Effects of Brazilian green propolis and artepillin C on collagen metabolism and fibroblast behaviors: Implications for skin wound healing.

Osteoarthritis (OA) is a aging-associated degenerative joint disease. The objective was to determine relative senescence gene expression in joints and leukocytes of OA horses toward considering senotherapeutics to manage OA. To define local (joint) and systemic (peripheral blood mononuclear cells [PBMCs]) senescence burden, synovial fluid cell single-cell RNA sequencing and PBMC mRNA sequencing datasets (n = 65 samples) were examined. Differential analyses were conducted using limma to compare OA versus control. A custom 3,043-gene senescence set curated from published metadata was applied to differential analyses to investigate senescence-specific pathways. Senescence genes were divided into 8 categories; scores were calculated with fast gene set enrichment analysis with P value computed via permutation and log2 fold change ranks. Synovial fluid single-cell RNA sequencing data revealed cell type-specific heterogeneity in senescence gene expression. Fast gene set enrichment analysis pathway analysis confirmed enrichment/upregulation in inflammatory and stress-induced senescence in dendritic, cycling, CD8 T, and gamma delta T cells. Senescence-associated secretory phenotype pathways were predominantly represented in cycling cells. Senescence genes aryl hydrocarbon receptor (AHR), IL-1 receptor antagonist (IL1RN), heme oxygenase 1 (HMOX1), plasminogen activator, urokinase receptor (PLAUR), and tissue inhibitor of metalloproteinase 1 (TIMP1) were upregulated in multiple synovial fluid cell types. In contrast, genes in most senescence categories were downregulated in PBMCs. Senescence pathways were differentially expressed in aged horses with OA, with upregulation of senescence genes in the joint and downregulation in PBMCs. Therapeutic strategies targeting senescent cells may be a disease-modifying strategy to treat equine OA.

Stroke remains a significant global cause of death and long-term disability, categorized into hemorrhagic stroke, ischemic stroke, and transient ischemic attack. Recovery outcomes can vary greatly, often influenced by genetic factors. This review highlights key genes involved in stroke recovery, including brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), insulin-like growth factor-1 (IGF-1), vascular endothelial growth factor (VEGF), C-X-C motif chemokine ligand 12 (CXCL12), hypoxia-inducible factor 1-alpha (HIF1A), nuclear factor erythroid 2-related factor 2 (NRF2), nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing 3 (NLRP3), sirtuin 1 (SIRT1), and tissue inhibitor of metalloproteinase 3 (TIMP3). These genes play important roles in neurotrophic support and neuronal survival, angiogenesis, inflammation and immune modulation, and extracellular matrix remodeling after stroke. Insights from both human and animal studies underscore the potential of these genetic markers as prognostic indicators and therapeutic targets for stroke recovery. Understanding these factors may lead to more personalized rehabilitation strategies, and future research is needed to explore gene-environment interactions and translate genetic findings into effective stroke recovery therapies.

Matrix metalloproteinases and tissue inhibitors of metalloproteinases as the biomarkers of Alzheimer's disease: A meta-analysis.

Background/Objectives: Early detection of pancreatic ductal adenocarcinoma (PDAC) can improve patient survival and biomarkers to facilitate this are greatly needed. We recently reported a serum biomarker signature comprising tissue inhibitor of metalloproteinase 1 (TIMP1), intercellular adhesion molecule 1 (ICAM1), cathepsin D (CTSD), thrombospondin 1 (TSP1/THBS1), and carbohydrate antigen 19-9 (CA 19-9), that detected Stage I and II PDAC with high sensitivity and specificity. In this assay, CA 19-9 is measured with a commercial instrument and individual ELISAs were developed to measure TIMP1, ICAM1, CTSD, and THBS1. Here, we report the analytical performance of these four analytes in their ELISA formats. Methods: Biomarker precision, linearity, algorithm precision, matrix effects, hook effect, method comparison, interference, and analyte stability were evaluated against acceptance criteria per CLSI guidelines. Results: High, medium, and low concentrations of each biomarker met acceptance criteria for inter- and intra-day precision (%CVs < 14%) and for linearity (%CVs < 11%). Matrix effects did not impact quantitation of any analyte nor was hook effect present. All analytes met acceptance criteria for accuracy and stability (all biases < 11.2% and <16.5%, respectively). For interference, two CTSD measurements and one ICAM1 measurement in HAMA-spiked samples showed 20.7-29% biases, falling slightly outside of acceptance criteria (<20% bias). All other analyte concentrations met interference acceptance criteria. In total, 94.1% of all diagnostic calls were made with 100% certainty, indicating high precision of the assay's algorithm. Conclusions: All analytes demonstrated acceptable analytical precision, linearity, accuracy, and stability, showing high overall analytical performance of each analyte.

CRIP1 exacerbates osteoarthritis progression by recruiting UBE3A to induce the ubiquitination-mediated degradation of MFGE8.

Neuroprotective efficacy of royal jelly and propolis against cadmium-induced dysregulation of neurogenesis and neurotransmitters in the brain of rats: Molecular mechanisms and ultrastructure investigations.

ABSTRACT Beta-glucans (βTGs) are a class of dietary fibers and biologically active polysaccharides derived from natural sources, known for their diverse bioactive properties. Their documented effects include anti-tumor, anti-inflammatory, prebiotic, anti-obesity, anti-allergic, anti-microbial, antiviral, anti-osteoporotic, and immunomodulating activities. Despite these well-established benefits, the role of βTG in dehydroepiandrosterone (DHEA)-induced polycystic ovary syndrome (PCOS) remains largely unexplored. This study investigated the protective effects of βTG treatment on PCOS and its potential to reverse PCOS-induced changes. Female Sprague-Dawley (SD) rats were randomly divided into four groups (n = 8 each): control, PCOS, PCOS+βTG, and βTG. We assessed biochemical markers related to oxidative stress, antioxidant status, inflammation, cytokines, and hormone levels. Additional analyses included immunohistochemistry and histopathology. Membrane array analysis was used to profile growth factors, cytokines, and chemokines. However, βTG normalized deviations in the estrous cycle caused by PCOS and positively affected the reproductive system (p < 0.05). It also reduced the inflammatory response in PCOS rats by decreasing inflammatory cytokines (p < 0.05). Furthermore, oxidative stress was significantly reduced, and antioxidant enzyme activities were markedly elevated in the βTG group (p < 0.05). Histopathological alterations were prevented by βTG, which also induced the expression of essential proteins such as beta-nerve growth factor (bNGF), tissue inhibitor of metalloproteinase-1 (TIMP-1), Agrin, cytokine-induced neutrophil chemoattractant-1 (CINC-1), brain-derived neurotrophic factor (BDNF), and basic fibroblast growth factor (FGF-2/bFGF) (p < 0.05). In conclusion, βTG treatment effectively protects against oxidative stress, inflammation, hormone imbalance, and histopathological damage in ovarian tissue caused by PCOS.

Tamoxifen remains a first-line treatment for estrogen receptor-positive breast cancer. Emerging evidence indicates that aberrant 5-methylcytosine (m5C) modification of RNAs contributes to chemotherapeutic resistance in various types of cancer, however their role in determining tamoxifen resistance in breast cancer remains elusive. We measured global m5C RNA methylation and expression of its regulating enzymes in tamoxifen-sensitive (MCF-7) versus tamoxifen-resistant cells (MCF-7 Tam1). Expression of the most significantly dysregulated enzyme NOP2/Sun RNA methyltransferase family member 7 (NSUN7), the key m5C writer methyltransferase, was depleted using siRNA-mediated knockdown. Functional assays were performed to measure sensitivity to tamoxifen, cell migration, and colony-forming potential. RNA sequencing followed by enrichment and network analysis identified NSUN7-regulated pathways and hub genes. Finally, the prognostic relevance of hub genes was assessed using the Gene Expression Profiling Interactive Analysis platform. Tamoxifen-resistant cells exhibited a significant elevation in global m5C levels and expression of NSUN7. siRNA-mediated reduction of NSUN7 significantly restored sensitivity to tamoxifen, reducing the half-maximal inhibitory concentration by ~50%, and significantly inhibited cell migration and colony-forming potential. Transcriptomic profiling and enrichment analysis identified that NSUN7 targets were enriched in crucial pathways, including mitogen-activated protein kinase pathway, phosphatidylinositol signaling, cell cycle, and focal adhesion. Notably, NSUN7 depletion caused dysregulation in the expression of genes implicated in tamoxifen resistance, such as brain acid-soluble protein 1 (BASP1), tissue inhibitor of metalloproteinase 3 (TIMP3), ajuba LIM protein (AJUBA), S-phase kinase-associated protein 2 (SKP2) and yes-associated protein 1 (YAP1). Network analysis further identified NSUN7-regulated hub genes significantly associated with disease prognosis. Our study identified that NSUN7 regulates key oncogenic pathways associated with tamoxifen resistance and metastasis. Its suppression enhanced tamoxifen sensitivity and reduced metastatic potential, collectively highlighting NSUN7 as a novel driver of tamoxifen resistance and a potent therapeutic target in estrogen receptor-positive breast cancer.

The pathogenesis of aortic aneurysm (AA) is characterized by chronic inflammation of the aortic wall, the associated accumulation of macrophages, and degradation of the extracellular matrix, including elastin. Colchicine (COL) has long been known for its anti-inflammatory effects, so in this study we investigated its effects on AA. In vitro, tumor necrosis factor (TNF)-α-stimulated macrophages and vascular smooth muscle cells (VSMCs) were treated with and without COL for 24 h. Unstimulated cells were used as controls. COL significantly reduced interleukin (IL)-1β, TNF-α, monocyte chemotactic protein (MCP)-1, nuclear factor kappa B (NF-κB), matrix metalloproteinase (MMP)-9, and activated caspase-1 in macrophages, and increased lysyl oxdase (Lox) and tissue inhibitor of metalloproteinase (TIMP)-2 expression in VSMCs. In vivo, aged male apolipoprotein E-deficient (ApoE-/-) mice were infused with angiotensin II (Ang II) for 28 days. The mice received either normal saline (NS) or COL orally. The control group of ApoE-/-mice did not receive Ang II infusion or treatment. COL significantly suppressed aortic enlargement and reduced AA incidence by preserving elastin and decreasing IL-1β, TNF-α, MCP-1, NLRP3 inflammasome, neutrophil elastase, and myeloperoxidase expression. No significant differences were observed in the enzymatic activities of MMP-2 and MMP-9 between the 2 groups. The results suggested that COL prevents AA progression in a clinically relevant model and is expected to be a novel preventive agent for AA.

Inguinal hernia represents a multifactorial condition driven by extracellular matrix (ECM) dysregulation, collagen imbalance, and oxidative stress. Across studies, a consistent reduction in the collagen I:III ratio, coupled with altered expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), underpins weakened fascia and hernia susceptibility. Aging further impairs ECM remodeling through fibroblast senescence, cross-linking deficits, and elastic fiber attrition, while oxidative stress and inflammation amplify tissue degradation and impair repair mechanisms. Evidence from clinical and experimental studies underscores the interplay between surgical technique, mesh choice, redox balance, and recurrence risk. Understanding the combined impact of aging and oxidative stress provides a mechanistic framework for targeted therapeutic and surgical strategies aimed at preventing hernia development and recurrence.

Novel heart failure (HF) pharmacotherapies, including angiotensin receptor-neprilysin inhibitor (ARNI) and sodium-glucose cotransporter 2 inhibitors (SGLT2is), may confer cardiovascular benefits by attenuating myocardial fibrosis. However, direct evidence from human failing myocardial samples is limited. Thus, we assessed the associations of ARNI and SGLT2i therapies with fibrosis on myocardial samples from advanced HF patients undergoing heart transplantation (HTX). Ninety-three patients receiving stable combined HF pharmacotherapy (uninterrupted use of β-blockers, mineralocorticoid receptor antagonists, angiotensin-converting enzyme inhibitors or ARNI, and optionally SGLT2is) without the need for pharmacological or mechanical circulatory support for at least three months pre-HTX, were enrolled. Standardized regions of the left ventricular anterior wall from explanted hearts were analysed. The primary outcome was the percentage of interstitial myocardial fibrosis area measured via histology. The expression of pro-fibrotic and pathological remodelling markers was quantified with qRT-PCR. Both ARNI and SGLT2i therapies were associated with reduced interstitial collagen accumulation in adjusted analyses. None of the tested clinical parameters (sex, age, serum creatinine, presence of hypertension or diabetes) interacted with the effects of ARNI or SGLT2i on fibrosis. Additionally, no interaction was observed between ARNI and SGLT2i use regarding collagen content. Among the examined genes, ARNI was linked to a decreased beta-to-alpha myosin heavy chain expression ratio, while in SGLT2i-treated patients, reduced mRNA levels of matrix metalloproteinase 9 (MMP9) and tissue inhibitor of metalloproteinases 2 (TIMP2) were observed in adjusted models. ARNI and SGLT2i therapies might exert anti-fibrotic effects in advanced HF.

Wound healing is a complex process involving coordinated actions of multiple cell types. Therefore, when developing therapeutics to promote wound healing, it is essential to consider the synergistic contributions of various cells at different stages of the healing process. In this study, we evaluated the potential of different extracts of Botryocladia leptopoda as wound-healing agents by examining their effects on antioxidant activity, cytotoxicity, cell migration, anti-inflammatory properties, and expressions of specific biomarkers associated with wound healing. Results indicated that the ethanol extract (FE) and hexane extract (HE) exhibited the highest DPPH radical scavenging activity, reaching up to 94%. The alkaline extract (AE) showed the strongest antioxidant ability in the FICA assay, with a maximum of 99%. In addition, the FE and AE provided anti-inflammatory actions that inhibited tumor necrosis factor (TNF)-α and interleukin (IL)-6 in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Further analyses suggested that the FE and AE enhanced cell proliferation (210% and 112%) and migration (442.2% and 535.6%) and regulated wound healing-related genes, including matrix metalloproteinase 2, MMP9, and tissue inhibitor of metalloproteinase 2 (TIMP2) to avoid scar formation and accelerate wound healing. Lastly, the identification of potential compounds within the extract using the UHPLC system further supports its prospective medical applications. Taken together, these findings indicated that the FE and AE from B. leptopoda exhibited remarkable in vitro wound-healing properties, highlighting their potential for applications in pharmaceutical industries and health food development.

BackgroundPolypharmacy, defined as taking ≥ 5 different daily medications, is common in older adults and has been linked with neuropsychiatric/neurological and other health conditions. To clarify the potential molecular implications, we tested the hypothesis that polypharmacy may influence DNA methylation (DNAm) patterns in aging, in a longitudinal Italian cohort (N = 1,098; mean (SD) age at recruitment: 58.8 (5.6) years, 51.3% women; median (IQR) follow-up 12.6 (1.1) years).ResultsWe tested associations of polypharmacy with several DNAm aging clocks (Hannum, Horvath, GrimAge, DNAmPhenoAge, DunedinPACE), through linear mixed models incrementally adjusted for age, sex, education, prevalent health conditions and lifestyles, leukocyte counts and residual batch effects. This revealed significant positive associations of GrimAge acceleration and DunedinPACE with the switch to polypharmacy status during follow-up (Beta (SE): 0.024 (0.008) and0.0012 (0.0004)). While the association of GrimAge was driven by a DNAm-based surrogate of tissue inhibitor metalloproteinase 1 (TIMP-1), no significant association was detected for component CpGs of DunedinPACE. When we tested associations of polypharmacy with 668,413 CpGs epigenome-wide, we observed no statistically significant findings (top hit: cg07675998; chr11q13.1; Beta (SE) = 0.009 (0.002); p = 1.5 × 10–6). However, these showed significant enrichments of several biological functions and pathways related to renal tissue, lipoproteins, inflammatory and immune response.ConclusionsThese findings suggest an influence of polypharmacy on accelerated epigenetic aging and on altered methylation patterns in the genome, suggesting a potential implication of pathways related to renal tissue development, lipoproteins and cholesterol homeostasis, inflammatory and immune response, in line with previous proteomic analyses of polypharmacy mouse models. These observations also suggest potential targets for mitigating disruptive effects of polypharmacy on elderly health.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13148-025-02007-7.

Abstract Glioblastoma patients in Japan can receive bevacizumab (Bev) from the time of initial diagnosis—a unique clinical strategy rarely adopted worldwide. This enables sequential molecular analyses using paired tumor samples from same patients obtained before, during, and after Bev exposure. To investigate resistance mechanisms to anti-angiogenic therapy, we performed qPCR-based gene expression analysis for Annexin A2 (ANXA2), metalloproteinase-2 (MMP2), metalloproteinase-9 (MMP9), tissue inhibitor of metalloproteinases-1 (TIMP1) and plasminogen activator inhibitor-1 (PAI-1) using RNA extracted from 68 Formalin-fixed Paraffin-embedded tumor samples from 33 patients with glioblastoma, including 32 paired specimens (initial surgery and refractory samples). We first focused on the paired analysis. In patients who received neoadjuvant Bev (Effective Bev) before initial surgery, comparison between primary and refractory tumors revealed significantly elevated PAI-1 expression in refractory specimens (p = 0.005), with a trend toward TIMP1 upregulation (p = 0.129). This molecular shift was not observed in pairs without neoadjuvant Bev exposure (Naïve Bev). ANXA2 and MMP9 expression negatively correlated with Bev cycle number (ρ = -0.456, p = 0.009; ρ = -0.395, p = 0.026). ANXA2 expression was higher in tumors treated with fewer than 15 Bev cycles (p = 0.037), and MMP9 was upregulated in T2 circumscribed MRI pattern (p = 0.030). Moreover, it was statistically shown that the expression level of MMP9 influences PFS (0.008). These findings suggest that prolonged Bev exposure might affect resistance by inducing invasive gene expression and distinct imaging phenotypes. Paired analyses further revealed that these molecular changes were more evident in patients who received neoadjuvant Bev. Integrating paired molecular profiles with treatment and imaging data may inform future therapeutic strategies for recurrent glioblastoma by identifying biomarkers of resistance and tailoring duration for anti-angiogenic treatment.

Background: Chronic kidney disease (CKD) has been linked to elevated inflammatory biomarkers, yet few studies have evaluated these associations using iothalamate-based measures of kidney function, highlighting the need for further research. Research question: Is CKD associated with elevated inflammatory markers when defined by measured glomerular filtration rate (mGFR) and urinary albumin-to-creatinine ratio (UACR)? Methods: Participants from the Genetic Epidemiology Network of Arteriopathy (GENOA) cohort were included if they had data available for at least one inflammatory marker at visit 2. 17 inflammatory markers were assessed, including C-reactive protein (CRP), interleukin-6 (IL-6), IL-18, tumor necrosis factor receptors 1 and 2 (TNFR1, TNFR2), serum amyloid A, intercellular and vascular adhesion molecules (ICAM, VCAM), E-selectin, P-selectin, monocyte chemoattractant protein (MCP), myeloperoxidase (MPO), receptor for advanced glycation end products (RAGE), matrix metalloproteinases 2 and 9 (MMP-2, MMP-9), and tissue inhibitors of metalloproteinases 1 and 2 (TIMP-1, TIMP-2). Kindey function was assessed using the urinary clearance of iothalamate. CKD was defined as mGFR &lt; 60 mL/min/1.73 m 2 or UACR ≥ 30 mg/g. To further assess whether associations persisted without albuminuria, we also defined CKD solely by mGFR &lt; 60 mL/min/1.73 m 2 . Generalized estimating equations (GEE) were used to assess associations between CKD status and inflammatory markers, accounting for familial clustering. Exponentiated beta coefficients from GEE models were used to present geometric mean (GM) ratios of each biomarker by CKD status, adjusted for demographics, lifestyle factors, cardiovascular risk factors, and medication use. Results: Among 1,010 participants (mean age 61 ± 10 years; 67% women; 48% Black adults), 207 were classified as having CKD. After adjusting for potential confounders, individuals with CKD had significantly higher levels of hsCRP, VCAM, TNFR1, and TNFR2 compared to those without CKD (GM ratios &gt; 1; p &lt; 0.05), while the remaining inflammatory markers showed no significant differences ( Figure 1 ). Importantly, these elevations remained significant when CKD was defined by mGFR alone ( Figure 2 ). Conclusion: Our findings suggest that CKD, defined by mGFR, is associated with specific inflammatory biomarkers, underscoring the need for mechanistic studies to investigate these associations.

Background: Activation of hepatic stellate cells (HSCs) in response to liver injury increases extracellular matrix (ECM) deposition and upregulation of tissue inhibitor of metalloproteinase-2 (TIMP-2), promoting fibrogenesis, a key feature of metabolic dysfunction-associated fatty liver disease (MAFLD). This study was conducted to determine the diagnostic potential of serum TIMP-2 levels in patients with MAFLD/nonalcoholic fatty liver disease (NAFLD).Methods: This case-control study included 100 MAFLD patients and 100 healthy controls. Demographic profiling, ultrasonography, and FibroScan with controlled attenuation parameter (CAP) scoring were conducted. Serum TIMP-2 was quantified using enzyme-linked immunosorbent assay (ELISA), and routine biochemical parameters (liver function test, kidney function test, fasting plasma glucose, postprandial blood sugar, HbA1c, total cholesterol, triglycerides, and GGT) were assessed using fully automated chemistry analyzers. Receiver operating characteristic (ROC) curve analysis was performed to assess the diagnostic utility of TIMP-2.Results: Serum TIMP-2 levels were significantly higher in MAFLD patients (p < 0.001) compared with controls. Serum TIMP-2 showed a significant positive correlation with ALT (rho = 0.569, p < 0.001), GGT (rho = 0.551, p < 0.001), fasting plasma insulin (rho = 0.562, p < 0.001), and FibroScan scores (r = 0.560, p < 0.001). ROC analysis yielded an AUC of 0.933 (95% CI: 0.889-0.963) with a TIMP-2 cut-off value of 8.909, showing a sensitivity of 91% and specificity of 99%.Conclusion: Serum TIMP-2 levels are significantly elevated in MAFLD and show strong correlations with liver enzymes and CAP scores, suggesting a relationship with hepatic fat accumulation and liver injury. Additionally, associations with lipid levels, white blood cell count, and neutrophils imply a link to systemic inflammation and liver fibrosis, highlighting the promising role of TIMP-2 as a noninvasive, blood-based biomarker for early detection and monitoring of liver fibrosis. Further studies are needed to substantiate the diagnostic potential of serum TIMP-2 in MAFLD patients.

IntroductionMatrix metalloproteinases (MMPs) are enzymes participating in tumorigenesis and tumor progression through their proteolytic and cell-signaling properties. They are regulated mostly by endogenous tissue inhibitors of metalloproteinases (TIMPs). The expression of both MMPs and TIMPs is often altered in cancers. Many studies investigated their potential as circulating cancer biomarkers, with confounding results, which might be induced by preanalytical issues, particularly by using serum instead of plasma. The study aims to investigate plasma levels of selected MMPs and TIMPs in association with the diagnosis and prognosis of colorectal cancer.MethodsThe clinico-pathological data of 148 patients operated for colorectal cancer were collected from the medical records system at the University Hospital Pilsen. Sixty-eight age-matched healthy subjects were included as controls. Plasma levels of MMP-2, -7, -8, -9, -10, and TIMP-1, -2, -3, and -4 were assessed with multiplex immunoassays with the technology xMAP.ResultsMMP-8 and -9 levels were significantly elevated in patients (P= 0.0002 and 0.0009, respectively), TIMP-2 levels were significantly decreased in colorectal cancer patients (P = 0.0016). When comparing the early colorectal cancer (stage I and II) with advanced colorectal cancer (stage III and IV), MMP-8 and TIMP-1 were significantly increased in advanced colorectal cancer (P = 0.0173 and <0.0001, respectively). The area under the curve and the receiver operating characteristic were between 0.680 and 0.530 for all studied biomarkers. In univariate analysis, overall survival was significantly elevated in patients with MMP-7, -8, or TIMP-1, which was higher than cut-offs (hazard ratio = 4.57, 2.03, and 7.64, respectively).ConclusionThese findings suggest that plasma MMP-7, MMP-8, and TIMP-1 are potential prognostic biomarkers for colorectal cancer . None of the investigated biomarkers revealed diagnostic potential.

TIMP1: A novel immune-related signature associated with invasiveness and inhibition of pituitary adenoma.

Multifunctionality of tissue inhibitor of metalloproteinase-1 in central nervous system diseases.