Search Results (221)

Collagen is the primary protein in the extracellular matrix of human cells and the body and is essential for cell structure and function. Here, for the first time, we report a method for producing recombinant triple-helical collagen type III (rhCOL3) in transgenic tobacco as a bioreactor. We constructed a pMDV-COL3A1 vector containing the human type III collagen gene COL3A1, as well as a pMDV-COL3A1:5E vector that coexpressed COL3A1 and the enzymes required for its posttranslational modification. These two vectors were used to transform tobacco genetically. The COL3A1 gene was successfully coexpressed in tobacco plants with four enzymes that promote its posttranslational modification. The transcriptional level of COL3A1 in the transgenic lines coexpressing posttranslational modification genes was greater than that in the transgenic lines expressing only COL3A1. The enzyme-modified recombinant collagen was subsequently purified from a COL3A1:5E transgenic line. Our experimental results demonstrated that the terminal propeptides of plant-derived rhCOL3 can be correctly cleaved through the enzymatic hydrolysis of procollagen by coexpressed procollagen C proteinase (PCP) and procollagen N proteinase (PNP). The plant-derived rhCOL3 was thermally stable because the purified peptide chains can form a triple helix structure. Experiments have shown that plant-derived rhCOL3 has biological activity. In this study, functional recombinant full-length mature type III collagen with a triple-helix structure was successfully expressed in tobacco, providing a foundational plant-made material for future applications of collagen in human skin and bone repair in regenerative medicine. Full article

Background: Collagen XIα1, encoded by the COL11A1 gene, is a minor fibrillar collagen that is overexpressed in various human cancers, in which its presence correlates with tumor aggressiveness and progression. Methods: In this study, we developed two novel mouse monoclonal antibodies (mAbs)—anti-colXIα1 clone 3 and anti-colXIα1 clone 9—that target the putative C-telopeptide of human collagen XIα1. These antibodies target the RRHTEGMQA sequence, a unique nine-amino-acid stretch within the putative C-telopeptide of human collagen XIα1. Results: Corresponding to nearly identical V(D)J gene segments and complementarity-determining regions (CDRs), the antibodies specifically bound the RRHTEGMQA epitope in ELISAs but did not react with the C-propeptide. This specificity was further confirmed with the purified anti-colXIα1 clone 9 mAb, which demonstrated strong reactivity against recombinant proteins containing the RRHTEGMQA sequence in both ELISAs and Western blot assays. This sequence seems to behave as a linear B-cell neoepitope, in which the RRHT motif is crucial for epitope recognition. Otherwise, no immunodetections were observed, either in cultures and lysates from the COL11A1-highly expressing A204 human cell line or on tissue sections from specimens of human pancreatic ductal adenocarcinoma (PDAC), with strong desmoplastic reactions. Conclusions: Given the lack of precise knowledge of the characteristics of the putative C-telopeptide of human collagen XIα1, the presented antibodies could enhance our understanding of the processing of human procollagen XIα1 and contribute to better characterization of the tumor microenvironment of COL11A1-expressing cancers. Full article

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection results in dysregulated inflammatory and coagulation pathways that drive immunothrombosis and contribute to adverse clinical outcomes. While individual cytokines and endothelial biomarkers have been associated with disease severity and mortality, the prognostic relevance of combined inflammatory and endothelial signatures remains incompletely characterised. To identify inflammatory cytokines and markers of endothelial activation associated with mortality in patients with severe COVID-19 requiring supplemental oxygen. This retrospective observational study included 73 consecutive adults admitted to a dedicated supplemental oxygen unit with severe COVID-19. Plasma concentrations of IL-1α, IL-1β, IL-6, IL-8, IL-10, TNF-α, von Willebrand factor (VWF) antigen and propeptide, ADAMTS13 antigen and activity, and ADAMTS13 autoantibodies were measured on admission using ELISA-based assays. Associations with mortality were assessed using non-parametric analyses, age-adjusted logistic regression, multivariable logistic regression, and receiver operating characteristic (ROC) curve analysis. Increasing age was independently associated with mortality. After adjustment for age, higher IL-1α concentrations were associated with increased odds of death, whereas a higher IL-6/IL-10 ratio was independently protective. In multivariable models, including non-ratio variables, ADAMTS13 autoantibody levels remained independently associated with mortality. In ratio-based multivariable analysis, both the ADAMTS13 activity/autoantibody ratio and the IL-6/IL-10 ratio were independently protective, while age was no longer significant. IL-10 and ADAMTS13 autoantibodies demonstrated moderate discriminative performance for mortality prediction (AUC ~0.70). A combined biomarker model incorporating IL-1α, IL-8, IL-10, and ADAMTS13 autoantibodies yielded very high predicted mortality probabilities. Our findings highlight IL-1α and ADAMTS13 autoantibodies as independent predictors of mortality in severe COVID-19, reflecting the interplay between inflammatory and endothelial pathways. Biomarker ratios capturing immune and endothelial balance—particularly the ADAMTS13 activity/autoantibody ratio—may enhance risk stratification and support integrated prognostic models. Full article

Background and Objectives: People living with HIV (PLWH) have excess osteoporosis and fractures not fully captured by dual-energy X-ray absorptiometry (DXA). We evaluated whether trabecular bone score (TBS), calcaneal quantitative ultrasound (QUS) and bone turnover markers improve vertebral fracture risk assessment beyond areal bone mineral density (BMD) in PLWH. Methods: In this cross-sectional study, 87 antiretroviral-treated adults undergoing DXA had lumbar spine TBS and calcaneal QUS. Morphometric vertebral fractures were identified, correlates of degraded TBS were analyzed using multivariable regression, and sequential logistic models quantified the incremental contribution of TBS and CTX to discriminate for prevalent morphometric vertebral fractures. Results: Low BMD (osteopenia/osteoporosis) was present in 62% of participants, degraded TBS in 37% and morphometric vertebral fractures in 17%. Degraded versus normal TBS was associated with older age (49.1 vs. 39.7 years), longer HIV duration and lower nadir CD4+ count, as well as more frequent tenofovir disoproxil fumarate exposure (66% vs. 52%; all p ≤ 0.04). In multivariable analysis, age (per 10-year increase; adjusted odds ratio [aOR] 1.78; 95% CI 1.13–2.83) and nadir CD4+ < 200 cells/mm³ (aOR 2.29; 95% CI 1.06–4.97) independently predicted degraded TBS. In sequential cross-sectional models for prevalent morphometric vertebral fractures, the area under the curve increased from 0.71 (clinical variables) to 0.79 after adding lumbar spine T-score and to 0.85 after adding TBS; adding CTX yielded 0.87 without a statistically significant incremental gain. Conclusions: In PLWH, TBS captures bone quality deficits and improves vertebral fracture risk discrimination beyond BMD, supporting its integration alongside DXA in routine HIV care. Full article

Signaling mediated by CLAVATA3/EMBRYO SURROUNDING REGION-RELATED (CLE) peptides and their receptors is essential for plants to adapt to abiotic stress. To address the global issue of drought-induced growth inhibition and mortality in poplar (Populus spp.), this study investigated the function of the PtrCLE1A gene from Populus trichocarpa Torr. et Gray in drought tolerance regulation. We employed gene cloning, expression vector construction, and genetic transformation of poplar, combined with bioinformatics analysis, subcellular localization, phenotypic observation, physiological index measurement, and gene expression analysis. The results demonstrated that both PtrCLE1A and PtrCLE1B encode pre-propeptides containing a signal peptide, with an identical mature peptide sequence (RLSPGGPDPRHH), and their putative receptors are PtrCLV1/2. Furthermore, the PtrCLE1A pre-propeptide was localized around the plasma membrane in tobacco (Nicotiana benthamiana Domin) mesophyll cells, consistent with its predicted function. PtrCLE1A and PtrCLE1B are primarily expressed in the roots and xylem of P. trichocarpa. Additionally, only the PtrCLE1A promoter contained drought-responsive cis-elements, and its expression was induced by drought stress in root, xylem, and leaf tissues of P. trichocarpa. Overexpression of the PtrCLE1A gene in Populus tomentosa Carrière (triploid) significantly increased adventitious root length under osmotic stress. Overexpression lines exhibited 22.00% to 22.92% longer adventitious roots than EV lines at 50/100 mM mannitol, and 65.12% to 73.17% longer at 150 mM mannitol. The OE lines also exhibited higher photosynthetic capacity and instantaneous water use efficiency (iWUE), along with reduced membrane damage under drought conditions, indicating enhanced drought resistance. This study provides new genetic resources and a theoretical foundation for molecular breeding of drought-tolerant poplar. Full article

Osteoarthritis (OA) is the most common multifactorial joint disease characterized by progressive cartilage degradation and impaired tissue repair. Osteochondral defects represent a major clinical challenge within OA, as damage to cartilage and underlying bone can initiate degenerative changes and contribute to joint deterioration. The Wnt/β-catenin signaling pathway plays an important role in OA pathogenesis, and its dysregulation contributes to chondrocyte catabolism and cartilage loss. NOTUM, an extracellular Wnt inhibitor, has emerged as a potential therapeutic modulator capable of restoring signaling balance and promoting cartilage homeostasis. This study aimed to evaluate the efficacy of NOTUM compared with hyaluronic acid (HA), human adipose-derived mesenchymal stromal cells (hAd-MSCs), and Colchicine in a rabbit osteochondral defect model relevant to osteoarthritis. Twenty-seven New Zealand White rabbits underwent standardized femoral condyle injury and received single-dose treatments. Serum levels of cartilage biomarkers—Procollagen Type IIA N-terminal Propeptide (PIIANP) and Cartilage Oligomeric Matrix Protein (COMP)—were measured by ELISA at 4, 6, and 8 weeks post-surgery, and histological repair at week 12 was assessed using the modified O’Driscoll scoring system. NOTUM treatment significantly increased PIIANP and decreased COMP levels compared with HA, indicating enhanced cartilage synthesis and reduced degradation. Histological scores confirmed superior surface morphology and tissue composition in NOTUM-treated joints. These findings suggest that NOTUM performs a protective and regenerative effect through Wnt/β-catenin modulation, supporting the conclusion that it enhances osteochondral defect repair and motivating further studies of NOTUM as an OA therapy. Full article

Glucose-dependent insulinotropic polypeptide (GIP) was the first incretin hormone identified, best known for promoting glucose-stimulated insulin secretion. Increasing evidence has expanded its physiological relevance beyond glucose metabolism, revealing a significant role for GIP in the gut–bone axis. In vitro studies demonstrate that GIP inhibits osteoclast differentiation and activity while promoting osteoblastic bone formation. Findings from genetic animal models and human variant analyses further support the essential role of endogenous GIP signaling in maintaining bone mass and quality. Exogenous administration of GIP suppresses the bone-resorption marker C-terminal telopeptide of type I collagen (CTX) and increases the bone-formation marker procollagen type I N-terminal propeptide (P1NP) in healthy individuals, reflecting an acute shift toward reduced bone resorption and enhanced bone formation. Moreover, GIP confers protection against bone deterioration in multiple pathological conditions, including postmenopausal osteoporosis, inflammatory bone loss, obesity, and diabetes, etc., suggesting therapeutic potential beyond physiological contexts. Recent evidence also shows that GIP attenuates orthodontic tooth movement by limiting mechanically induced osteoclast activity, highlighting its broader skeletal actions. In this review, we summarize recent advances regarding the role of GIP in bone metabolism, integrating evidence from cellular studies, animal models and human investigations, and discuss future directions for GIP-based interventions. Full article

Background/Objectives: Orthodontic tooth movement is a biological process involving coordinated bone resorption and formation in response to mechanical stimulation. The aim of this study was to evaluate the temporal changes in C-terminal telopeptide of type I collagen (CTX), procollagen type I N-terminal propeptide (PINP), and vitamin D-binding protein (VDBP) levels in gingival crevicular fluid (GCF) and saliva during fixed orthodontic treatment, as well as to assess the relationships among these biomarkers. Methods: The study included a total of 27 systemically and periodontally healthy individuals comprising 14 males and 13 females. Clinical periodontal parameters were assessed at three time points: before treatment (T0), at 24–48 h (T1), and on day 40 (T2). GCF and saliva samples were collected at the same time points. Levels of CTX, PINP and VDBP in GCF and saliva were quantified using enzyme-linked immunosorbent assay. The data were analyzed using both parametric and non-parametric statistical tests. Temporal changes across the three time points were evaluated using mixed-effects models, differences between GCF and saliva biomarker levels were assessed using paired tests, and correlations were examined using Spearman correlation analysis. Results: GCF and salivary CTX levels demonstrated a significant increase from T0 to T1, while PINP levels exhibited a substantial rise from T1 to T2 (p < 0.001). Levels of VDBP in both GCF and saliva did not demonstrate significant temporal changes (p > 0.05). Higher VDBP levels in both fluids were found to be negatively associated with increases in CTX and positively associated with increases in PINP (p < 0.05). Furthermore, salivary CTX and VDBP levels exhibited a consistent increase compared to those measured in GCF at all time points (p < 0.05). Conclusions: Fixed orthodontic forces elicit sequential resorptive and formative responses in both GCF and saliva. The potential of VDBP to function as a local modulator is indicated, with the capacity to influence the balance between osteoclastic and osteoblastic activity. The evaluation of these biomarkers in non-invasive biological samples may offer a valuable approach for monitoring bone metabolism throughout orthodontic treatment. Full article

Background: Impaired bone health is a recognized complication of type 1 diabetes. This study evaluated the effects of low-carbohydrate (LC) and Mediterranean (MED) diets on bone turnover markers in adolescents and young adults. Methods: In a 24-week randomized controlled trial, 40 individuals aged 12–21 years, with type 1 diabetes, were assigned to an LC or MED intervention (20 participants per group). C-terminal telopeptide (CTX) and procollagen type 1 N-terminal propeptide (P1NP) were measured at baseline and 24 weeks. Results: The groups had similar baselines. At 24 weeks, the between-group difference in delta glucose time in range was not statistically significant; median daily carbohydrate intake was 86 g (68–95) in LC and 130 g (102–173) in MED (p < 0.001). Comparing LC to MED, the median BMI z-score was lower (−0.1 [−0.3 to −0.1] vs. 0.0 [−0.1 to −0.1], p = 0.10), and calcium (p = 0.035) and magnesium intakes (p = 0.030) were lower. These associations did not remain statistically significant after false-discovery-rate correction. The median-adjusted alkaline phosphatase level decreased significantly in the LC group (p = 0.009). The median CTX changed following LC from 395 pg/mL (232–591) to 423 pg/mL (289–591) (p = 0.278); and following MED, from 357 pg/mL (244–782) to 296 pg/mL (227–661) (p = 0.245). P1NP changed in LC from 95 ng/mL (68–112) to 88 ng/mL (62–97) (p = 0.056) and in MED from 76 ng/mL (54–198) to 71 ng/mL (55–122) (p = 0.594). Conclusions: Exploratory analyses of bone turnover markers showed insignificant differences following LC and MED diets. Full article

Objective: The aim was to evaluate the effects of two high-resistance training (RT) protocols combined with curcumin supplementation on antioxidant capacity, systemic inflammation, bone and muscle health, and body composition. Methods: Eighty-one apparently healthy older adults [(68.2 ± 4.6 years (57% women); BMI 26.4 ± 4.8 kg/m²; minimally active according to IPAQ] were randomly allocated to accentuated eccentric (Aecc), maximal strength (Max), or a non-training control (C). Additionally, participants received either a bio-optimized curcumin formulation (Cur) or a placebo (Pla), resulting in six study groups: Aecc-Cur, Aecc-Pla, Max-Cur, Max-Pla, C-Cur, and C-Pla. Participants underwent pre- and post-intervention assessments of oxidative stress, inflammation, and bone health parameters, whole-body composition, and muscle function. Aecc and Max performed six familiarization sessions and a 16-week intervention. Participants in the curcumin groups received 500 mg/day of a bio-optimized curcumin formulation (Cursol^TM; 2 × 250 mg capsules per day, corresponding to 10.50 mg/day of curcumin) throughout the intervention. Data were analyzed using three-way repeated-measures ANOVA/ANCOVA with time (pre–post) as the within-subject factor and training group and supplementation as between-subject factors, with Least Significant Difference post hoc comparisons and effect sizes (Hedges’ g, ηp²) reported, and the significance level set at p < 0.05. Results: Aecc was the most effective in improving antioxidant capacity (glutathione; F = 25.57, p ≤ 0.001, ηp² = 0.262) and bone biomarkers (serum-procollagen type I N-propeptide—P1NP, p ≤ 0.001, ηp² = 0.504; serum beta C-terminal cross-linked telopeptide of type I collagen—β-CTX—p = 0.022, ηp² = 0.074, and their ratio—P1NP/β-CTX—p ≤ 0.001, ηp² = 0.605). Interleukin-6 (IL-6) decreased more in Aecc (p ≤ 0.001, ηp² = 0.584) and tumor necrosis factor-alpha (TNF-α) in Max (p ≤ 0.001, ηp² = 0.471). Both groups similarly improved body composition and muscle function. Bone mineral density was generally unchanged. Overall, curcumin supplementation enhanced the benefits of high-RT programs (further glutathione increase in Aecc [Hedge’s g: 0.49]; IL-6 decrease in both modalities [Hedge’s g: 0.48–1.27]; decrease in TNF-α in controls [Hedge’s g: 0.47]; better outcomes in P1NP/β-CTX in all groups [Hedge’s g: 0.46–1.46]; among others). Conclusions: Aecc is recommended for supporting antioxidant capacity and bone health, while the choice between Aecc and Max may depend on the individual’s inflammatory profile. Curcumin supplementation further amplifies the benefits of both RT protocols across most outcome variables. Full article

The present study aimed to determine the acute effects of high-intensity dynamic resistance training (HI-DRT) and whole-body electromyostimulation (WB-EMS) on markers of bone formation and resorption in young healthy women. Using a crossover design, 17 students of dentistry (26.5 ± 4.0 years, 21.5 ± 2.5 kg/m²) were randomly assigned to begin either with HI-DRT (five exercises, three sets to repetition maximum) or 20 min of non-superimposed, low-frequency (85 Hz), intermitted (6 s impulse/4 s impulse break) WB-EMS. The study outcome parameters were total Procollagen Type-1 N-Terminal Propeptide (P1NP) and Type-I Collagen Cross-Linked C-Telopeptide (CTX), which were sampled immediately prior to and 15 min post intervention. ANCOVA was applied to determine the main effects, i.e., differences in pre–post changes in CTX and P1NP between the interventions. No participant was lost to follow-up or reported adverse effects related to the exercises. Briefly, we observed significant differences (p = 0.019, d′ = 1.19) for changes in P1NP that were maintained in the HI-DRT (p = 0.446) and decreased in the WB-EMS group (p = 0.002). In contrast, we did not observe differences for HI-DRT- vs. WB-EMS-induced CTX changes (p = 0.509; d′ = 0.134). In summary, while HI-DRT provides significantly more favorable effects on bone formation markers compared to WB-EMS, the clinical significance of this finding in predicting the general effectiveness of an exercise protocol on bone strength remains to be determined. (Clinical trials.gov; registration date: 2025-02-06; ID: NCT06813092.) Full article

There are 25 crystal structures of Lipase B from Candida antarctica (CalB) that have been previously reported. In this study, we report the first CalB crystal structure that shows the assumed pro-peptide region at the N-terminus (Ala19–Arg25). This 1.45 Å structure shows that this segment of seven amino acids is an extension of the N-terminal loop and that it does not interact with or effect conformational changes in the flexible lid domain, which covers the active site of the enzyme. As such, this region is unlikely to be a classical pro-peptide. Full article

Maintaining muscle health is essential for preserving mobility, independence, and quality of life with age. As muscle mass and function decline, the risk of frailty, chronic disease, and disability increases. Sarcopenia, characterized by the progressive loss of muscle mass, strength, and function, is a major contributor to these adverse outcomes in older adults. Early identification and monitoring of sarcopenia are critical for timely intervention to prevent irreversible decline. Muscle biomarkers offer a promising approach for detecting muscle deterioration and guiding treatment strategies. This review explores key biomarkers—including insulin-like growth factor 1 (IGF-1), myostatin, interleukin-6 (IL-6), irisin, interleukin 15 (IL-15), and procollagen type III N-terminal propeptide (P3NP)—that reflect underlying processes such as muscle anabolism, inflammation, metabolism, and remodeling. Alterations in these markers are associated with muscle health status. Furthermore, hormonal status, biological sex, and nutritional factors all modulate biomarker levels, emphasizing the need for personalized assessments. Integrating biomarker analysis into clinical practice has the potential to enhance early diagnosis, inform personalized interventions, and ultimately promote healthy aging by maintaining muscle function and reducing disability risk. Full article

Background: Atrial fibrillation (AF) is common complication of heart failure with preserved ejection fraction (HFpEF) that sufficiently intervenes in the prognosis. The aim of the study is a) to investigate the possible discriminative value of adropin for newly onset AF in patients with HFpEF without a previous history of AF and who are being treated in accordance with conventional guideline and b) to compare it with predictive potencies of conventionally used predictors. Methods: A total of 953 patients with HFpEF who had sinus rhythm on ECG were enrolled in the study. The course of the observation was 3 years. Echocardiography and assessment of conventional hematological, biochemical parameters and biomarker assay including N-terminal brain natriuretic pro-peptide (NT-proBNP), high-sensitivity cardiac troponin T, tumor necrosis factor-alpha, high-sensitivity C-reactive protein (hs-CRP), galectin-3, interleukin-6, soluble suppressor tumorigenisity-2 (sST2) and adropin, were performed at baseline. Results: Incident atrial fibrillation was found in 172 patients with HFpEF, whereas 781 had sinus rhythm. In unadjusted rough Cox regression model, age ≥ 75 years, type 2 diabetes mellitus, chronic kidney disease (CKD) stages 1–3, left atrial volume index (LAVI) ≥ 40 mL/m², NT-proBNP ≥ 1440 pmol/mL, hs-CRP ≥ 5.40 mg/L, adropin ≤ 2.95 ng/mL, sST2 ≥ 15.5 ng/mL were identified as the predictors for new onset AF in HFpEF patients. After adjusting for age ≥ 75 years, a presence of type 2 diabetes mellitus and CKD stages 1–3, the levels of NT-proBNP ≥ 1440 pmol/mL and adropin ≤ 2.95 ng/mL were independent predictors of new onset AF in patients HFpEF. We also found that discriminative value of adropin was superior to NT-proBNP, while adding adropin to NT-proBNP did not improve predictive information of adropin alone. Conclusions: adropin ≤ 2.95 ng/mL presented more predictive information than NT-proBNP ≥ 1440 pmol/mL alone for new cases of AF in symptomatic patients with HFpEF, whereas the combination of both biomarkers did not improve the predictive ability of adropin alone. Full article

Independent studies reported metabolic alterations in connective tissues of hernia patients, especially involving collagen fibers, compared to healthy controls. In the present work, we evaluated plasma concentrations of metalloproteinases (MMPs) and lysyl oxidase (LOX), enzymes involved in collagen metabolism, and peptides produced during collagen biosynthesis (PINP, PIIINP, and PIVNP) as potential biomarkers for the estimation of hernia risk. Zymography and ELISA assays were performed with plasma samples of 51 patients with primary or recurrent inguinal hernia and 42 healthy controls. A reduction in PINP (p = 0.007) and a concomitant increase in PIIINP (p < 0.001) were observed in patients. In controls, PINP levels were inversely related to age, whereas in patients PIIINP levels increased with age. Body mass index (BMI) showed a strong positive correlation with PIIINP plasma levels in controls but not in patients (p < 0.001). Moreover, patients with larger lesions had the lowest PINP/PIIINP ratio (p = 0.003). PIVNP collagen did not differ between controls and hernia patients. Plasma MMP-9 was reduced in patients (p = 0.015), while MMP-2 and LOX were unchanged. However, MMP-2 concentrations appeared lower in patients with familial history of hernia compared to those without. In regression analysis, the PINP/PIIINP ratio was inversely related to hernia risk, and a cut-off value of 0.948 was found by ROC analysis which classified hernia patients with a sensitivity of 82.9% and a specificity of 77.1%. In conclusion, our findings identified the PINP/PIIINP ratio as the most relevant molecular predictor of inguinal hernia risk. Full article