Substantial Necrosis Research Articles

Anlotinib treatment for rapidly progressing pediatric embryonal rhabdomyosarcoma in the maxillary gingiva: a case report

BackgroundEmbryonal rhabdomyosarcoma (ERMS) is a highly aggressive form of soft-tissue sarcoma that predominantly affects children. Due to limited benefits and resistance to therapy, there is an unmet need to explore alternative therapeutic strategies.Case presentationIn this report, we present a rare case of pediatric ERMS located on the right side of the maxillary gingiva. A composite reference guide integrating clinical, radiographic, and histopathologic findings was used for a definitive diagnosis. Targeted next-generation sequencing of tumor biopsy was performed to identify genetic alterations. A 12-year-old female was admitted to the Pediatric Intensive Care Unit (PICU) and underwent a tracheotomy to relieve asphyxiation caused by a 5.5 cm diameter mass compressing the tongue root and pharyngeal cavity. Hematoxylin and eosin staining revealed a hybrid morphology characterized by clusters of round and spindle cells. Further immunohistochemistry assays indicated positive immunoreactivity for desmin, myogenin, and MyoD1. Various genetic alterations were identified, including mutations in GNAS, HRAS, LRP1B, amplification of MDM2 and IGF1R, and two novel IGF1R fusions. Negative PAX-FOXO1 fusion status supported the clinical diagnosis of ERMS. Initial treatment involved standard chemotherapy; however, the tumor persisted in its growth, reaching a maximum volume of 12 cm × 6 cm × 4 cm by the completion of treatment. Subsequent oral administration of anlotinib yielded a significant antitumor response, characterized by substantial tumor necrosis and size reduction. Following the ligation of the tumor pedicle and its removal, the patient developed a stabilized condition and was successfully discharged from PICU.ConclusionsOur study highlights the importance of accurate diagnosis established on multifaceted assessment for the effective treatment of ERMS. We present compelling evidence supporting the clinical use of anlotinib as a promising treatment strategy for pediatric ERMS patients, especially for those resistant to conventional chemotherapy.

MR Molecular Image Guided Treatment of Pancreatic Cancer with Targeted ECO/miR-200c Nanoparticles in Immunocompetent Mouse Tumor Models.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by desmoplasia due to increased deposition of extracellular matrix (ECM) proteins. This work investigates the efficacy of targeted ECO/miR-200c nanoparticles (ELNP) on ECM remodeling in PDAC and tumor proliferation with MR molecular imaging (MRMI) with MT218 in immunocompetent mouse models. The miR-200c mediated regulation of EMT markers was measured in PDAC cells in vitro. Wild-type mice bearing mutated KRAS-driven KPC subcutaneous or orthotopic tumors were dosed weekly with RGD-ELNP/miR-200c at 1mg-RNA/kg for a total of 4 doses. We utilized MT218-MRMI to non-invasively monitor the alteration of tumor ECM EDN-FN levels by miR-200c and tumor response to the treatment. The changes were also validated by posthumous histopathology. Transfection of PDAC cells with ELNP/miR-200c downregulated the expression of FN1 and EDB-FN and some mesenchymal markers, inhibiting 3D spheroid formation and migration of KPC PDAC cells. RGD-ELNP/miR-200c treatment resulted in significant signal reduction in the MT218 enhanced MRMI images of both subcutaneous and orthotopic KPC tumors compared to those prior to treatment and treated with a non-specific control. MT218-MRMI results were suggestive of EDB-FN downregulation in tumors, which was later confirmed by immunohistochemistry. Tumor growth in subcutaneous tumors was significantly attenuated with RGD-ELNP/miR-200c and was an observed trend in orthotopic tumors. Substantial necrosis and remodeling were observed in both models treated with RGD-ELNP/miR-200c based on H&E staining. These results demonstrate the feasibility of RGD-ELNP/miR-200c in modulating PDAC ECM and restraining tumor growth and the utility of MT218-MRMI for non-invasively monitoring miR-200c efficacy.

Engineered nanovesicles mediated cardiomyocyte survival and neovascularization for the therapy of myocardial infarction

Myocardial infarction (MI) leads to substantial cellular necrosis as a consequence of reduced blood flow and oxygen deprivation. Stimulating cardiomyocyte proliferation and angiogenesis can promote functional recovery after cardiac events. In this study, we explored a novel therapeutic strategy for MI by synthesizing a biomimetic nanovesicle (NV). This biomimetic NVs are composed of exosomes sourced from umbilical cord mesenchymal stem cells, which have been loaded with placental growth factors (PLGF) and surface-engineered with a cardiac-targeting peptide (CHP) through covalent bonding, termed Exo-P-C NVs. With the help of the myocardial targeting effect of homing peptides, NVs can be enriched in the MI site, thus improve cardiac regeneration, reduce fibrosis, stimulate cardiomyocyte proliferation, and promote angiogenesis, ultimately resulted in improved cardiac functional recovery. It was demonstrated that Exo-P-C NVs have the potential to offer novel therapeutic strategies for the improvement of cardiac function and management of myocardial infarction.

Immune cell populations in the tumour environment following calcium electropora-tion for cutaneous metastasis: a histopathological study.

Calcium electroporation (CaEP) involves injecting calcium into tumour tissues and using electrical pulses to create membrane pores that induce cell death. This study assesses resultant immune responses and histopathological changes in patients with cutaneous metastases. The aimed cohort comprised 24 patients with metastases exceeding 5 mm. Tumours were treated once with CaEP (day 0) or twice (day 28). Biopsies were performed on days 0 and 2, with additional samples on days 7, 28, 30, 35, 60, and 90 if multiple tumours were treated. The primary endpoint was the change in tumour-infiltrating lymphocytes (TILs) two days post-treatment, with secondary endpoints evaluating local and systemic immune responses via histopathological analysis of immune markers, necrosis, and inflammation. Seventeen patients, with metastases primarily from breast cancer (14 patients), but also lung cancer (1), melanoma (1), and urothelial cancer (1), completed the study. Of the 49 lesions treated, no significant changes in TIL count or PD-L1 expression were observed. However, there was substantial necrosis and a decrease in FOXP3-expression (p = 0.0025) noted, with a slight increase in CD4+ cells but no changes in CD3, CD8, or CD20 expressions. Notably, four patients showed reduced tumour invasiveness, including one case of an abscopal response. This exploratory study indicates that CaEP can be an effective anti-tumour therapy potentially enhancing immunity. Significant necrosis and decreased regulatory lymphocytes were observed, although TIL count remained unchanged. Several patients exhibited clinical signs of immune response following treatment.

Ameliorative delivery of docetaxel and curcumin using PEG decorated lipomers: A cutting-edge in-vitro/ in-vivo appraisal

The development of a PEG-decorated lipid-polymer hybrid system camouflaged with natural and synthetic chemotherapeutic moieties is an influential approach melding the biomimetic properties of long-circulating vesicles to utilize different mechanisms to dwindle the tumor growth. Therefore, a safe and efficient lipid-coated nano-particulate system (LCNPs) was proposed to investigate the in-vitro, ex-vivo and in-vivo demeanors of such amalgamation.Docetaxel loaded PLGA nanoparticles (DTX-NPs) were prepared by solvent evaporation while curcumin liposomes were mapped out using the film hydration method. Physicochemical characterizations were executed in terms of size, surface morphology, differential scanning calorimetry (DSC) and fourier-transform infrared spectroscopy (FTIR). In-vitro cytotoxicity was effectuated using MCF-7 cell line. Hemolysis, erythrocyte aggregation and acute in-vivo toxicity were carried out to establish the biocompatibility. The hydrodynamic diameters of samples were in the nano-range and corresponded to the findings of scanning electron microscopy (SEM) and atomic force microscopy (AFM). The absence of distinctive peaks of DTX-NPs in FTIR and DSC analysis of LCNPs depicts the shielding of the lipid bilayer over the nanoparticle. Cytotoxicity induced by the LCNPs represented the efficient delivery of cargo to the tumor cells. LCNPs also exhibited the least toxicity under ex-vivo and in-vivo circumstances compared to free drugs. Additionally, histological studies showed no evidence of substantial necrosis. Additionally, histological studies showed no evidence of notable necrosis.

A new model of ischemia/reperfusion injury using precision-cut liver slices

Background: Primary cells isolated from the liver are widely used to investigate hepatic pathophysiology. However, these cells do not represent cell-to-cell interactions, three dimensional architecture, and zonal structure in the liver. Furthermore, characterization of cellular and molecular events during ischemia/reperfusion (I/R) is challenging in liver biopsies. Thus, a new model of I/R injury resembling the native liver is needed. Methods: We generated precision-cut liver slices (PCLS) from mouse and human livers using a Krumdieck automatic tissue slicer (Alabama R & D, Munford, AL). Thickness of individual PLCL was 200 microns. To simulate anoxia, nutrient depletion, and tissue acidosis during ischemia, PCLS were incubated in Krebs-Ringer-N-(2-Hydroxyethyl)piperazine-N-(2-ethanesulfonic acid) (HEPES) solution at pH 6.2 in the anaerobic chamber (Coy Laboratory, Grass Lake, MI) for 30 min. To simulate normoxia, nutrient repletion, and restoration to normal pH during reperfusion, ischemic PCLS were aerobically incubated in Dulbecco's modified eagle medium (DMEM) at pH 7.4 for up to 4 h. Changes in tissue viability were assessed with the release of lactate dehydrogenase (LDH) into the extracellular medium. Some PCLS were labeled with green fluorescent rhodamine-123 (Rd-123, mitochondrial membrane potential indicator) and red fluorescent propidium iodide (PI, necrosis marker). Confocal images of PCLS were collected after I/R. Results: LDH release assay revealed that while PCLS from chow diet (CD)-fed lean mice tolerated I/R injury well, those from high fat diet (HFD)-fed steatotic mice were susceptible to I/R injury. Confocal microscopy showed substantial mitochondrial depolarization (loss of Rd-123) and necrosis (nuclear labeling of PI) in the HFD group, which was, however, minimally observed in the CD group. Of importance, mitochondrial dysfunction and cell death after reperfusion occurred predominantly in hepatocytes. Heightened I/R injury in steatotic livers was also confirmed in primary hepatocytes isolated from either CD- or HFD-fed mice. To compare the similarities and differences between mouse and human livers, PCLS were generated from discarded human livers deemed unsuitable for transplant and subjected to 30 min of in vitro ischemia. The loss of mitochondrial membrane potential and hepatocyte death progressively increased after I/R in discarded human livers. At 1 h after reperfusion, diffusive fluorescence of Rd-123 and nuclear labeling of PI became evident, indicative of mitochondrial dysfunction and tissue injury after I/R. SUMMARY: LDH assay and confocal analysis of PCLS demonstrated heightened susceptibility of steatotic livers to I/R injury. In contrast to PCLS from lean livers, those from steatotic livers rapidly developed mitochondrial depolarization and onset of necrosis, cardinal features of I/R injury, even after short ischemia. The early phase of reperfusion injury in steatotic livers occurred predominantly in parenchymal cells. CONCLUSION: In vitro I/R with PCLS is a model suitable for hepatic I/R injury. This work (J-S K) was funded by the National Institutes of Health (DK079878), Mid-America Transplant Foundation (07201903), Foundation for Barnes-Jewish Hospital (4776, 5153), and Washington University Department of Surgery Pilot Grant. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Effects of aquaculture effluents on the slender sea pen Virgularia mirabilis

This study aims to assess in situ the impact of effluents originating from an Atlantic salmon (Salmo salar) farm on a nearby slender sea pen (Virgularia mirabilis) field. We evidenced (1) the presence and persistence of emamectin residues (i.e. a common chemotherapeutants used for treating ectoparasites in salmons) in V. mirabilis tissue 56 days after treatment and (2) lethal and sublethal responses of V. mirabilis to effluents discharged by the salmon farm. Particularly, sea pens near the fish farm exhibited significant overproduction of mucus, contraction of polyps’ tentacles, and disappearance of associated fauna. Furthermore, sea pens located directly underneath the farm showed substantial tissue necrosis and, in the most severe case, complete tissue loss and mortality. Our results suggest that lethal damages on sea pens occur directly below the farm, and that sublethal effects are visible up to 500 m from the farm. However, the presence of V. mirabilis below the studied farm, which has been active for more than twenty years, suggests that V. mirabilis population possesses the capacity to recover from the impacts of the farm, thereby preventing the complete disappearance from the area. In this context, it would be particularly interesting to run a temporal survey following the health state of V. mirabilis during an entire production cycle to have a more precise overview of fish farm impacts on this species, including during and after the post-production fallowing period.

Pyrroloquinoline quinone protects against murine hepatitis virus strain 3-induced fulminant hepatitis by inhibiting the Keap1/Nrf2 signaling

Fulminant hepatitis (FH) is a life-threatening clinical liver syndrome characterized by substantial hepatocyte necrosis and severe liver damage. FH is typically associated with severe oxidative stress, inflammation, and mitochondrial dysfunction. Pyrroloquinoline quinone (PQQ), a naturally occurring redox cofactor, functions as an essential nutrient and antioxidant and reportedly inhibits oxidative stress and exerts potent anti-inflammatory effects. In the present study, we aimed to evaluate the therapeutic efficacy of PQQ in murine hepatitis virus strain 3 (MHV-3)-induced FH and examined the underlying mechanism. An MHV-3-induced FH mouse model was established for in vivo examination. Liver sinusoidal endothelial cells (LSECs) were used for in vitro experiments. Herein, we observed that PQQ supplementation significantly attenuated MHV-3-induced hepatic injury by suppressing inflammatory responses and reducing oxidative stress. Mechanistically, PQQ supplementation ameliorated MHV-3-induced hepatic damage by down-regulating the Keap1/Nrf2 signaling pathway in vivo and in vitro. Furthermore, Nrf2 small interfering RNA targeting LSECs abrogated the PQQ-mediated protective effects against MHV-3-related liver injury. Our results deepen our understanding of the hepatoprotective function of PQQ against MHV-3-induced liver injury and provide evidence that alleviating oxidative stress might afford a novel therapeutic strategy for treating FH.

Determination of Maximum Tolerable Cold Ischemia Time in a Mouse Model of Cervical Heterotopic Uterus Transplantation.

Uterus transplantation (UTx) is an emerging treatment for uterine factor infertility. Determining the maximum tolerable cold ischemia time is crucial for successful UTx. However, the limit for cold ischemia in the uterus is unclear. This study aimed to examine cold ischemia's effects on mouse uteri and identify the maximum cold ischemia duration that uteri can endure. We systematically assessed the tolerance of mouse uteri to extended cold ischemia, 24 h, 36 h, and 48 h, using the cervical heterotopic UTx model. Multiple indicators were used to evaluate ischemia-reperfusion injury, including reperfusion duration, macroscopic examination, oxidative stress, inflammation, and histopathology. The function of transplants was evaluated through estrous cycle monitoring and embryo transfer. Mouse uteri subjected to 48 h of cold ischemia exhibited significant delays and insufficiencies in reperfusion, substantial tissue necrosis, and loss of the estrous cycle. Conversely, uteri that underwent cold ischemia within 36 h showed long survival, regular estrous cycles, and fertility. Our study demonstrated that mouse uteri can endure at least 36 h of cold ischemia, extending the known limits for cold ischemia and providing a pivotal reference for research on the prevention and treatment of cold ischemic injury in UTx.

Effectiveness of Rhenium(I)-diselenoether Low Doses in a Triple-negative Breast Cancer Chicken Embryo Model.

Rhenium(I)-diselenoether (Re-diSe) is a promising anticancer agent composed of one rhenium and two selenium atoms. Its effectiveness was established in inhibiting cancer cells while maintaining low toxicity toward normal cells at a 5 μM dose for 120 hours in MDA-MB-231 cells. In MDA-MB-231 breast tumor-bearing mice, anti-tumor and anti-metastatic effects were observed at a 10 mg/kg dose. However, contradictory results were observed in the 4T1 breast cancer model, where a dose of 60 mg/kg had a pro-tumor effect. To address these discrepancies, the efficacy of Re-diSe at the effective 10 mg/kg dose was validated in a transplanted MDA-MB-231 breast tumor model using the chicken chorioallantoic membrane assay. MDA-MB-231 cancer cells were xenografted onto the chicken chorioallantoic membrane (CAM), and daily drug administration was carried out for nine days at doses of 0.1, 1, and 10 mg/kg. At the study's conclusion, a standard histological analysis was conducted. The low dose of 0.1 mg/kg showed a significant reduction in tumor weights compared to controls. The 1 mg/kg dose resulted in an increased inflammation score but did not induce a significant difference in tumor weights compared to the 0.1 mg/kg dose. Notably, at the 10 mg/kg dose, six out of 11 treated embryos displayed no visible signs of tumors. These tumors exhibited extensive tumor necrosis and significant infiltration by inflammatory cells. In this particular model, the anticancer efficacy of Re-diSe was achieved at the low dose of 0.1 mg/kg. The higher dose of 10 mg/kg, while eliminating visible tumors, might have immune-mediated effects, as indicated by substantial tumor necrosis and infiltration by inflammatory cells. Overall, this study successfully demonstrated the effectiveness of Re-diSe as an anticancer agent.

Phonozen-mediated photodynamic therapy comparing two wavelengths in a mouse model of peritoneal carcinomatosis.

This study assessed the therapeutic efficacy of intraperitoneal photodynamic therapy (PDT) using photosensitizer activation at two different wavelengths, 405 and 664nm, in a mouse model of peritoneal carcinomatosis. The dark and light cytotoxicity of chlorin e6-polyvinylpyrrolidone (Phonozen) were measured in vitro under 402 ± 14 and 670 ± 18nm LED activation in bioluminescent human gastric cancer cells, MKN45-luc. Cell viability was measured at 6h after irradiation using the PrestoBlue assay. Corresponding in vivo studies were performed in athymic nude mice by intraperitoneal injection of 1 × 106 MKN45-luc cells. PDT was performed 10 d after tumor induction and comprised intraperitoneal injection of Phonozen followed by light irradiation at 3h, delivered by a diffusing-tip optical fiber placed in the peritoneal cavity and coupled to a 405 or 664nm diode laser to deliver a total energy of 50J (20 mice per cohort). Whole-body bioluminescence imaging was used to track the tumor burden after PDT out to 130days, and 5 mice in each cohort were sacrificed at 4h post treatment to measure the acute tumor necrosis. Photosensitizer dose-dependent photocytotoxicity was higher in vitro at 405 than 664nm. In vivo, PDT reduced the tumor growth rate at both wavelengths, with no statistically significant difference. There was substantial necrosis, and median survival was significantly prolonged at both wavelengths compared with controls (46 and 46 vs. 34 days). Phonozen-mediated PDT results in significant cytotoxicity in vitro as well as tumor necrosis and prolonged survival in vivo following intraperitoneal light irradiation. Blue light was more photocytotoxic than red in vitro and had marginally higher efficacy in vivo.

Revolutionizing RCC Immunotherapy to Unlock Predictive Biomarkers

Revolutionizing RCC Immunotherapy to Unlock Predictive Biomarkers

Magnetophoretic Intranasal Drug-loaded Magnetic Nano-aggregates as a Platform for Drug Delivery in Status Epilepticus.

Status epilepticus is associated with substantial morbidity and neuronal necrosis, and the duration of the seizure would affect its following complications. Eliminating the duration would have valuable outcomes; however, the presence of BBB is an obstacle. The purpose of the current study was to achieve a nose-to-brain magnetic drug delivery system to accelerate the onset of action, and to reduce the mucociliary clearance via implementing the magnetic field. The drug-entrapped magnetic nanoaggregates were prepared via a 2-step method, synthesis of the magnetic nanoparticles and drug loading. Optimization of the variables, including ammonium hydroxide:water ratio, beta-cyclodextrin%, duration of the mixing time, amount of Pluronic, and drug:magnetic nanoaggregates mass ratio was performed according to particle size, PDI, zeta potential, release profile and entrapment efficiency. The efficacy of optimized formulation was assessed in the animal model. According to the analysis performed by the software, drug-to-nanoparticle ratio and the duration of mixing time were found to be significantly effective (p < 0.05) for entrapment efficiency and particle size distribution, respectively. The optimum formulation with an approximate average size of 581 nm and 61% entrapment efficiency was obtained, which released about 80% of its drug content within the first 20 minutes. The in vivo efficacy was significantly improved (p < 0.05) by administration of magnetic nanoaggregates in the presence of a simple external magnet placed on the glabellar region of the animals, compared to the control groups. This drug delivery system could be suggested as a fast-acting alternative for seizure cessation in status epilepticus emergencies.

Abstract PD11-02: PD11-02 A Phase II Randomized Window of Opportunity Trial Evaluating Cytotoxic and Immunomodulatory effects of Intratumoral INT230-6 in Early Stage Breast Cancer: the INVINCIBLE Trial

Abstract Background: The majority of breast cancers outside of the triple negative subtype are considered immunological quiescent and are therefore minimally responsive to immunotherapies. One potential method to combat this is through local therapies that induce cell death, thereby exposing tumor antigens, providing adjuvants for anti-tumor immune priming, and potentially increasing responsiveness to immunotherapies. We have conducted a randomized, Phase 2 presurgical Window-Of-Opportunity trial for intratumoral (IT) INT230-6 (comprising VINblastine (VIN) Cisplatin (VIN)) evaluating clinical and BioLogical Effects in patients with early-stage operable Breast Cancer (the INVINCIBLE trial- https://clinicaltrials.gov/ct2/show/NCT04781725). INT230-6 contains a dispersion enhancer molecule (SHAO) with the cytotoxic agents and is designed to cause tumor necrosis by dispersion throughout the tumor and diffusion into cancer cells. Previous in vitro studies have demonstrated that INT230-6 halts cancer cell replication and induces cell death recruiting dendritic cells and T-cells to the tumor microenvironment. In this trial, IT injections of INT230-6 are conducted to 1) exploit the potential of regional cytotoxic chemotherapy on breast cancer in vivo and 2) assess the immune response within the tumor, microenvironment and systemically in the host blood prior to surgical resection. Methods: Women with newly diagnosed and awaiting surgery for early-stage intermediate or high-grade T1-T2 invasive breast cancers were recruited to the trial. The study has two parts. Part I was a randomized (2:1) open label trial comparing 1-3 doses of INT230-6 injected weekly versus no treatment prior to surgery to evaluate safety, feasibility, and optimal drug dosing. Part II was a double-blinded randomized (2:1) trial where patients received one IT dose of INT230-6 vs saline injection. The primary objective was to estimate the proportion of patients with tumor necrosis and complete cell cycle arrest (CCCA) at the time of surgery compared to control. In addition, we performed targeted sequencing and proteomic profiling in tumour samples from the INT230-6 clinical trial. Results: The study recruited 90 patients with age ranges of 40-77 yrs (mean = 60 yrs) with tumors ranging from 1.5-4.3 cm (mean = 2.4cm). No surgeries were delayed or altered as a result of trial participation and the most common (&amp;gt;10%) AEs were injection site pain, injection site reaction and nausea/vomiting. Compared to the control group, up to 95% tumor necrosis was present in varying biologic subtypes and histologies, including invasive lobular carcinoma. Preliminary gene expression analysis showed significant differential gene expression between the baseline biopsy and surgical specimens. Pathway analysis identified genes associated with TCR signaling, B cells, T cells, chemokine signaling and NF-κB signaling were significantly changed in the post treatment samples. There was a relative increase in CD4 and CD8 T cells and B and NK cells within the tumor and in the tumour microenvironment. Conclusion: Preliminary evidence shows that a single dose of INT230-6 can cause substantial tumor necrosis and stimulate an immune response in breast cancers prior to surgery with minimal adverse effects and good tolerability. This window of opportunity clinical trial demonstrates that INT230-6 injection is a novel and simple method to convert traditionally immune quiescent breast cancers into immunogenic tumors. This can open the door to future potential immunotherapeutic options in early stage breast cancer. Citation Format: Angel Arnaout, Susan Robertson, Kianoosh Keyhanian, Megan Hopkins, Linda Liao, Vida Talebian, Arif Awan, John MS Bartlett, Gregory R. Pond, Lazlo Radvanyi, Lewis H. Bender, Ian B. Walters, Vanessa Lopez Ozuna, Melanie Spears. PD11-02 A Phase II Randomized Window of Opportunity Trial Evaluating Cytotoxic and Immunomodulatory effects of Intratumoral INT230-6 in Early Stage Breast Cancer: the INVINCIBLE Trial [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr PD11-02.

Differences between cardiac troponin I vs. T according to the duration of myocardial ischaemia.

Cardiac troponin T (cTnT) and troponin I (cTnI) are expressed as an obligate 1:1 complex in the myocardium. However, blood levels of cTnI often rise much higher than that of cTnT in myocardial infarction (MI), whereas cTnT is often higher in patients with stable conditions such as atrial fibrillation. Here we examine high-sensitive (hs) cTnI and hs-cTnT after different durations of experimental cardiac ischaemia. hs-cTnI, hs-cTnT, and the hs-cTnT/hs-cTnI ratio were measured in plasma samples from rats before and at 30 and 120 min after 5, 10, 15, and 30 min of myocardial ischaemia. The animals were killed after 120 min of reperfusion, and the infarct volume and volume at risk were measured. hs-cTnI, hs-cTnT, and the hs-cTnT/hs-cTnI ratio were also measured in plasma samples collected from patients with ST-elevation myocardial infarction (STEMI). hs-cTnT and hs-cTnI increased over 10-fold in all rats subjected to ischaemia. The increase of hs-cTnI and hs-cTnT after 30 min was similar, resulting in a hs-cTnI/hs-cTnT ratio around 1. The hs-cTnI/hs-cTnT ratio was also around 1 in blood samples collected at 120 min in rats subjected to 5 or 10 min of ischaemia where no localized necrosis was observed. In contrast, the hs-cTnI/hs-cTnT ratio at 2 h was 3.6-5.5 after longer ischaemia that induced cardiac necrosis. The large hs-cTnI/hs-cTnT ratio was confirmed in patients with anterior STEMI. Both hs-cTnI and hs-cTnT increased similarly after brief periods of ischaemia that did not cause overt necrosis, whereas the hs-cTnI/hs-cTnT ratio tended to increase following longer ischaemia that induced substantial necrosis. A low hs-cTnI/hs-cTnT ratio around 1 may signify non-necrotic cTn release.

Tumor metabolism derived from 18F-FDG PET/CT in predicting the macrotrabecular-massive subtype of hepatocellular carcinoma

The recently described pathological subtype of hepatocellular carcinoma (HCC), named macrotrabecular massive (MTM), is associated with an unfavorable prognosis. This study aimed to evaluate the potential for tumor metabolism obtained by β-2-[18F] fluoro-2-deoxy-D-glucose positron emission tomography/computed tomography (18F-FDG PET/CT) to be used as a preoperative imaging indicator for predicting MTM-HCCs. This study was designed to be cross-sectional. Patients who underwent preoperative 18F-FDG PET/CT and who had surgically-diagnosed HCC between June 2015 and June 2021 were retrospectively included. Tumor metabolism was determined by the tumor-to-normal liver standardized uptake value ratio (TLR) of the primary tumor as shown on 18F-FDG PET/CT. Clinical, pathological, and PET/CT characteristics were compared between non-MTM-HCCs and MTM-HCCs. Univariate analyses were used to screen the predictive factors of MTM-HCCs, then multivariate binary logistic regression analyses were performed. A regression-based diagnostic model was then established. Substantial necrosis was assessed to compare the predictive performance between traditional imaging and TLR measured on 18F-FDG PET/CT. The receiver operating characteristic (ROC) curve analyses and the DeLong test were used to assess the predictive performance. A total of 93 patients (mean age, 52.6±11.3 years; 81 male) with 36 MTM-HCCs were included. Multivariate binary logistic regression analyses identified higher platelet count [PLT; ≥118.5×103/µL; odds ratio (OR), 3.63; 95% confidence interval (CI), 1.13-12.87; P=0.035], higher aspartate transaminase (AST; ≥52 IU/L; OR, 4.15; 95% CI: 1.34-14.33; P=0.017), and larger TLR (≥2.2; OR, 5.55; 95% CI: 1.90-17.56; P=0.002) as independent predictors of MTM-HCCs. A TLR ≥2.2 helped to identify 72.2% of the MTM-HCCs with a specificity of 75.4%. The AUC of the regression-based diagnostic model for predicting MTM-HCCs was 0.835 (95% CI: 0.746-0.923), with a sensitivity of 80.6% and a specificity of 78.9%. Substantial necrosis enabled the identification of MTM-HCCs with 52.8% sensitivity and 87.7% specificity, with an AUC of 0.702 (95% CI: 0.588-0.817). There was no statistical difference between TLR and substantial necrosis in predicting MTM-HCCs using the DeLong test (P>0.05). Tumor metabolism determined by TLR on 18F-FDG PET/CT is a valuable imaging indicator for MTM-HCCs. Noninvasive prediction of this subtype can achieve good sensitivity and excellent predictive performance based on the regression model of AST, PLT, and TLR.

RETRACTED: The heterogeneity of PD-L1 protein in gastric cancer: expression and distribution characteristics

Programmed death receptor ligand 1 (PD-L1) is expressed at different levels in tumour tissues and tumour-infiltrating monocytes (TIMCs). The interpretation of PD-L1 expression in gastric cancer (GC) is more difficult because of its heterogeneity. The PD-L1 immunohistochemistry (IHC) by E1L3N assay was performed in GC tissues. The level and distributed characteristics of PD-L1 expression were observed to illustrate its heterogeneity both in the cancer tissues and TIMCs. The relationship between PD-L1 level and necrotic features of tumor cells, the number of TIMCs, the distribution of tertiary lymphoid tissue (TLS) in the stroma, and other clinicopathological factors were analysed. A Cox regression model was used to assess the prognostic value of PD-L1 expression. Of the 110 GC samples, not only more cases (51/110 cases) could be detected by combined positive score (CPS) for PD-L1 expression compared the other two, tumour positive score (TPS), and mononuclear immune-cell density score (MIDS), but also there were more cases with the high level of PD-L1 expression by CPS, even if with good consistency among them (P < 0.05). The tumour cells with high expression of PD-L1 was prone to show a diffuse distributing, whereas mottled type in the low level. It was noteworthy that the strongly colored tumor cells tended to exhibit a mossy pattern which were distributed along the border between cancer nests and stroma, and the same pattern happened to occur in the positive mesenchymal cells contacting the tumor border, essentially lymphocytes and macrophages. The substantial necrosis in the tumour and the number of TIMCs was analyzed statistically significant correlated with CPS (P < 0.05), while other clinicopathological factors such as histological type, tumour size, invasion depth, TNM stage were uncorrelated. The number and distribution of TLS in the tumour and para-tumoural stroma indirectly affected PD-L1 in GC by associating with the quantity and pattern of TIMCs. Cox regression analysis revealed that the prognosis was poor when PD-L1 was positive. CPS is the best indicator for PD-L1 expression in GC, which tend to be increased expression following a large number of TIMCs and substantial tumour necrosis appeared. Heterogeneity was reflected in the different distributed pattern of PD-L1 expression, especially the mossy-like pantten of the staining tumor cell in the interface between tumour nests and stroma, regardless of the amount and intensity of PD-L1 expression. TLS is valuable for observing microscopic images to influence the quantity and pattern of TIMCs. CPS can be used as an independent prognostic factor for GC.

MRI-based preoperative markers combined with narrow-margin hepatectomy result in higher early recurrence

PurposeTo investigate the applicability of MRI-based preoperative risk markers in assisting clinicians to define an appropriate surgical margin width for patients with solitary hepatocellular carcinoma. MethodsPatients who underwent preoperative MRI with hepatectomy were randomly divided into development (65%) and internal validation (35%) datasets between January 2015 and January 2019. Multivariate logistic analysis was used to evaluate MRI-based markers of early recurrence (≤2 years) in the development dataset. Independent factors in the development dataset were investigated using a multivariable Cox analysis. The multivariable logistic and Cox models were verified using the risk score system in the validation dataset. Recurrence-free survival (RFS) was calculated according to MRI-based preoperative markers together with a narrow or wide margin in all datasets. ResultsA narrow resection margin was identified as an independent risk factor for early postoperative recurrence (P < 0.001) according to multivariable Cox analysis. RFS was significantly shorter in patients with narrow resection margins than that of those with wide resection margins (P < 0.005). Patients with the three MRI-based preoperative markers (tumour size > 5 cm, substantial necrosis, and non-smooth margins) combined with a narrow resection margin had a shorter RFS than that of those with a wide margin (P < 0.005). Patients without these markers also benefitted from a wide margin (P < 0.05). ConclusionsMRI-based preoperative risk markers in combination with narrow resection margins were associated with a higher recurrence rate compared with wide resection margins, and hence, such patients may benefit from a wide-margin hepatectomy.

Differential impact of doxorubicin dose on cell death and autophagy pathways during acute cardiotoxicity

Doxorubicin (DOX) is an effective anthracycline used in chemotherapeutic regimens for a variety of haematological and solid tumors. However, its utility remains limited by its well-described, but poorly understood cardiotoxicity. Despite numerous studies describing various forms of regulated cell death and their involvement in DOX-mediated cardiotoxicity, the predominate form of cell death remains unclear. Part of this inconsistency lies in a lack of standardization of in vivo and in vitro model design. To this end, the objective of this study was to characterize acute low- and high-dose DOX exposure on cardiac structure and function in C57BL/6 N mice, and evaluate regulated cell death pathways and autophagy both in vivo and in cardiomyocyte culture models. Acute low-dose DOX had no significant impact on cardiac structure or function; however, acute high-dose DOX elicited substantial cardiac necrosis resulting in diminished cardiac mass and volume, with a corresponding reduced cardiac output, and without impacting ejection fraction or fibrosis. Low-dose DOX consistently activated caspase-signaling with evidence of mitochondrial permeability transition. However, acute high-dose DOX had only modest impact on common necrotic signaling pathways, but instead led to an inhibition in autophagic flux. Intriguingly, when autophagy was inhibited in cultured cardiomyoblasts, DOX-induced necrosis was enhanced. Collectively, these observations implicate inhibition of autophagy flux as an important component of the acute necrotic response to DOX, but also suggest that acute high-dose DOX exposure does not recapitulate the disease phenotype observed in human cardiotoxicity.

NIMG-54. RADIOMIC FEATURES FROM LESION HABITAT PREDICT RESPONSE TO COMBINATION OF NIVOLUMAB AND BEVACIZUMAB IN PATIENTS WITH RECURRENT GLIOBLASTOMA: A FEASIBILITY STUDY

Abstract PURPOSE The use of immunotherapy in glioblastoma management is under active investigation. Glioblastomas are “cold” tumors, meaning that they have inactivated or fewer tumor infiltrative lymphocytes in addition to substantial tumor necrosis, attributing to their poor response to immunotherapy. A significant challenge is the apriori identification of Glioblastoma patients who will respond favorably to immunotherapy. In this work, we evaluated the ability of computerized MRI-based quantitative features (radiomics) extracted from the lesion habitat (including enhancing lesion, necrosis, and peritumoral hyperintensities) to predict response and progression-free survival (PFS) in recurrent GBM patients treated with combination of Nivolumab and Bevacizumab. METHODS Immunotherapy response assessment in neuro-oncology (iRANO) criteria along with PFS were used to analyze n=50 patients enrolled in a randomized clinical trial where patients received Nivolumab with either standard or low dose Bevacizumab. These patients were assessed to see if they had complete response, partial response, stable disease (i.e. responders, n=31), or disease progression (i.e. non-responders, n=19). Lesion habitat constituting necrotic core, enhancing tumor, and edema were delineated by expert radiologist on Gd-T1w, T2w and FLAIR MRI scans. COLIAGE radiomic features from each of the delineated regions were selected using minimum redundancy maximum relevance (mRMR) via cross-validation, to segregate non-responder patients from responders. A multivariable cox proportional hazard model was used to predict survival (PFS). RESULTS CoLlAGe correlation, sum average, and sum variance features (capture local heterogeneity) from the lesion habitat, were found to segregate non-responder patients from responders with an accuracy of 86%, followed by 80% using features from peritumoral hyperintensities and 78% from enhancing tumor. In our survival analysis, C-index of 0.688 was obtained using features from the entire lesion habitat, followed by peritumoral hyperintensities (0.675) and enhancing tumor (0.656). CONCLUSION Radiomic features from the lesion habitat may predict response to combination of Nivolumab and Bevacizumab in recurrent Glioblastomas.