Search Results (7,716)

Purpose: Castration-resistant prostate cancer (CRPC) responds poorly to conventional chemotherapy. We evaluated a cell-based enzyme–prodrug therapy using adipose-derived stem cells (ADSCs) engineered to express cytosine deaminase (CD) or carboxylesterase 2 (CE2), paired with their respective prodrugs 5-fluorocytosine (5-FC) or irinotecan (CPT-11), to compare their antitumor efficacy. Materials and Methods: Human telomerase reverse transcriptase (hTERT)-immortalized ADSCs were transduced with CD or CE2, and transgene expression and stem cell phenotype were confirmed. CD expression was verified at the transcript level and by functional 5-FC-to-5-fluorouracil (5-FU) conversion, whereas CE2 expression was verified by transcript analysis and immunoblotting. Tumor tropism toward PC3 prostate cancer cells was tested using migration assays and analysis of chemoattractant ligand/receptor expression. Prodrug-induced self-killing and bystander tumor cell killing were assessed through viability assays and co-culture with PC3 cells. For the CE2/CPT-11 system, SN-38 was not directly quantified; functional activity was inferred from prodrug-dependent cytotoxicity and in vivo efficacy. In vivo efficacy was evaluated in nude mice with PC3 tumors treated systemically with engineered ADSCs plus prodrug. Results: CD- and CE2-expressing ADSCs were successfully established and retained mesenchymal stem cell (MSC) characteristics. Both cell types exhibited significant migration toward PC3 cells. The CE2/CPT-11 system produced stronger prodrug-mediated cytotoxicity than CD/5-FC, with CE2-modified ADSCs showing higher sensitivity to CPT-11 and inducing greater apoptosis in co-cultured PC3 cells. In vivo, both treatments suppressed tumor growth, but CE2/CPT-11 achieved greater inhibition (tumor volume ~26% of control vs. ~32% for CD/5-FC at day 14). No overt clinical toxicity was observed based on body weight and daily clinical monitoring; however, hematology/serum chemistry were not assessed. Conclusions: Engineered ADSCs home to CRPC tumors and enable local prodrug activation, producing significant antitumor effects. Within the constraints of our in vitro assays and subcutaneous xenograft model, CE2/CPT-11 demonstrated stronger efficacy outcomes than CD/5-FC. Mechanistic attribution to intratumoral SN-38 exposure should be confirmed by direct metabolite measurements in future studies. Full article

Lung cancer progression is regulated by multiple factors, including ferroptosis and gut microbiota-mediated butyrate metabolism. This study investigates the anti-tumor effects of ginsenoside Rh4 on lung cancer cells via ferroptosis mechanisms in vitro and in vivo. In vitro, ginsenoside Rh4 inhibited the proliferation of Lewis lung carcinoma (LLC) and A549 cells and triggered ferroptosis, effects that were suppressed by the ferroptosis inhibitor Ferrostatin-1 (Fer-1). In vivo, tumor-bearing mouse models were established and treated with 100 mg/kg ginsenoside Rh4 for 21 days. Tumor growth, ferroptosis markers, gut microbiota, and butyrate were analyzed, with in vitro validation of butyrate’s pathway effects. Ginsenoside Rh4 induced ferroptosis in LLC cells both in vitro and in vivo, inhibiting tumor growth. It promoted ferroptosis by disrupting iron homeostasis through elevated Fe²⁺ and transferrin receptor (TFRC), and impaired antioxidant defense via depletion of glutathione (GSH) and reduction in ferritin heavy chain 1 (FTH1), solute carrier family 40 member 1 (SLC40A1), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4). Additionally, ginsenoside Rh4 enhanced lipid peroxidation, indicated by increased lipid peroxides (LPO) and malondialdehyde (MDA). In vivo, it suppressed the KEAP1/NRF2/HO-1 pathway, reducing antioxidant enzyme activity. Gut microbiota modulation and butyrate production further amplified ferroptosis by activating transcription factor 3 (ATF3)-mediated GPX4 suppression. Ginsenoside Rh4 induces ferroptosis by inhibiting the KEAP1/NRF2/HO-1 pathway and remodeling the gut microbiota to increase butyrate levels, which synergistically enhance tumor cell ferroptosis sensitivity through ATF3 activation and suppression of GPX4. Full article

Background: Myocardial infarction (MI) is the leading cause of morbidity and mortality globally. A major pathological progression of MI is the excess generation of reactive oxygen species (ROS), which results in oxidative stress and damage to the ischemic heart. Because damage to the myocardium is irreversible, the development of new therapeutic agents that can decrease the degree of ischemic damage following MI is crucial. The traditional Chinese medicine formulation, Qishen Yiqi dropping pills (QSYQ), has been clinically used in the treatment of various cardiovascular diseases; however, the precise mechanisms underlying its therapeutic effects remain unelucidated. Methods: In this study, we established murine models of MI via coronary artery ligation to investigate the protective effects and mechanisms of QSYQ following MI. Results: The administration of QSYQ significantly improved cardiac function, reduced infarct size, and attenuated ventricular remodeling in mice that underwent MI. Moreover, MI-induced oxidative stress and downregulated levels of antioxidant enzymes were prevented in mice administered QSYQ via upregulating the SIRT3/FOXO3a signaling pathway. Importantly, pretreatment with a selective SIRT3 inhibitor 3-TYP abolished the cardioprotective effects of QSYQ. Conclusions: Our findings elucidate the role and mechanism of QSYQ in protecting against oxidative damage and restoring redox homeostasis following myocardial infarction. This study provides support for the therapeutic potential of QSYQ in the clinical treatment of myocardial ischemic diseases. Full article

Cadmium (Cd) pollution has caused severe environmental hazards and human health risks. Phytoremediation, a green and sustainable approach, has emerged as a promising solution for Cd-contaminated soil remediation. Sedum plumbizincicola, a typical Cd hyperaccumulator, can efficiently uptake Cd from soil and translocate it to above-ground tissues, making it an ideal model for studying Cd tolerance mechanisms. Our preliminary studies demonstrated that the Rubber elongation factor (SpREFl) enhances Cd tolerance in S. plumbizincicola, and yeast one-hybrid screening identified SpNAC089 (NCBI accession number: PV553670.1) as a potential upstream transcription factor of SpREFl. In this study, we systematically investigated the regulatory mechanism of the SpNAC089 transcription factor on SpREFl. Subcellular localization assays showed that SpNAC089 is exclusively localized in the cell nucleus, and yeast transcriptional activation experiments confirmed its intrinsic transcriptional autoactivation activity. Transgenic S. alfredii overexpressing SpNAC089 exhibited significantly enhanced cadmium tolerance—with milder leaf yellowing and growth inhibition under Cd stress—and reduced Cd accumulation in roots, stems, and leaves compared to wild-type (WT) plants. Further mechanistic analyses revealed that SpNAC089 directly binds to the 1901–1950 bp region of the SpREFl promoter, which contains cis-acting elements (MBS and TCA motifs). This binding activates SpREFl transcription, thereby upregulating the activities of antioxidant enzymes (superoxide dismutase, SOD; peroxidase, POD) and reducing malondialdehyde (MDA) content under Cd stress, ultimately mitigating oxidative damage. These findings uncover a novel transcriptional regulatory pathway (SpNAC089-SpREFl) underlying Cd tolerance in S. plumbizincicola and highlight SpNAC089 as a candidate gene for optimizing phytoremediation strategies of Cd-polluted soils. Full article

Background/Objectives: Lafora disease is a fatal and progressive neurodegenerative disorder characterized by the accumulation of insoluble polyglucosan inclusions, known as Lafora bodies, due to impaired glycogen metabolism. Therapeutic strategies aimed at reducing intracellular glycogen accumulation represent a promising approach to mitigating disease progression. This study aimed to evaluate the feasibility of promoting Lafora body degradation by increasing the protein levels of human pancreatic amylase, a glycogen-degrading enzyme, through the SINEUP approach. Methods: Two SINEUP constructs specifically targeting human pancreatic amylase were designed and tested in continuous tumor-derived cell lines of central nervous system origin, as well as in primary fibroblasts obtained from a patient with Lafora disease. Human pancreatic amylase protein and mRNA levels were assessed to determine the specificity of SINEUP-mediated regulation. Enzymatic activity assays were performed to evaluate functional protein upregulation, and intracellular glycogen content was measured in patient-derived fibroblasts. Results: Both SINEUP constructs significantly increased human pancreatic amylase protein expression without affecting mRNA levels, confirming a post-transcriptional mechanism of action. The elevated protein levels were associated with a significant increase in enzymatic activity. In primary fibroblasts derived from a Lafora disease patient, enhanced amylase expression correlated with a marked reduction in intracellular glycogen content. Conclusions: These findings provide proof of concept that SINEUP-mediated upregulation of glycogen-degrading enzymes may represent a viable therapeutic strategy to counteract Lafora body accumulation. Further studies are warranted to assess the efficacy, safety, and translational potential of this approach, particularly in relevant animal models of Lafora disease. Full article

Fructokinase (FRK) initiates fructose phosphorylation, channeling carbon into central metabolic pathways, yet its functional diversity and regulatory networks in C₄ cereals remain poorly understood. Here, we performed a comprehensive pan-genome analysis of the FRK gene family in foxtail millet (Setaria italica), identifying 697 SiFRKs across 110 accessions and revealing extensive presence–absence variation shaped by evolution and domestication. Among nine characterized members in the reference genome, SiFRK4 exhibited broad and high expression, a diurnal rhythm, and substantial natural variation. Biochemical assays confirmed its fructokinase activity in vitro. We discovered a novel physical interaction between SiFRK4 and the key photoreceptor Phytochrome C (SiPhyC), which co-localized in the cytoplasm. Functional analysis of SiPhyC mutants demonstrated that loss of SiPhyC disrupted carbohydrate homeostasis, elevating fructose while depleting sucrose and starch. Our findings reveal a physical and genetic link between the light-signaling component SiPhyC and the metabolic enzyme SiFRK4, suggesting their interaction influences carbon partitioning. This study provides foundational insights into the sugar metabolism network of a resilient C₄ model crop and identifies potential targets for metabolic engineering and breeding. Full article

The proliferation of generative artificial intelligence (AI) as an autonomous recommendation agent fundamentally challenges traditional paradigms of marketing communication. As AI systems increasingly mediate consumer–brand relationships, understanding how artificial agents construct persuasive discourse—distinct from human communicators—becomes critical for developing effective dual-channel marketing strategies. Grounded in Source Credibility Theory and the Computers Are Social Actors (CASA) paradigm, this study investigates the semantic and structural divergence between AI-generated product recommendations and human influencer marketing messages in social commerce contexts. Employing a mixed-methods computational approach integrating term frequency analysis, TF-IDF weighting, Latent Dirichlet Allocation (LDA) topic modeling, and BERT-based contextualized semantic embedding analysis (KR-SBERT), we examined 330 Instagram influencer posts and 541 AI-generated responses concerning inner beauty enzyme products—a hybrid category combining functional health claims with hedonic beauty appeals—in the Korean social commerce market. AI-generated responses were collected through a systematically designed query protocol with empirically grounded prompts derived from actual consumer search behaviors, and analytical robustness was verified through sensitivity analyses across multiple parameter thresholds. Our findings reveal a fundamental divergence in persuasive architecture: human influencers construct experiential narratives exhibiting message characteristics typically associated with peripheral-route cues (sensory descriptions, emotional testimonials, social context), while AI recommendations employ systematic, evidence-based discourse exhibiting message characteristics typically associated with central-route argumentation (functional mechanisms, ingredient specifications, objective criteria). Topic modeling identified four distinct thematic clusters for each source type: human discourse centers on embodied experience and relational consumption, whereas AI discourse organizes around informational utility and rational decision support. Jensen–Shannon Divergence analysis (JSD = 0.213 bits) confirmed moderate distributional divergence, while chi-square testing (χ² = 847.23, p < 0.001) and Cramér’s V (0.312, indicating a medium-to-large effect) demonstrated statistically significant and substantively meaningful differences. These findings extend CASA theory by demonstrating that AI recommendation agents develop a characteristic “AI communication signature” distinguishable from human persuasion patterns. We propose an integrated Dual-Agent Persuasion Proposition—synthesizing CASA, ELM, and Source Credibility perspectives—suggesting that AI and human recommenders serve complementary functions across different stages of the consumer decision journey—a proposition whose predictions regarding sequential persuasive effectiveness and consumer processing routes await experimental validation. These findings carry implications for AI content strategy optimization, platform design, and emerging regulatory frameworks for AI-generated content labeling. Full article

Plants have a remarkable ability to regenerate tissues and organs from single cells, a property that underpins in vitro protoplast regeneration. Efficient protoplast-to-plant regeneration remains a major bottleneck for genome engineering in many crop species, including broccoli (Brassica oleracea var. italica). In this study, we established and optimized a regeneration system for broccoli cv. Claremont by evaluating enzyme composition, light quality, and culture media at successive stages of development. Among the tested enzyme mixtures, 1.5% Cellulase R-10 combined with 0.4% Macerozyme R-10 yielded the highest protoplast viability and recovery. Alginate-embedded protoplasts were cultured under control (dark), blue, and red + far-red LED illumination. Red + far-red treatment significantly enhanced microcolony formation, plating efficiency, and shoot regeneration compared with blue light, whereas blue illumination consistently reduced regenerative performance. The inclusion of activated charcoal in the regeneration medium further increased shoot production. The generalized linear model analyses identified light quality as a significant predictor of both shoot number and regeneration. To our knowledge, this study provides one of the first demonstrations of LED-assisted enhancement of protoplast regeneration in broccoli. The optimized protocol enables whole-plant recovery within approximately 5 months and offers a practical platform for CRISPR-based genome editing and advanced breeding applications in B. oleracea. Full article

Accurate and physiologically relevant in vitro models are needed to predict nutrient digestibility and hindgut fermentation in pigs, as conventional in vivo trials are resource-intensive and raise animal welfare concerns. This study evaluated and compared the predictive performance of three in vitro digestion approaches—shaking (S), dialysis (D), and a combined shaking plus dialysis (SD) method—for estimating in vivo apparent total tract digestibility (ATTD) and fermentation characteristics across weaning, growing, and finishing pigs. Commercial diets were subjected to simulated gastric and small-intestinal digestion using S, D, or SD, followed by fecal inoculation to model hindgut fermentation for 12 and 48 h. During the gastrointestinal phase, crude protein digestibility was highest with D (>75%), intermediate with SD, and lowest with S (50–60%), indicating that product removal by dialysis mitigated enzyme inhibition from metabolite accumulation. After 48 h of fermentation, all methods showed strong linear correlations with in vivo ATTD (r > 0.93), but only D achieved high absolute agreement (Lin’s CCC > 0.95 for dry matter and crude protein). Moreover, D and SD at 48 h closely reflected in vivo fecal profiles of skatole, indole, and microbial enzyme activities, with D at 12 h showing an especially strong correlation for protease (r = 0.98). While D provided the most precise predictions of absolute values, the SD method offered an optimal balance between physiological relevance and operational efficiency, supporting its use as a robust, high-throughput platform for porcine feed evaluation and fecal nitrogenous odorant prediction. Full article

Short-chain acyl-CoA dehydrogenase (SCAD) is a critical enzyme in mitochondrial fatty acid β-oxidation, catalyzing the initial dehydrogenation of short-chain acyl-CoAs. Mutations in the ACADS gene cause SCAD deficiency (SCADD), a disorder with remarkably heterogeneous clinical presentation. However, the molecular mechanisms underlying substrate specificity and the pathogenicity of most ACADS variants remain poorly understood. Here, we present high-resolution cryo-EM structures of human SCAD in complex with its physiological substrate butyryl-CoA (C₄) and the longer substrate hexanoyl-CoA (C₆). The butyryl-CoA-bound structure at 2.1 Å resolution details a pre-catalytic geometry ideal for hydride transfer, with Glu392 positioned as the catalytic base. We systematically characterized nineteen disease-associated mutations, which we classify into three functional categories: those disrupting FAD binding, those impairing substrate binding, and those compromising protein folding and stability. In addition, using the W177R mutant as a representative model, we demonstrate that folding-defective mutations provoke protein aggregation, leading to proteotoxicity, oxidative stress, and apoptosis, revealing a pathogenic mechanism beyond mere catalytic loss. In brief, our integrated findings elucidate the structural determinants of substrate specificity and catalytic mechanism in SCAD, and provide mechanistic insights into the functional impairments caused by mutations linked to SCADD. Full article

Oxidative stress in intestinal epithelial cells has been increasingly recognized as a key factor in various intestinal disorders. Gastrodia elata polysaccharide-2 (GEP-2), a water-soluble polysaccharide known for its antioxidant properties, has shown potential against intestinal injury. However, its effects on intestinal epithelial cells and the molecular mechanisms involved are not yet fully understood. In this study, we established a hydrogen peroxide (H₂O₂)-induced oxidative stress model using human colonic epithelial cells (NCM460) to evaluate the protective effects of GEP-2. We assessed cell viability, antioxidant enzyme activities, reactive oxygen species (ROS) levels, and mitochondrial membrane potential (MMP). The results demonstrated that GEP-2 pretreatment significantly improved the viability of NCM460 cells subjected to H₂O₂ damage. Additionally, it could enhance the antioxidant defense, reduce the levels of ROS, malondialdehyde (MDA), and maintain the MMP. Transcriptomic analysis identified 169 differentially expressed genes upregulated in the glutathione metabolism. JAK-STAT pathway and downregulated in inflammation. Furthermore, it was shown that GEP-2 treatment activated the Nuclear factor erythroid 2-related factor 2 (Nrf2)/quinone oxidoreductase 1 (NQO1)-mediated antioxidant response and promoted the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway. Therefore, GEP-2 exerts multi-targeted cell protection by coordinating the Nrf2/NQO1 antioxidant axis and the JAK/STAT survival signaling pathway, providing a theoretical basis for the development of novel antioxidants. Full article

In desert regions, frequent aeolian dust events lead to rapid dust accumulation on greenhouse films, critically compromising light transmittance and inhibiting crop growth. To address this challenge, this study integrated Computational Fluid Dynamics–Discrete Phase Model (CFD-DPM) simulations with field experiments to conduct a comprehensive investigation spanning from microscopic deposition mechanisms to macroscopic physiological responses. Particle characterization revealed a distinct aerodynamic sorting effect, wherein fine particles (<65 μm) preferentially adhered to film surfaces driven by airflow, contrasting sharply with the gravitational settling of coarse ground particles. Numerical simulations further confirmed that as wind speeds increased from 2 to 7 m/s, dust deposition rates exhibited a significant exponential reduction, with accumulation predominantly concentrated in the windward and wake zones. The dust layer covering the film induced a substantial reduction in the indoor daily light integral (DLI), which leads to influence tomato growth that stunted plant height and suppressed the net photosynthetic rate. Physiologically, antioxidant enzyme activities exhibited an initial surge followed by a decline, reflecting photosynthetic constraints and oxidative stress. Consequently, a high-frequency cleaning interval of 7–14 days is recommended to significantly enhance photosynthetic capacity and stress resilience. Full article

Leucine-rich repeat-containing 8A (LRRC8A; also known as SWELL1), the essential subunit of volume-regulated anion channels (VRACs), is amplified in multiple malignancies and has been implicated in tumor progression and therapeutic resistance. Three-dimensional (3D) cancer spheroids have been well-established as in vitro models that recapitulate characteristics of tumor stemness and intrinsic drug resistance. In the present study, spheroid formation in human breast cancer cell lines, YMB-1 and MDA-MB-468, conferred resistance to multiple anticancer drugs, including doxorubicin (DOX), gemcitabine (GEM), and 5-fluorouracil (5-FU), thereby mimicking the characteristic properties of breast cancer stem-like cells. LRRC8A expression was upregulated in 3D spheroids compared with adherent 2D monolayers, and its pharmacological inhibition induced membrane hyperpolarization accompanied by intracellular Cl⁻ accumulation. Inhibition of LRRC8A significantly sensitized spheroids to DOX, GEM, and 5-FU. Spheroid formation increased the expression of multidrug resistance-related protein 3 (MRP3) and the drug-metabolizing enzyme cytochrome P450 3A4 (CYP3A4), whereas LRRC8A inhibition suppressed their expression. The transcriptional upregulation of MRP3 and CYP3A4 was mediated through the NRF2–CEBPB/D transcriptional axis. Collectively, these findings suggest that LRRC8A inhibition may represent a therapeutic strategy to overcome chemoresistance by repressing MRP3 and/or CYP3A4 expression in breast cancer stem cells. Full article

Sepsis-induced liver injury (SILI) stands as an independent prognostic factor for mortality among patients diagnosed with sepsis. Maresin-1 (MaR1) is a proresolving lipid mediator. However, its significance in SILI is uncertain. The current study sought to investigate MaR1’s effectiveness in treating SILI, as well as its molecular mechanism. In male C57BL/6J mice, we generated a SILI model by using cecal ligation and puncture (CLP). We further investigated how MaR1 influences inflammation, hepatic autophagy and apoptosis. We showed that treatment with MaR1 ameliorates SILI-induced hepatic injury, as reflected in decreased blood level of the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzymes, as well as better appearance of liver tissues. Furthermore, this medication markedly reduced the expression of inflammatory mediators. Importantly, MaR1 inhibited hepatocyte apoptosis by regulating the Bax/Bcl-2 ratio, decreasing cleaved caspase-3 expression, lowering apoptotic cell count, and increasing autophagy. The findings demonstrated that MaR1 treatment reduced p62 protein expression while raising Beclin1 levels and the LC3-II/LC3-I ratio, and facilitated autophagosome formation. The observed effects were most likely due to the stimulation of PI3K/Akt signaling, which was completely prevented by LY294002 (LY), a specific PI3K inhibitor. MaR1’s protective effect in SILI may be mediated via stimulation of the PI3K/Akt pathway, which reduces inflammation and regulates apoptosis and autophagy. Our results give additional experimental evidence of the potential therapeutic uses of MaR1 in the treatment of SILI. Full article

Background: Fibromyalgia (FM) and eating disorders (ED) represent distinct clinical entities traditionally managed within separate medical specialties, yet emerging evidence suggests significant comorbidity and potential shared pathophysiological mechanisms. Both conditions disproportionately affect women, involve complex multifactorial etiologies and substantially impair quality of life. Despite documented clinical overlaps, the mechanistic connections linking these conditions remain poorly characterized, and integrated treatment approaches are lacking. Objective: This narrative review examines the role of oxidative stress and nuclear factor erythroid 2-related factor 2 (Nrf2) pathway dysfunction as a unifying molecular mechanism connecting fibromyalgia and eating disorders, with emphasis on implications for integrated rehabilitation strategies. Methods: We synthesized current evidence on oxidative stress pathophysiology in fibromyalgia and eating disorders, focusing on Nrf2-Keap1 pathway function, clinical comorbidity patterns and rehabilitation interventions targeting antioxidant defense mechanisms. In PubMed, representative search strings included “(fibromyalgia [MeSH] OR fibromyalgia [Title/Abstract]) AND (“eating disorders” [MeSH] OR “anorexia nervosa” [MeSH] OR “bulimia nervosa” [MeSH])” and “fibromyalgia AND (“oxidative stress” OR Nrf2 OR “redox”)”. Articles in English published through December 2025 were considered, with additional records identified by manually screening reference lists. Results: Fibromyalgia patients exhibit elevated oxidative stress markers, impaired antioxidant enzyme function and compromised Nrf2 activity correlating with disease severity, with studies reporting approximately 30–50% reductions in coenzyme Q10 levels compared with healthy controls. Similarly, eating disorders demonstrate mitochondrial dysfunction and oxidative stress dysregulation, though patterns differ across eating disorder phenotypes. Nrf2 serves as the master regulator of cellular antioxidant defense, coordinating expression of over 500 genes involved in detoxification, cytoprotection, inflammation modulation and metabolic regulation. Evidence suggests Nrf2 activity is regulated by energy balance, potentially linking nutritional status with cellular stress responses. Rehabilitation interventions, including graduated exercise and nutritional optimization with Nrf2-activating foods (cruciferous vegetables, polyphenols, omega-3 fatty acids), offer mechanism-based therapeutic approaches through hormetic Nrf2 activation and direct Keap1 modification. Conclusions: Multidisciplinary rehabilitation programs integrating physical therapy, exercise prescription and nutritional strategies targeting Nrf2 activation offer evidence-based, mechanism-driven approaches to address shared oxidative stress pathophysiology. Nrf2 pathway dysfunction represents a promising and biologically plausible molecular target that may help to unify our understanding of fibromyalgia and eating disorders pending confirmation from prospective clinical studies in comorbid populations. Future research should prioritize prospective clinical trials testing Nrf2-targeted interventions in comorbid populations and collaborative patient-centered care models. Full article