Search Results (884)

Background/Objectives: Fluoroquinolones cause Achilles tendinopathy within a narrow, early therapeutic window, yet preventive strategies given alongside these antibiotics remain largely untested. This exploratory pilot study assessed whether four mechanistically distinct adjuvants attenuated histopathological tendon change in a rabbit model of levofloxacin exposure. Methods: Twenty-one male New Zealand White rabbits received oral levofloxacin (100 µg/kg/day, seven days). A contralateral-limb design yielded 42 tendons (two cohorts provided paired limb-specific comparisons and a third cohort received the systemic intervention). Animals were allocated to various groups, receiving platelet-rich plasma (PRP), injectable porcine collagen, oral vitamin E + selenium, topical essential oils with augmented soft-tissue mobilization (ASTM), or a levofloxacin-only control. The main outcome was a modified Bonar composite (tenocyte morphology, cellularity, vascularity, paratendinous fibrosis; 0–12) at three months; additional outcomes were the individual domains and ultrasonographic tendon thickness. Results: Composite scores were low throughout (group medians 2.0–4.0) and did not differ between groups (H = 3.663, p = 0.45); no domain differed, and the untreated control did not show the greatest change. Within animals, collagen-treated tendons scored higher than the paired essential-oil tendons (median difference +3.0, nominal p = 0.031, Bonferroni-adjusted p = 0.062). Tendon thickness increased by day 14 only with vitamin E + selenium (+14.1%; p = 0.003, and p = 0.006 adjusted for baseline); this finding arose in a separate cohort with thinner baseline tendons, cannot be confidently attributed to the intervention, and was unrelated to histopathology (ρ = −0.02, p = 0.91). Macroscopic findings differed between groups (nominal p = 0.031) without predicting tissue grade. Conclusions: No statistically detectable difference in the study-specific four-domain composite was identified among the treatment conditions; the study lacked a levofloxacin-free control and could not provide a definitive efficacy assessment. The model produced only mild pathology, limiting its power, and neither imaging nor gross inspection predicted tissue-level change. These preliminary observations are hypothesis-generating and are intended to inform the design of an adequately powered confirmatory study. The findings are inconclusive for efficacy; the design cannot distinguish lack of adjuvant efficacy from failure of the model to induce measurable disease. Full article

Background: Patients with stage IIB–IIIB cutaneous melanoma remain at substantial risk of recurrence despite complete surgical resection. Accurate risk stratification is essential to optimize patient selection for adjuvant therapy and surveillance strategies. We aimed to evaluate the clinicopathological and molecular factors associated with recurrence in a real-world cohort of high-risk melanoma patients. Methods: We retrospectively analyzed 97 patients with stage IIB–IIIB cutaneous melanoma treated at a single tertiary referral center. Clinicopathological characteristics, molecular features, treatment-related variables, and sentinel lymph node (SLN) pathological characteristics were evaluated. Recurrence-free survival (RFS) was estimated using the Kaplan–Meier method. Independent prognostic factors were identified using multivariable Cox proportional hazards regression. Results: After a median follow-up of 88 months, 31 patients (32.0%) developed recurrence. Stage IIC showed the highest recurrence rate in the cohort (52.4%), compared with 25% in stage IIIA and 31.4% in stage IIIB. Consistently, in the multivariable analysis, compared with stage IIB, only stage IIC remained independently associated with worse recurrence-free survival (hazard ratio [HR] = 3.57, 95% confidence interval [CI]: 1.08–11.82; p = 0.037), whereas neither stage IIIA nor stage IIIB differed significantly from stage IIB. Female sex was independently associated with a lower risk of recurrence (HR 0.33, 95% CI 0.14–0.79; p = 0.012). Although BRAF V600E-mutated tumors showed higher recurrence rates and shorter recurrence-free survival in univariable analyses, BRAF V600E status was not independently associated with recurrence in the multivariable model (p = 0.250). Higher lymphoid infiltration density was associated with improved RFS in univariable analysis, although this finding was limited by substantial missing data and non-standardized pathological assessment. Conclusions: In this exploratory retrospective analysis, female sex and stage IIC were independently associated with RFS. These findings should be interpreted cautiously given the small sample size, limited number of recurrence events, and incomplete availability of some clinicopathological and molecular variables. The prognostic associations observed for BRAF V600E status and lymphoid infiltration, as well as the analyses according to adjuvant treatment, should be considered exploratory and hypothesis-generating. The unfavorable outcomes observed in stage IIC disease support growing evidence that this subgroup represents a biologically aggressive form of melanoma despite the absence of regional nodal involvement. Our findings do not challenge the validity of the AJCC staging system but suggest that clinically relevant heterogeneity may exist within established AJCC stage groups. Full article

Background: Antimicrobial resistance (AMR) is among the most pressing challenges in modern infectious medicine, driving progressive failure of standard antibacterial therapy and rising mortality from infections caused by multidrug-resistant (MDR) pathogens. Antibiotic potentiation through adjuvant compounds capable of restoring the activity of existing drugs represents a promising strategy to overcome resistance without developing fundamentally new antibacterial molecules. This study aimed to evaluate the antibiotic-potentiating activity of a dextrin/polyvinyl alcohol/iodine complex (D/PVA/I-1), developed at the Scientific Center for Anti-Infectious Drugs JSC (Almaty, Kazakhstan), against clinically relevant MDR reference strains. Methods: Nine reference strains, Staphylococcus aureus (ATCC 33591, BAA-39), Escherichia coli (ATCC BAA-196, BAA-2523), Klebsiella pneumoniae (ATCC BAA-2524, 700603), Acinetobacter baumannii (ATCC BAA-1790), Streptococcus pneumoniae (ATCC BAA-660), and Haemophilus influenzae (ATCC 33930), and two clinical isolates, P. aeruginosa SCAID PHRX1-2019 and E. coli SCAID WND1-2021, were tested. Antibiotic-potentiating activity was assessed by checkerboard assay with calculation of the fractional inhibitory concentration index (FICI); bactericidal kinetics were evaluated by time-kill analysis. The effect of D/PVA/I-1 on cytoplasmic membrane permeability was investigated using a crystal violet uptake assay. Results: Of 45 D/PVA/I-1–antibiotic combinations tested across nine antibiotics, synergy (FICI ≤ 0.5) was demonstrated in 44.4% of cases, partial synergy in 48.9%, and additive effects in 6.7%; no antagonistic interactions were detected. The most pronounced potentiating effect occurred against Gram-positive pathogens, particularly MRSA strains (66.7% synergistic combinations). Time-kill analysis confirmed suppression of the regrowth phenotype characteristic of MDR strains under monotherapy and restoration of bactericidal activity against antibiotics to which strains exhibited intrinsic resistance. D/PVA/I-1 induced a dose- and time-dependent increase in membrane permeability in both Gram-positive and Gram-negative organisms. Conclusions: D/PVA/I-1 is an effective broad-spectrum antibiotic potentiator and represents a promising basis for combination therapy regimens against MDR infections. Full article

The present study was designed to evaluate the structural and biochemical efficacy of Omega-3 fatty acids in preventing renal injury in streptozotocin (STZ)-induced diabetic kidney injury and to correlate systemic oxidative stress parameters with histological and ultrastructural parameters. Twenty-eight male Wistar Albino rats were randomly divided into four groups (n = 7) as follows: Control, Omega-3 (500 mg/kg, orally), Diabetes Mellitus (DM; 50 mg/kg STZ, i.p.), and DM+Omega-3 (STZ, followed by 500 mg/kg Omega-3 for 42 days). Biochemical analysis of renal function (blood urea nitrogen (BUN) and creatinine) and oxidative status (malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), total antioxidant status (TAS), total oxidant status (TOS) and oxidative stress index (OSI)) was measured by ELISA. Histopathological scoring and Caspase-3 staining were performed by Hematoxylin and Eosin (H&E) and immunohistochemically, respectively. Transmission Electron Microscopy (TEM) was used to examine the renal tissues. Our results showed that the DM group presented with severe hyperglycemia, uremia, and a significant oxidative shift, as evidenced by increased MDA/TOS levels and depleted antioxidant defenses. Light microscopy showed extensive glomerular damage, tubular degeneration and inflammatory infiltration in the kidneys of diabetic rats. Immunohistochemical analysis revealed strong Caspase-3 expression in the tubular and glomerular compartments. TEM showed severe podocyte effacement and damage to the glomerular basement membrane (GBM). Notably, Omega-3 supplementation significantly reversed renal dysfunction, restored the pro-oxidant/antioxidant balance and reduced histopathological injury scores. Moreover, Omega-3 treatment efficiently blocked Caspase-3-mediated apoptotic signaling and preserved the ultrastructural integrity of the glomerular filtration barrier (GFB) and tubular mitochondria. Our results demonstrate the potent nephroprotective effects of Omega-3 fatty acids in reducing oxidative stress, preventing programmed cell death and maintaining the stability of the renal parenchyma’s microarchitecture. These findings suggest that Omega-3 fatty acids could be an effective adjuvant therapeutic agent for the management of diabetic nephropathy. Full article

Background: Immune-checkpoint blockade (ICB) is a recent addition to the treatment options for breast cancer, especially the triple-negative (TNBC) subset. Inhibiting immunosuppressive factors in the tumor microenvironment (TME) and preventing or reversing T cell exhaustion may increase the likelihood of a response to ICB. We have shown that decitabine and guadecitabine, DNA methyltransferase inhibitors (DNMTi), prevent the systemic and TME accumulation of myeloid-derived suppressor cells (MDSCs), which are immunosuppressive. Results: Here, we show that adding DNMTi to the neoadjuvant + adjuvant ICB-based treatment of ICB-resistant 4T1 and E0771 murine tumors in Balb/C and C57Bl/6 mice, respectively, effectively reduced the tumor burden, with 52% of 4T1 tumors completely regressing across several studies. DNMTi-based therapy overcame ICB resistance (ICBR) in a selected subline of E0771, and treated tumors showed a reduction in MDSCs. We also show that, in E0771, DNMTi can overcome ICBR to multiple checkpoint inhibitors, and in 4T1, it can modulate anti-tumor immunity by enhancing central memory T cell (Tcm) formation and reducing T cell exhaustion. Conclusion: These pre-clinical findings support the further investigation of incorporating DNMTi as a new immunotherapy modality for TNBC. Full article

Purpose: This study aimed to determine the optimal temporal threshold for distinguishing “early” from “late” liver metastasis in patients who developed liver metastasis after colorectal cancer (CRC) surgery, and to evaluate whether KRAS and BRAF^V600E mutations, along with other clinicopathological factors, are associated with the timing of liver metastasis. Methods: This retrospective study utilized clinical and pathological data from patients who developed liver metastasis after radical CRC surgery at two centers from 2019 to 2023. X-tile software was used to identify the optimal temporal threshold. Logistic regression analysis was applied to determine if KRAS/BRAF^V600E mutations and other potential factors are independently associated with the time to onset of liver metastasis. Results: X-tile analysis identified 11 months post-surgery as the optimal cutoff for distinguishing early metachronous liver metastasis (EMLM) from late metachronous liver metastasis (LMLM), classifying 114 cases into the EMLM group and 72 into the LMLM group. Comparative analysis indicated statistically significant differences between the two groups in lymphovascular tumor emboli, perineural invasion, and postoperative adjuvant therapy (p < 0.05). Logistic regression analysis revealed that neither KRAS mutation (OR, 1.185; 95% CI: 0.641–2.190; p = 0.587) nor BRAF^V600E mutation (OR, 2.836; 95% CI: 0.302–26.642; p = 0.363) was independently associated with the timing of liver metastasis. In contrast, postoperative adjuvant therapy showed a statistical association with a likelihood of LMLM (OR, 0.253; 95% CI: 0.105–0.611; p = 0.002). Conclusions: This study identified 11 months post-CRC surgery as the optimal cutoff for differentiating EMLM versus LMLM. In this cohort, no statistically significant association was observed between KRAS/BRAF^V600E mutations and the timing of liver metastasis, whereas postoperative adjuvant therapy was statistically correlated with the likelihood of LMLM. This stratification may guide personalized surveillance strategies and provide valuable insights for future mechanistic investigations into the temporal heterogeneity of post-surgical liver metastasis. However, the interpretation and generalization of the findings require external validation in prospective cohorts. Full article

Neonatal calf diarrhea (NCD) is one of the most important problems of calf breeding across the world. It causes deaths in calves in the first 10 days of their life, and it is mainly caused by Escherichia coli(E. coli), Bovine Rotavirus (BRV) and Bovine Coronavirus (BCoV). The lack of vaccines with consistently high protective efficacy against the main causes of NCD makes disease control highly challenging. The current study aims to design a multi-epitope mRNA-based vaccine targeting the major pathogens responsible for NCD using immunoinformatic tools and molecular modeling approaches. BRV capsid protein VP6, BCoV Spike glycoprotein and E. coli F5 fimbrial protein were used as antigenic proteins to predict potential epitopes. Fifteen selected epitopes were linked with suitable linkers and conjugated with a built-in adjuvant, resulting in the design of a stable, antigenic and non-allergenic vaccine candidate against NCD pathogens. Furthermore, molecular docking analysis shows strong binding affinity between the vaccine candidate and the bovine toll-like receptors TLR2 and TLR4 at low energy and high stability. Based on these findings, the proposed multi-epitope vaccine represents a promising approach for the prevention and control of neonatal calf diarrhea and provides a solid scientific foundation for future experimental studies to validate its efficacy and safety in vivo. Full article

Cyclin-dependent kinase 4/6 inhibitors have fundamentally changed the management of hormone receptor-positive, human epidermal growth factor receptor 2-negative breast cancer. However, these drugs are not interchangeable and the field is moving away from the notion of a uniform “class effect.” In early breast cancer, adjuvant abemaciclib and ribociclib improve invasive disease-free survival in patients at a high risk of recurrence, whereas palbociclib does not. This difference likely stems from agent-specific pharmacological profiles, differences in trial design, and patient selection, rather than simply dosing nuances. In metastatic breast cancer, all three agents prolong progression-free survival when combined with endocrine therapy, but only ribociclib and potentially abemaciclib have shown an overall survival advantage. In addition, resistance remains a major obstacle in clinical practice. We propose that resistance mechanisms can be meaningfully grouped into two categories: target-driven (e.g., RB1 loss, CDK6 amplification) and bypass-driven (e.g., ESR1 mutations, PI3K/AKT pathway activation, APOBEC3-mediated mutagenesis). Distinguishing between these classes helps in the design of rational sequencing algorithms and combinatorial regimens. Emerging strategies, such as next-generation protein degraders, oral selective estrogen receptor degraders, antibody–drug conjugates, and inhibition of autophagy, are promising methods for overcoming resistance. Moving forward, the greatest need in breast cancer treatment will be not simply developing additional agents but using current therapies more intelligently by refining biomarker-guided patient selection, tailoring treatment duration, and ensuring broad global access. Full article

Alzheimer’s disease (AD) is a severe neurodegenerative disorder whose pathological progression is closely associated with the reduced binding affinity of apolipoprotein E ε4 (ApoE4) for amyloid-β (Aβ), which impairs Aβ clearance. Existing computational molecular design approaches are largely limited to single-target optimization and therefore lack the capacity for synergistic dual-target modulation. Herein, we proposed EvoPlay-MuZero, a hybrid computational framework incorporating a dual-peptide bridging (DPB) strategy. The framework adopted latent-state planning in MuZero reinforcement learning to enhance exploration and sequence-generation efficiency in high-dimensional sequence spaces. For the first time, it enabled the automated design of bispecific peptides targeting ApoE4 and Aβ, thereby forming a synergistic molecular bridge via a flexible linker. A full-process pipeline for structural and energetic evaluation was established by integrating AlphaFold3 and PDBePISA. Benchmark experiments on the 1SSC, 2CNZ, and 3R7G datasets demonstrated that EvoPlay-MuZero substantially outperformed the vanilla EvoPlay in convergence speed and the yield of valid generated sequences. Specifically, the optimal DPB molecule (15 × 25-3A) achieved a calculated interfacial solvation energy score (${\Delta }^{i}G$) of −29.5 kcal/mol, demonstrating substantially enhanced interface-stabilization properties compared to the native baseline control. This study provides a novel molecular intervention strategy for adjuvant therapy in AD and highlights the considerable potential of reinforcement learning in multi-target drug design. Full article

The 27th annual Western Canadian Gastrointestinal Cancer Consensus Conference (WCGCCC) was held in Calgary, Alberta, on 26–27 September 2025. The WCGCCC is an interactive multidisciplinary conference that was attended by healthcare professionals from across Western Canada (British Columbia, Alberta, Saskatchewan, and Manitoba) who are involved in the care of patients with colorectal cancer. Specialists from the fields of medical and radiation oncology, pathology, surgery, and a family physician in oncology participated in presentations and discussions for the purpose of developing the recommendations presented here. This consensus statement addresses recent advances in the management of colorectal cancer in a Western Canadian context, with respect to adjuvant exercise, as per the CHALLENGE trial, adjuvant Aspirin and PI3K testing, as per the ALASCCA trial, adjuvant immunotherapy, as per the ATOMIC trial, the use of encorafenib and an EGFR inhibitor with chemotherapy, as per the BREAKWATER trial, the use of combination immunotherapy as per the CHECKMATE 8HW trial and optimal strategies for omitting radiation and non-operative management of non-metastatic rectal cancer. Full article

The ubiquitin ligase protein tripartite-motif containing 56 (TRIM56) positively regulates Toll-like receptor-3 (TLR3) signaling by forming a complex with Toll-Interleukin-1 receptor domain-containing adapter protein inducing interferon (IFN)-beta (TRIF), independent of its E3 ligase activity. Whether TRIM56 modulates other TLR pathways in innate antiviral immunity, however, is unclear. Herein, we show ectopic expression of TRIM56 augments activation of IFN regulatory factor-3 (IRF3)-dependent promoters following stimulation by lipopolysaccharide (LPS) in HEK293-TLR4-MD2-CD14 cells while leaving activation of NF-κB-dependent promoter unaffected, suggesting TRIM56 specifically promotes immune signaling through the TLR4-TRIF axis but not the MYD88 arm downstream of this TLR. Confirming its impact on endogenous antiviral responses in immune sentinel cells naturally harboring the TLR4 pathway, we demonstrated enforced expression of TRIM56 enhanced LPS-induced expression of IFN-beta and IFN-stimulated genes (ISGs) and establishment of an antiviral state in bone marrow-derived macrophages. Importantly, depletion of endogenous TRIM56 impaired LPS-induced antiviral gene expression and cellular antiviral defense. Altogether, these data add to understanding of the role of TRIM56 in TLR-mediated innate immune responses. Given that TRIM56 is an ISG and that many immune adjuvants and some viral proteins activate TLR4, the findings of this study could have implications for designing immunotherapies, especially those against viral infections. Full article

Transcranial magnetic stimulation (TMS) is a non-invasive brain stimulation method with neuromodulatory capacity in neurodegenerative diseases such as multiple sclerosis (MS). Its therapeutic value is linked to its activity against oxidative stress by activation of antioxidant defenses. The sex hormones, estrogens (E), progesterone (P) and testosterone (T), have demonstrated their power as adjuvants to TMS, improving cortical excitability. The aim of this study was to evaluate the effect of these hormones as adjuvants to extremely low-frequency electromagnetic fields (ELFEFs) in the treatment of experimental autoimmune encephalomyelitis (EAE), the experimental model of MS. The effect of these hormones as replacement therapy was also evaluated in ovariectomized rats treated with ELFEFs. Sixty-five female Dark Agouti rats were divided into 13 groups (5 rats/group), in which biomarkers of oxidative stress and the glutathione redox cycle in non-nervous organs (kidney, liver, heart, intestines and blood) were analyzed. The results show that ELFEFs alone are more effective against oxidative stress. However, P and E were more effective than ELFEFs, both as adjuvants and in hormone replacement therapy, in activating the glutathione system. Therefore, it could be concluded that sex hormones play an important role against MS, enhancing the antioxidant effect of ELFEFs. Full article

This review summarizes the development of an oral inactivated enterotoxigenic Escherichia coli (ETEC) vaccine, ETVAX, describing its progression from preclinical evaluation of vaccine components in animal models to clinical assessment of safety, immunogenicity and, ultimately, protective efficacy in first- and second-generation formulations. Multiple Phase 1 and Phase 2 clinical trials conducted, initially in Swedish adults and subsequently in adults and children in Bangladesh and several African countries (Egypt, Zambia and The Gambia), have shown that the candidate vaccines are well tolerated, safe and capable of inducing robust intestinal or intestine-derived immune responses against the included antigens in most vaccine recipients. ETVAX, developed based on the results gained from studies of the first- and second-generation candidate vaccines, comprises four inactivated E. coli strains engineered to overexpress the most prevalent colonization factors combined with a toxoid and a mucosal adjuvant. Recent trials have shown that ETVAX confers significant protection against moderate–severe ETEC-associated diarrhea in young children in The Gambia and in travelers to Benin in Africa, underscoring its potential value both for endemic populations and travelers. Full article

Background: Lipopolysaccharide (LPS) functions as a Toll-like receptor 4 (TLR4) agonist that triggers innate immunity; however, structural variations between pathogenic and commensal bacteria distinctly influence its immunostimulatory profile. This study evaluated the immunostimulatory activity of LPS derived from the commensal bacterium Hafnia alvei and explored its potential as an exploratory vaccine adjuvant. Methods: Cytokine induction was evaluated in immune cells across diverse host species, and receptor activation was assessed via reporter assays. To investigate in vivo immunogenicity and preliminary tolerability, H. alvei LPS was formulated into a prototype oil-in-water (O/W) emulsion utilizing ovalbumin (OVA) as a model antigen. Results: LPS from H. alvei strain BA2000346 exhibited immunostimulatory activity comparable to that of Escherichia coli, while inducing greater TNF-α expression than pathogenic Salmonella and Pseudomonas strains. Distinct from E. coli LPS, it demonstrated the capacity to activate both TLR4 and the mannose-recognizing Dectin-2 receptor in reporter systems. This cytokine induction was consistent across various strains and host species. Furthermore, the prototype O/W emulsion formulation enhanced antigen-specific humoral and cellular immune responses while demonstrating preliminary tolerability based on body-weight monitoring and visual clinical observation. Conclusions: H. alvei-derived LPS exhibits TLR4 and Dectin-2 agonist activity in vitro. When synergized with an O/W emulsion delivery system, it provides a preliminary indication of cross-species stimulatory potential and supports further investigation as a hypothesis-generating platform for future vaccine adjuvant development. Full article

Antimicrobial resistance represents a major global health challenge, highlighting the urgent need for alternative bioactive compounds from natural sources. This study investigated the phytochemical composition and antimicrobial potential of saponin-enriched extracts from the trunk bark of Argania spinosa (L.) Skeels, collected from two contrasting Algerian regions: the coastal area of Stidia (ES) and the Saharan region of Tindouf (ET). Extraction yields were comparable (approximately 12.6%). UHPLC-MS analysis revealed distinct phytochemical profiles, with ES enriched in oleanane-type saponins and flavonoids, whereas ET showed a higher abundance of bayogenin-type derivatives. Key compounds included arganine C, E, and J, as well as catechin and quercetin. Antimicrobial activity was evaluated using agar well diffusion and broth microdilution assays against clinically relevant microorganisms, including the reference strains Staphylococcus aureus and Listeria innocua, together with Staphylococcus epidermidis, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, Proteus mirabilis, and Candida albicans. Both extracts exhibited broad-spectrum antimicrobial activity, although ES consistently showed lower Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal, Fungicidal Concentration (MBC)/(MFC) values than ET. MIC values ranged from 25 to 50 mg/mL for ES and from 50 to 100 mg/mL for ET. Synergistic interactions were observed between ES and gentamicin against S. aureus and between both extracts and kanamycin against K. pneumoniae. Membrane permeability assays demonstrated that both extracts increased bacterial membrane permeability, with ET producing a stronger permeabilizing effect. Atomic force microscopy of ES-treated cells revealed marked alterations in bacterial surface morphology, while molecular docking supported strong interactions of mi-saponin B and arganine derivatives with key bacterial targets. Collectively, these findings highlight the potential of A. spinosa bark saponins as natural antimicrobial agents and promising antibiotic adjuvants against multidrug-resistant pathogens. Full article