Combination strategy is crucial for enhancing cancer therapeutic efficacy, but co-delivery of multiple active pharmaceutical ingredients (APIs) remains challenging. Although nanomedicines address spatiotemporal co-delivery of APIs, it typically requires extensive excipients and formulation screening, which hampers drug discovery efficiency. Herein, we have developed a data-driven design workflow aided by ML to accelerate the rational design of ICG-templated API co-assembled nanoparticles for immunomodulation and antitumor synergistic therapy. The machine learning models can identify the important physicochemical properties that affect co-assembly of APIs and ICG, and may provide references for the design of nanoparticles. To illustrate the practicality of this workflow, compounds with lysosomal activity and drugs with synergistic effects were respectively explored to prepare nanomedicines for immunomodulation or synergistic anti-tumor effects. In one case, the models predicted co-assembled nano-adjuvants from lysosomal toll-like receptor 7/8 agonists to enhance antitumor immunity. In another case, the models launched multi-drug co-assembled nanomedicines including AZD7762 (AZD) and camptothecin (CPT) for synergistic cancer therapy. Both of the nanomedicines showed superior immune activation and antitumor efficiency than the free APIs. This study provides a ML-aided workflow to accelerate the design of co-assembled nanomedicines and offers feasible strategies to enhance the efficacy of anti-tumor therapies.
Alzheimer's therapy remains limited by poor drug targeting and multifactorial pathology. The therapeutic potential of SGLT-2 inhibitors like empagliflozin (EGZ) is constrained by poor brain bioavailability. Current study investigates the potential of EGZ-nanostructured lipid carrier (ENLC) for brain delivery via nasal route. The ENLC were prepared using hot melt emulsification technique followed by probe sonication and optimized using Box-Behnken design. ENLC were incorporated into poloxamer 407-chitosan in situ gel (ENPCG) to improve nasal retention, controlled release, and direct brain transport via olfactory and trigeminal uptake. ENPCG demonstrated a sustained drug release of 56.36 ± 3.37 % and enhanced nasal permeation. Nasal kinetics revealed high Cmaxmucosa (48.2 ± 1.42 µg/cm2) relative to plain EGZ-suspension (15.9 ± 0.7 µg/cm2) in goat nasal mucosa. ENPCG significantly improved cognitive memory in sporadic AD model, as confirmed by behavioural, biochemical, and histopathological assessments in Wistar rats. Pharmacokinetic study in Sprague Dawley rats revealed a 4.5-fold increase in AUC0-t of intranasal ENPCG (30.56 ± 0.45 μg/mL*h) relative to intravenous ENLC (6.73 ± 0.15 μg/mL*h). ENPCG showed 95.31 ± 3.89 % drug targeting potential. Furthermore, a strong point-to-point ex vivo-in vivo correlation (R2 = 0.9952) was observed, suggesting a non-invasive potential of ENPCG for translating AD interventions.
1,3,4-Oxadiazoles are privileged heterocyclic scaffolds with broad bioactivities, holding significant importance in pharmaceutical and agrochemical discovery. Herein, we report a novel, efficient, and simple synthetic route to 2-amino-1,3,4-oxadiazoles via NaOH-mediated desulfurative cyclization of hydrazides and isothiocyanates under mild reaction conditions. This developed methodology features broad substrate tolerance, excellent yields, operational simplicity, and mild conditions, providing a straightforward pathway for versatile syntheses of valuable 1,3,4-oxadiazole derivative. Consequently, the present reaction opens an alternative path for the preparation of 1,3,4-oxadiazoles via regioselective cyclization hydrazides and isothiocyanates.
Prolonged infusion (extended [EI, 2-4 h] or continuous [CI, 24 h] extended) of beta-lactam antibiotics is considered to have advantages for patients with severe infection compared with intermittent bolus (IB). However, the choice of EI and CI is unclear due to the lack of direct comparison. We aimed to compare the EI and CI of beta-lactams in patients with severe infections using a network meta-analysis method. We systematically searched PubMed, Embase, Web of Science, Cochrane Library, CNKI, Wanfang Database, and Weipu Database for randomized controlled trials (RCTs) comparing EI, CI, or IB with beta-lactams in adults with severe infections. The primary outcome was all-cause mortality. A frequentist network meta-analysis with a random-effects model was performed. Risk of bias was assessed using the Cochrane RoB 2 tool. Thirty-five RCTs (10,627 patients) were included. Risk of bias was moderate to high in most studies. For mortality, EI ranked highest (SUCRA 74.20%) with numerically lower rates versus IB (EI: OR 0.80, 95% CI 0.55-1.17; CI: OR 0.86, 95% CI 0.62-1.02). Both EI and CI significantly improved clinical cure rates versus IB (EI: OR 1.58, 95% CI 1.13-2.23; CI: OR 1.35, 95% CI 1.05-1.85), and EI ranked first (SUCRA 87.72%). For microbiological success, CI ranked highest (SUCRA 83.03%), followed by EI (SUCRA 42.98%) and IB (SUCRA 23.99%), but no significant difference was found. For hospital stay, EI was associated with a reduction of borderline statistical significance (MD -3.49 days, 95% CI -6.79 - -0.08), whereas CI did not show a significant reduction (MD 1.11 days, 95% CI -1.24 - 3.63), and EI ranked best (SUCRA 98.09%). No significant adverse event differences were observed. Subgroup analyses showed variable treatment rankings across categories, with no statistically significant subgroup effects. In patients with severe infections, both EI and CI improved clinical cure versus IB, whereas mortality did not differ significantly. Indirect evidence suggests EI may be more effective than CI in most outcomes except microbiological response. EI trended to shorten hospital stay but the difference was of borderline significance. Considering its practical feasibility, EI appears to be a favorable option based on current evidence. However, this finding is based on indirect evidence and requires confirmation in head-to-head trials. PROSPERO CRD420251242437.
Lablab purpureus (hyacinth bean) is a multifaceted legume traditionally integrated into diverse food systems and sustainable agricultural practices, particularly in Asia and Africa. Known for its adaptability to harsh environments, nitrogen-fixing ability, and use as both food and fodder, this crop holds untapped potential for addressing modern nutritional and health challenges. Recent scientific investigations have brought L. purpureus into the spotlight due to its impressive phytochemical profile, which includes phenolics (phenolic acids and flavonoids). These constituents are associated with a wide range of therapeutic properties, such as antioxidant, antimicrobial, anti-inflammatory, antidiabetic, and hepatoprotective activities. Nutritionally, L. purpureus is rich in proteins, essential amino acids, dietary fiber, vitamins, and minerals, making it a valuable candidate for combating malnutrition and enhancing food security, particularly in regions with limited access to diverse food sources. Furthermore, its functional properties suggest promising applications in nutraceutical and pharmaceutical product development. However, despite these attributes, L. purpureus remains largely underutilized and insufficiently represented in mainstream agricultural, food, and biomedical research. This review consolidates and synthesizes current knowledge on the phytochemical composition, nutritional value, and therapeutic potential of L. purpureus, while also identifying critical knowledge gaps and research priorities. By drawing attention to this overlooked legume, the review encourages greater scientific interest and investment in its study and application. As global demand grows for plant-based functional foods and sustainable crop alternatives, L. purpureus presents itself as a viable, eco-friendly, and health-promoting resource worthy of renewed focus and broader utilization across multiple sectors.
The increasing prominence of stereogenic-at-sulfur motifs in drug discovery and catalysis has created a growing demand for versatile synthetic methods. We describe here an Fe(TIBSPDP)-catalyzed enantioselective NH2-amination of sulfenamides to access chiral sulfinamidines. This method accommodates a broad range of sulfenamides, enabling direct access to diverse nitrogenated stereogenic-at-sulfur architectures. The synthetic utility is demonstrated by the efficient preparation of pharmaceutically relevant compounds, such as an aza-analog of drug candidate LY181984 and a PP2A modulator. Moreover, this reaction represents the first example of asymmetric NH2 transfer using the widely employed bioinspired Fe(PDP)-type catalysts.
Purification in the industry often requires repeated purification of large volume sample using a fixed chromatographic process. Therefore, this study aimed to design an automated chromatographic system which is able to process virtually unlimited sample volume using the minimalism principle. An automated repetitive purification liquid chromatography (ARP-LC) system was developed using active-pump-based sampling and cyclic operational control. The system demonstrated exceptional precision, with injection reproducibility yielding RSD ≤ 1.32% in peak area and retention time stability of ≤ 0.69% RSD. When applied to the purification of coumarins from Cortex Fraxini extract, the system efficiently processed 4 L of the sample, yielding 11 distinct fractions in only seven consecutive runs. Subsequent activity-guided purification identified several high-purity coumarin compounds exhibiting significant antioxidant (EC₅₀ values as low as 5 μg/mL) and antibacterial activities. The ARP-LC system showcases high robustness, scalability, and compatibility with medium- to high-pressure chromatographic modes, providing an efficient and reproducible platform for the large-scale purification of complex natural products with clear implications for pharmaceutical development and industrial applications.
Diclofenac (DCF) is a frequently detected pharmaceutical pollutant in wastewater and poses ecological risks due to its persistence during conventional treatment. This study investigates the electro-oxidation (EO) of DCF using three commercially available anode materials (Ti/IrO2, Pt/Ti, and mixed metal oxide (MMO)) under varying operational conditions to identify an optimal balance between oxidation capacity and energy efficiency. The effects of current density (2-20 mA/cm2), initial pH (3.5-9), and initial DCF concentration (5-40 mg/L) on degradation performance, kinetics, and specific energy consumption (SEC) were evaluated. Among the tested anodes, Pt/Ti showed the highest overall performance, achieving 71.2% DCF removal under the optimum conditions (10 mA/cm2, pH 7, and initial DCF concentration 10 mg/L). Under these conditions, the SEC was calculated as 19.7 kWh/g DCF (140 kWh/m3), indicating an effective balance between degradation efficiency and energy consumption. Kinetic analysis revealed that while increasing current density significantly enhanced degradation rates, it also led to a rise in energy demand. The initial DCF concentration was found to have a relatively minor effect on degradation kinetics. The Pt/Ti anode demonstrated a stable performance across a wide pH range, with neutral pH providing slightly more favorable conditions. The overall results indicate that Pt/Ti anodes can provide stable electrochemical performance with moderate energy consumption and represent a feasible electrode option for DCF removal in EO-based treatment systems.
Protopanaxadiol (PPD) is a bioactive ginsenoside with significant anti-inflammatory potential; however, its low natural abundance and dependence on inefficient intestinal microbial bioconversion hinder pharmaceutical development. To overcome these supply and bioavailability constraints, we developed a metabolically engineered rice variety, DJ-PPD, capable of directly biosynthesizing the aglycone PPD. This study investigated the anti-inflammatory and antioxidant mechanisms of DJ-PPD extract in lipopolysaccharide (LPS)-stimulated BV2 cells. DJ-PPD treatment significantly reduced nitric oxide (NO) production, pro-inflammatory cytokines (IL-1β, IL-6, TNF-α), and the expression of iNOS and COX-2. Its efficacy surpassed conventional ginseng extract and was comparable to synthetic PPD (S-PPD). Mechanistically, DJ-PPD inhibited NF-κB, MAPKs, and Akt phosphorylation while activating the NRF2/HO-1 antioxidant pathway. These findings demonstrate that DJ-PPD simultaneously inhibits pro-inflammatory cascades and reinforces intrinsic antioxidant defences. By effectively bypassing the need for gut microbiota metabolism, this genetically engineered rice represents a sustainable, bioavailable, and commercially viable multi-target therapeutic candidate for neuroinflammatory conditions.
Ensuring the quality, safety, and functional performance of stem cell cultures remains a critical challenge in biomedical research, drug development, and emerging cell-based therapies, as early metabolic disturbances can compromise outcomes and therapeutic efficacy. Glucose (Glu) consumption and L-lactate (LA) production serve as central indicators of cellular bioenergetic state, stress, and viability. However, conventional analytical approaches rely on discontinuous, labor-intensive assays that disrupt workflows and limit early intervention. Here, we report a miniaturized electrochemical biosensing platform for serial time-resolved analysis of Glu and LA in human induced pluripotent stem cell (hiPSC) culture media using a micropillar array (MPA)-based microfluidic electrochemical device (MED). The device exhibits linear detection ranges of 0.5-35 mM for Glu and 0.5-40 mM for LA, with limits of detection (LODs) of 0.18 ± 0.01 mM and 0.19 ± 0.01 mM, respectively. Using serially collected culture media, the MED quantifies dose-dependent metabolic responses to mitochondrial inhibition with oligomycin, revealing suppressed metabolic turnover and altered Glu-LA coupling prior to observable morphological changes. By enabling paired metabolic readouts from small-volume, undiluted culture-media samples, this platform provides a practical analytical route toward earlier recognition of culture-state deviations and more informed monitoring of stem-cell culture quality and drug-induced metabolic responses.
Administrative health data are vital for investigating medication safety during pregnancy. In Taiwan, while the National Health Insurance Research Database (NHIRD) and Birth Certificate Application (BCA) capture pregnancy data, no standardized approach exists for integrating these sources to identify episodes and assign gestational age (GA). This study aimed to develop a hierarchical pregnancy-identification algorithm tailored to Taiwan's linked claims and birth registry data. We adapted an ICD-10-CM/PCS-based algorithm to the Taiwanese coding environment, incorporating clinician input, local billing practices, and birth registry information. The algorithm refines pregnancy outcome classification, estimates pregnancy start dates, and assigns GA. As a proof of concept, we applied the final algorithm to 2016-2022 data to summarize outcome distributions. We developed a seven-step hierarchical algorithm that uses claims codes to classify pregnancy outcomes, groups records into episodes using spacing rules, assigns pregnancy start dates from ordered prenatal services, validates and refines GA via linkage to the BCA database, determines the number of pregnancies and fetuses, and checks the plausibility of GA and inter-pregnancy intervals. A total of 1,696,229 pregnancies contributed by 1,169,779 women were identified; outcome distributions (69.3% live birth, 16.3% spontaneous abortion, 5.7% elective abortion, 6.0% ectopic pregnancy, 1.4% delivery with unknown outcome, 1.1% stillbirth, 0.2% trophoblastic and other abnormal products of conception) were consistent with national statistics and prior literature. This hierarchical algorithm provides a transparent, reproducible framework for identifying pregnancies and estimating GA in Taiwan. It establishes a critical foundation for future real-world studies of medication safety during pregnancy.
The blood-brain barrier (BBB) remains the most formidable obstacle in CNS drug development, severely hindering the delivery of therapeutic agents to the brain. While many natural compounds, such as the flavonoid luteolin (Lut), possess potent anti-neuroinflammatory properties, their clinical potential is restricted by poor pharmacokinetic profiles and minimal BBB permeability. To address this systemic challenge, we developed a versatile, brain-targeting nanoplatform utilizing mesenchymal stem cell-derived extracellular vesicles (MSC-EVs). For active CNS targeting, these EVs were functionalized with a chimeric RVG-CP05 peptide via modular, non-covalent anchoring and subsequently loaded with Lut. This RVG@EV-Lut nanocomposite was characterized for its physicochemical properties and evaluated using a Transwell-based in vitro BBB model. Therapeutic efficacy and biodistribution were assessed in a C57BL/6J mouse model of LPS-induced neuroinflammation. RVG functionalized EV exhibited dynamic stability in vitro, significantly increased cellular uptake by both endothelial cells and microgliaand and enhanced the active transport of Lut across the BBB in vitro. Compared to free Lut and non-targeted vesicles, the RVG@EV-Lut platform demonstrated superior brain accumulation and prolonged retention in vivo imaging. This targeted delivery resulted in a robust suppression of cerebral pro-inflammatory cytokines, reduced neuronal apoptosis, and preservation of hippocampal cytoarchitecture. Critically, these effects were translated into a marked restoration of spatial memory and cognitive performance in the treated mice. Our findings demonstrate that the RVG@EV-Lut platform effectively overcomes the BBB to deliver therapeutic payloads directly to the CNS. This modular engineering strategy provides a scalable and broadly applicable solution for enhancing the brain delivery of compounds with poor pharmacokinetics.
OptoH3R is an artificial fusion protein that combines the photosensitive elements of rhodopsin with the signaling domain of the histamine H3 receptor (H3R), enabling light-controlled activation of downstream H3R pathways. Although our previous study demonstrated that OptoH3R mimics the acute effects of H3R activation on neuronal activity, whether this tool can also recapitulate the long-term receptor desensitization and internalization processes associated with prolonged H3R activation remains unclear. In this study, primary cortical neurons and HeLa cells were employed to investigate the alterations in subcellular localization of OptoH3R upon sustained photoactivation, with histamine-stimulated wild-type (WT) H3R serving as a control. Furthermore, the role of β-arrestin in this process was explored. Time-lapse fluorescence imaging revealed that the number of puncta progressively increased over time following laser stimulation. Subsequent co-staining experiments with endosome marker EEA1 showed that 75.5% of light‑induced puncta were EEA1‑positive. Notably, the increase in OptoH3R-positive vesicles within neuronal cells was attenuated by the β-arrestin inhibitor barbadin, a pattern consistent with the internalization observed in histamine-stimulated WT H3R. Collectively, our findings demonstrate that OptoH3R recruits β-arrestin signaling upon sustained optical stimulation, thereby recapitulating H3R desensitization dynamics. This establishes OptoH3R as a useful tool for dissecting the spatiotemporally specific functions of H3R, including both its acute signaling and long-term β-arrestin-related mechanisms.
Mucopolysaccharidosis type I (MPS-I) is a rare, multisystemic lysosomal storage disease (LSD) caused by mutations in the IDUA gene, which encodes the enzyme alpha-L-iduronidase. Current treatments include hematopoietic stem cell transplantation and enzyme replacement therapy (ERT), administered via weekly intravenous infusions. ERT is of limited efficacy owing to its inability to reach critical tissues such as the brain and bone. To address these limitations, this study explores a novel method to improve drug delivery to target organs and simplify administration: oral administration of enzyme encapsulated within nanostructured lipid carriers (NLC). Encapsulation of ERT within NLC enabled effective oral administration. In vitro analysis showed that our NLC formulation was as effective as intravenous ERT in correcting enzyme activity and reducing glycosaminoglycan (GAG) accumulation in fibroblasts from MPS-I patients, when administered periodically. Permeability studies confirmed passage across the intestinal barrier. Proteomic analyses demonstrated normalization of protein expression in energetic pathways related to hexose metabolism, and significant improvements in protein dysregulation in the cytoskeleton, cellular trafficking, lysosomal function, GAG biosynthesis and degradation, and the extracellular matrix. Furthermore, in vivo studies in MPS-I knockout (KO) mice demonstrated biodistribution of NLC-encapsulated enzymes to all tissues affected by the disease, including passage across the blood-brain barrier and access to poorly vascularized bone. These findings suggest that oral administration of ERT via NLC encapsulation represents a significant advancement in MPS-I treatment, enabling drug delivery to previously inaccessible areas. This study opens important avenues of research for future therapeutic strategies targeting LSDs.
Patient involvement within academic and for-profit medicines and medical technology research and development is happening across organisations and functions. Frameworks exist to conceptualise involvement activities along a spectrum based on patient contributions and their share of voice and decision-making. Examples range from Informing and Education of patients through to Co-design and Co-Production e.g., in research and development. Varying levels of patient expertise profiles also exist based on a spectrum of experience and expertise - from Patient by Experience through to Patient Key Opinion Leader (KOL) - though researchers often lack clarity on the most relevant skills and knowledge of patient partners suited to the task at hand. Here, we propose a conceptual matrix model combining these two concepts into a simple, pragmatic framework intended for patient involvement professionals in research and development, but with broader relevance for other contexts. As authors with a mix of academic, for-profit industry, and patient advocacy backgrounds, we utilised intellectual exchange and consensus formation in three workshops to develop the model. We have developed a conceptual matrix model and applied illustrative case study examples from among our collective experiences and network to demonstrate the model in action. Trends in sample sizes, patient diversity, project longevity, and share of influence exist along the matrix axes. For example, greater numbers and diversity of patients are typically seen at the lower levels of involvement, where there is greater capacity for breadth of involvement. Our aim for this model is to support and encourage patient involvement professionals to better determine appropriate levels of involvement and patient expertise needed for their projects and activities, enabling them to facilitate more appropriate, fair, effective, and sustainable patient involvement. WHAT IS THE MATRIX MODEL?: Universities and for-profit organisations regularly involve patients when they research and develop medicines and medical technologies. Patients can be involved in different types of projects. For example, they can review research plans or patient materials. Different patient involvement activities can be thought of in terms of levels on a spectrum. Patient expertise can also be thought of as levels on a spectrum. Some patients may have expertise in their personal experience. Some patients may have more backgrounds and types of expertise, such as in research processes. So far, these two spectra have been used separately. Researchers may not be aware of the different skills and knowledge that patients may have. So, we took these two spectra – the level of patient involvement and the level of patient expertise – and combined them into a single practical tool using a matrix model (graph). HOW DID WE CREATE IT?: Our group has a mix of expertise, including university researchers, for-profit industry professionals, and patients. We met over three workshops to develop our matrix. We also chose real-life case study examples of patient involvement activities to show how the matrix can be used. HOW CAN IT BE USED?: The matrix is designed for people working in patient involvement at universities and for-profit organisations to consider what levels of involvement and expertise are best for their projects. It might also be helpful for researchers from other types of organisations. We intend for this to create more useful and effective patient involvement.
Cataract remains the most prevalent cause of visual impairment and irreversible blindness worldwide. It is characterized by lens opacification arising from crystallin denaturation, leading to variable degrees of vision loss and poor quality of life. Although crystallin denaturation is the primary pathological basis of cataract, oxidative stress and inflammation also play significant roles. It was first confirmed that nattokinase (NK) could degrade cataractous tissue and alleviate the opacification using human cataract phacoemulsification fluid, thus supporting the in vivo anti-cataract efficacy of NK-based nanocomposites (NNs). Herein, we developed minimalist NNs as eye drops, through electrostatic self-assembly between NK and natural macromolecules. The NNs exert three-pronged therapeutic effects (antioxidant, anti-inflammatory, and anti-crystallin denaturation) that disrupt the pathological cycle in cataract. The in vitro cellular studies demonstrated that the NNs were internalized via receptor-mediated endocytosis and micropinocytosis, and efficiently achieved a synergistic scavenging effect of reactive oxygen species (ROS). In ultraviolet B (UVB)-induced cataract mice, the NNs successfully suppressed oxidative stress and inflammation, while protecting crystallins through enhanced intracorneal retention, ultimately ameliorating lens opacity index. Moreover, further analysis revealed that NNs effectively inhibited ROS accumulation by up to 15.78-fold, along with the ROS/NLRP3/pyroptosis axis and the denaturation of crystallins, thereby offering new insights into therapeutic regimens for cataract. Consequently, the NNs alleviated lens opacification by breaking the cataract vicious cycle through a three-pronged inhibition of oxidative stress, inflammation, and crystallin denaturation. Our findings pave the way for utilizing NNs as a novel strategy for reversing cataract, as well as associated ocular disorders.
The spike glycoprotein of SARS-CoV-2, particularly its receptor-binding domain (RBD), is a key target for therapeutic monoclonal antibodies (mAbs). Epitope mapping is therefore critical for the development of effective antiviral therapeutics. In this study, diethylpyrocarbonate covalent labeling mass spectrometry (DEPC CL-MS) was applied to map epitopes on the beta (B.1.351) and omicron (B.1.1.529) variants of the SARS-CoV-2 RBD, as well as on the original SARS-CoV-2 spike S1 subunit, in complex with anti-SARS-CoV-2 mAbs. Combined with bottom-up LC-MS/MS, DEPC labeling enabled site-specific identification of residues exhibiting significant modification changes. Clustering of these residues on the protein surface identified potential epitopes for the 1D1 mAb on the RBDs and for the 1D3 mAb on the C-terminal domain of the S1 subunit. Structural interpretation was supported by available experimental data, with AlphaFold models providing supplementary context where needed. Thus, "theory guides, but experiment decides": AlphaFold aided epitope identification when high-resolution antigen-antibody complex structures were unavailable, but conclusions were ultimately resolved experimentally. Together, these findings establish DEPC CL-MS as a useful and complementary approach for epitope mapping, providing residue-level insights into antigen-antibody interactions and advancing structural understanding for antiviral mAb development.
Inflammatory markers such as high-sensitivity C-reactive protein (hs-CRP) and the neutrophil-to-lymphocyte ratio (NLR) have been individually associated with cardiovascular risk in patients undergoing percutaneous coronary intervention (PCI). However, their combined prognostic value remains incompletely defined. Consecutive patients undergoing PCI between 2012 and 2022 were included and stratified according to baseline hs-CRP (≥2 mg/L vs <2 mg/L) and NLR (≥75th vs <75th percentile). The primary endpoint was 1-year major adverse cardiovascular events (MACE). Multivariable Cox proportional hazards models were used to assess associations after adjustment for confounders. Among 7,303 included patients, those with concomitantly elevated NLR and hs-CRP exhibited the highest incidence of overall MACE, followed by patients with only one elevated inflammatory marker, while the lowest event rates were observed in those with both markers below threshold. No significant interaction between NLR and hs-CRP was observed (p=0.451). Elevated NLR was associated with a higher risk of MACE in both hs-CRP strata (hs-CRP ≥2 mg/L: HR 1.83, 95% CI 1.42-2.35, p<0.001; hs-CRP <2 mg/L: HR 1.55, 95% CI 1.10-2.17, p=0.012). After multivariable adjustment, this association remained significant only in patients with hs-CRP ≥2 mg/L, driven primarily by all-cause mortality and myocardial infarction. In patients undergoing PCI, elevated NLR identifies a gradient of increasing risk, particularly when combined with hs-CRP ≥2 mg/L. A dual-marker inflammatory approach may improve post-PCI risk stratification beyond either biomarker alone.
Alpine snowpacks provide valuable archives of atmospheric deposition, yet the southern slopes of the Greater Caucasus remain underrepresented in high-altitude geochemical monitoring networks. This study investigated the hydrochemical composition of fresh snow, annual snowpack, and perennial snowfields across an altitudinal transect from 1838 to 2857 m a.s.l. within the Caucasian State Nature Biosphere Reserve to characterize depositional sources and evaluate how snow archive type and landscape structure influence geochemical signal preservation. Major ions were determined by titrimetric and turbidimetric methods, while trace elements and rare earth elements were quantified by ICP-AES and ICP-MS. Complementary soil and vegetation samples assessed landscape controls on elemental accumulation. Marine-associated ions, including Na, Mg, Cl, and Sr, remained detectable across the transect at sites approximately 50 km inland from the Black Sea, indicating orographic transport and scavenging of maritime aerosols during south-westerly circulation. Forest canopy interception reduced sub-canopy snowpack elemental concentrations by approximately 70 to 80 percent relative to open alpine environments, establishing montane forest sites as the most conservative local deposition baseline. Rare earth element concentrations exceeded upper crustal Clarke values by factors of 2 to 5 and corresponded to REE-enriched Paleogene volcanic lithologies of the Mzymta basin, supporting their use as robust lithogenic tracers. Because fresh snow, annual snowpack, and perennial snowfields preserve atmospheric chemistry over fundamentally different temporal scales, they cannot be treated as interchangeable monitoring archives. A spatially distributed annual snowpack sampling design anchored by montane forest reference sites is recommended for integrated seasonal atmospheric monitoring in topographically complex mountain terrain.
One of the main problems in the treatment of ovarian cancer is resistance to the main drug (cisplatin). Thiostrepton (TST), a thiazole antibiotic, helps weaken cancer cells and enhance the efficacy of platinum-based therapy. This study examines the combination of TST and cisplatin in the SKOV3/CDDP model of resistance, focusing on associated changes in oxidative stress, mitochondrial function, genotoxicity, apoptosis, inflammatory mediators, and Wnt/β-catenin output. SKOV3/CDDP cells were treated with cisplatin at an IC50 concentration, TST at concentrations (1, 2.5, 5 µM), and the combination of these two substances for 24 h. Cell viability, intracellular energy, and membrane damage were measured by MTT and ATP assays and LDH release, respectively. Apoptosis was assessed using caspase-3/7 and TUNEL activity.DNA damage was analyzed using the Comet and CBMN assays. Oxidative stress and antioxidant defense were assessed by measuring ROS, MDA, GSH/GSSG ratio, and SOD, catalase, and GPx activities. Mitochondrial dysfunction (ΔΨm, NAD+/NADH), cytokines (IL-6, IL-8, TNF-α, IL-1β), and Wnt/β-catenin target genes (c-MYC, AXIN2) were evaluated. The combination of cisplatin and TST was much more potent than when either was used alone. This combination increased oxidative stress processes, increased DNA damage, and decreased antioxidant capacity in cancer cells. Inflammatory cytokine levels were also increased, and c-MYC and AXIN2 expression were reduced, which was accompanied by the attenuation of β-catenin-dependent transcription. Overall, TST increased the sensitivity of resistant ovarian cancer cells to cisplatin. Its anticancer effect was enhanced by oxidative-mitochondrial stress and DNA damage pathways, along with reduced Wnt/β-catenin transcription.