C-terminal Propeptide Research Articles

Utilizing cluster analysis of biomarkers alongside echocardiographic deformation parameters to forecast the outcome of atrial fibrillation post-ablation

Abstract Background Atrial fibrillation (AF) management often involves catheter ablation, but its recurrence necessitates careful clinical management and potential adjustments in medication or further ablation procedures. This study aims to investigate the predictive value of biomarkers and echocardiographic parameters for early AF recurrence. Methods In the prospective study, we enrolled patients with symptomatic paroxysmal or persistent atrial fibrillation undergoing catheter ablation. Pulmonary vein isolation was performed in every index procedure, with additional ablation or substrate modification as per physician preference. Blood samples were collected from both the atrium and peripheral vein to assess levels of tumor endothelial marker 1 (TEM1), galectin 3, connective tissue growth factor (CTGF), transforming growth factor-beta (TGF-β), C-terminal propeptide of procollagen type I (PICP), and Smad3. Pre-procedural echocardiographic parameters, including the E/e’ ratio, peak atrial longitudinal strain (PALS), and peak atrial contractile strain (PACS), were recorded. Clustering models with sparse canonical association analysis (CCA) were employed to develop predictive models integrating biomarker and echocardiographic data for early atrial fibrillation recurrence. Results Out of 80 prospectively enrolled patients, 68 completed the study, with six cases of early atrial fibrillation recurrence observed. Following dimension reduction and clustering, we found that the combined use of biomarkers and echocardiography significantly predicted atrial fibrillation recurrence (HR: 9.44, p=0.05, 95% CI: 1.0-88.9). Notably, among these parameters, TEM1 biomarker sampled from the peripheral vein (p=0.02), left atrium dimension (p=0.07), and left ventricle dimension (p=0.02) exhibited the most significant predictive power. Conclusions Our study highlights the effectiveness of integrating biomarkers and echocardiography parameters in predicting early recurrence of atrial fibrillation post-ablation in patients undergoing catheter ablation.main_figure

Insights for possible association and impact of thyroidectomy to osteoarthritis

Background and aim of studyThyroidectomy and osteoarthritis have drawn more attention in last decades due to increase various local and systemic risk factors. This study is aimed to determine the association and impact between thyroidectomy and osteoarthritis by serological measurement of most specific related markers.ResultsMeasurement of thyroid markers showed the level of thyroid-stimulating hormone (TSH) was significantly increased, while parathyroid hormone (PTH), triiodothyronine (T3), and thyroxine (T4) levels were decreased in osteoarthritis subjected to thyroidectomy group (OTG) when compared to hyperthyroidism subjected to thyroidectomy group (TG), osteoarthritis group (OG), and healthy control group (CG). Detection the activity of bone markers showed the level of R-factor was significantly elevated concomitant with significant reduction in Dickkopf related protein 1 (DKK1), human hyaluronan-binding protein 2 (HABP2), osteocalcin (OC) in OG and OTG groups, while osteopontin (OPN) and procollagen I C-terminal propeptide (PICP) were significantly increased and decreased in TG and OTG. Furthermore, the level of S100 Calcium binding protein (S100CBP) showed significant decreased in patient’s groups, while TG with OTG groups exhibited significant reduction in sclerostin (SOST) concentration. Regarding the inflammatory markers, the levels of interleukin-1 (IL-1) was increased in the OTG, while the level of interleukin-10 (IL-10) was increased in OG and TG groups, and reduced in OTG. While, the level of transforming growth factor-beta (TGF-β) was decreased in OG and TG associated with significant increases in tumor necrosis factor-alpha level (TNF-α) in OTG. Measurement of oxidant and antioxidant activity markers showed the levels of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were significantly reduced in all patient’s groups compared to control, except the level of CAT in TG, whereas, malondialdehyde (MDA) level was increased in OG and OTG patients. Furthermore, the levels of Alkaline phosphatase (ALP), C-Reactive protein (CRP), and erythrocyte sedimentation rate (ESR) were increased in all patient groups compared to control, while fatty acid-binding protein (FABP) level was increased in OTG only.ConclusionThis unique study in Iraq is identified the interaction effect and impact of thyroidectomy to osteoarthritis according to the results that showed various changes and degree of correlation of study biomarkers in all patient groups, however more depth of specific quantitative and qualitative studies are required to support this association and the impact claim at molecular level.

Using targeted proteomics-based detection of collagen propeptides to quantify fibrillar collagen biogenesis in vitro

The collagen superfamily, as the major structural component of the extracellular matrix, encompasses 28 distinct subtypes, with type-I and –III forming fibrils crucial for the matrix scaffold. During collagen biogenesis, trimers of type-I and –III procollagen are secreted into the extracellular matrix. The N- and C-terminal propeptides of these trimers are proteolytically cleaved from procollagen during secretion, initiating collagen fibril formation. The propeptides are released into extracellular space and, therefore, have been used to quantify collagen biogenesis. But high-throughput methods for the quantification of these biomarkers are still lacking. This study presents a state-of-the-art multiplexed approach for the simultaneous quantification of PINP, PICP, PIIINP and PIIICP from cell culture supernatants. The ability of targeted proteomics to quantify these propeptides from cell culture samples was assessed in this study. Using tryptic digestion and solid phase extraction, we were able to accurately quantify precollagen propeptides in a range of 3–1000 ng/mL. The assay showed an average inter-assay variance of 6.86 % with an overall recovery ranging from 92 to 98 %. The assay was validated using recombinant protein standards diluted in surrogate matrix and tested using transforming growth factor β1 mediated induction of normal human dermal fibroblasts. In summary, the assay presented in this paper offers a novel, robust, and precise high-throughput method for measuring human collagen propeptides in cell culture supernatants, empowering researchers to assess collagen biogenesis effectively in in vitro experiments.

Asprosin as a potential new paracrine regulator in the heart

Abstract Funding Acknowledgements None. Background Asprosin was recently described as a novel hormone that is released by white adipose tissue upon fasting. However, a functional role of asprosin in human cardiovascular tissues has not been yet investigated at the tissue level. Asprosin is the C-terminal propeptide of the extracellular matrix (ECM) glycoprotein fibrillin-1 and is cleaved off from the pro-fibrillin-1 precursor by the endopeptidase furin. Fibrillin-1 is expressed in the heart and aorta, and FBN1 mutations are causative for aortic aneurysm formation and cardiomyopathy. Whether asprosin is also present in cardiac tissues and utilized by cardiac resident cells remains unknown. Purpose In the present study, we investigated the localization and distribution of asprosin protein in human atrial and ventricular heart tissue. Methods Human heart tissue was obtained from patients undergoing aortic valve replacement. In addition, we used ventricular tissue from mice. Cryosections from human (n=8) and mouse (n=8) heart samples were analysed by immunohistochemistry. The anti-asprosin antibodies were raised against human or mouse asprosin respectively and applied in combination with anti-CD31/PECAM, an endothelial cell marker, or anti-sarcomeric alpha-actinin, a Z-line marker. The immunofluorescence of incubated specimens was examined by confocal microscopy. Results Our data suggest that asprosin is localized in myocardium in different cell types: cardiomyocytes (Figure 1 a-d), arterial smooth muscle cells (Figure 1 a), micro- and vascular endothelial cells (Figure 1 d), and with the lowest intensity, in periarterial located epicardial adipocytes (Figure 1 e). In cardiomyocytes, incubation with anti-asprosin resulted in a high signal intensity. Asprosin immunoreactivity was observed: in intercalary discs (Figure 1 b-d), as striated pattern in the sarcoplasm, and as weak signal in the perinuclear area (Figure 1 b, c). In human tissue, the anti-asprosin signal was about four times higher in intercalated discs than in the sarcoplasm. The distribution pattern in ventricular cardiomyocytes of mice were similar. Conclusion Our results show that in human myocardium, asprosin localises mainly intracellular. Asprosin accumulations were found in intercalary discs. In short, asprosin was found at locations that are known to be crucial for regulation of cardiac function. Intracellular presence of asprosin may indicate cellular utilization by uptake from extracellular asprosin pools. Whether these pools are replenished exclusively by resident cells or by other tissues such as WAT is currently unknown. However, further research is needed to prove this hypothesis. Figure. 1. Confocal microscopy of asprosin localization in human myocardium. Cryosections incubated with anti-asprosin (red) or combined with anti-alpha actinin (sarcomeric) or anti-CD31/PECAM (green) antibodies and DAPI (blue, nuclei). SM: vascular smooth muscle cells, BV: blood vessels, Arrowheads: Intercalated disks.

Effect of antidepressants on bone mineral density and bone metabolism in ovariectomized depressed rats

Purpose: To investigate the effect of antidepressants on bone mineral density and bone metabolism in ovariectomized rats with depression. Methods: 48 female rats were randomly and equally assigned to six groups, namely, sham (Sn), ovariectomized depression (OD), ovariectomized depression rats treated with sertraline (ODs), citalopram (ODx), reboxetine (ODr), and venlafaxine groups (ODw). Behavioral alterations and bone-related parameters were evaluated before and after treatment. Results: After treatment, ODs, ODx, ODw, and ODr groups had higher levels of horizontal and vertical exercise scores, sugar water consumption, osteoclasts, and serum CTX-1 (p < 0.05) compared to OD group. Osteocytes, serum serotonin (5-HT), osteocalcin (BGP), and type I procollagen N-terminal propeptide (PINP) were significantly decreased (p < 0.05) after treatment in ODs, ODx, ODw, and ODr groups. A positive correlation was observed between 5-HT, BGP, PINP, and whole-body bone mineral density (r = 0.931, 0.907, and 0.843, p < 0.05) respectively, while a negative correlation was observed between collagen type I C-terminal propeptide (CTX-I) and bone mineral density (r = -0.855, p < 0.05). Conclusion: Antidepressants reduce bone mineral density and osteocyte proliferation, while increasing osteoclast proliferation. These effects are associated with reductions in 5-HT, PINP and BGP levels, and increase in CTX-I.

The crosslinking sites and molecular conformation of gelatin hydrogel modified by transglutaminase

This work aimed to investigate the covalent crosslinking sites induced by transglutaminase (TGase, EC 2.3.2.13) in gelatin hydrogel and their impact on the gelatin molecular conformation. LC-MS/MS results demonstrated that Lys644 and Gln1351 of α1 chain and Gln19 of α2 chain were the main sites. The results of molecular dynamic (MD) simulation indicated that at the moderate degree of covalent crosslinking (13.55%), Radius of gyration (Rg) values decreased from 11.5 to 10.0 nm at 4 °C and led to a more compact structure, therefore enhancing structural tightness and gel strength. As the degree of crosslinking rose, the number of crosslinking sites gradually increased, with a predominant distribution in the C-terminal pro-peptide of gelatin chain. At the high crosslinking degree (37.48%), Rg value decreased at 100 °C and gelatin chains were connected through a “head to tail” linkage configuration, resulting in a significant reduction in the triple helix-like structure. The structure maintained the gel-forming ability at high temperature while lowering the aggregation energy of water molecules from −2.359 × 105 to −2.455 × 105 kJ/mol, leading to thermal irreversibility of gelatin hydrogel.

Proteomic profiles of left atrial volume and its influence on response to spironolactone: Findings from the HOMAGE trial and STANISLAS cohort.

High left ventricular filling pressure increases left atrial volume and causes myocardial fibrosis, which may decrease with spironolactone. We studied clinical and proteomic characteristics associated with left atrial volume indexed by body surface area (LAVi), and whether LAVi influences the response to spironolactone on biomarker expression and clinical variables. In the HOMAGE trial, where people at risk of heart failure were randomized to spironolactone or control, we analysed 421 participants with available LAVi and 276 proteomic measurements (Olink) at baseline, month 1 and 9 (mean age 73 ± 6 years; women 26%; LAVi 32 ± 9 ml/m2). Circulating proteins associated with LAVi were also assessed in asymptomatic individuals from a population-based cohort (STANISLAS; n = 1640; mean age 49 ± 14 years; women 51%; LAVi 23 ± 7 ml/m2). In both studies, greater LAVi was significantly associated with greater left ventricular masses and volumes. In HOMAGE, after adjustment and correction for multiple testing, greater LAVi was associated with higher concentrations of matrix metallopeptidase-2 (MMP-2), insulin-like growth factor binding protein-2 (IGFBP-2) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) (false discovery rates [FDR] <0.05). These associations were externally replicated in STANISLAS (all FDR <0.05). Among these biomarkers, spironolactone decreased concentrations of MMP-2 and NT-proBNP, regardless of baseline LAVi (pinteraction > 0.10). Spironolactone also significantly reduced LAVi, improved left ventricular ejection fraction, lowered E/e', blood pressure and serum procollagen type I C-terminal propeptide (PICP) concentration, a collagen synthesis marker, regardless of baseline LAVi (pinteraction > 0.10). In individuals without heart failure, LAVi was associated with MMP-2, IGFBP-2 and NT-proBNP. Spironolactone reduced these biomarker concentrations as well as LAVi and PICP, irrespective of left atrial size.

Cardiac remodeling in patients with heart failure with mildly reduced ejection fraction and metabolic disorders: association with biomarkers and autonomic nervous system parameters

Aim. The high prevalence of obesity in a cohort of patients with heart failure and mildly reduced ejection fraction (HFmrEF) determines the relevance of clarifying the role of biomarkers and autonomic imbalance in myocardial remodeling, taking into account metabolic risk factors.Material and methods. We examined 19 men with postinfarction cardiosclerosis and class II HFmrEF (median age 62 years), overweight/class I-II obesity, type 2 diabetes in 53/47%, 48% of cases, respectively, who received therapy. The biomarker panel included N-terminal pro-brain natriuretic peptide (NT-proBNP), galectin-3, pro-collagen I C-terminal propeptide (PICP), N-terminal propeptide of procollagen type III (PIIINP), C-terminal telopeptide of type I collagen, matrix metalloproteinase-9 (MMP-9), tissue inhibitor of matrix proteinase-1 (TIMP-1), leptin and adiponectin. Heart rate variability (HRV) and turbulence were obtained using 24-hour Holter monitoring. We assessed the time and frequency domains of HRV (24 h) and 5 min recordings of wakefulness at rest, calculated TO (turbulence onset) and TS (turbulence slope).Results. Significant positive associations of leptin and TIMP-1 levels with left ventricular hypertrophy markers were confirmed. Positive correlations of peak e' with following HRV indicators were revealed: SDNN (r=0,68; p=0,02) and RMSSD (r=0,69; p=0,003). Lower TS values were associated with higher index parameters of left ventricular mass (p&lt;0,05 for all). Associations of biomarkers with autonomic nervous system (ANS) were observed: MMP-9 with RMSSD (r=0,54) and pNN50 (r=0,51); TIMP-1 with TO (r=0,46); PICP/PIIINP ratio with HFn (5 min) (r=-0,49); NT-proBNP/adiponectin ratio with SDNN (r=-0,49); leptin level with TS (r=-0,54) (p&lt;0,05 for all).Conclusion. In patients with HFmrEF of ischemic origin and additional metabolic risks, serum biomarkers of fibrosis, adipokines, and ANS parameters are associated mainly with markers of increased left ventricular filling pressure. The study results predetermine the further search for potential risk-stratification markers of unfavorable myocardial remodeling and prognosis in large samples of patients with metabolic deviations and HF with EF &gt;40% against the background of modern drug therapy.

Markers of bone metabolism and overall survival in men with bone-metastatic hormone sensitive prostate cancer (HSPC): A subset analysis of SWOG S1216, a phase III trial of androgen deprivation with or without orteronel.

Circulating biomarkers of bone metabolism are significantly associated with overall survival (OS) in men with advanced prostate cancer. In the SWOG S1216 phase III trial, we showed that elevated bone biomarkers are significantly associated with an increased risk of death in hormone-sensitive prostate cancer (HSPC) regardless of the status of bone metastases, identifying three risk groups with differential OS outcomes based on bone biomarker status. Here we report the association of bone biomarkers with OS in men with HSPC and documented skeletal metastases as part of a planned subset analysis of S1216. Bone resorption [C-telopeptide (CTx); Pyridinoline (PYD)] and bone formation markers [C-terminal collagen propeptide (CICP); bone alkaline phosphatase (BAP)] were assessed in blood from men with bone metastatic HSPC. Patients were randomly divided into training (n = 238) and validation (n = 475) sets. In the training set, recursive partitioning that maximizes discrimination of OS was used to identify the dichotomous cut-point for each biomarker and for a combination of biomarker split points to define prognostic groups. In the validation set, Cox proportional hazards models were used to assess the impact of biomarkers on OS, adjusted for patient and tumor characteristics. Of 1279 men, 713 had both baseline bone metastases and evaluable bone biomarkers. Patient characteristics were similar between the overall population and the subset with bone metastases. Elevated levels of CICP, CTX, and PYD were strongly prognostic for OS. Hazard ratios (95% CI) for OS adjusted for treatment arm and baseline clinical variables were: BAP-1.31 (0.93, 1.84), p = 0.12; CICP-1.58 (1.09, 2.29), p < 0.02; CTx - 1.55 (1.12, 2.15), p = 0.008; and PYD-1.66 (1.27, 2.217), p = 0.0002. There was no evidence of interaction between elevated biomarkers and treatment (all p > 0.2). Recursive partitioning algorithms identified four groups of patients with differential OS outcomes based on bone biomarkers, adjusted for baseline clinical variables, with median OS ranging from 2.3 years (highest risk group) to 7.5 years (lowest risk group). In this planned S1216 subset analysis of men with HSPC and bone metastases, elevated serum markers of bone metabolism were significantly associated with worse OS. Bone biomarker levels alone and in combination with patient and tumor characteristics identify unique subsets of men with differential OS outcomes. NCT01809691.

Daily supplement of sesame oil prevents postmenopausal osteoporosis via maintaining serum estrogen and aromatase levels in rats

Estrogen deficiency is one of the main causes of postmenopausal osteoporosis in elderly women. Hormone replacement therapy has been employed to manage postmenopausal osteoporosis; however, it has raised concerns related to heart attacks and breast cancer. Sesame oil has been reported to affect sex hormone status. The aim of the present study is to evaluate the effect of sesame oil supplement on postmenopausal osteoporosis in rats. We used female Sprague Dawley rats that underwent bilaterally ovariectomy (OVX) as an experimental postmenopausal osteoporosis animal model. These rats were orally administrated sesame oil (0.25 or 0.5 mL/kg/day) for four months as the therapeutic group. We assessed bone mineral density (BMD) and the levels of osteocalcin, procollagen-I C-terminal propeptide (PICP), collagen cross-linked N-telopeptide (NTx), estradiol, and aromatase in the sera. The daily supplementation of sesame oil significantly increased BMD, serum osteocalcin levels, and trabecular areas in the OVX-treated rats. Sesame oil also elevated serum PICP levels and decreased NTx levels in these rats. Furthermore, sesame oil effectively maintained serum estradiol and aromatase levels in the OVX-induced osteoporosis rats. In conclusion, daily supplementation of sesame oil prevents postmenopausal osteoporosis by maintaining serum estrogen and aromatase levels, while also modulating the imbalance between bone formation and resorption in osteoporosis rats.

Prospective Assessment of Bone Metabolism Biomarkers and Survival in Metastatic Castration-resistant Prostate Cancer Patients Treated with Radium-223: The PRORADIUM Study

BackgroundRadium-223 is an active therapy option for bone metastatic castration-resistant prostate cancer (mCRPC). The lack of adequate biomarkers for patient selection and response assessment are major drawbacks for its use. ObjectiveTo assess the prognostic value of bone metabolism biomarkers (BMBs) in ra-223–treated mCRPC patients. Design, setting, and participantsA prospective cohort study of mCRPC patients treated with Ra-223 (PRORADIUM study: NCT02925702) was conducted. Outcome measurements and statistical analysisThe main objective of the study was to evaluate the association between high (≥median) baseline values in at least three bone formation (bone alkaline phosphatase [BAP] and C-terminal type-I collagen propeptide) and bone resorption (N-terminal telopeptide and pyridinoline) biomarkers, and survival. The independent prognostic value of each BMB was also assessed. The association with time to radiographic, clinical, and prostate-specific antigen (PSA) progression; time to skeletal-related events; and PSA response were secondary objectives. Multivariable (MV) Cox-regression models were evaluated. Results and limitationsA total of 169 patients were included. Of the patients, 70.4% received Ra-223 in second/third line; 144 (85.2%) were Eastern Cooperative Oncology Group 0–1, 126 (74.6%) were in pain, and 80 (47.5%) had more than ten bone metastases. Sixty-seven (39.6%) patients had elevation in at least three BMBs. The median overall survival was 12.1 mo (95% confidence interval [CI]: 10–14.7). No association was observed with other treatment-related secondary outcome parameters. Patients with high values in three or more BMBs had significantly worse survival (9.9 vs 15.2 mo; hazard ratio [HR]: 1.8 [95% CI: 1.3–2.5]; p < 0.001) in the univariate analysis, but not independent in the MV analysis (HR: 1.33; 95% CI: 0.89–2; p = 0.181). High baseline BAP was the only biomarker associated with survival in the MV model (HR: 1.89; 95% CI: 1.28–2.79; p = 0.001). Addition of BAP to the MV clinical model increased the area under the receiver operating characteristic curve 2-yr value from 0.667 to 0.755 (p = 0.003). ConclusionsBiomarkers of bone formation, especially BAP, have prognostic value in mCRPC patients treated with radium-223. Its predictive value remains to be assessed, ideally in prospective, adequately powered, randomised clinical trials. Patient summaryIn this study, we evaluate the role of bone metabolism biomarkers to help improve the use of radium-223 as therapy for advanced prostate cancer. We found that bone alkaline phosphatase may be a suitable tool.

Riboflavin deficiency reduces bone mineral density in rats by compromising osteoblast function

Insufficient riboflavin intake has been associated with poor bone health. This study aimed to investigate the effect of riboflavin deficiency on bone health in vivo and in vitro. Riboflavin deficiency was successfully developed in rats and osteoblasts. The results indicated that bone mineral density, serum bone alkaline phosphatase, bone phosphorus, and bone calcium were significantly decreased while serum ionized calcium and osteocalcin were significantly increased in the riboflavin-deficient rats. Riboflavin deficiency also induced the reduction of Runx2, Osterix, and BMP-2/Smad1/5/9 cascade in the femur. These results were further verified in cellular experiments. Our findings demonstrated that alkaline phosphatase activities and calcified nodules were significantly decreased while intracellular osteocalcin and pro-collagen I c-terminal propeptide were significantly increased in the riboflavin-deficient osteoblasts. Additionally, the protein expression of Osterix, Runx2, and BMP-2/Smad1/5/9 cascade were significantly decreased while the protein expression of p-p38 MAPK were significantly increased in the riboflavin-deficient cells compared to the control cells. Blockage of p38 MAPK signaling pathway with SB203580 reversed these effects in riboflavin-deficient osteoblastic cells. Our data suggest that riboflavin deficiency causes osteoblast malfunction and retards bone matrix mineralization via p38 MAPK/BMP-2/Smad1/5/9 signaling pathway.

Breed-specific evaluation of serum biochemical markers in canine hip dysplasia observed in a Tornjak dog population.

Canine hip dysplasia (CHD) is a global disease and an improved diagnostic approach to this orthopedic condition is needed, with an emphasis on inexpensive and minimally invasive techniques. This research aimed to determine the differentiating potential of selected biochemical markers in serum between healthy dogs and dogs with hip dysplasia in a breed-specific study that involved the Tornjak dog population. 99 Tornjak dogs radiographically categorized (Federation Cynologique Internationale procedure and scoring scheme) between December 2019 and April 2021, as having no sign of hip dysplasia or near normal hip joints (nondysplastic group; n = 51) vs mild, moderate, or severe hip dysplasia (dysplastic group; 48). Serum concentrations and enzyme activity of the biochemical markers hyaluronic acid (HA), procollagen II C-terminal propeptide (PIICP), and metalloproteinase 9 (MMP9) were compared among adult dogs with or without radiographic hip dysplasia. Statistical tests used to examine the differentiating potential of biochemical markers in Tornjak dog groups were assessed using the Mann-Whitney U test, logistic regression, and receiving operating characteristics (ROC) analysis. Tornjak dogs with radiographic CHD had significantly lower serum concentrations of HA and higher concentrations of PIICP and MMP9 activity compared to dogs with radiographically normal hips (P < .05). Selected biochemical markers could distinguish dogs with radiographic CHD from those without CHD with high sensitivity and specificity. These data suggest that the diagnostic technique of measuring serum concentrations of HA, PIICP, and MMP9 activity has a selective ability to distinguish dogs with dysplastic from dogs with normal hips.

Link between fibrosis-specific biomarkers and interstitial fibrosis in hypertrophic cardiomyopathy.

Cardiac fibrosis is a hallmark of hypertrophic cardiomyopathy (HCM) and has proven unfavorable clinical significance. Replacement fibrosis is better known, and has already been studied on a larger scale, whereas interstitial fibrosis is less explored. We aimed to analyze relationship between serum biomarkers and interstitial fibrosis, as assessed with cardiac magnetic resonance (CMR) in HCM. We performed 3T CMR scans in 50 HCM patients to assess interstitial fibrosis as expressed by extracellular volume (ECV). In all patients, we determined levels of serum cardiac-specific (troponin T [TnT], N-terminal prohormone of brain natriuretic peptide [NT-proBNP]) and fibrosis-specific (procollagen I C-terminal propeptide, procollagen III N-terminal propeptide, transforming growth factor β1, galectin 3) biomarkers. Patients were divided based on their median value of ECV. The final study population consisted of 49 patients. The median value of ECV in our cohort was 28.1%. Patients stratified according to median ECV differed in terms of several variables: body mass index, late gadolinium extent, NT-proBNP and galectin 3 levels (all P < 0.05). Cardiac biomarkers (TnT and NT-proBNP) and galectin 3 were significantly correlated with ECV (rS = 0.34; P = 0.02; rS = 0.39; P = 0.006; rS = 0.43; P = 0.002, respectively). Galectin 3 and body mass index were found to be independent predictors of ECV (odds ratio [OR], 2.29 [1.07-4.91]; P = 0.03; OR, 0.81 [0.68-0.97]; P = 0.02, respectively). Galectin 3 was an independent predictor of interstitial fibrosis in HCM patients expressed as elevated ECV values. The other fibrosis-specific biomarkers measured were not useful in detecting interstitial fibrosis in HCM. In addition, there was a positive correlation between classical cardiac biomarkers and interstitial fibrosis in HCM patients.

Effects of Etanercept and Adalimumab on Serum Levels of Cartilage Remodeling Markers in Women with Rheumatoid Arthritis.

Tumor necrosis factor α inhibitor (TNFαI) therapy is associated with a significant inhibition of radiographic progression, resulting in improved physical function and quality of life among patients with rheumatoid arthritis (RA). The mechanism by which TNFαI prevent joint destruction is still unknown. In this study, the effect of 15-month anti-TNF-α therapy in combination with methotrexate on circulating levels of biochemical markers of cartilage turnover in female RA patients was assessed. Serum levels of collagen type II C-terminal cleavage neoepitope (C2C), C-terminal propeptide of type II collagen (PIICP), cartilage oligomeric matrix protein (COMP), and matrix metalloproteinase-3 (MMP-3) were evaluated using immunoassays at baseline and 15 months after the start of TNFαI treatment. Baseline COMP, C2C, and MMP-3 levels and C2C/PIICP ratios were significantly higher in women with RA compared with those observed in the healthy subjects. No differences in PIICP levels between the controls and the women with RA were observed. After 15 months of TNFαI treatment, serum levels of C2C, COMP, and MMP-3 decreased, whereas the levels of PIICP increased but were still not different from those of the controls. These changes were accompanied by significantly reduced C2C/PIICP ratios. Before the start of TNFαI therapy, serum levels of COMP significantly correlated with the patients' ages (p < 0.05) and their 28-joint disease activity score values based on their erythrocyte sedimentation rates (DAS28-ESR; p < 0.05). Moreover, multiple linear regression analysis showed that baseline COMP levels retained a significant association with DAS28-ESR value (β = 287.74, p = 0.022, R2 model = 0.25) after model adjustments. The largest area under the ROC curve was obtained for C2C/PIICP ratios (AUC: 0.830, 95% CI: 0.727-0.932, p < 0.001). Our results suggest that long-term anti-TNF-α therapy combined with MTX has a beneficial effect on cartilage remodeling that is associated with clinical improvement among RA patients. Serum C2C/PIICP ratios may help to monitor the effectiveness of anti-TNF-α treatment among RA patients.

Correlation between Serum Myosin Light Chain 4 Levels and Recurrence after Radiofrequency Ablation in Patients with Atrial Fibrillation.

Atrial fibrillation (AF) is the most common arrhythmia that is harmful to human health. This study aims to explore the relationship between myosin light chain 4 (MYL4) and AF recurrence after radiofrequency ablation (RFA). Patients with AF (n = 85) were enrolled, and healthy subjects (n = 90) with normal sinus rhythm and no previous history of AF were selected as controls. The serum levels of MYL4, transforming growth factor (TGF) -β1, and procollagen type-I C-terminal propeptide (PICP) were determined. The correlation between MYL4 and atrial fibrosis remodeling indicators (TGF-β1/PICP) and left atrial diameter (LAD) was analyzed. The influence of MYL4 on AF recurrence after RFA was evaluated, and the independent correlation between them was assessed. Patients with AF and the controls showed no significant differences in age, gender, body mass index, systolic blood pressure, diastolic blood pressure, left ventricular ejection fraction, triglycerides, total cholesterol, high-density lipoprotein, low-density lipoprotein, white blood cell count, neutrophil/lymphocyte ratio, brain natriuretic peptide, and history of smoking, drinking, hypertension, and diabetes (P > 0.05), but with increased LAD in patients with AF (P < 0.01). Serum MYL4 level was reduced in patients with AF (0.6 ± 0.2) compared with that of controls (0.1 ± 0.6) (P < 0.01), and it was negatively correlated with TGF-β1, PICP, and LAD (r = -0.2389, P < 0.05; r = -0.5174, P < 0.01; r = -0.3191; P < 0.01). Low levels of MYL4 increased the risk of AF recurrence after RFA (χ2 = 16.64; P < 0.0001). A low MYL4 level in patients with AF showed a poorer prognosis. Serum MYL4 level and AF type were independent risk factors affecting AF recurrence after RFA.

An anti-biofilm cyclic peptide targets a secreted aminopeptidase from P. aeruginosa

Pseudomonas aeruginosa is an opportunistic pathogen that causes serious illness, especially in immunocompromised individuals. P. aeruginosa forms biofilms that contribute to growth and persistence in a wide range of environments. Here we investigated the aminopeptidase, P. aeruginosa aminopeptidase (PaAP) from P. aeruginosa, which is highly abundant in the biofilm matrix. PaAP is associated with biofilm development and contributes to nutrient recycling. We confirmed that post-translational processing was required for activation and PaAP is a promiscuous aminopeptidase acting on unstructured regions of peptides and proteins. Crystal structures of wild-type enzymes and variants revealed the mechanism of autoinhibition, whereby the C-terminal propeptide locks the protease-associated domain and the catalytic peptidase domain into a self-inhibited conformation. Inspired by this, we designed a highly potent small cyclic-peptide inhibitor that recapitulates the deleterious phenotype observed with a PaAP deletion variant in biofilm assays and present a path toward targeting secreted proteins in a biofilm context.

The viable bioengineered allogeneic cellularized construct StrataGraft® synthesizes, deposits, and organizes human extracellular matrix proteins into tissue type-specific structures and secretes soluble factors associated with wound healing

BackgroundStrataGraft® (allogeneic cultured keratinocytes and dermal fibroblasts in murine collagen–dsat) is an FDA-approved viable bioengineered allogeneic cellularized construct for adult patients with deep partial-thickness burns requiring surgery. We characterized the structural and functional properties of StrataGraft to improve product understanding by evaluating extracellular matrix (ECM) molecule distribution and secreted protein factor expression in vitro. MethodsECM protein expression was determined using indirect immunofluorescence on construct cross sections using commercial antibodies against collagen III, IV, VI, laminin-332, and decorin. Human collagen I expression was verified by enzyme-linked immunosorbent assay (ELISA) for collagen I C-terminal propeptide. Soluble protein factor secretion was quantified by multiplex biomarker assays and singleplex ELISA in conditioned media from meshed constructs. ResultsStrataGraft cellular components produced collagen I, collagen III, collagen VI, and decorin in patterns indicating an organized ECM. Distributions of collagen IV and laminin-332 indicated formation of basement membranes and dermal-epidermal junctions. Soluble protein factors were observed in the pg/cm2/h range from 1 h to the experiment end at 168 h. ConclusionsThe organization of the ECM proteins was like human skin and the viable cellular components provided sustained secretion of soluble wound healing factors, making StrataGraft an attractive option for treating severe burns.

Bone biomarkers and subsequent survival in men with bone-metastatic hormone sensitive prostate cancer (HSPC): Results from the phase III SWOG S1216 trial of androgen deprivation +/- orteronel.

5048 Background: We previously reported that circulating serum biomarkers of bone metabolism were significantly associated with overall survival (OS) in men with advanced prostate cancer, either in the hormone sensitive (Lara, et al. ASCO 2022) or castration resistant state (Lara, et al. JNCI 2016). In S1216, we showed that elevated bone biomarkers were significantly associated with an increased risk of death in HSPC regardless of bone metastases (mets). We also identified three risk groups (low, intermediate, and poor) with differential OS outcomes (median OS: 8.2, 5.1, and 2.1 years, respectively) based on combinations of bone biomarkers in these patients. Here we report the association of bone biomarkers with OS in men with HSPC and documented skeletal mets as part of a planned analysis of S1216. Methods: Bone resorption [C-telopeptide (CTx) and Pyridinoline (PYD)] and bone formation markers [C-terminal collagen propeptide (CICP) and bone alkaline phosphatase (BAP)] were assessed from patient sera. Patients were randomly divided into training (n=238) and validation (n=475) sets. In the training set, recursive partitioning that maximizes discrimination of OS was used to identify the dichotomous cut-point for each biomarker and for a combination of biomarker split points to define prognostic groups. In the validation set, Cox proportional hazards models were used to assess the impact of biomarkers on OS, adjusted for patient and tumor characteristics. Results: Of 1,279 men in S1216, 713 had both baseline bone mets and evaluable bone biomarkers. Patient characteristics were similar between the overall population and the subset with bone mets. Elevated levels of CICP, CTX, and PYD were strongly prognostic for OS, but not for BAP. Hazard ratios (95% CI) for OS adjusted for treatment arm and baseline clinical variables were: BAP – 1.31 (0.93, 1.84), p=0.12; CICP – 1.58 (1.09, 2.29), p&lt;0.02; CTx – 1.55 (1.12, 2.15), p=0.008; and PYD – 1.66 (1.27, 2.217), p=0.0002. There was no evidence of interaction between elevated biomarkers and treatment (all p&gt;0.2). Recursive partitioning algorithms identified four groups of patients with differential OS outcomes based on bone biomarkers, adjusted for baseline clinical variables, with median OS ranging from 2.3 years (highest risk group) to 7.5 years (lowest risk group). Conclusions: In this subset analysis of men with HSPC and bone mets in S1216, elevated serum markers of bone metabolism were significantly associated with worse OS. Bone biomarker levels alone and in combination with patient and tumor characteristics identify unique subsets of men with differential OS outcomes. Clinical trial information: NCT01809691 .

Bone Biomarkers and Subsequent Survival in Men with Hormone-sensitive Prostate Cancer: Results from the SWOG S1216 Phase 3 Trial of Androgen Deprivation Therapy with or Without Orteronel

BackgroundBone biomarkers are strongly prognostic for overall survival (OS) in men with castration-resistant prostate cancer but not fully established for hormone-sensitive prostate cancer (HSPC). ObjectiveBone biomarkers in HSPC were prospectively evaluated as part of a phase 3 study of androgen deprivation therapy ± the CYP17 inhibitor orteronel. Design, setting, and participantsPatients were randomly divided into training (n = 316) and validation (n = 633) sets. Recursive partitioning and Cox proportional hazard models were employed. Outcome measurements and statistical analysisBone resorption (C-telopeptide and pyridinoline) and bone formation markers (C-terminal collagen propeptide and bone alkaline phosphatase) were assessed from patient sera. Results and limitationsOf 1279 men, 949 had evaluable baseline bone biomarkers. Optimal cutoffs were identified to define elevated levels of each of the four biomarkers (all p < 0.05) that were associated with worse OS. After adjusting for clinical risk factors in the validation set, elevated bone biomarkers were statistically significantly associated with an increased risk of death (hazard ratios ranging from 1.37 to 1.92). Recursive partitioning algorithms applied to the training set identified three risk groups (low, intermediate, and poor) with differential OS outcomes (median OS: 8.2, 5.1, and 2.1 yr, respectively) based on combinations of bone biomarkers. These results were confirmed in the validation set. ConclusionsIn men with HSPC initiating androgen deprivation therapy, bone biomarkers are strongly and independently prognostic for OS. Bone biomarker levels alone or in combination with clinical covariates identify unique subsets of men with differential OS outcomes. These results validate the clinical value of bone biomarker assessment in the HSPC state, extending bone biomarker utility beyond the castration-resistant state. Patient summaryIn men with newly diagnosed metastatic prostate cancer, high levels of bone turnover biomarkers are associated with a shorter lifespan.