ARF GTPase protein 1 (GIT1) is a scaffold protein that is overexpressed in hepatocellular carcinoma (HCC) and colorectal cancer (CRC). GIT1 forms a complex with methionine adenosyltransferase 2B (MAT2B) that activates RAS-RAF-MEK-ERK signaling in HCC and CRC to enhance tumorigenicity. Here, we investigated in a proof-of-concept study whether a small molecule that disrupts GIT1-MAT2B interaction can be effective in HCC and CRC treatment. Since the GIT1 crystal structure is unavailable, we developed a molecular model and used computer-based drug discovery approach to screen for small molecules targeting the GIT1 ankyrin repeat domain, the region closest to where MAT2B interacts that is accessible. Of nine compounds tested, compound 3 (C3) selectively interacts with GIT1 and shows an anti-cancer effect in a GIT1-dependent manner. C3 is antiproliferative, induced apoptosis and G2/M cell cycle arrest while inhibiting colony formation and migration in liver and colon cancer cells. C3 lowered interaction between GIT1 and MAT2B, and with downstream effectors cRAF, MEK and ERK, lowering MEK activity and cyclin D1 expression. Unexpectedly, C3 stabilized GIT1 interaction with cyclin B1 while weakening cyclin B1’s interaction with components of the anaphase promoting complex, concomitant with sustained cyclin B1 expression and mitosis arrest. In mice, C3 administration was well tolerated and inhibited murine CRC growth and liver metastasis in immune competent mice and human CRC growth in the livers of nude mice. In conclusion, a small molecule inhibitor that disrupts GIT1’s normal interactome is a promising new approach to treating liver and colon cancers.

We previously reported that the heptamethine cyanine dye-conjugated artesunate (DZ-1-ART) induces apoptosis in monolayer (2D) cell culture models. However, in 2D cultures, cells grow on flat, rigid plastic surfaces that fail to recapitulate the three-dimensional architecture of tumor tissues. This artificial environment alters cell polarity, morphology, and mechanical signaling, leading to non-physiological behavior. To overcome these limitations, we developed patient-derived tumoroids to assess the tumoricidal efficacy of the DZ-1-ART conjugate. In this study, tumoroids were established from fresh tissue samples of a malignant neoplasm of the sigmoid colon. The anticancer activity of DZ-1-ART was evaluated in these tumoroids. Propidium iodide staining confirmed DZ-1-ART-induced cytotoxicity, while TUNEL and immunoblotting assays demonstrated that this cytotoxicity was mediated by apoptosis. Furthermore, MitoTracker staining and near-infrared fluorescence indicated mitochondrial localization of DZ-1-ART. The JC-1 assay showed disruption of mitochondrial membrane potential following DZ-1-ART treatment. Additionally, deferoxamine and MitoTEMPO pretreatment revealed that DZ-1-ART induced mitochondria-mediated reactive oxygen species (ROS) generation in tumoroids. Collectively, these findings suggest that DZ-1-ART acts as a potent mitochondria-targeting anticancer agent with potential for precision therapy.

Microplastics (MPs) are environmental contaminants with sizes of the order of less than 5 mm that can enter the human body through inhalation and ingestion. Studies have shown that MPs can pose a threat to human health, and thus evaluation of the presence and potential adverse effects of MPs in tissues is critical. Typical MP studies include enzymatic or chemical tissue digestion that can lead to the loss of some MPs. Moreover, digestion does not allow mapping the location of the contaminant within the tissue architecture. This study aimed to develop a method to evaluate the presence of MPs in histological (thin) sections of tissues without digestion using optical photothermal infrared (O-PTIR) microspectroscopy at sub-micron (500 nm) spatial resolution. Tissue phantoms containing specific amounts and types of MPs and biological tissues were evaluated using polarized light microscopy (PLM) and O-PTIR, and several data analysis approaches were employed to detect MPs in non-digested samples. MPs of sizes from 3 to 85 µm were detected and characterized in tissue phantoms. Furthermore, we detected MPs related to the breakdown products of nylon and cellulose particles in thin sections of biological tissues and discussed obstacles related to the use of database spectra for comparison with O-PTIR spectra, demonstrating the potential of this novel approach and the associated challenges.

Abstract Background: Resistance to androgen-deprivation therapy leads to metastatic castration-resistant prostate cancer (mCRPC), a lethal stage with limited treatment options. Artemisinin (ART) derivatives exert anti-tumor activity through reactive oxygen species (ROS) generation and mitochondrial disruption but lack tumor selectivity. Our laboratory identified a near-infrared (NIR) heptamethine carbocyanine dye (HMCD) that preferentially accumulates in tumors via organic anion-transporting polypeptides (OATPs) upregulated under hypoxia, enabling tumor-specific delivery. Methods: To achieve subcellularly targeted chemotherapy, four HMCD-ART conjugates with distinct linkers were synthesized: HMCD-ART1 (artesunate linker), HMCD-ART2 (carbamate linker), HMCD-ART3 (ether linker), and HMCD-ART4 (ester linker). These were evaluated in androgen-independent prostate cancer models (22Rv1, PC-3, C4-2B, and resistant derivatives). Cytotoxicity, selectivity, and chemosensitization were analyzed in vitro, while subcellular localization, ROS generation, and cell-death mechanisms were examined by fluorescence microscopy, flow cytometry, and Western blotting. Tumor targeting and therapeutic efficacy were determined by NIR imaging and 22Rv1 xenograft models. Results: Among the conjugates, HMCD-ART1 showed the greatest potency and selectivity, effectively killing taxane-, abiraterone-, and enzalutamide-resistant prostate cancer cells while sparing normal prostate epithelial cells. HMCD-ART1 enhanced sensitivity to chemotherapeutics, inhibited colony formation and migration, and demonstrated prolonged tumor retention for up to eight weeks. Mechanistic analyses revealed mitochondrial and lysosomal co-localization, ROS induction, cytochrome c release, DNA damage (pATM, γH2AX), and mitochondrial-fission-dependent necrotic death. In vivo, HMCD-ART1 significantly suppressed xenograft tumor growth without observable systemic toxicity. Conclusion: HMCD-ART1 integrates NIR-guided tumor targeting with mitochondrial-specific cytotoxicity, providing a novel therapeutic platform to overcome drug resistance in mCRPC. This strategy represents a next-generation precision chemotherapeutic approach combining targeted delivery, subcellular selectivity, and enhanced safety. Citation Format: Yan Ou, Adrian Lim, Mouad Edderkaoui, Ruoxiang Wang, Qiang Wang, Stephen J. Pandol, Yi Zhang. Subcellular targeted therapy using HMCD-artemisinin conjugates for metastatic castration-resistant prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 409.

Hepatic colorectal metastases, also termed colorectal liver metastases (CRLM), remain a major clinical challenge, despite advances in surgery and chemotherapy. The complexity of CRLM pathology necessitates novel therapeutic approaches, yet current preclinical models often fail to accurately recapitulate the human tumor microenvironment. To overcome these limitations, we utilized patient-derived three-dimensional (3D) CRLM tumoroids to evaluate the efficacy of DZ-1-DHA, a novel conjugate consisting of the heptamethine carbocyanine dye DZ-1 linked to the anti-malarial derivative dihydroartemisinin (DHA). Data from TUNEL and immunoblotting assays revealed that treatment of CRLM tumoroids with DZ-1-DHA led to significant tumor cell death, accompanied by apoptotic signaling. Fluorescence imaging with MitoTracker, 2',7'-dichlorofluorescin diacetate, MitoSOX, and JC-1 showed that DZ-1-DHA accumulates in mitochondria, where it induces generation of cytotoxic reactive oxygen species (ROS) and causes mitochondrial membrane depolarization. Furthermore, data from treatment with deferoxamine or MitoTEMPO indicated that DZ-1-DHA promotes mitochondrial ROS production through a Fenton-like mechanism. These findings demonstrate that DZ-1-DHA triggers apoptosis through mitochondrial stress and apoptotic signaling pathways. Also, DZ-1-DHA represents a promising second-line therapeutic strategy for CRLM. By inducing selective tumor cell death through mitochondrial-targeted apoptosis in a clinically relevant 3D model. This promising approach needs in vivo validation for safety and efficacy.

Abstract Introduction: STAT1 mediates interferon signaling and immune responses with diverse roles in cancer. In colorectal cancer (CRC), plasma cells are critical immune mediators, and STAT1 expression in these cells may reflect their functional status. Lower STAT1-expressing plasma cells have been linked to a weakened anti-tumor immune response that facilitates tumor escape. Understanding this relationship could reveal novel prognostic biomarkers and therapeutic targets for CRC. Materials and Methods: Thirty CRC samples underwent G4X spatial profiling, each with a manually selected 4.5 × 4.5 mm2 capture area targeting key histological features, resulting in spatial gene expression profiles for a 360-gene panel for 3,088,252 cells. Tissue sections were processed following the G4X spatial transcriptomics protocol enabling RNA detection at subcellular resolution. Manual pathology annotation was performed by a collaborating pathologist, identifying regions including cancer, dysplasia/cancer in situ, benign epithelium, interface, muscularis, and stroma. Cancer-associated cells were defined by two criteria: (1) spatial localization within annotated cancer regions, and (2) absence of canonical epithelial and malignant epithelial markers (CEACAM5, EPCAM, MUC12). Differential expression analyses identified associated cell markers, and the proportions of cell types expressing these markers were quantified. Cell-level regression models incorporated random patient intercepts and clinical covariates (age, gender, microsatellite status, T and N stage), followed by sample-level univariate analyses across all 30 samples to assess associations between STAT1-positive cell clusters and clinical recurrence. Results: Differential expression analysis identified STAT1 as the top upregulated gene in cancer-associated cells (log2 fold-change = 2.11; adjusted p < 0.001). Clustering of STAT1-positive cancer-associated cells revealed 10 distinct cell clusters, including 5 cancer-associated fibroblast subtypes, tumor-associated macrophages, mixed T cells, 2 epithelial-like cells, and plasma cells. Regression analyses consistently indicated a negative association between proportion of STAT1-positive plasma cells and colorectal cancer recurrence (OR = 0.19; 95% CI = 0.03-1.13; p = 0.068). Conclusion: STAT1-expressing plasma cells in the colorectal cancer microenvironment are significantly associated with recurrence risk. These findings support further validation efforts through immunohistochemistry in a larger study cohort, demonstrating the promise of leveraging spatial transcriptomic insights to inform routine pathology for patient stratification. Citation Format: I-Chuang Liao, Minh-Khang Le, Vivek Pujara, Haley Jun, Devanshi Pratiher, Jane C. Figueiredo, Nathalie Nguyen, Chintda Santiskulvong, Angie Laguna, Yi Zhang, Keluo Yao, Joshua Jay Levy. Cell-level spatial transcriptomics detection of microenvironmental STAT1-expressing plasma cells in colorectal cancer recurrence by G4X [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 4235.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy. In this paper, we present DZ-ART1, a first-in-class, dual-function therapeutic. This innovative agent is composed of a tumor-targeting heptamethine carbocyanine dye conjugated to artemisinin (ART). Near-infrared imaging demonstrated precise tumor localization of DZ-ART1 in mice. DZ-ART1 accumulated 5- to 10-fold more in cancer cells compared to normal cells. DZ-ART1 significantly decreased the survival of eight PDAC cell lines with little effect on normal cells. It increased the lethal effect of chemotherapies in vitro and in vivo. Functional assays confirmed DZ-ART1’s ability to disrupt mitochondrial bioenergetics, deplete ATP, and induce reactive oxygen species production. Mitochondrial depletion of cancer cells decreased DZ-ART1 uptake and cytotoxicity, highlighting its mechanistically unique, mitochondria-dependent action. Transcriptomic profiling revealed DZ-ART1’s broad reprogramming of PDAC pathways related to cell survival, cancer stemness, and metastasis. In three rigorously validated preclinical models — Krasþ/LSLG12D;Trp53þ/LSLR172H;Pdx-1-Cre (KPC) transgenic, syngeneic, and patient-derived xenografts (PDX) — DZ-ART1 markedly suppressed tumor growth and metastasis and prolonged survival, all without toxicity to normal tissues. DZ-ART1 represents a new class of anti-cancer therapeutics that uniquely combines diagnostic imaging and cancer cell organelle-specific targeting.

Background Cancer remains a leading cause of death worldwide, and many therapies are limited by poor tumor specificity, systemic toxicity, and drug resistance. Tumor-targeted strategies that enhance drug accumulation while minimizing off-target effects are therefore essential. We hypothesized that conjugating dihydroartemisinin (DHA) to the tumor-targeting near-infrared dye DZ-1 would improve tumor-selective accumulation, promote mitochondrial localization, and enhance reactive oxygen species (ROS)-mediated cytotoxicity. We also investigated how linker chemistry influences drug stability and therapeutic efficacy. Methods Three types of linkers (ether, ester, and carbamate) were used to conjugate DZ-1 with dihydroartemisinin (DHA). The tumoricidal efficacy of these DZ-1-DHA conjugates was evaluated in HCT116 and BxPC3 cells. Cytotoxicity was assessed using the trypan blue exclusion assay. Apoptosis was analyzed by TUNEL staining and immunoblotting. Drug accumulation was compared among the conjugates. Mitochondrial membrane potential was assessed using JC-1 staining, and ROS generation was measured using DCF, MitoSOX, and Mito-TEMPO assays. Results Ester-linked DZ-1-DHA showed the most potent, dose-dependent cytotoxicity in both HCT116 and BxPC3 cells. Apoptosis was confirmed by immunoblotting and TUNEL assays. Fluorescence microscopy revealed that the ester-linked conjugate effectively accumulated in mitochondria, efficiently generated reactive oxygen species (ROS), and disrupted mitochondrial membrane potential. Conclusions Our findings demonstrate that linker chemistry critically influences DZ-1-DHA accumulation and mitochondrial ROS generation, which in turn determine tumoricidal efficacy. Among the conjugates, the ester-linked DZ-1-DHA exhibited superior anticancer activity. These results provide mechanistic insight and highlight linker optimization as a key design principle for the development of next-generation tumor-targeted artemisinin therapeutics.

ABSTRACT A 15‐year‐old female adolescent was previously in good health. Ultrasound examination revealed an 84 × 45 mm heterogeneous echo mass in the left inner lobe of the liver, exhibiting clear boundaries and a regular shape. Additionally, multiple small cysts were observed within the heterogeneous echo mass, and occasionally, a strong spot‐like echo with a comet tail sign was noted. Imaging results indicated that the left inner lobe of the liver exhibited a lamellar structure with an anomalous signal shadow, suggesting a high probability of bile duct hamartoma.

Objective Inflammation is a key driver of disc herniation, a major cause of back pain and disability. Heterogeneous macrophages infiltrated at disc hernia sites, yet their role in disease pathology and pain remains unclear. This study investigates the role of CX3CR1⁺ macrophages and microglia in local inflammation and pain using transgenic mouse models and surgically induced disc herniation model. Method A lumbar annular puncture model was applied to three global transgenic mouse strains: CCR2RFP/+/CX3CR1GFP/+ dual-reporter mice to trace monocyte infiltration, and CX3CR1GFP/+ and CX3CR1GFP/GFP mice to investigate CX3CR1-contribution to disc herniation induced local inflammation and neuroinflammation in dorsal root ganglia (DRG) and spinal cord. Behavioral assays were used to evaluate mechanical sensitivity. Results Disc herniation induced peak inflammatory cell infiltration at hernia sites at 1-week post injury, reduced by 4 weeks in both CX3CR1GFP/+ and CX3CR1GFP/GFP mice. CX3CR1GFP/+ mice displayed more CD11b+ and F4/80+ macrophages, less disc height and higher mechanical sensitivity than CX3CR1GFP/GFP mice. Both genotypes exhibited increased microglial activation in the ipsilateral dorsal horn of spinal cord compared to contralateral sides by 1 week, while CX3CR1GFP/+ microglia showed higher lysosomal activity and changes in morphology than CX3CR1GFP/GFP microglia. Ipsilateral DRGs from CX3CR1GFP/+ mice showed elevated CX3CR1+ and F4/80+ cells, substance P, and calcitonin gene-related peptide, compared to CX3CR1GFP/GFP at 1 week. Conclusions These findings indicate a mechanistic role of CX3CR1 in mediating inflammation and pain in disc herniation. Targeting CX3CR1 may disrupt the signal interactions between disc, DRG, and spinal cords.

STUDY DESIGN:Retrospective cohort study. OBJECTIVE:To investigate postoperative medical and surgical outcomes of 3-level cervical disk arthroplasty (CDA) in comparison to 3-level anterior cervical discectomy and fusion (ACDF). SUMMARY OF BACKGROUND DATA:Cervical disk arthroplasty has emerged as a noninferior alternative to anterior cervical discectomy and fusion. However, few studies investigated the clinical outcomes of 3-level CDA. At present, CDAs beyond 2-level are performed as off-labeled procedures across the United States and can be deemed experimental by insurance companies. METHODS:Three-level CDA and 3-level ACDF patients between 18 and 84 years old from 2010 to 2020 were identified within the PearlDiver database. Ninety-day postoperative medical and surgical complications, emergency department visits, and readmission were measured. Two-year complications and 4-year rate of revisions were also studied. The Pearson χ 2 test was used to assess demographics and pre-existing comorbidities. Independent effects of CDA and ACDF on the postoperative outcomes were determined with multivariable logistic regression after adjusting for demographic factors and pertinent comorbidities. PearlDiver Bellwether was used to conduct all statistical analyses via its research query interface. RESULTS:After matching 2901 three-level CDA patients with 14,378 ACDF patients, multivariate analysis showed that the CDA group had a significantly lower rate of 90-day dysphagia, pneumonia, wound complications, and surgical site infections. CDA patients also showed a reduced rate of 90-day ED visits and readmission. At the 2-year follow-up, CDA patients had a lower rate of instrumentation failure. At 4 years, 786 CDA patients and 3890 ACDF patients remained, and the CDA group had a lower posterior revision rate and a less overall revision rate. CONCLUSIONS:The current study represents the largest comparative study examining clinical outcomes following 3-level CDA. CDA established itself as a safe and noninferior procedure to ACDF. However, the safety and efficacy of 3-level CDA require more long-term data to validate.

Heptamethine cyanine dyes and anticancer agents based conjugates are being developed for enhanced targeting and killing of cancer cells. DZ-1 dye conjugated agents induced cytotoxicity and mechanism of action have been shown in previous studies. In this study, a conjugated form of DZ-1 and artesunate (DZ-1-ART) was used to evaluate its cytotoxicity and elucidate the mechanism of actions in various cancer cell lines. Cells survival assays indicated dose-dependent cytotoxic activities of DZ-1-ART in HCT116, BxPC-3, and OVCAR-3 cell lines. Immunoblotting and terminal deoxynucleotidyl transferase dUTP nick-end labeling assay confirmed involvement of apoptosis in DZ-1-ART-induced cytotoxicity. To elucidate the anticancer mechanism of the action of DZ-1-ART, MitoTracker and JC-1 assay were used. The results showed that translocation of DZ-1-ART in the mitochondria was followed by disruption of mitochondrial outer membrane potential. Dichlorofluorescin diacetate assay confirmed the generation of reactive oxygen species (ROS) in DZ-1-ART treated cancer cells. An antioxidant, N-acetyl cysteine treatment with DZ-1-ART showed reduction in cell death as well as suppression of ROS generation. When compared to HCT116 wild-type cells, Bak and Bax-deficient HCT116 cells also showed similar levels of cytotoxicity of DZ-1-ART. Taken together, this study's results reported that DZ-1-ART could induce mitochondria-mediated, ROS-generated, and Bak and Bax-independent apoptosis in cancer cells.

Objectives: Pretargeting strategies enhance the specificity and safety of radiopharmaceuticals by separating tumor targeting from radionuclide delivery. To address the rapid clearance and systemic exposure of directly labeled small-molecule agents, a DZ-1–based pretargeting system was developed, utilizing its broad-spectrum tumor-targeting characteristics. Methods: Three DZ-TCO precursors (DZ-1-TCO, DZ-Lys-TCO, and DZ-Lys-PEG4-TCO) were synthesized and evaluated by near-infrared fluorescence imaging in HeLa and U87MG tumor-bearing mice. Two tetrazine probes (methyl-tetrazine and mono-substituted tetrazine) were labeled with 68Ga to yield 68Ga-DOTA-Me-Tz and 68Ga-DOTA-H-Tz, whose stability was assessed in PBS and serum. Pretargeted PET imaging was performed using different precursor/probe combinations and pretargeting intervals (24, 48, and 72 h). Results: All precursors exhibited tumor accumulation peaking at 24 h and signal retention up to 96 h. Both 68Ga-DOTA-Me-Tz and 68Ga-DOTA-H-Tz maintained >85% radiochemical stability after 4 h. PET imaging identified DZ-Lys-TCO as the most effective precursor (1.98 ± 0.72 %ID/g, T/M 3.86 ± 0.91). Using 68Ga-DOTA-H-Tz, the 48 h interval achieved optimal uptake (3.24 ± 0.95 %ID/g) with the highest tumor-to-muscle ratio (8.30 ± 3.39). Biodistribution confirmed rapid renal clearance, low off-target accumulation, and peak tumor uptake of 3.53 ± 1.76 %ID/g (T/M 10.9 ± 0.3 at 30 min). Conclusions: The DZ-TCO/68Ga-DOTA-Tz pretargeting system enables high-contrast tumor imaging with low background. The combination of DZ-Lys-TCO and 68Ga-DOTA-H-Tz at a 48 h interval provides optimal performance, representing a promising platform for precise and safe radiopharmaceutical imaging.

e15132 Background: Certain heptamethine cyanine dye derivatives like MHI-148 (1a) and IR-783 exhibit strong affinity for cancer cells. These dyes selectively target and accumulate within cancer cells due to overexpression of organic anion-transporting polypeptides (OATPs) and hypoxia-induced factors. In contrast, chlorin e6 (Ce6, 4), a commonly used photosensitizer in cancer treatment, lacks effective cancer targeting, leading to limited therapeutic efficacy and non-specific tissue damage. Methods: To enhance Ce6's tumor specificity, researchers developed novel tumor-targeting photosensitizer-carrier-conjugates (PSCs) by attaching Ce6 to heptamethine cyanine dyes. Results: These PSCs showed improved cancer cell uptake and retention compared to Ce6 alone, as demonstrated in breast cancer cells (MCF7) both in vitro and in mice xenograft models. The synthesis of PSCs involved three steps starting from Ce6 anhydride, yielding moderate results. The conjugates were complexed with either cold Cu for reference or radioactive 64Cu for quantitative studies and PET imaging. Both complexed and non-complexed PSCs were effective for in vitro and in vivo imaging studies. NIRF imaging revealed enhanced cancer cell uptake of PSCs compared to Ce6 alone, while PET imaging provided quantitative tumor visualization. PSCs demonstrated superior tumor imaging efficiency compared to 18F-FDG in mice xenografts. Conclusions: These findings highlight the potential of PSCs as multifunctional theranostic agents for image-guided photodynamic and radiotherapeutic cancer treatment.

IntroductionWe previously reported the identification of circulating fatty objects (CFOs), abnormal entities found in peripheral blood samples from cancer patients. CFOs are large spherical objects with high contents of heavy cholesterol lipids, posing a risk of vessel embolization and circulation occlusion, common complications in clinical cancers. Initial characterization suggested that CFOs bear resemblance to bile salts, prompting further investigation into the potential link between bile and cancer-associated vascular occlusion.MethodsTo explore this connection, we analyzed portal blood samples from patients diagnosed with pancreatic ductal adenocarcinoma (PDAC), as CFOs were most frequently detected in this type of cancer (20.4% incidence). Portal vein samples were collected via transhepatic endoscopic ultrasound-guided fine-needle aspiration and treated with ammonium chloride hemolysis to facilitate enumeration and characterization of CFOs in the nucleated blood cell fraction.ResultsCFOs were observed in 14 out of the 16 portal samples, indicating a significantly higher incidence compared to peripheral samples. Despite this, portal CFOs exhibited similar characteristics to peripheral CFOs. In a parallel study, a portion of the portal samples was filtered through an Exthera’s Seraph 100 column before hemolysis, resulting in no detection of CFOs.DiscussionThese findings suggest that CFOs originate from bile. It is probable that CFOs are insoluble bile lipids that have ebbed into the portal vein and subsequently shunted to the systemic circulation due to PDAC-induced hepatobiliary system abnormalities. It appears that hemofiltration can effectively remove CFOs from circulation.

Retrospective cohort study. To investigate postoperative medical and surgical outcomes of 3-level cervical disk arthroplasty (CDA) in comparison to 3-level anterior cervical discectomy and fusion (ACDF). Cervical disk arthroplasty has emerged as a noninferior alternative to anterior cervical discectomy and fusion. However, few studies investigated the clinical outcomes of 3-level CDA. At present, CDAs beyond 2-level are performed as off-labeled procedures across the United States and can be deemed experimental by insurance companies. Three-level CDA and 3-level ACDF patients between 18 and 84 years old from 2010 to 2020 were identified within the PearlDiver database. Ninety-day postoperative medical and surgical complications, emergency department visits, and readmission were measured. Two-year complications and 4-year rate of revisions were also studied. The Pearson χ2 test was used to assess demographics and pre-existing comorbidities. Independent effects of CDA and ACDF on the postoperative outcomes were determined with multivariable logistic regression after adjusting for demographic factors and pertinent comorbidities. PearlDiver Bellwether was used to conduct all statistical analyses via its research query interface. After matching 2901 three-level CDA patients with 14,378 ACDF patients, multivariate analysis showed that the CDA group had a significantly lower rate of 90-day dysphagia, pneumonia, wound complications, and surgical site infections. CDA patients also showed a reduced rate of 90-day ED visits and readmission. At the 2-year follow-up, CDA patients had a lower rate of instrumentation failure. At 4 years, 786 CDA patients and 3890 ACDF patients remained, and the CDA group had a lower posterior revision rate and a less overall revision rate. The current study represents the largest comparative study examining clinical outcomes following 3-level CDA. CDA established itself as a safe and noninferior procedure to ACDF. However, the safety and efficacy of 3-level CDA require more long-term data to validate.

Intervertebral disc degeneration is a leading cause of back and leg pain and a major contributor to disability worldwide. Despite its prevalence, treatments remain limited due to incomplete understanding of its pathology. In vivo models pose challenges for controlled conditions, while in vitro cell cultures lack key cell–cell and cell–matrix interactions. To address these limitations, we developed a novel tissue slice culture model of mouse discs, in which intact mouse discs were sliced down to 300 μm thickness with a vibratome and cultured ex vivo at various time points. The cell viability, matrix components, structure integrity, inflammatory responses, and macrophage interactions were evaluated with biochemistry, gene expression, histology, and 3D imaging analyses. Disc slices maintained structural integrity and cell viability, with preserved extracellular matrix in the annulus fibrosus (AF) and mild degeneration in nucleus pulposus (NP) by day 5. Interleukin-1 (IL-1) induced disc degeneration manifested by increased glycosaminoglycan release in media and reduced aggrecan and collagen II mRNA levels in disc cells. Cultured disc slices promoted macrophages towards pro-inflammatory phenotype with elevated mRNA levels of il-1α, il-6, and inos. Macrophage overlay and 3D imaging demonstrated macrophage infiltration into the NP and AF tissues up to ~100 µm in depth. The disc tissue slice model captures key features of intervertebral discs and can be used for investigating mechanisms of disc degeneration and therapeutic evaluation.

Ulcerative colitis (UC), a major form of inflammatory bowel disease (IBD), is marked by chronic mucosal inflammation and epithelial barrier disruption. The complement system, a central component of the immune system, is a key modulator of UC pathogenesis. Recent studies from clinical observations and dextran sulfate sodium (DSS)-induced murine colitis models reveal that complement activation plays a context-dependent role in UC. Complement components such as C3a, C5a, and the membrane attack complex (MAC) exacerbate mucosal inflammation and epithelial injury by promoting neutrophil recruitment and cytokine storms, while initiator molecules C1q and mannose-binding lectin (MBL) support protective functions, including immune regulation, apoptotic cell clearance, and tissue repair. This review synthesizes current clinical and experimental evidence on the bidirectional roles of the complement system in UC, with emphasis on the regulatory balance between protective and pathogenic pathways. Emerging therapeutics, ranging from C5aR antagonists to colon-targeted complement inhibitors, highlight the translational potential of modulating complement activity in UC. Understanding these mechanisms provides a framework for developing complement-targeted therapies and precision treatment strategies in IBD.

Tumor cellularity (TC) in lung adenocarcinoma slides submitted for molecular testing is important in identifying actionable mutations, but lack of best practice guidelines results in high interobserver variability in TC assessments. An artificial intelligence (AI)-based pipeline developed to assess TC in hematoxylin and eosin (H&E) whole slide images (WSIs) and in tumor areas (TAs) within WSIs includes a new model (CaBeSt-Net) trained to mask cancer cells, benign epithelial cells, stroma in H&E WSIs using immunohistochemistry-restained slides, and a model to detect all cell nuclei. High masking accuracy (>91%) by CaBeSt-Net computed using 1024 H&E regions of interest and intraclass correlation coefficient >0.97 assessing TC assessments reliability by one pathologist and AI in 20 test regions of interest supported the pipeline's applicability to TC assessment in 50 study H&E WSIs. Using the pipeline, TCs assessed in TAs and WSIs were compared with those by three pathologists. Reliabilities of these ratings by the pathologists supported by the pipeline were good (intraclass correlation coefficient >0.82, P < 0.0001). The consistency of sample categorizations as inadequate or adequate (TC ≤ 20% cut point) for molecular testing among the pathologists assessing TCs without AI support was moderate in TAs (κ = 0.410, P < 0.0001) and slight in WSIs (κ = 0.132, nonsignificant). With AI support, the consistency was substantial in both WSIs (κ = 0.602, P < 0.0001) and TAs (κ = 0.704, P < 0.0001). By visualizing cancer and measuring TC in the sample, this novel AI-based pipeline assists pathologists in selecting samples for molecular testing.

BACKGROUND CONTEXT:The L4-S1 region contributes the most to lumbar lordosis, and adjacent segment disease (ASD) is a common complication following lumbar interbody fusion. To date, no study has directly compared sagittal alignment parameters and incidence of ASD between ALIF and TLIF for the treatment of L4-S1 degenerative disc disease. PURPOSE:This study aims to evaluate and compare the efficacy of anterior lumbar interbody fusion (ALIF) versus transforaminal lumbar interbody fusion (TLIF) in restoring L4-S1 lordosis and the impact on the development of ASD. STUDY DESIGN:A retrospective cohort study. PATIENT SAMPLE:A total of 102 TLIF patients and 53 ALIF patients were identified from a single-center database. OUTCOME MEASURES:Complications, revisions, and radiological sagittal parameters. METHODS:We conducted a retrospective study involving patients who underwent L4-S1 fusion for degenerative lumbar disease from January 2017 to December 2021. Patients were categorized into TLIF and ALIF groups, with demographic, surgical, and radiographic data collected. Radiographic parameters were assessed preoperatively, postoperatively, and at final follow-up (45.8 ± 13.2 months). RESULTS:A total of 155 patients were analyzed (102 TLIFs and 53 ALIFs). Both groups exhibited significant changes in L4-S1 lordosis postoperatively; however, ALIF resulted in a more pronounced increase in L4-S1 lordosis (6.5° vs 1.3°) and reduced compensatory changes at L3/4 compared with TLIF (-2.4° vs 0.2°). ALIF also maintained 72% of the lordosis restoration over time. Moreover, ALIF had lower incidences of ASD (3.8% vs 15.7%, p=.034) and significantly fewer reoperations (7.5% vs 25.5%, p=.009). A binary logistic regression analysis revealed that an increase in postoperative L4-S1 lordosis (OR 0.903 [95% CI: 0.820-0.994]) and a decrease in delta L3/4 (OR 0.757 [95% CI: 0.597-0.961]) were significantly associated with the reduced risk of ASD. CONCLUSIONS:ALIF demonstrated superior effectiveness compared to TLIF in restoring and maintaining L4-S1 lordosis, mitigating compensatory lordosis, and decreasing the incidence of ASD. LEVEL OF EVIDENCE:III.