Biomedicines

Objective: This study aims to investigate the role of thrombospondin-1 (THBS1) in polycystic ovary syndrome (PCOS) pathogenesis and its mechanism in regulating granulosa cell (GC) function. Methods: Follicular fluid and granulosa cells from 21 PCOS patients and 21 age-matched non-PCOS controls were analysed for THBS1 expression and clinical correlations. A dehydroepiandrosterone (DHEA)-induced PCOS rat model with adeno-associated virus serotype 9 (AAV9)-mediated THBS1 knockdown was used to assess phenotypic changes. The KGN human granulosa-like cell line was employed to evaluate THBS1 overexpression effects on proliferation, apoptosis, and steroidogenesis. Mechanistic studies included RNA sequencing with Gene Set Enrichment Analysis (GSEA), co-immunoprecipitation, molecular docking against the latent TGF-β1 crystal structure (PDB 9VJJ), molecular dynamics simulation, an active/total TGF-β1 ELISA, and pharmacological TGF-β receptor inhibition. Results: THBS1 was elevated in PCOS follicular fluid and granulosa cells and correlated positively with serum AMH and LH after Benjamini–Hochberg FDR correction. AAV9-mediated ovarian THBS1 knockdown (37.4% protein reduction, p = 0.006) ameliorated cystic morphology, restored estrous cyclicity, and normalised serum AMH/LH/T. In KGN cells, THBS1 overexpression suppressed proliferation, induced apoptosis and inflammatory cytokines, and dysregulated steroidogenic enzymes. Transcriptome analysis revealed upregulation of canonical TGF-β/Smad pathway components (SERPINE1, SMAD7, TGFB2, INHBA, CCN2, COL1A1/2). Molecular docking and 100-ns dynamics simulation supported a stable interaction between THBS1 and latent TGF-β1 (ΔG_TOTAL ≈ −120 kcal·mol⁻¹). Co-immunoprecipitation confirmed physical association in cells, and ELISA showed elevated TGF-β1 in PCOS follicular fluid and rat serum, both attenuated by THBS1 knockdown. Pharmacological TGF-β receptor inhibition with SB-431542 rescued THBS1-induced cellular dysfunction. Conclusions: THBS1 is associated with PCOS-related granulosa cell dysfunction through the TGF-β/Smad pathway and represents a candidate biomarker and exploratory therapeutic target that warrants validation in independent multicentre cohorts. Full article

Background/Objectives: Cigarette smoking is a major risk factor for periodontitis, but the salivary host-response profile associated with smoking-related periodontal inflammation remains incompletely characterized. This study compared salivary NETosis-related and oxidative-inflammatory biomarkers among current smokers, former smokers, and never-smokers with periodontitis. Methods: This cross-sectional study included 159 systemically healthy adults with periodontitis (53 per group: current smokers, former smokers, never-smokers). Individuals with systemic diseases or concomitant medications that could interfere were excluded. Unstimulated whole saliva was analyzed for NETosis-related biomarkers (MPO-DNA complexes, citrullinated histone H3, neutrophil elastase, cell-free DNA) and oxidative-inflammatory markers (MMP-8, IL-1β, IL-6, TNF-α, 8-OHdG, total antioxidant capacity). Results: Salivary MPO-DNA complexes differed significantly among groups (current smokers 33.52 ± 9.96, former smokers 26.90 ± 8.38, never-smokers 19.20 ± 7.50 ng/mL; p < 0.001, η² = 0.317). The composite NETosis score (η² = 0.702) and oxidative-inflammatory score (η² = 0.718) showed the same graded pattern. Biochemical verification confirmed clear group separation (salivary cotinine: current smokers 312.3 ± 77.0, former smokers 9.7 ± 5.1, never-smokers 3.2 ± 1.4 ng/mL). Smoking exposure was positively correlated with biomarker levels and the severity of periodontal disease. Smoking status remained independently associated with MPO-DNA complexes and the NETosis score after covariate adjustment. Conclusions: Current smoking was associated with an enhanced salivary NETosis-related and oxidative-inflammatory phenotype. Former smokers displayed an intermediate profile. Salivary MPO-DNA complexes and composite biomarker scores warrant further investigation as candidate non-invasive indicators of smoking-associated periodontal inflammatory burden, pending diagnostic performance analyses and prospective validation. Full article

Background: Short-term mortality stratification in sepsis remains clinically challenging, particularly because outcome is influenced by acute inflammation, coagulation abnormalities, organ dysfunction, and baseline comorbidity burden. This study evaluated the clinical performance of the BIO-S and BIO-SC composite bioscores for 28-day mortality stratification in adults with sepsis and septic shock. Methods: We conducted a prospective observational monocentric cohort study including 572 adult patients admitted between January 2022 and December 2024. BIO-S integrated procalcitonin (PCT), neutrophil-to-lymphocyte ratio (NLR), International Normalized Ratio (INR), and Sequential Organ Failure Assessment (SOFA) score, while BIO-SC extended this model by adding the Charlson Comorbidity Index (CCI). Prognostic performance was assessed using receiver operating characteristic (ROC) curve analysis, DeLong comparisons, bootstrap validation, calibration analysis, Kaplan–Meier survival curves, and Cox proportional hazards models. Results: The cohort included 418 patients with sepsis and 154 patients with septic shock. Overall 28-day mortality was 31.5% and was significantly higher in septic shock than in sepsis, 77.9% versus 14.4%, p < 0.001. BIO-S and BIO-SC showed strong discriminatory ability for 28-day mortality, with areas under the curve (AUCs) of 0.889 and 0.897, respectively. BIO-SC had the highest AUC, although the difference between BIO-SC and BIO-S was not statistically significant by the DeLong test, p = 0.328. At the optimal thresholds, BIO-S showed 97.8% sensitivity and 69.4% specificity, while BIO-SC showed 89.4% sensitivity and 77.8% specificity. Both bioscores stratified observed mortality across predefined risk categories and remained significantly associated with 28-day mortality in adjusted Cox models. Conclusions: BIO-S and BIO-SC showed clinically relevant performance for 28-day mortality stratification in adults with sepsis and septic shock. BIO-SC provided a numerically higher AUC and slightly better calibration, suggesting that comorbidity burden may improve prognostic characterization, although further independent multicenter validation is needed before broader clinical implementation. Full article

Background/Objectives: Impaired quality of life (QoL) represents one of the most important clinical determinants in heart failure with preserved ejection fraction (HFpEF). This study aimed to evaluate the incremental explanatory value of functional performance and inflammatory biomarkers for QoL in a clinically stable HFpEF cohort. Methods: A single-center observational study enrolled 110 consecutive patients with stable HFpEF. Functional capacity was assessed using the six-minute walk test (6MWT), expressed mainly as percentage of predicted distance. Health-related QoL was measured using the EQ-5D-5L utility index (primary outcome). Circulating IL-6, CRP, and NT-proBNP were obtained from peripheral blood. Hierarchical multivariable linear regression was applied to quantify the incremental contribution of clinical variables, functional capacity, and biomarkers. Results: The median age was 72 years, and 52.7% of the participants were women. The median 6MWT distance was 340 m (75.9% of predicted), and the median EQ-5D index was 0.76. The baseline clinical regression model (age, sex, atrial fibrillation, and glomerular filtration rate) explained 23.5% of EQ-5D variance. The addition of functional capacity increased explained variance to 45.2% (ΔR² = +0.217). The inclusion of IL-6 and NT-proBNP provided a modest additional increase (R² = 0.468; ΔR² = +0.042 in addition to Model 2). In the fully adjusted model, functional capacity (β = 0.376, p < 0.001) and IL-6 (β = −0.185, p < 0.05) remained independent predictors, whereas NT-proBNP lost significance. Conclusions: In stable HFpEF, objective functional capacity represents the dominant determinant of QoL, while inflammatory activation provides an independent but smaller contribution. Functional assessment may therefore be central to patient-centered phenotyping and therapeutic targeting. Full article

Background/Objectives: Kangaroo Mother Care (KMC) has been proposed as a protective intervention that may modulate the epigenetic regulation of stress-related genes, such as SLC6A4, which encodes the serotonin transporter. Few studies have explored this association in humans. This study aimed to evaluate whether KMC affects the methylation status of SLC6A4 in preterm newborns. Methods: This longitudinal observational study included preterm infants with birth weight ≤ 1800 g and gestational age between 25 and 34 weeks. Blood samples were collected at birth, Neonatal Intensive Care Unit discharge, and hospital discharge. Methylation levels at 13 CpG sites within the SLC6A4 promoter region were quantified by bisulfite conversion and pyrosequencing. Methylation dynamics were analyzed using linear mixed-effects models adjusted for clinical covariates. Results: 75 preterm infants were analyzed (51 KMC; 24 non-KMC). Methylation levels ranged from 0.78% to 10.76% across all CpG sites and remained stable over time. At hospital discharge, the KMC group exhibited lower methylation at CpG6 than the non-KMC group (median = 0.96% vs. 1.21%, p = 0.021), but this difference was not statistically significant after correction for multiple testing. No significant differences were observed at other sites or in longitudinal methylation trajectories between groups. Conclusions: KMC was not associated with major longitudinal changes in SLC6A4 methylation during the neonatal period. The nominal difference at CpG6 should be interpreted as exploratory and warrants further investigation. Larger, multicenter studies with long-term follow-up are needed to clarify the epigenetic mechanisms linking early caregiving experiences with stress regulation and neurodevelopmental outcomes in preterm infants. Full article

Background: In patients with type 2 diabetes mellitus (T2DM), angiotensin-converting enzyme inhibitors (ACE-Is) and angiotensin receptor blockers (ARBs) are first-line antihypertensive treatments with important cardiovascular benefits, but their impacts on coronary-specific inflammation are unknown. Pericoronary adipose tissue (PCAT) attenuation, as assessed by coronary computed tomography angiography (CCTA), serves as a specific biomarker for coronary inflammation. Here, we aim to assess whether treatment with ACE-I or ARB is correlated with lower PCAT attenuation. Methods: In this retrospective observational study, we analyzed 223 patients with T2DM and coronary atherosclerosis who underwent CCTA from 1 January 2017 to 1 September 2024 at our institution. PCAT attenuation was measured in the proximal right coronary artery. Propensity score matching and multivariate linear regression analyses were performed for comparisons. Results: Of the 223 patients (mean age of 64.9 ± 8.8 years, 69.1% male), 122 patients were on ACE-I or ARB (ACE-I/ARB). ACE-I/ARB users had similar PCAT attenuation as their counterparts after propensity score matching (−72.1 ± 7.5 and −71.7 ± 8.1 HU, respectively; p = 0.722). Subgroup analysis in patients with glomerular filtration rate (GFR) < 90 mL/min revealed lower PCAT attenuation in ACE-I/ARB users (−74.8 ± 6.6 vs. −71.4 ± 7.1 HU; p = 0.038), with a significant interaction between these two factors in the multivariate analysis (p = 0.047). Other antihypertensive treatments (beta blockers, dihydropyridine calcium channel blockers, and thiazides) were not linked with lower coronary inflammation. Conclusions: In T2DM patients with coronary atherosclerosis, we did not find an association between ACE-I/ARB treatment and lower coronary inflammation as defined by PCAT attenuation, although such a relationship may exist in those with reduced GFRs. Full article

The gut microbiome plays a key role in the pathogenesis of cardiovascular disease through systemic inflammation, impaired lipid metabolism, and proatherogenic gut metabolites like trimethylamine N-oxide. Gut dysbiosis contributes to decreased level of microbial metabolites such as short-chain fatty acids, bile acids, coprostanol, and phenylacetylglutamine, as well as increased intestinal permeability and platelet hyper-reactivity, and exacerbating cardiovascular risk. New microbiome-focused treatments such as probiotics, prebiotics, synbiotics, and fecal microbiota transplantation are showing potential to help reduce cardiovascular diseases. However, bringing these therapies into clinical settings is difficult because they vary by strain and individual response. The gut–heart connection offers an innovative approach to preventing and treating heart condition, but additional research is needed to ensure lasting effectiveness and safety. Full article

Background: Complex Regional Pain Syndrome type 1 (CRPS-1) is a multifactorial disorder characterized by persistent pain, neuroinflammation, and tissue remodeling following trauma in the absence of overt nerve injury. Despite advances in understanding its pathophysiology, the mechanisms underlying the transition to chronic pain remain incompletely defined, and reliable circulating biomarkers are lacking. Piezo-type mechanosensitive ion channel component 1 (Piezo 1), a mechanosensitive ion channel that transduces mechanical stimuli into intracellular calcium signaling, has emerged as a regulator of inflammation, extracellular matrix remodeling, and cellular stress responses. Experimental evidence indicates that Piezo 1 activation can modulate cytokine production and mechanotransduction pathways relevant to chronic pain and inflammatory conditions. Methods: In this study, we evaluated circulating Piezo 1 levels in CRPS-1 patients and explored their association with clinical parameters and response to neridronate treatment. Results: Although Piezo 1 levels were significantly altered compared to controls, no associations were observed with pain intensity or therapeutic response. Conclusions: These findings suggest that, despite its biological relevance, circulating Piezo 1 is not a clinically informative biomarker in CRPS-1. The results support a predominantly local role of Piezo 1-mediated mechanotransduction in processes relevant to chronic inflammation and nociceptive sensitization. Full article

Major depressive disorder remains a leading cause of disability, and decades of monoamine-centered pharmacology have yielded delayed and often incomplete relief. Rapid-acting antidepressants reshaped the field by linking swift symptom improvement to glutamatergic plasticity, yet durable benefit depends on how newly reconfigured circuits are stabilized and tuned. This review synthesizes evidence that antidepressant efficacy arises from the coordinated engagement of synaptic plasticity, spanning induction and consolidation, and intrinsic excitability, which provides gain control, and proposes an integrated framework to guide future discovery. It first outlines induction through N-methyl-D-aspartate receptors (NMDARs) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs), exemplified by ketamine and esketamine, followed by consolidation mediated by tropomyosin receptor kinase B (TrkB) signaling, translational disinhibition via eukaryotic elongation factor 2 kinase (eEF2K), and presynaptic stabilization indexed by synaptic vesicle glycoprotein 2A (SV2A); together, these processes transform transient potentiation into persistent network change. It then highlights intrinsic excitability, emphasizing voltage-gated potassium channel subfamily Q (Kv7), hyperpolarization-activated cyclic nucleotide-gated (HCN), and G protein-gated inwardly rectifying potassium (GIRK) channels as circuit-level governors that normalize firing and limit relapse-prone hyperexcitability. Finally, it presents the Induction–Consolidation–Maintenance (ICM) framework as a hypothesis-generating roadmap for future studies, with SV2A positron emission tomography (PET), electroencephalography (EEG), and functional magnetic resonance imaging (fMRI) biomarkers discussed as candidate tools rather than validated guides for treatment timing or patient selection. The proposed contribution is not another list of plasticity pathways, but a phase-specific model that links synaptic induction, consolidation, and excitability-based maintenance to distinct therapeutic windows, biomarkers, and relapse-prevention strategies. Full article

Background and Objectives: Multidrug-resistant (MDR) uropathogens are reshaping the empirical management of acute pyelonephritis, particularly in Eastern European centers. We aimed to describe MDR patterns, identify admission-level predictors, including renal impairment/renal-failure status at presentation and major healthcare exposure variables, and derive a bedside risk score (PYELO-MDR-Risk) for adult pyelonephritis at a Romanian tertiary hospital. Methods: We retrospectively analyzed 129 consecutive culture-confirmed acute pyelonephritis admissions at “Victor Babeș” University Hospital, Timișoara (March 2022–March 2025). MDR was defined as non-susceptibility to ≥1 agent in ≥3 antimicrobial categories. We compared MDR and non-MDR cases on demographics, microbiology, time-to-effective therapy (TTE), and outcomes; multivariable logistic regression identified independent predictors and was the basis for a points-based score with bootstrap-based internal validation (1000 resamples). Results: Fifty-four patients (41.9%) had MDR pyelonephritis. Escherichia coli remained the dominant uropathogen (55.8%) but was less prevalent in the MDR group (40.7% vs. 66.7%; p = 0.003), whereas Klebsiella pneumoniae and Pseudomonas aeruginosa were enriched. Independent predictors of MDR were antibiotic exposure ≤90 days (aOR 5.7, 95% CI 2.4–13.6), recurrent UTI (aOR 3.4, 1.4–8.2), recent hospitalization (aOR 3.1, 1.2–8.0), and renal impairment/renal-failure status at admission (aOR 2.4, 1.0–6.2). Immunosuppression, prior urinary tract instrumentation, and nephrolithiasis/urolithiasis were evaluated as candidate predictors but did not independently improve the final point score after adjustment. MDR was associated with delayed effective therapy (28.4 vs. 9.7 h; p < 0.001), longer hospitalization (13.7 vs. 8.9 days; p < 0.001), and higher 30-day readmission (20.4% vs. 8.0%; p = 0.038). The PYELO-MDR-Risk score (range 0–12) achieved an optimism-corrected AUC of 0.84 with adequate calibration (Hosmer–Lemeshow p = 0.624). Conclusions: MDR drives a substantial fraction of pyelonephritis admissions in Western Romania and tracks closely with prior antibiotic and healthcare exposure. The PYELO-MDR-Risk score offers a transparent bedside tool for empirical-therapy decisions in the local setting, pending national and international external validation. Full article

Current diagnostic methods for OSA and CSA are costly, unreliable, and the therapeutic options are limited, with varying efficacy across patients. As the prevalence of sleep apnea and cardiovascular disease continue to rise, identifying innovative therapies through advances in biomedicine and personalized medicine has become increasingly critical. This review discusses the mechanistic links between sleep apnea and subsequent cardiovascular outcomes from a technical perspective, focusing on innovations currently applied to the diagnosis and treatment of sleep apnea, and opportunities for further advancement in the field. Full article

Background: Zinc homeostasis regulated by ZIP transporters is critical for tumor glycolytic reprogramming and progression, yet the role of specific ZIP family members in lung adenocarcinoma (LUAD) remains unclear. This study aimed to identify the key ZIP transporter in LUAD and elucidate its molecular mechanisms and therapeutic value. Methods: siRNA-based functional screening of the ZIP family was performed in A549 and PC9 cells. A combination of in vitro cellular assays, in vivo animal models, clinical sample analysis and bioinformatics was used to validate the function of ZIP7 and explore its regulatory mechanisms. Results: ZIP7 (SLC39A7) was identified as a critical driver of glycolysis and proliferation in LUAD. It was significantly upregulated in LUAD tissues and cell lines. Mechanistically, ZIP7 increased inhibitory phosphorylation of GSK3β at Ser9 to stabilize NRF2, maintained low intracellular ROS levels, and sustained mTOR signaling to promote glycolytic flux. ZIP7-induced lactate secretion also drove M2-like macrophage polarization and PD-L1 upregulation to establish an immunosuppressive microenvironment. Notably, genetic or pharmacological inhibition of ZIP7 markedly enhanced the antitumor efficacy of anti-PD-1 therapy in vivo. Conclusions: ZIP7 is a pivotal oncogenic zinc transporter in LUAD that drives tumor progression via metabolic reprogramming and immune remodeling. Targeting ZIP7 represents a promising strategy to improve the efficacy of anti-PD-1 immunotherapy for LUAD. Full article

Background: Polyamide 6 microplastics (PA6-MPs), as emerging environmental pollutants, have attracted increasing attention due to their potential health risks. Their accumulation in human intervertebral disc tissue (86.4 particles/g) suggests a possible role in intervertebral disc degeneration (IVDD). However, direct evidence and mechanistic understanding remain limited. This study aimed to investigate the association between PA6-MPs exposure and IVDD, based on the hypothesis that PA6-MPs promote IVDD progression by targeting key regulatory molecules and disrupting cellular homeostasis. Methods: Potential PA6-related targets were predicted using multiple public databases, and IVDD-related differentially expressed genes were obtained from the GEO database. Overlapping targets were identified and analyzed through protein–protein interaction (PPI) network construction, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to screen core targets and pathways. Molecular docking was performed to evaluate PA6–protein binding. In vitro validation was conducted using primary human nucleus pulposus cells exposed to PA6-MPs, with cell viability, proliferation, and phenotypic changes assessed by CCK-8, EdU, live/dead staining, and immunofluorescence (IF). Results: A total of 222 PA6-related targets and 1035 IVDD-associated genes were identified, yielding 10 overlapping targets. Four core targets, including NR3C1 and HDAC1, were selected. Molecular docking and experiments demonstrated stable binding and concentration-dependent inhibition of cell viability and proliferation. Conclusion: PA6-MPs may accelerate IVDD progression in a concentration-dependent manner by targeting key molecules and perturbing inflammatory homeostasis. These findings link environmental exposure to IVDD and provide a basis for future risk assessment and targeted intervention strategies. Full article

Background: Perivascular adipose tissue (PVAT) plays a significant role in the atherosclerotic processes of arteries and aneurysmal occurrence and can be assessed by increased PVAT density in Hounsfield units (HU) on MSCT. Our objectives were to examine possible differences and associations of PVATs in several locations of aneurysmal occurrence (abdominal aorta, iliac, and lower extremity arteries), measured on the aneurysmal site and the same location on an unaffected artery. Also, we examined the possible association of aneurysmal PVAT with visceral fat parameters. Methods: In this retrospective and cross-sectional study, a total of 198 patients were included and divided into four groups: single abdominal aortic aneurysm (AAA, n = 55), single iliac artery aneurysm (n = 36), single lower extremity (leg) artery aneurysms of femoral or popliteal arteries (n = 43), and multiple aneurysms in one individual (AAA and iliac artery, n = 64). PVAT density measurements were performed at the level of the widest lumen of the aneurysm and at the same level on the contralateral artery. Aneurysmal diameter and thrombus width were also measured. The volume and the mean attenuation of visceral fat were automatically calculated by manually designating the targeted segmentation anatomical item. Results: A difference in PVAT densities among the first three groups of patients with a single aneurysm at different location was found (p = 0.027), as well as at the contralateral side (p < 0.001). In patients with multiple aneurysms, higher PVAT density was found at the iliac site compared to the AAA site (p = 0.002, paired test). Aneurysm diameter correlated to thrombus width, but not with PVAT densities. Comparison of PVAT densities with visceral fat densities showed moderate positive correlation (r = 0.446, Pearson correlation coefficient; ρ = 0.387, Spearman rank correlation; p ≤ 0.001), that was significant in the case of abdominal and iliac artery aneurysms, but not with leg aneurysms. Conclusions: Increased densities of PVAT were found on the aneurysmal site, suggesting its probable role in aneurysmal occurrence. PVAT density varied according to aneurysm location. Visceral fat may be associated with increased PVAT density and aneurysm development in abdominal and iliac regions, whereas this association was not observed in leg aneurysms. Full article

Background/Objectives: Retinitis pigmentosa (RP) is characterized by progressive degeneration of photoreceptors, with increasing evidence supporting the involvement of inflammation in disease progression. Aqueous humor (AH) reflects the intraocular microenvironment and represents an accessible source for biochemical analysis. This study aimed to characterize the profile of the main inflammatory proteins and bioactive lipids in the AH of RP patients and to compare it with healthy subjects. Methods: The AH was analyzed for cytokines using multiplex immunoassays and for lipid species using liquid chromatography–mass spectrometry. The concentrations of the analyzed molecules were compared between RP patients and the control group and then correlated with age and ellipsoid zone (EZ) width in RP patients. Results: A total of 26 RP patients and 13 controls were recruited. Significantly elevated levels of the pro-inflammatory IL-6 and a significant decrease in vascular endothelial growth factor (VEGF) were found in RP patients compared to controls. In RP patients, lipidomic analysis demonstrated significant increases in medium- and long-chain sphingomyelins (SMs) and very-long-chain unsaturated phosphatidylcholines (PCs). Higher levels of Cer 16:0, PC 32:0, and PC 34:0 were significantly associated with greater EZ preservation in RP patients. Additionally, in RP patients, VEGF and GM-CSF levels increased significantly with age, while IL-8 showed a non-significant decreasing trend. Conclusions: By integrating proteomic and, for the first time, lipidomic analyses of AH, we identified significant alterations in pro-inflammatory cytokines and bioactive lipid species in RP patients compared to controls, further highlighting a link between inflammatory activity, patient age, and disease stage. These preliminary findings need further validation in larger longitudinal cohorts to confirm the clinical utility of these bioactive mediators as potential disease biomarkers. Full article

Background: The main “peptic” cells of the gastric glands provide the body’s only source of pepsinogen synthesis and incretion. Small amounts of endogenously supplied pepsinogen in biological fluids play an important role in anabolic processes in the mother and newborn. The aim: This work aimed to analyze the dynamics of pepsinogen activity in the blood serum, saliva, urine, and coprofiltrate in pregnant women in each trimester of pregnancy and in the postpartum period, taking into account the body weight of the newborn—normal weight, underweight, and overweight. Methods: Data from studies involving non-pregnant (n = 45) and pregnant (n = 152) women with newborns with different weights were analyzed. There were 86 women with a normal-weight newborn, 34 women with an underweight newborn, and 32 women with an overweight newborn. Total proteolytic activity in biological fluids was determined using the spectrophotometric tyrosine (tyr) Kunitz–Northrop method modified by Korot’ko. A 2% solution of dry plasma was used as a substrate. Outcomes: In non-pregnant women, the blood proteolytic activity was 58.1 ± 1.4 tyr U/mL, and saliva at 1520.9 ± 112.2 tyr U/mL, urine at 4520.3 ± 154.3 tyr U/mL, and coprofiltrate at 442.2 ± 20.5 tyr U/mL. We established that the pepsinogen activity during pregnancy is distributed unevenly, taking into account the body weight of the newborn, and changes significantly in women with an underweight or overweight newborn. Conclusions: Pepsinogen homeostasis in pregnant women is maintained by renal and extrarenal pathways, and an important role is played by the salivary glands, with the most significant changes occurring in women with overweight and underweight newborns. Full article

Background: Glucagon-like peptide-1 receptor (GLP1R) agonists, particularly semaglutide, show neuroprotective effects in genetic models of Alzheimer’s disease (AD). However, their delayed and long-term effects in sporadic AD, such as the intracerebroventricular streptozotocin (STZ) injection, remain insufficient. It is unclear how long the effects of GLP1R agonists persist after discontinuation and whether a single course can suppress progressive neurodegeneration. This study aimed to evaluate the delayed effects of semaglutide administration on morphological changes in neurons and glial cells in the hippocampus associated with cognitive impairment in an STZ-induced rat model of AD. Methods: Rats received bilateral intracerebroventricular STZ injections (3 mg/kg) followed by a 5-week course of intraperitoneal administration of semaglutide (0.1 mg/kg, every other day), and were euthanized 60 days after discontinuation of semaglutide administration. Immunomorphological methods were used to detect neuronal, astrocytic and microglial alterations. A novel object recognition test was performed to assess behavioral effects. Results: STZ-treated animals demonstrated cognitive impairments, ventriculomegaly, a significant increase in p-tau protein fluorescence intensity (p = 0.02), a decrease in CA1–CA3 field area (by 23%, p = 0.008), and reduced hippocampal neuronal density. Decreases in TOMM20 (mitochondrial marker) and synaptophysin levels were accompanied by significant glial activation in the hippocampal CA3 field. Semaglutide administration significantly reduced the enlarged ventricular lumen (by 43.5%), decreased p-tau fluorescence intensity, reduced vimentin-positive reactive astrocytes (by 68.4%), and increased synaptophysin fluorescence intensity. Furthermore, it reduced microglial activation (decreasing IBA1 cell density and elongation) and alleviated the disrupted AQP4 distribution. However, semaglutide did not completely halt the neurodegenerative process and showed no effect on the number of doublecortin-positive cells in the dentate gyrus. Conclusions: Hippocampal changes assessment revealed that course administration of semaglutide exerts prolonged effects, attenuating the severity of pathomorphological alterations and behavioral changes in a sporadic AD model after drug discontinuation. Full article

Cytokines are key regulators of immune responses and tissue remodeling, playing a central role in physiological homeostasis and pathological inflammation. Dysregulation of cytokine signaling networks has been implicated in a wide range of diseases, where persistent inflammatory activation leads to progressive tissue destruction and impaired repair mechanisms. In the oral environment, cytokines critically influence the balance between tissue resorption and regeneration, particularly in processes involving dentin and alveolar bone remodeling. Pathological root resorption (PRR) represents a clinically significant model of cytokine-driven tissue destruction, characterized by the loss of dental hard tissues mediated by osteoclast-like cells within a dysregulated inflammatory microenvironment. Although mechanical, infectious, and iatrogenic factors are well-established triggers, they alone do not fully explain the variability in clinical outcomes, suggesting an important role for host-related factors. New research highlights the relationship between inflammatory signaling pathways, genetic susceptibility, and molecular biomarkers in shaping the onset and progression of PRR. In particular, the RANK/RANKL/OPG axis, cytokine networks, and gene polymorphisms have been identified as key determinants of osteoclast activation and resorptive activity. At the same time, advances in salivary and gingival crevicular fluid biomarker research provide new opportunities for early detection and real-time monitoring. Despite these advances, current knowledge remains fragmented, with heterogeneous study designs, inconsistent genetic associations, and a lack of standardized diagnostic protocols, all of which limit clinical translation. Therefore, a comprehensive and integrative synthesis of cytokine-mediated mechanisms in PRR is needed. This review aims to provide an updated and critical overview of cytokine and chemokine involvement in PRR, integrating molecular pathways, genetic determinants, and emerging biomarkers within a unified framework while highlighting translational implications for precision dentistry. Full article

Background/Objectives: Chronic low back pain (CLBP) affects approximately 20% of the global population and is a leading cause of years lived with disability. Long-term, real-world evidence for inhaled cannabis in patients refractory to conventional multimodal therapy remains scarce. We assessed the five-year efficacy and safety of inhaled cannabis in CLBP patients who had documented failure of ≥1 year of opioid analgesics, anticonvulsants, antidepressants, NSAIDs, and physiotherapy, with each patient serving as their own historical control. Methods: We analyzed prospectively collected clinical data from 241 consecutive adults with treatment-refractory CLBP (mean age 49.3 ± 14.9 years; 37.8% female; mean pain duration 15.1 years) initiated on inhaled medical cannabis (predominantly smoking, THC 4–22%, CBD 2–22%) in a single-center tertiary orthopedic clinic between 2020 and 2025 (Hasharon Hospital, Rabin Medical Center, Israel; IRB protocols 0807-21-RMC and 0634-25-RMC). Year-0 outcomes during conventional therapy were compared with outcomes at Years 1–5 on cannabis. Primary outcomes were the Numeric Rating Scale (NRS), Oswestry Disability Index (ODI), and Brief Pain Inventory severity/interference (BPI-S/BPI-I). Concomitant-medication trajectories were a secondary outcome. The primary analysis was a mixed model for repeated measures (MMRM) with random intercept and slope, REML estimation, and time as a categorical fixed effect. Multiple imputation (MAR, m = 20, Rubin’s rules) was the primary missing-data approach; complete-case and tipping-point pattern-mixture sensitivity analyses were used. A multivariate Hotelling T² provided a joint test across the four correlated PROMs. Concomitant-medication discontinuation was modeled with GEE logistic regression and exact McNemar tests. Time to discontinuation was estimated by Kaplan–Meier and Cox regression. The Bonferroni-adjusted significance threshold for the four primary outcomes was α = 0.0125. BioWell gas-discharge-visualization (GDV) parameters were exploratory only. Results: Of 241 patients, 238 (98.8%) provided Year-5 data and 224 (92.9%) remained on cannabis at Year 5; only five patients (2.1%) discontinued for adverse events or inefficacy. All four primary PROMs improved markedly and durably. MMRM-estimated Year-5 minus Year-0 changes were: NRS −5.36 (95% CI −5.65, −5.07), ODI −17.68 (95% CI −19.73, −15.63), BPI-S −6.73 (95% CI −6.99, −6.47), and BPI-I −3.41 (95% CI −3.65, −3.16); all four contrasts had |z| ≥ 16.9 and p < 10⁻²⁰. MI-pooled estimates were within 0.05 of MMRM (FMI < 0.03 for all outcomes). Hotelling T² was F(4, 232) = 872.8, p < 10⁻²⁰. At Year 5, 89.2% achieved ≥30% NRS reduction, 77.2% ≥ 50%, and 93.4% met the NRS minimum clinically important difference (MCID); ODI MCID 65.6%, BPI-S MCID (≥1 pt) 98.3%, BPI-I MCID (≥1 pt) 91.3%. Concomitant opioid use fell from 100% at baseline to 4.6% at Year 5 (within-patient absolute risk reduction 95.4%, McNemar exact p = 1.16 × 10⁻⁶⁹), NSAID from 100% to 7.1%, SSRI/SNRI from 80.5% to 5.4%, and gabapentinoid from 38.6% to 2.5%. The ARR-derived NNT for opioid discontinuation was 1.05; this NNT is referenced to each patient’s own documented maximal-conventional-therapy state and is not equivalent to a between-arm randomized-trial NNT. Cannabis dose × time interaction was consistent with no pharmacological tolerance (β = −0.0044 per gram-month per year, p = 0.074). Across 1205 patient-years of cannabis exposure (calculated as 241 patients × 5 follow-up years from Year 1 through Year 5; baseline Year 0 represents pre-cannabis state and is not included in person-time on cannabis), 1338 organ-system AE events were recorded at 1.110/patient-year (Poisson 95% CI 1.05–1.17); 99.8% of graded events were mild (grade 1), with ocular (476 events, 0.40/PY), cognitive (460, 0.38/PY), and gastrointestinal (368, 0.31/PY) reactions predominating. The Year-3 retention dip reflected a documented telemedicine-clinic phenomenon during 2022–2024, with patients returning to in-person follow-up by Year 4–5. BioWell GDV discriminated NRS ≥ 4 only at chance level (BWS AUC 0.574, 95% CI 0.54–0.60; BWV AUC 0.51). Conclusions: In a treatment-refractory CLBP cohort with five-year longitudinal follow-up, inhaled cannabis was associated with large, sustained, and statistically robust improvements in pain, disability, and pain interference, accompanied by near-total displacement of opioids, NSAIDs, antidepressants, and gabapentinoids. These observational associations, although mechanically less susceptible to bias for the binary medication-discontinuation outcomes than for self-reported PROMs, cannot be interpreted causally in the absence of a concurrent randomized control arm and may reflect a combination of pharmacological effect, regression to the mean from a high pre-treatment baseline, expectancy and self-selection effects intrinsic to an actively chosen open-label therapy, and secular trends in pain reporting. The within-patient benefit-risk profile—ARR-derived NNT ≈ 1 for opioid sparing against a predominantly mild adverse-event burden—supports consideration of cannabis as a potentially clinically meaningful, opioid-sparing option in patients who have failed multimodal conventional therapy, pending confirmation in randomized comparative trials. Full article