis (HR = 4.40; p = 0.023), and the presence of LVI (HR = 3.84; p = 0.047) were strong independent predictors of mortality. While PD-L1 status alone did not directly dictate overall survival, mean overall survival was markedly reduced in the low TILs cohort (32.2 months) compared to the high TILs group (114.2 months). Conclusions: The high prevalence of PD-L1 expression supports routine screening for immunotherapy eligibility in MBC. Our findings suggest that PD-L1-negative cases represent a high-risk biological subset driven by alternative immune evasion mechanisms. Integrating TIL density with conventional pathological parameters provides a more robust prognostic framework, enabling personalized therapeutic strategies for this challenging malignancy. Full article

Background/Objectives: Total neoadjuvant therapy (TNT), integrating systemic chemotherapy and radiotherapy before surgery or surveillance, has become a standard approach for locally advanced rectal cancer (LARC). However, optimal sequencing strategies and long-term outcomes of watch-and-wait (W&W) following sandwich TNT remain insufficiently characterized. We evaluated oncologic outcomes and treatment response in patients treated with an institutional sandwich TNT protocol. Methods: We conducted a retrospective cohort study of consecutive patients with LARC treated with sandwich TNT (induction chemotherapy followed by hypofractionated intensity-modulated radiotherapy with simultaneous integrated boost [IMRT-SIB] chemoradiotherapy and consolidation chemotherapy) at the Institute of Oncology Ljubljana between 2016 and 2023. The primary endpoint was an overall complete response (CR; pathological [pCR] and clinical [cCR]). Secondary endpoints included tumor regression grade (TRG), major pathological response (MPR), R0 resection rate, organ preservation, overall survival (OS), and disease-free survival (DFS). Results: Among 205 patients (median age 61 years), overall CR was 29.5% (pCR 19.3% and cCR 10.2%). Major pathological response (TRG 3–4) occurred in 37.6%. R0 resection was achieved in 94.5%. In the W&W cohort (n = 21), local regrowth occurred in 33.3% (95% CI, 14.6–57.0%) over a median follow-up of 4.96 years. Total mesorectal excision (TME)-free survival at 5 years was 73.1% (95% CI, 55.0–97.2%). Estimated 5-year OS was 81.1% (95% CI, 75.5–87.2%) and 5-year DFS was 75.2% (95% CI, 69.0–82.0). In multivariable analysis, non-R0 resection (HR 6.06, 95% CI, 1.99–18.42), MRI circumferential resection margin positivity (HR 3.11, 95% CI, 1.53–6.33), and MRI extramural vascular invasion positivity (HR 1.97, 95% CI, 1.05–3.91) remained independent predictors of DFS. Conclusions: Institutional sandwich TNT yields meaningful tumor response and durable survival in MRI-defined high-risk LARC. Structured W&W offers organ preservation with acceptable oncologic control under intensive surveillance. Full article

Objectives: This study aimed to investigate the impact of TDs on the survival of patients with locally advanced rectal cancer (LARC). Additionally, we propose a novel staging method that combines TDs and lymph node metastases (LNMs) to enhance prognostic accuracy. Methods: Patients with LARC were retrospectively identified from the Surveillance, Epidemiology, and End Results (SEER) database and a Sun Yat-sen University (SYSU) cohort. Propensity score matching (PSM) was utilized to minimize selection bias when evaluating TDs. We quantitatively stratified TDs counts and integrated them with regional LNMs to formulate a novel tumor node metastasis (TNM) staging system. Furthermore, a prognostic nomogram incorporating TDs was constructed and validated to predict survival. Results: Overall, 19,991 patients were included in the SEER database, with 2667 (13.3%) TDs-positive and 17,324 (86.7%) TDs-negative tumors. After PSM, multivariate Cox analysis reveals that TDs are an independent adverse prognostic factor (HR = 1.521, 95% CI: 1.366–1.693, p < 0.001). Patients with high-risk group (TDs > 4) at any TNM stage exhibit OS comparable to or worse than that of stage IIIC disease. For patients staged as T4N2M0, the high-risk group (TDs > 4) demonstrates OS equivalent to stage IV disease. The nomogram achieved C-indices of 0.713 (training cohort, n = 8586) and 0.789 (external validation cohort, n = 304), with AUCs of 0.774 (3-year) and 0.710 (5-year). Conclusions: The presence of TDs is associated with poorer OS, and integrating TDs with LNMs improves the accuracy of TNM staging. The nomogram (C-index = 0.789) provides enhanced prognostic stratification and survival prediction. Full article

Background/Objectives: Proton pump inhibitors (PPIs) are known to alter gut microbiota composition; however, their association with disease courses and outcomes in patients with inflammatory bowel disease (IBD) remains uncertain. Our aims were to evaluate the association between PPI use and treatment escalation, Clostridioides difficile infection, and healthcare utilization in IBD. Methods: We conducted a retrospective cohort study on the TriNetX platform. IBD patients with PPIs or histamine-2 receptor antagonists (H2RAs) were matched one-to-one using propensity scores. Outcomes included initiation of corticosteroids, biologic therapy, Clostridioides difficile (C. difficile) infection, and healthcare utilization. Outcomes were assessed during the 0–12-month and 3–12-month follow-up windows. Associations were estimated using odds ratios (ORs) and hazard ratios (HRs) with 95% confidence intervals. Results: After matching, 12,808 patients were included in each group. During 0–12 months of follow-up, PPI use was associated with higher odds of systemic corticosteroid exposure (OR 1.56, 1.35–1.79), biologic therapy initiation (OR 1.99, 1.72–2.29), C difficile infection (OR 1.42, 1.18–1.70), and healthcare utilization (OR 1.18, 1.03–1.36) compared with H2RA use. Time-to-event analyses showed persistent associations with systemic corticosteroid exposure (HR 1.50, 1.31–1.72) and biologic therapy initiation (HR 1.91, 1.66–2.19), with attenuation of associations for infection and healthcare utilization in 3–12-month lag-time analyses. Similar patterns were observed in ulcerative colitis and Crohn’s disease subgroups. Conclusions: PPI was associated with higher risks of treatment escalation and C. difficile compared with H2RA in IBD. These findings highlight the importance of individualized selection and periodic reassessment of acid suppression therapy as part of personalized management strategies in IBD. Full article

Myocardial ischemia–reperfusion injury (MIRI) significantly compromises the clinical benefits of revascularization and constitutes a central pathological mechanism worsening prognosis in myocardial infarction patients. Accordingly, dissecting the molecular mechanisms underlying MIRI and formulating effective therapeutic interventions are of great clinical significance. Lycium barbarum polysaccharide (LBP), the primary active constituent of Lycium barbarum, has garnered considerable attention in the prevention and treatment of cardiovascular diseases due to its anti-inflammatory, antioxidant, vasomotor function-improving, and antithrombotic properties. This study aims to investigate the ability of LBP to alleviate MIRI, with a specific focus on its role in modulating the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome. Myocardial ischemia/reperfusion (I/R) models in rats and hypoxia/reoxygenation (H/R) models in H9c2 cells were established. Myocardial injury and the therapeutic effect of LBP were evaluated by 2,3,5-Triphenyl tetrazolium chloride (TTC) staining, Hematoxylin-eosin (H&E) staining, Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) staining, and Enzyme-linked immunosorbent assay (ELISA). To elucidate the specific mechanism underlying LBP against MIRI, an Nrf2-overexpressing cell line was generated in H9c2 cells, and pharmacological inhibition of Nrf2 with ML385 was applied for complementary validation. The effects of LBP on H/R-induced oxidative stress, inflammatory response (IL-18, IL-1β), and pyroptosis-related protein expression (NLRP3, apoptosis associated speck-like protein containing a CARD (ASC), cysteine-dependent aspartate-specific proteases (caspase)-1, Gasdermin D (GSDMD)) were systematically evaluated. LBP administration conferred robust cardioprotection in I/R rats, as evidenced by a significant reduction in myocardial infarct size, improved preservation of myocardial fiber architecture, and attenuated leakage of cardiac injury biomarkers (lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB)). Mirroring these in vivo findings, LBP pretreatment effectively shielded H9c2 cardiomyocytes from H/R insult, markedly enhancing cell viability while curtailing reactive oxygen species (ROS) accumulation and apoptotic activation. A pivotal finding was the pronounced downregulation of Nrf2 in the H/R group, a deficit that was conclusively reversed by LBP treatment. To decisively establish a causal role for Nrf2, we employed a loss-of-function approach; Nrf2 inhibition completely abrogated the protective benefits of LBP, culminating in exacerbated tissue damage, a surge in ROS, and the upregulation of key pyroptosis effectors (NLRP3, ASC, caspase-1, GSDMD). Conversely, a complementary gain-of-function experiment demonstrated that Nrf2 overexpression alone was sufficient to mimic LBP’s effects, significantly blunting H/R-induced ROS production and pyroptosis. LBP alleviates MIRI by inhibiting pyroptosis through activating the Nrf2/NLRP3 axis, thus representing a promising therapeutic candidate for ischemic heart disease with the potential to improve patient outcomes. Full article

Background: Syndesmotic screw breakage is a well-recognized mechanical complication following ankle fracture fixation. Although several studies have investigated patient-related and technical factors associated with screw breakage, the temporal pattern of screw failure and implant survival remains less clearly defined. Therefore, this study aimed to evaluate one-year syndesmotic screw survival using time-to-event analysis and to identify factors associated with screw breakage. Materials and Methods: A total of 132 patients with unstable AO-Weber 44-B/C ankle fractures treated with syndesmotic screw fixation were retrospectively analyzed. Patients were followed for a minimum of 12 months or until screw breakage occurred. Screw survival was evaluated using Kaplan–Meier analysis and Cox proportional hazards regression was performed to identify factors associated with screw breakage. Demographic variables, fracture type, and screw-related parameters were analyzed. Receiver operating characteristic (ROC) analysis was used to assess the discriminative ability of age. Results: Screw breakage occurred in 31 patients (23.5%) during follow-up. Kaplan–Meier analysis demonstrated significantly lower screw survival in Weber C fractures compared with Weber B fractures (log-rank p < 0.001). Cox regression analysis identified younger age (HR: 0.965, 95% CI: 0.937–0.993, p = 0.016) and Weber C fracture type (HR: 1.811, 95% CI: 1.260–2.602, p = 0.001) as independent predictors of screw breakage. ROC analysis showed that age had moderate discriminative ability (AUC: 0.719, 95% CI: 0.612–0.816), with a cut-off value of 35.5 years. Conclusions: Younger age and Weber C fracture type are associated with an increased risk of syndesmotic screw breakage and Weber C fractures also demonstrating reduced screw survival. These findings may assist in patient counseling; however, the clinical implications of screw breakage remain uncertain. Full article

Radiotherapy (RT) is one of the main treatments for breast cancer, but response varies between patients. Tumour hypoxia and intrinsic radiosensitivity are major determinants of response to RT. Using TCGA-BRCA, a 563-gene hypoxia meta-signature was built by combining curated hypoxia gene sets from MSigDB with published hypoxia metagenes (Buffa, Winter, Elvidge, Fardin, and related sets). After Cox screening and penalised regression, a simple three-gene hypoxia score (CP, GPC3, STC1) was derived. In parallel, based on DSB-repair factors highlighted by Mladenov et al. as key regulators of intrinsic radiosensitivity, a four-gene radiosensitivity (RS) signature (ATR, RPA2, BLM, MRE11A) was trained using only RT-treated patients. In TCGA, both signatures were prognostic and showed significant interaction with RT status in Cox models. The hypoxia score was strongly associated with worse outcomes in RT-untreated patients, but this effect was much weaker in RT-treated patients (Hypoxia × RT HR = 0.009, p = 0.044). The RS score showed a similarly strong interaction with RT (RS × RT HR = 0.011, p = 0.003). When we combined both signatures into one interaction model, it gave the best performance (C-index = 0.785), and both interaction terms stayed independently significant. The hypoxia score was then validated externally in METABRIC (N = 1979; 1143 events), where it remained associated with overall survival, although more weakly than in TCGA (HR = 1.34, 95% CI: 1.10–1.63; p = 0.0042). Overall, these results suggest that hypoxia and DSB-repair capacity capture two complementary sides of radiosensitivity and RT-modified survival patterns, and they support further prospective testing and validation in independent datasets with strong RT annotation. Full article

Background/Objectives: Cirrhosis is associated with increased mortality. In this study, we aimed to investigate the prognostic relevance of 6-min-walk-distance- and cardiopulmonary exercise testing (CPET)-derived peak oxygen uptake (VO₂) as estimates of exercise capacity in outpatients with cirrhosis. Methods: Patients underwent a comprehensive clinical characterization including cardiopulmonary exercise testing, six-minute-walking-test-derived distance, and echocardiography. We stratified the cohort using established prognostic thresholds for the six-minute-walking-test-derived distance (440 m) and peak VO₂ (65% predicted) and Child–Pugh class (A vs. B/C). Competing risk analyses were performed using cumulative incidence functions and subdistribution hazard models to assess the impact of baseline variables on mortality, accounting for liver transplantation (LT) as a competing event and for age and sex. The prognostic value of exercise performance was analyzed first, followed by the stepwise inclusion of additional variables; multicollinearity precluded a full multivariable model. Results: We enrolled 197 patients in Child–Pugh Class A, B, and C (N = 92, N = 80, N = 25 patients; male N = 146, age: 56 ± 9 years). During the observation time of 85 (25–105) months, 48 patients underwent a liver transplant, and 88 died. Both the six-minute-walking-test-derived distance ≤ 440 m (p = 0.002, sHR: 0.996 95% CI: 0.993–0.998) and peak VO₂ ≤ 65% predicted (p = 0.023, sHR: 0.987 95% CI: 0.976–0.998) were strong independent predictors of mortality. While the six-minute-walking-test-derived distance consistently remained significant across most models, the peak VO₂ retained significance only when adjusted for creatinine. Combining exercise capacity and the Child–Pugh classification identified patients at a particularly high mortality risk. Conclusions: In patients with liver cirrhosis outside the liver transplant setting, the impaired six-minute-walking-test-derived distance and peak VO₂ serve as predictors of mortality and may help to identify patients at a particularly high mortality risk. These results suggest that functional capacity provides complementary information to established liver disease severity scores and could be considered in a multidimensional risk assessment approach in patients with liver cirrhosis. Full article

Toxoplasma gondii establishes latent infection in a substantial proportion of the global population, yet the long-term health consequences of this infection remain incompletely characterized. We conducted a retrospective observational matched cohort study using longitudinal electronic health record data from a nationwide integrated healthcare provider, including members aged 18–45 years who underwent routine Toxoplasma serologic screening, typically performed in obstetric evaluation, excluding patients with clinical toxoplasmosis, immunosuppression, or HIV. Seropositive individuals were matched 1:1 without replacement to seronegative controls to align demographic, temporal, and socioeconomic variables. Time-to-event associations with predefined medical conditions were evaluated using Cox proportional hazards models with false discovery rate correction. The final cohort included 19,443 seropositive individuals and 19,443 matched controls (96.7% female), with a tight baseline balance of demographic and temporal characteristics. During follow-up, seropositivity was associated with increased risks of tobacco dependence (aHR 1.65), alcohol dependence (2.32), suicide attempt (1.82), motor vehicle accidents (1.22), and work accidents (1.27), as well as multiple infectious conditions, including hepatitis B (1.55), hepatitis C (2.15), and syphilis (2.43), with an overall trend toward increased all-cause mortality (1.32, 95% CI [1.00–1.74]). These findings suggest that asymptomatic Toxoplasma infection in young adults is associated with increased long-term behavioral and medical comorbidity. Full article

Background: Adrenocortical carcinoma (ACC) is a rare and aggressive endocrine malignancy, for which population-level evidence regarding prognostic factors and survival conditions is limited. The available data mostly represent single-institution series, limiting their applicability. This study, therefore, assesses clinicopathological features and determines independent predictive variables of overall survival (OS) in patients with ACC using a population-based cohort. Methods: This retrospective observational cohort study used data from the Surveillance, Epidemiology, and End Results (SEER) Program between 2000 and 2022, initially identifying 1176 patients with ACC. Adult patients (≥18 years) with histologically confirmed ACC were identified using ICD-O-3 histology code 8370/3 and primary site code C74.0. Cases with zero-month survival, missing survival data, or identified only through autopsy or death certificate were excluded. To ensure dataset harmonization, patients with missing or indeterminate tumor grade and unknown stage were also excluded. After applying these inclusion and exclusion criteria, the final analytic cohort consisted of 267 patients. Data on demographic factors, stage of the disease, and treatment (surgery, chemotherapy, radiotherapy) were extracted. OS was evaluated using the Kaplan–Meier method, and independent prognostic factors were identified using Cox proportional hazards regression analysis. Results: The median OS was 54 months [95% confidence intervals (CI): 36–85]. The estimated 1-, 3-, and 5-year OS rates were 77%, 57%, and 48%, respectively. Survival differed significantly according to tumor grade, stage, and surgical treatment. In multivariable Cox regression analysis, increasing age [Hazard ratio (HR): 1.03, 95% CI: 1.02–1.04; p < 0.001], high tumor grade (HR: 2.21, 95% CI: 1.43–3.41; p < 0.001), and distant-stage disease (HR: 3.24, 95% CI: 1.95–5.38; p < 0.001) were independently associated with an increased risk of mortality, whereas surgical treatment was associated with improved survival (HR 0.53, 95% CI 0.30–0.93; p = 0.028). Chemotherapy and radiotherapy were not significantly associated with mortality. Conclusions: In this SEER-based cohort of patients with adrenocortical carcinoma, older age, high tumor grade, and distant-stage disease were independently associated with worse OS, whereas documented receipt of surgery was associated with longer OS. Treatment-related associations should be interpreted cautiously in view of the inherent limitations of registry-based data. Further prospective multicenter studies are needed to confirm these findings. Full article

Growth differentiation factor 15 (GDF15) is a stress-responsive cytokine strongly associated with aging, multimorbidity, and cardiovascular disease. Although prior studies have established its prognostic value in high-risk populations, its role in the general population remains less defined. The aim of this study was to determine if there is an association between plasma GDF15 levels, heart disease and mortality in a representative population-based cohort. We analyzed 1532 participants (mean age 55 years; 54.6% women) with available baseline plasma GDF15 concentrations. Participants were stratified according to an optimal cutoff of 1081 pg/mL, derived from ROC curve analysis for mortality. Associations with prevalent heart disease were assessed using multivariable logistic regression models adjusted for cardiovascular risk factors and NT-proBNP. Mortality was analyzed using Cox proportional hazards models, with model performance evaluated by C-index and time-dependent ROC curves. Individuals with GDF15 > 1081 pg/mL were older and exhibited a more adverse cardiometabolic profile with higher prevalence of comorbidities. Elevated GDF15 was independently associated with ischemic cardiomyopathy (OR 3.34, 95% CI: 1.38–8.11), particularly in men (OR 4.26, 95% CI: 1.40–12.96), but not in women. No independent associations were observed with arrhythmias, valvulopathy, or heart failure after adjustment for NT-proBNP. During a median follow-up of 6.2 years, 51 deaths occurred. Elevated GDF15 independently predicted all-cause mortality (HR 2.47, 95% CI: 1.19–5.13), though the effect was attenuated after adjustment for NT-proBNP. GDF15 improved model discrimination (ΔC-index = +0.01; LRT p = 0.011) and showed robust time-dependent predictive ability, with AUCs of 0.76, 0.82, and 0.85 at 2, 4, and 6 years, respectively. In this population-based cohort, elevated GDF15 identified individuals with an adverse health profile, was independently associated with ischemic cardiomyopathy in men, and predicted mortality. Although its incremental predictive value over NT-proBNP was modest, GDF15 could provide complementary biological information and may enhance multimarker strategies for cardiovascular risk stratification in the general population. Full article

The growing need for remote patient monitoring, accelerated by the global pandemic and an aging population, necessitates the development of advanced non-contact technologies for measuring vital signs. In this study, an integrated, non-contact system for accurately measuring heart rate (HR) and body temperature (BT) is developed and validated. The proposed system combines a 60 GHz radar sensor and infrared (IR) sensor for HR and BT measurements, respectively, enhanced with advanced signal processing and an AI-based computer vision algorithm. A Window Filter and a Peak Uniformity algorithm were applied to the raw radar signal to mitigate noise and motion artifacts. For Temp measurement, an IR sensor with a narrow five-degree field of view (FOV) was integrated with a YOLO Pose-based tracking system using a camera and servo motors to automatically orient the sensor towards the user’s face. The system was validated with 30 healthy adult participants, benchmarked against a MAX30102 PPG sensor and Braun ThermoScan 7 for BT and BT measurements, respectively. The advanced signal processing reduced the HR Mean Absolute Error from 13.73 BPM to 5.28 BPM (p = 0.002), while the AI-guided IR sensor reduced the BT MAE from 4.10 °C to 1.64 °C (p < 0.001). These findings demonstrate that integrating 60 GHz radar with AI-driven tracking provides a promising approach for home-based trend monitoring. Full article

The rising demand for sustainable and circular approaches in the agro-industrial sector has generated interest in repurposing herbal tea residues as sources of high-value bioactive compounds. This work focusses on recovering phytochemicals from Rosa canina L. peel and seed dust (by-products of processing of herbal tea in filter tea bags) using green extraction techniques. Two environmentally friendly technologies were used: ultrasound-assisted extraction (UAE) with a sonotrode and subcritical fluid extraction (SBFE). The extracts were qualitatively profiled using (HR) LC-ESI-QToF-MS/MS and quantified using HPLC-PDA. Both by-products contained phenolic substances, including gallic acid derivatives, ellagic acid, and flavonoids such as quercetin and quercetin-3-O-glucoside (only in the peel). Additionally, Folin–Ciocalteu’s assay was used to determine Total Phenolic content (TP). The extraction efficiency was considered in terms of phenolic compound recovery and total phenolic content obtained under the respective experimental conditions. The maximum TP for SBFE was reported in samples extracted with ethanol–water (48:52) at 180 °C, producing 3876.67 GAE mg/L for peel and 1648.57 GAE mg/L for seeds. In the UAE, extraction with ethanol–water (48:52) for 10 min yielded the maximum TP of 2773.81 GAE mg/L for peel and 957.86 GAE mg/L for seeds. These findings highlight the potential of R. canina infusion by-products as long-term sources of bioactive compounds for use in nutraceutical, cosmetic, and pharmaceutical industries. Full article

RipP1 is a well-characterized avirulence effector that induces a hypersensitive response (HR) in three tobacco species. However, the molecular mechanisms by which host proteins recognize RipP1 to activate a defense response and modulate host–pathogen interactions remain largely unknown. In this study, we screened a Nicotiana benthamiana cDNA library via yeast two-hybrid assay and identified FRIGIDA-like protein 4a (FRL4a) as a host protein interacting with RipP1. Secondary structure analysis of FRL4a and construction of serial mutants revealed that the ClyA-like domain of FRL4a is the key region mediating its interaction with RipP1. Using virus-induced gene silencing (VIGS) and quantitative real-time PCR (qPCR) analysis, we found that the ability of RipP1 to induce HR was significantly attenuated in FRL4a-silenced plants, and RipP1 no longer suppressed the ethylene signaling pathway. Pathogenicity tests by inoculating R. solanacearum on N. benthamiana with different FRL4a expression levels showed enhanced bacterial wilt resistance in FRL4a-silenced plants but increased susceptibility in FRL4a-overexpressing plants. Collectively, these findings demonstrate that RipP1 suppresses the ethylene pathway through its interaction with FRL4a, and FRL4a acts as a negative regulator of tobacco resistance to bacterial wilt. Full article