Background: Healthy non-pharmacological lifestyle factors, such as regular physical exercise and dietary supplementation, have been shown to significantly improve cognitive outcomes over time compared to a more sedentary lifestyle and poor diet. Furthermore, exercise may serve as a potential protective factor in attenuating the effects associated with cognitive decline that are characteristic of neurodegenerative disorders, such as Alzheimer's disease (AD). Evidence indicates that certain dietary interventions can also attenuate the effects of neurodegeneration and positively impact longevity. Supplementation with polyphenols such as ellagic acid (EA), which is abundant in pomegranate juice, may help provide neuroprotective and longevity benefits.

Methods: This study examined the potential protective potential of EA and exercise and provides insight into the combined use of a polyphenol-rich diet and exercise to enhance behavioral outcomes and lifespan in a transgenic Drosophila melanogaster (fruit fly) model of AD with the Aβ₄₂ gene.

Results: Fruit flies subjected to a 120-minute exercise regimen performed better on a climbing assay than flies that did not exercise. Conversely, flies that exercised for 30 min passed marginally more trials on a learning and memory assay using an aversive stimulus than flies that did not exercise, whereas both groups performed better than flies subjected to the more intense exercise condition.

Conclusion: These results suggest a hormetic effect of exercise regarding memory performance. Finally, flies fed a low dose of dietary EA (0.24 mg/mL) lived significantly longer than flies fed the control diet or higher concentrations of EA, again suggesting a hormetic effect of EA on longevity.

Background: Alzheimer's disease (AD) represents the most prevalent neurodegenerative disorder, with mitochondrial dysfunction being observed in both AD patients and mouse models. Nonetheless, further investigation is required to elucidate the pathogenic genes associated with AD and to develop early diagnostic methodologies centered on mitochondrial function.

Methods: In this study, the dataset GSE132903 was retrieved from the GEO database, encompassing both non-demented (ND) control and AD samples. Through the combination of differential expression gene analysis, weighted gene co-expression network analysis, and intersection with mitochondrial database gene sets, four hub genes associated with AD were identified. These four hub genes were subsequently validated in APP/PS1 and 5xFAD mouse models using molecular biology techniques.

Results: The hub genes identified through bioinformatics analysis include SYNJ2BP, VDAC1, NUBPL, and COX19. Within the GSE132903 dataset, the expression levels of SYNJ2BP, NUBPL, and COX19 were significantly elevated in the AD group compared to the non-demented (ND) group, whereas VDAC1 expression was reduced in the AD group relative to the ND group. Furthermore, in the hippocampus of APP/PS1 and 5xFAD mouse models, the expression patterns of SYNJ2BP and NUBPL were consistent with the bioinformatics analysis results.

Conclusion: Hub genes identified here through bioinformatics and molecular biology may help early diagnosis of AD patients and may also help build new AD models to explore its pathogenesis.

Background: This research aimed to examine the impact of co-transplantation of ethyl stearate (PubChem CID: 8122) and neural stem cells (NSCs) on endogenous NSCs in Parkinson's disease (PD) model rats, and also determine the impact of ethyl stearate on quiescent neural stem cells (qNSCs) and explore its possible mechanism.

Methods: The induction of PD rats was achieved through the injection of 6-hydroxydopamine (6-OHDA) into the right striatum (STR) and substantia nigra (SN). Transplant treatment was then followed for 6 weeks in the right STR. Apomorphine (APO)-induced rotation behavior and the pole climbing test were used to assess the effects of transplant treatment in each group. Western blotting (WB) and immunofluorescence staining were used to observe changes in endogenous NSCs. The effect of ethyl stearate on qNSCs was examined in vitro, with detection of associated indicators achieved through WB, immunofluorescence, flow cytometry and qRT-PCR.

Results: Co-transplantation of NSCs with ethyl stearate significantly ameliorated motor deficits in PD rats and upregulated the expression of tyrosine hydroxylase (TH), Sox2, and Nestin. We established an in vitro system for NSC quiescence and activation. In qNSCs, ethyl stearate treatment increased Ki67 positivity and reduced the proportion of cells in the G0–G1 phase of the cell cycle, whereas withdrawal of ethyl stearate exerted the opposite effects. Ethyl stearate also downregulated EphB2 protein levels and upregulated p-AKT and CyclinD1 in qNSCs; these effects were reversed upon its removal.

Conclusion: These findings imply that co-transplantation of NSCs and ethyl stearate may help ameliorate motor impairment in PD rats by activating endogenous NSCs and promoting their development into dopaminergic neurons. Ethyl stearate may regulate the entry and exit of NSCs from quiescence through the EphB2/AKT/CyclinD1 pathway.

Background: The absence of effective animal models for sporadic Alzheimer's disease (AD) remains a pivotal barrier to therapy development. Because methanol metabolism produces endogenous formaldehyde, a neurotoxic agent linked to cognitive decline, this study investigated whether chronic, low-dose methanol exposure could recapitulate AD-like pathology and cognitive deficits in rhesus monkey, thereby establishing a nonhuman primate animal model driven by this environmental-metabolic insult.

Methods: Adult rhesus monkeys received low-concentration methanol for 9 months. Behavioral tests for cognition, locomotion, sleep, and vision were conducted. Postmortem analyses involved histopathological examination, immunohistochemistry, immunofluorescence, and Western blot to evaluate neuronal integrity, microglial activation, and the expression of key proteins associated with AD (amyloid-β [Aβ], phosphorylated tau, TAR DNA-binding protein 43 [TDP-43]) and cellular stress (synaptic markers, mitochondrial fission, autophagy, and apoptosis-related proteins).

Results: Chronic methanol exposure led to progressive cognitive and memory impairment without significant motor or visual deficits. Neuropathology revealed brain atrophy, neuronal loss, synaptic damage, microglial activation, and mitochondrial structural disorganization. Critically, the exposed animals exhibited hallmark AD-like molecular alterations, including increased Aβ deposition, tau hyperphosphorylation, and TDP-43 dysregulation. Furthermore, neurotoxicity was associated with elevated urinary formaldehyde, enhanced mitochondrial fission, increased autophagy, and elevated apoptosis.

Conclusion: Chronic low-dose methanol exposure in rhesus monkeys recapitulates progressive cognitive deficits and AD-like neuropathological features. This model, driven by endogenous formaldehyde toxicity, effectively mimics key aspects of sporadic AD. Our findings shed light on the neurotoxic mechanisms of methanol and propose a reproducible and translationally relevant nonhuman primate model for studying AD pathogenesis and evaluating potential therapeutics.

The incidence of benign airway stenosis (BAS) is on the rise, and current treatment options are associated with a significant risk of restenosis. Therefore, there is an urgent need to explore new and effective prevention and treatment methods. Animal models serve as essential tools for investigating disease mechanisms and assessing novel therapeutic strategies, and the scientific rigor of their construction and validation significantly impacts the reliability of research findings. This paper systematically reviews the research progress and evaluation systems of BAS animal models over the past decade, aiming to provide a robust foundation for the optimized construction of BAS models, intervention studies, and clinical translation. This effort is intended to facilitate the innovation and advancement in BAS prevention and treatment strategies.

Cynomolgus macaques, a species of Old World primate native to southeastern and eastern Asia and the island of Mauritius, are one of the most important nonhuman primate models for infectious disease. Although the closely related rhesus macaque is classified into subspecies based on geographic origin, no such subdivision exists for cynomolgus macaques, and they continue to be used interchangeably in infectious disease research, reducing the comparability of data produced from these studies. Research into the population genetics of cynomolgus macaques has found significant differences between macaques native to different areas, including their genetic diversity, with macaques from insular populations such as Mauritius and the Philippines exhibiting highly restricted heterozygosity compared to mainland populations native to Indonesia or Cambodia. In the context of infectious disease studies, research into pathogens, including Ebola virus, Crimean-Congo hemorrhagic fever virus, and Mycobacterium tuberculosis have found differences in study outcomes, survival times, and immune cell responses between different populations of macaques. This review provides an overview of the differences between cynomolgus macaque populations in the context of genetic diversity, and in response to infection, and highlights the need for clear reporting of geographic origin of primates used in research. This will improve data comparison between studies and help to further refine this important animal model.

Background: The golden Syrian hamster is a valuable animal model for studying carcinogenesis, metabolic disorders, cardiovascular diseases, and viral infections due to its biological and pathological similarities to humans. However, the development of genetically engineered hamsters has lagged behind that of mice and rats, largely because of an embryonic development block at the two-cell stage in vitro. Although CRISPR/Cas9-mediated gene knockout has been achieved in hamsters, precise DNA fragment insertion or conditional knockout (cKO) models have not previously been reported, likely due to technical limitations in embryo manipulation and insufficient efficiency of homology-directed repair (HDR).

Methods: In this study, we generated conditional alleles of the ApoF gene in golden Syrian hamsters. A two-cut strategy was applied using Cas9 protein, two sgRNAs, and a single donor plasmid containing exon 2 flanked by loxP sites and two ~0.8 kb homology arms. A mixture of Cas9 protein, sgRNAs, and the donor plasmid was microinjected into the pronuclei of one-cell stage hamster embryos.

Results: The efficiency of CRISPR/Cas9-mediated loxP knock-in reached up to 27%, and the genetically modified floxed alleles were successfully transmitted through the germline. The functionality of the inserted loxP sites was validated by in vivo Cre-mediated recombination following local administration of AAV vectors, including AAV-cTnT-Cre in the heart and AAV-CMV-Cre in the brain.

Conclusions: To our knowledge, this work represents the first successful establishment of a conditional knockout model in the golden Syrian hamster, providing a valuable tool for mechanistic studies of gene function and disease modeling.

Background: Integrins facilitate binding to the extracellular matrix and other cells. Their subunit β2 is exclusively expressed by leukocytes, binds to the intercellular cell adhesion molecule 1 (ICAM-1), and is pivotal for their recruitment to sites of inflammation such as the atherosclerotic plaque.

Methods: To investigate β2-integrin–mediated adhesiveness, a well-established assay for human whole blood was adapted for the analysis of murine T cell subsets. Changes in avidity and affinity were assessed by incubation of murine complexes ICAM-1 in murine whole blood and consecutive stimulation with PMA and Mg²⁺/EGTA. Underlying signaling pathways in β2-integrin–mediated adhesiveness upon chemokine stimulation with CCL-19 were identified by incubation with reducing substances, and a Ca²⁺ chelator and ROS and Ca²⁺ measurements were carried out.

Results: Incubation of murine whole blood with PMA leads to 30-fold and Mg^2+/EGTA to 65-fold increase in β2-integrin–mediated adhesiveness of T cells. Specificity of the assay was proven by preincubation of a blocking antibody, leading to a 60% reduction in adhesion capacity. ROS species and Ca²⁺ are crucial for chemokine-mediated β2-integrin activation. In vivo relevance was proven by induction of T cell adhesiveness in whole blood of mice upon myocardial infarction.

Conclusions: Our assay allows specific quantification of β2-integrin–mediated affinity and avidity of T cells in whole blood samples. In congruence to human adhesion, these mechanisms are ROS and Ca²⁺ dependent and significantly elevated after myocardial infarction. Our refined and robust assay may be of particular use in phenotyping involved mechanisms in T cell activation in atherosclerotic cardiovascular disease.

Background: The analgesic effects of multiple electroacupuncture (EA) sessions and single EA sessions differ significantly in pain management. Area 24b (A24b) of the anterior cingulate cortex (ACC) is crucial in pain processing. EA relieves pain by targeting and modulating the neuronal activity within this subregion. However, whether the cumulative effect of EA antinociception is connected to A24b mechanisms has remained unclear.

Methods: In our study, we used the Complete Freund's Adjuvant (CFA) model to induce inflammatory pain and the Spared Nerve Injury (SNI) model to induce neuropathic pain, and adult male C57BL/6, FosTRAP, and FosTRAP:Ai9 mice were used as experimental subjects to investigate the cumulative effect of EA antinociception and whether multiple EA sessions and a single EA session regulate different neuronal populations in the A24b.

Results: We observed that EA effectively alleviated pain in mice, with three EA sessions yielding superior analgesic effects compared to a single session. Using chemical genetics combined with FosCreER technology to activate EA-TRAPed cells in the A24b, we found that pain relief was more pronounced with three EA sessions. Moreover, chemogenetic inhibition of EA-TRAPed cells in the A24b reversed the analgesic effects of a single EA session but not those of three EA sessions. Fluorescent in situ hybridization results indicated that three EA sessions significantly increased the number of GABAergic neurons in the A24b compared with a single session. Additionally, retrograde tracing revealed that the A24b circuit that monosynaptically innervates EA-TRAPed cells included projections from the central lateral nucleus (CL), lateral mediodorsal thalamic nucleus (MDL), lateral habenula (LHb), dorsal raphe nucleus (DR), caudal linear nucleus of the raphe (CLi), dorsal tuberomamillary nucleus (DTM), periventricular hypothalamic nucleus (Pe) and hippocampal fields CA1, CA2, and CA3. These findings suggest that multiple EA sessions and single EA sessions activated different neuronal populations in the A24b. The enhanced analgesic effect of multiple EA sessions may be attributed to an increase in the proportion of GABAergic neurons within the A24b.Conclusions: Multiple and single EA sessions recruit distinct neuronal populations in A24b, with the stronger analgesic effect of repeated EA linked to a higher proportion of GABAergic neurons in this region.

Background: The development of relevant and robust large animal models of hepatocellular carcinoma is needed to test new therapeutic strategies for this disease. Transgenic approaches hold promise in addressing this complex problem. One such model, the Oncopig, has been reported to develop tumors of up to 4 cm in diameter within 7–14 days at sites of in situ vector inoculation. However, the resulting lesions reportedly contained an extensive inflammatory component that has not been evaluated in detail.

Methods: Herein, we describe our results from multiparametric characterization of the lesions generated using liver biopsy cores incubated in vector solution and replaced in the tissue. The study consisted of 3 animals in 3 cohorts (total of 9 animals) that were evaluated at 14, 21, and 28 days. CT imaging, immunohistochemistry, multiplex immunofluorescence, and comprehensive blood analyses were used to quantify composition of the hepatic masses that developed following AdCre inoculation.

Results: The tumors were hypovascular on CT and predominantly composed of CD45+ cells with a strong lymphohistiocytic component, with no carcinomas identified. Ki-67 staining showed proliferation of CD45+ immune cells but no neoplastic component. To provide further insight, the results are evaluated in the context of tumor growth kinetics.

Conclusion: While progress has been made in generating targetable lesions, achieving a robust large animal model of liver cancer that faithfully recapitulates the human disease remains a challenging goal.

Background: Chronic hyperuricemia is associated with complications such as gout and uric acid nephropathy, but uric acid also exhibits biological activities (e.g., antioxidant effects, potential neuroprotective properties against neurodegenerative diseases). Nonhuman primates are ideal models for studying neurodegenerative diseases; however, existing nonhuman primate hyperuricemia models cannot sustain long-term elevated serum uric acid levels, nor recapitulate the impaired uric acid excretion observed in clinical hyperuricemic patients.

Methods: First, we detected uricase expression in cynomolgus monkeys and compared it with that in mice. Then, we established a cynomolgus monkey hyperuricemia model by administering a mixture of potassium oxonate, hydrochlorothiazide, and adenine via fruits and vegetables. We further analyzed the regulatory effects of this model on uric acid metabolism (synthesis, degradation, and excretion) and the expression of uric acid transporter genes in the intestine and kidney.

Results: Cynomolgus monkeys express functional uricase, but at a lower level than mice. The established model maintained stable, long-term hyperuricemia by three mechanisms: increasing intestinal and renal uric acid excretion load, inhibiting hepatic uric acid degradation, and promoting uric acid synthesis. Additionally, the model downregulated the expression of intestinal/renal uric acid-secreting transporter genes, while upregulating uric acid-reabsorbing transporter genes.

Conclusions: This novel cynomolgus monkey hyperuricemia model provides a new tool for investigating the association between hyperuricemia and neurodegenerative diseases, and will help clarify the mechanism by which serum uric acid influences cognitive function.

Realistic models for cancer research representing disease progression that commensurately respond to therapeutics consistent with clinical observation are the holy grail for pre-clinical research and screening. Although such an ideal is elusive, well-characterized in vivo models facilitate our understanding of disease, progression, and therapeutic opportunities. Here, we characterize a commonly used syngeneic BALB/c mouse model of triple negative breast cancer (4T1) after establishing tumors in their flanks. Tumors developed at the subcutaneous injection site for all experimental mice and their volumes were monitored. We quantified a rare subset of breast cancer stem-like cells (CSCs), classified as CD44⁺/CD24⁻ phenotypes in in vitro and ex vivo cell populations. Chromosome numbers in ex vivo metaphase cells were greater than cells cultured in vitro (89.4 ± 3.4, range of 70–132 and 82.6 ± 1.1, range of 70–128; respectively). Further, we observed different types of chromosome aberrations, including gap, deletion, exchange, interstitial deletion, terminal deletion, ring, dicentric, and Robertsonian translocations. For both sources of cells, the number of aberrations was dominated by deletions, terminal deletions, and Robertsonian translocations. Ex vivo cells exhibited greater prevalence of deletions and terminal deletions, whereas in vitro cells displayed more ring aberrations and Robertsonian translocations. In conclusion, we successfully characterized cancer cells from a syngeneic mouse model of breast cancer in terms of rare CSC proportion and a variety of chromosomal aberrations, which is useful for understanding tumor traits associated with cancer development and therapeutic action. The data act as a valuable resource for other studies using the 4T1 BALB/c model.

Background: Humanized mouse models are essential for studying the human immune response and antibody development. However, conventional models show limited B cell maturation and antigen-specific humoral responses. To overcome these limitations, we used the NOG-EXL mice expressing human interleukin 3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) to enhance myeloid and B-cell lineage differentiation.

Methods: Human CD34⁺ hematopoietic stem cells (HSC) were transplanted into NOG-EXL mice to produce humanized immune systems. After immune cell reconstitution was confirmed across 12 weeks, the mice were immunized twice with inactivated severe fever with thrombocytopenia syndrome virus (SFTSV) antigens. Peripheral blood mononuclear cells and splenocytes were analyzed using multicolor flow cytometry to assess human immune cell subsets. Antigen-specific immunoglobulin G (IgG) production was quantified using enzyme-linked immunosorbent assay (ELISA), and virus-specific B cells were isolated using antigen-labeled recombinant protein probes.

Results: Twelve weeks after transplantation of HSCs into NOG-EXL mice, they exhibited robust engraftment of human leukocytes, including T, B, and dendritic cells, compared to NOG mice. Unlike NOG mice, humanized NOG-EXL mice exhibited an increase in human IgG levels, indicating the production of human antibody responses to antigens. Humanized NOG-EXL mice were immunized twice every 2 weeks with inactivated SFTSV, and antigen-specific human antibodies against the virus were detected in the mouse sera by ELISA. Sera from SFTSV-immunized humanized mice demonstrated neutralizing activity against SFTSV, confirming the induction of functional virus-specific neutralizing antibodies. Antigen-binding IgG-positive human B cells were isolated from mouse splenocytes using recombinant protein probes.

Conclusion: This model provides a valuable platform for evaluating humoral immunity and isolating B cells using high-affinity human monoclonal antibodies without genetic engineering.

Stroke induces profound neuroinflammation and systemic immune dysregulation, including disturbances in gut homeostasis. Experimental evidence suggests that intestinal barrier permeability (IBP) and bacterial translocation (BT) critically influence stroke outcomes. However, biological variability among commonly used rodent substrains has received limited attention. In this pilot study, we compared poststroke immune responses in two Wistar rat substrains obtained from different suppliers: RccHan (Envigo) and RjHan (Janvier). Naive animals (n = 4) and rats subjected to permanent cerebral ischemia (n = 8 per substrain) were evaluated 72 h after middle cerebral artery occlusion and stratified according to the presence or absence of BT. Immune cell populations in blood and bone marrow were analyzed using flow cytometry, and leukocyte infiltration into ischemic brain tissue was quantified using immunohistochemistry. Differences were considered statistically significant when p < 0.05. Both substrains developed significant infarcts and neurological deficits. RccHan rats exhibited larger infarct volumes and more extensive BT across multiple organs. In contrast, RjHan rats exhibited BT mainly confined to mesenteric lymph nodes but exhibited greater IBP. Although dissemination was broader in RccHan rats, overall bacterial burden was slightly lower compared with RjHan, and extraintestinal bacterial composition differed between groups. Particularly, RjHan rats exhibited stronger systemic and central immune activation, with significant alterations in lymphocyte and monocyte populations and enhanced granulocyte and T-cell infiltration within ischemic lesions. These findings demonstrate that substrain origin profoundly influences poststroke intestinal barrier integrity, bacterial dissemination, and immune responses considering substrain-related variability is essential to improve reproducibility and translational relevance in preclinical stroke research.

A lupus-like condition induced by intraperitoneal administration of pristane (2,6,10,14-tetramethylpentadecane) in mice is widely used as a model of systemic lupus erythematosus (SLE). Due to their phylogenetic distance from humans, murine models are not always suitable tool for studying the specific activity of therapeutic agents and the pathogenesis of SLE. In order to overcome species-specific limitations of murine models, this approach was tested in non-human primates—cynomolgus monkeys (Macaca fascicularis). Two intraperitoneal injections at a dose of 3.5 mL/kg, administered at weeks 1 and 23, recapitulated SLE features, including: production of antinuclear autoantibodies (ANA), membranoproliferative glomerulonephritis with immune complex (IC) deposition in the glomeruli. However, from week 27 five of eight pristane-treated monkeys developed progressive respiratory failure. Two of these died at week 28 and the remaining were euthanized at week 32. The histology of the monkey lungs suggested exogenous lipoid pneumonia. Thus, while pristane induced serological autoimmunity and characteristic renal manifestations in Macaca fascicularis, the consequent lipoid pneumonia limited the observation period and prevented comprehensive evaluation of SLE manifestations beyond 32 weeks.

Background: The traditional method of heterotopic abdominal heart transplantation (HTx) involves cross-clamping the inferior vena cava, which inevitably leads to bilateral lower limb ischemia (LI). This study first aimed to investigate the impact of LI on renal function in rats subjected to unilateral nephrectomy (UNx). Second, a modified method utilizing renal vessel–assisted anastomosis in rats with left UNx was compared with the traditional method for abdominal HTx.

Methods: Male Sprague–Dawley rats were utilized as subjects for both experimental phases. In experiment 1, the animals were divided into four groups: sham operation group; LI group—rats undergoing occlusion of the abdominal aorta and vena cava below the renal vessels; UNx group—rats with left UNx; and LI + UNx group. All operated animals were monitored for up to 7 days for biochemical markers, renal histopathology, and survival rates. In experiment 2, we introduced the renal vessel–assisted method as the experimental group and compared it against the traditional method as the control within rat heterotopic HTx models. We assessed operative characteristics, echocardiography results, histological findings, and graft survival.

Results: First, LI resulted in acute kidney dysfunction characterized by a decrease in 7-day survival rates and creatinine clearance rates in both the LI and LI + UNx groups compared to the sham operation and UNx groups. Particularly, histopathological damage in the kidney and liver did not exhibit significant effects during this period. Second, the implementation of the renal vessel–assisted method significantly reduced bleeding volume at suture sites and enhanced the 7-day survival rate compared to the traditional method.

Conclusion: Acute kidney injury was induced by LI postoperation in treated rats. The renal vessel–assisted method demonstrated its effectiveness as a superior alternative that mitigates complications associated with the traditional method.

Phrenic nerve stimulation (PNS) may preserve diaphragm activation and mitigate multiorgan injury during mechanical ventilation (MV); however, a minimal invasive rat model integrating PNS with MV is lacking. We established an omohyoid muscle-based PNS rat model combined with MV. Bilateral nerves were exposed within 20 ± 2 min by transection at the intermediate tendon of omohyoid muscle, minimizing trauma and bleeding. Threshold stimulation (0.6 ± 0.2 mA) correlated with body weight. Ventilator-synchronized stimulation increased compound muscle action potentials by ~30%, whereas histology confirmed intact nerve. Physiological parameters remained stable throughout ventilation. This model provides a safe and scalable platform for mechanistic and preclinical studies on PNS-mediated protection against MV-induced organ injury.

As cardiovascular disease is the leading cause of global mortality, innovative animal models are vital to demonstrating the translational value of experimental discoveries. Investigations focused on myocardial remodeling after ischemia reperfusion (I/R) are well suited to a porcine model, but the evolution of that injury and its impact on electrical conductivity or arrhythmia threshold have been difficult to monitor continuously. Multiple electrode telemetry devices may be fitted to the animals but are costly and prone to damage. Implantable telemetry devices are likewise expensive, carry surgical risk, and are often verified only for single use. Here, we report the utilization of the commercially available Fourth Frontier X2 external telemetry device for continuous monitoring of heart rate and rhythm after myocardial I/R injury in pigs, highlighting sustained monitoring across a 7-day study.