Markers Of Collagen Synthesis Research Articles

Urinary proteomic signature of mineralocorticoid receptor antagonism by spironolactone: evidence from the HOMAGE trial

ObjectiveHeart failure (HF) is characterised by collagen deposition. Urinary proteomic profiling (UPP) followed by peptide sequencing identifies parental proteins, for over 70% derived from collagens. This study aimed to refine...

Bone Turnover Markers during Growth Hormone Therapy for Short Stature Children Born Small for Gestational Age.

Growth hormone therapy (GHT) can improve growth velocity and final height, but can also accelerate the process of bone growth, which is related to structural bone modeling in both formation and resorption. This study evaluated the capacity of bone turnover markers to predict early growth response to one year of GHT in short stature children born small for gestational age (SGA). This study included 25 prepubertal children born SGA. We estimated P1NP (N-terminal procollagen type 1), CTX (C-terminal telopeptide of collagen type 1), P3NP (N-terminal procollagen type 3), NT-pro-CNP (amino-terminal C-type natriuretic peptide) and Ca-P metabolism using standard ECLIA (electrochemiluminescence), RIA (radioimmunoassay), and ELISA (enzyme-linked immunosorbent assay) methods. A statistically significant increase in bone resorption markers (CTX) was found at both 6 and 12 months. P1NP bone markers were increased at 6 months and after 12 months of therapy. The P3NP marker for collagen synthesis also increased after 12 months of therapy. We obtained significant increases in phosphorus levels at 6 and 12 months, and similar ALP (alkaline phosphatase) increases. We found a significant correlation between height (cm) and CTX after 6-12 months, as well as a P1NP/height (SD) correlation after 12 months. Calcium levels significantly correlated with height (SD) after 12 months. We found strong reactions of bone resorption and bone formation markers during growth hormone therapy, which may determine their selection as predictors of GHT outcome in children born SGA. However, the issue requires further research.

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.

Plasma levels of PRO-C3, a type III collagen synthesis marker, are associated with arterial stiffness and increased risk of cardiovascular death

Background and aimsThe N-terminal propeptide of type III collagen (PRO-C3) assay measures a pro-peptide released during type III collagen synthesis, an important feature of arterial stiffening and atherogenesis. There is a clinical need for improved non-invasive, cheap and easily accessible methods for evaluating individuals at risk of cardiovascular disease (CVD). In this study, we investigate the potential of using circulating levels of PRO-C3 to mark the degree of vascular stenosis and risk of cardiovascular events. MethodsBaseline plasma levels of PRO-C3 were measured by ELISA in subjects belonging to the SUrrogate markers for Micro- and Macro-vascular hard endpoints for Innovative diabetes Tools (SUMMIT) cohort (N = 1354). Associations between PRO-C3 levels with vascular characteristics, namely stiffness and stenosis, and risk of future cardiovascular events were explored. Subjects were followed up after a median of 35 months (interquartile range 34–36 months), with recorded outcomes cardiovascular death and all-cause mortality. ResultsWe found a correlation between PRO-C3 levels and pulse wave velocity (rho 0.13, p = 0.000009), a measurement of arterial stiffness. Higher PRO-C3 levels were also associated with elevated blood pressure (rho 0.07, p = 0.014), as well as risk of cardiovascular mortality over a three-year follow-up period (OR 1.56, confidence interval 1.008–2.43, p = 0.046). ConclusionsElevated circulating PRO-C3 levels are associated with arterial stiffness and future cardiovascular death, in the SUMMIT cohort, suggesting a potential value of PRO-C3 as a novel marker for declining vascular health.

Hemodynamics and plasma biomarkers in individuals with mild-to-moderate ascending aortic dilatation and tricuspid aortic valves

Abstract Background Ascending aortic (AA) dilatation is a silent disease that requires frequent follow-up measurements of the AA diameter. Unfortunately, the AA diameter is an insufficient predictor of future growth and complications. Hemodynamic and circulating biomarkers represent promising ways of improving the risk stratification in AA dilatation. Most patients with AA dilatation have tricuspid aortic valves (TAV). However, previous studies have mainly focused on AA dilation in individuals with bicuspid aortic valves (BAV) using either hemodynamic or circulating biomarkers separately. Purpose To assess the relationship between hemodynamics in the AA and circulating biomarkers in individuals with mild-to-moderate AA dilatation and age-and-sex matched controls with TAV. Methods The study cohort of individuals with mild-to-moderate AA dilatation were identified based on AA diameter on coronary CT angiography in the population-based Swedish CardioPulmonary BioImage Study (SCAPIS) in Linköping. 54 cases with AA diameter &amp;gt;= 40 mm and 50 controls with AA diameter &amp;lt;40 mm (mean age 59, 19% female) underwent 4D Flow cardiovascular magnetic resonance imaging (CMR). The 4D Flow CMR images were processed to compute mean and maximum velocities, wall shear stress (WSS), flow displacement (FD) at peak systole, and oscillatory shear index (OSI) in the AA. Markers of collagen synthesis and degradation; type I collagen α1 chain (COL1α1), matrix metalloproteinase (MMP)-1,-2,-3,-9,-12) and their inhibitors (TIMP-1,-2,-3,-4), were quantified in plasma (n=104). High sensitivity C-reactive protein and interleukin(IL)-6 were used to assess systemic inflammation. Results FD at the location of maximum dilatation, mean FD and mean OSI were higher in cases compared to controls. Mean velocity, mean WSS and maximum WSS were lower in cases compared to controls. None of the circulating biomarkers differed significantly between cases and controls. Interestingly, several circulating biomarkers correlated with hemodynamics markers in the whole cohort. COL1α1, MMP-2 and TIMP-2 correlated inversely with max OSI (r=-0.342, p=0.001; r=-0.277, p=0.006; r=-0.238, p=0.018, respectively) while MMP-3 correlated positively with mean velocity (r=0.274, p=0.006), mean WSS (r=0.270, p=0.007), max WSS (r=0.220, p=0.029), and inversely with FD at the location of maximum dilatation (r=-0.218, p=0.031). TIMP-4 correlated with mean OSI (r=-0.284, p=0.005). Conclusions Several hemodynamic markers obtained with 4D Flow CMR were altered in mild-to-moderate AA dilatation with TAV, while circulating biomarkers did not differ from controls. However, significant correlations were identified between several biomarkers and hemodynamic markers. We therefore speculate that the levels of circulating biomarkers may be altered as the disease progresses. Future studies should examine the implications of these novel findings along the trajectory of AA dilation and its association with complication risk.Methods figureHemodynamics markers table

Effects of Aging on Collagen in the Skeletal Muscle of Mice.

Aging affects several tissues in the body, including skeletal muscle. Multiple types of collagens are localized in the skeletal muscle and contribute to the maintenance of normal muscle structure and function. Since the effects of aging on muscle fibers vary by muscle fiber type, it is expected that the effects of aging on intramuscular collagen might be influenced by muscle fiber type. In this study, we examined the effect of aging on collagen levels in the soleus (slow-twitch muscle) and gastrocnemius (fast-twitch muscle) muscles of 3-, 10-, 24-, and 28-month-old male C57BL/6J mice using molecular and morphological analysis. It was found that aging increased collagen I, III, and VI gene expression and immunoreactivity in both slow- and fast-twitch muscles and collagen IV expression in slow-twitch muscles. However, collagen IV gene expression and immunoreactivity in fast-twitch muscle were unaffected by aging. In contrast, the expression of the collagen synthesis marker heat shock protein 47 in both slow- and fast-twitch muscles decreased with aging, while the expression of collagen degradation markers increased with aging. Overall, these results suggest that collagen gene expression and immunoreactivity are influenced by muscle fiber type and collagen type and that the balance between collagen synthesis and degradation tends to tilt toward degradation with aging.

The role of connective tissue metabolism disorders in the genesis of stress urinary incontinence in premenopausal women

Research objectives: to study the pathogenetic links in the development of stress urinary incontinence (SUI) by determining the state of connective tissue (CT), concentrations of trace elements, vitamins and estradiol in premenopausal women.Materials and methods. 189 women aged 45–55 years with SUI were examined. In addition to general clinical examination, concentrations of Total PІNP (type I collagen synthesis marker) and Total PІІІNP (type III collagen synthesis marker) and CT resorption marker Pyriliks-D, ionized magnesium, ionized calcium, total calcium, vitamins D and C, estradiol were studied in the blood serum in order to determine the state of fibrous CT.Results. In the structure of somatic morbidity were conditions defined as phenotypic manifestations of CT undifferentiated dysplasia. A decrease of the Total PІNP by 24.9% was found against the background of an increased Total PІІІNP by 46.7% and an increased Pyriliks-D by 54.1%. Women with SUI were characterized by a significant decrease in the average value of Mg2+ by an average of 16.7%, as well as a significant increase in the average value of Ca2+ by 5.3% and total Ca by 3.9%. The vitamin balance study shows a significant decrease of vitamin D by 40.6% and vitamin C by 37.0%. Estradiol concentration was at the minimum values of the physiological norm.Conclusions. Women in premenopausal age with SUI are characterized by a violation of the CT synthesis and resorption, which are realized by reducing the formation of the Total PІNP against the background of intensification of the Total PІІІNP and Pyriliks-D synthesis, combined with a disbalance of vitamins and trace elements. These features are highly likely to be considered as predictors of the SUI development in women of premenopausal age

Impact of essential amino acid intake, resistance exercise, and aging on the concentration of Achilles peritendinous amino acids and procollagen Iα1 in humans.

Recent studies have shown that consuming amino acid-rich compounds improves tendon collagen content and biomechanical properties. Yet, it is unclear if the consumption of amino acids alters local (peritendinous) amino acid concentrations. If aging or exercise influence local amino acid concentrations in conjunction with an amino acid bolus is also not known. We conducted two studies. In Study 1, young women (n = 7, 25 ± 2 years) completed two identical resistance training sessions with either essential amino acid (EAA) or placebo consumption. In Study 2, an EAA bolus identical to Study 1 was given to younger (n = 7; 27 ± 1year) and older adults (n = 6; 68 ± 2 years). Microdialysis was used to determine Achilles peritendinous amino acid and pro-collagen Iα1 (a marker of collagen synthesis) concentrations. In Study 1, amino acid consumption increased peritendinous concentrations of all EAA except histidine (p < 0.05). In Study 2, the peritendinous concentration of EAAs except for methionine, histidine, and lysine (p > 0.05) increased with time (p < 0.05). Further, the concentrations of most measured amino acids were greater in older adults (p < 0.05). Pro-collagen Iα1 concentration (p > 0.05) was unaffected by exercise, EAA, or aging (p > 0.05). Our findings demonstrate the following: (1) when not combined with exercise, an oral EAA bolus leads to only modest increases in Achilles peritendinous amino acid concentrations; (2) when combined with resistance exercise, EAA consumption resulted in greater peritendinous amino acid concentrations compared to no exercise; (3) the basal concentrations of most amino acids were greater in older adults, and (4) neither the EAA bolus nor exercise altered peritendinous pro-collagen concentrations.

Single-nucleus transcriptomics of IDH1- and TP53-mutant glioma stem cells displays diversified commitment on invasive cancer progenitors

Glioma is a devastating brain tumor with a high mortality rate attributed to the glioma stem cells (GSCs) possessing high plasticity. Marker mutations in isocitrate dehydrogenase type 1 (IDH1) and tumor protein 53 (TP53) are frequent in gliomas and impact the cell fate decisions. Understanding the GSC heterogeneity within IDH1- and TP53- mutant tumors may elucidate possible treatment targets. Here, we performed single-nucleus transcriptomics of mutant and wild-type glioma samples sorted for Sox2 stem cell marker. For the first time the rare subpopulations of Sox2 + IDH1- and TP53-mutant GSCs were characterized. In general, GSCs contained the heterogeneity root subpopulation resembling active neural stem cells capable of asymmetric division to quiescent and transit amplifying cell branches. Specifically, double-mutant GSCs revealed the commitment on highly invasive oligodendrocyte- and astroglia-like progenitors. Additionally, double-mutant GSCs displayed upregulated markers of collagen synthesis, altered lipogenesis and high migration, while wild-type GSCs expressed genes related to ATP production. Wild-type GSC root population was highly heterogeneous and lacked the signature marker expression, thus glioblastoma treatment should emphasize on establishing differentiation protocol directed against residual GSCs. For the more differentiated IDH1- and TP53-mutant gliomas we suggest therapeutic targeting of migration molecules, such as CD44.

Dyskalemia in people at increased risk for heart failure: findings from the heart 'OMics' in AGEing (HOMAGE) trial.

In people at risk of heart failure (HF) enrolled in the Heart 'OMics' in AGEing (HOMAGE) trial, spironolactone reduced circulating markers of collagen synthesis, natriuretic peptides, and blood pressure and improved cardiac structure and function. In the present report, we explored factors associated with dyskalaemia. The HOMAGE trial was an open-label study comparing spironolactone (up to 50mg/day) versus standard care in people at risk for HF. After randomization, serum potassium was assessed at 1 and 9months and was defined as low when ≤3.5mmol/L (hypokalaemia) and high when ≥5.5mmol/L (hyperkalaemia). Multivariable logistic regression models were constructed to identify clinical predictors of dyskalaemia. A total of 513 participants (median age 74years, 75% men, median estimated glomerular filtration rate 71mL/min/1.73m2 ) had serum potassium available and were included in this analysis. At randomization, 88 had potassium<4.0mmol/L, 367 had potassium 4.0-5.0mmol/L, and 58 had potassium>5.0mmol/L. During follow-up, on at least one occasion, a serum potassium<3.5mmol/L was observed in 6 (1.2%) and <4.0mmol/L in 46 (9%) participants, while a potassium>5.0mmol/L was observed in 38 (8%) and >5.5mmol/L in 5 (1.0%) participants. The median (percentile25-75 ) increase in serum potassium with spironolactone during the study was 0.23 (0.16; 0.29) mmol/L. Because of the low incidence of dyskalaemia, for regression analysis, hypokalaemia and hyperkalaemia thresholds were set at <4.0 and >5.0mmol/L, respectively. The occurrence of a serum potassium>5.0mmol/L during follow-up was positively associated with the presence of diabetes mellitus {odds ratio [OR]: 1.21 [95% confidence interval (CI) 2.14; 3.79]} and randomization to spironolactone (OR: 2.83 [95% CI 1.49; 5.37]). Conversely, the occurrence of a potassium concentration<4.0mmol/L was positively associated with the use of thiazides (OR: 2.39 [95% CI 1.32; 4.34]), blood urea concentration (OR: 2.15 [95% CI 1.34; 3.39] per 10mg/dL), and history of hypertension (OR: 2.32 [95% CI 1.02; 5.29]) and negatively associated with randomization to spironolactone (OR: 0.30 [95% CI 0.18; 0.52]). In people at risk for developing HF and with relatively normal renal function, spironolactone reduced the risk of hypokalaemia and, at the doses used, was not associated with the occurrence of clinically meaningful hyperkalaemia.

Evaluation of the complex approach effectiveness of atrophic colpitis treatment in menopausal aged women from the pathogenetic view

Objectives: to determine the status of connective tissue and ionized magnesium value before and after complex treatment of atrophic colpitis with the CO2 laser in menopausal women. Materials and methods. 160 women with atrophic colpitis took part in this prospective study and were divided into three groups depending on the therapy. The main group included 55 patients who underwent CO2 laser therapy. The comparison group included 40 women who underwent CO2 laser therapy and local hormonal treatment with suppositories containing estriol and magnesium saturation by magnesium lactate dihydrate. The control group consisted of 65 women who received only suppositories containing estriol. The duration of treatment was 12 months. In addition to general clinical studies, markers of collagen synthesis (Total PINP, Total PІІІNP), collagen resorption marker (Pyriliks-D), as well as the ionized magnesium value in blood serum were determined in all patients.Results. A significant increase and steady retention at the physiological level of the average value of the Total PINP in the comparison group was revealed. The average level of the Total PIІІNP in patients of this group underwent a significant decrease and remained steadily at the upper limit of the physiological norm. A significant decrease and stable retention at the physiological level of the Pyriliks-D was noted in the comparison group against the background of the proposed therapy. Correction of magnesium status in the comparison group leads to a reliable and stable increase in the Mg+ ions concentration in blood serum.Conclusions. The inclusion CO2 laser in combination with local hormonal therapy and long-term correction of mineral metabolism in the treatment of the comparison group led to the stable normalization and prolonged retention at the physiological level for 12 months of the procollagen type I N-terminal propeptide, procollagen type III N-terminal propeptide, collagen resorption marker Pyriliks-D and a significant increase in magnesium concentration. This is confirms the effectiveness of the proposed complex.

The association between markers of type I collagen synthesis and echocardiographic response to spironolactone in patients at risk of heart failure: findings from the HOMAGE trial.

Procollagen type I C-terminal propeptide (PICP) and procollagen type III N-terminal propeptide (PIIINP) are markers reflecting collagen synthesis in cardiac fibrosis. However, they may be influenced by the presence of non-cardiac comorbidities (e.g. ageing, obesity, renal impairment). Understanding the associations between markers of collagen synthesis and abnormalities of cardiac structure and function is important to screen for myocardial fibrosis and monitor the antifibrotic effect of medications. The HOMAGE (Heart 'OMics' in AGEing) trial showed that spironolactone decreased serum PICP concentrations and improved cardiac remodelling over 9 months in a population at risk of developing heart failure (HF). We evaluated the associations between echocardiographic variables, PICP, PIIINP and galectin-3 at baseline and during the course of the trial. Among 527 individuals (74 ± 7 years, 26% women), median serum concentrations of PICP, PIIINP and galectin-3 were 80.6 μg/L (65.1-97.0), 3.9 μg/L (3.1-5.0), and 16.1 μg/L (13.5-19.7), respectively. After adjustment for potential confounders, higher serum PICP was significantly associated with left ventricular hypertrophy, left atrial enlargement, and greater ventricular stiffness (all p &lt; 0.05), whereas serum PIIINP and galectin-3 were not (all p &gt; 0.05). In patients treated with spironolactone, a reduction in serum PICP during the trial was associated with a decrease in E/e' (adjusted-beta = 0.93, 95% confidence interval 0.14-1.73; p = 0.022). In individuals at high risk of developing HF, serum PICP was associated with cardiac structural and functional abnormalities, and a decrease in PICP with spironolactone was correlated with improved diastolic dysfunction as assessed by E/e'. In contrast, no such associations were present for serum PIIINP and galectin-3.

Bone Morphogenetic Protein-9 (BMP9) is Required for Survival and Limits Left Ventricular Matrix Metalloproteinase Activity After Acute Myocardial Infarction

<h3>Purpose</h3> Reorganization of extracellular matrix is a major determinant of short and long term wound healing after acute myocardial infarction (AMI), which includes central roles for matrix metalloproteinases (MMP) and collagen turnover. We recently identified that bone morphogenetic protein (BMP)-9 is a novel endogenous inhibitor of cardiac fibrosis in murine models of pressure overload induced heart failure. We hypothesized that BMP9 regulates MMP activity and is required for survival after AMI. <h3>Methods</h3> WT or BMP9-/- mice were subjected to left anterior descending artery ligation and animals recovered for 2 weeks. Left ventricular (LV) function was assessed using conductance catheter and LV tissue was harvested for molecular analysis in animals surviving to two weeks. To further assess a functional role for BMP9, in vitro studies were performed using human atrial fibroblasts (H-Atr-Fb). <h3>Results</h3> Compared to WT controls BMP9-/- mice had reduced survival (94.4% v 66.7%, p=0.04) and tissue necropsy showed a high rate of cardiac rupture among BMP9-/- mice (BMP9-/- 80%, WT 0%). Ejection fraction and stroke work were reduced and both LV end systolic and diastolic volumes were increased in WT and BMP9-/- mice after AMI. Compared to WT, SMAD3 signaling was increased (p=0.001) and SMAD1 signaling reduced (p=0.03) in BMP9-/- mice after AMI. Compared to WT, BMP9-/- mice had higher LV protein levels of endoglin, Type 1 collagen and periostin. We further observed significantly higher LV MMP9 levels and activity in BMP9-/- compared to WT sham (p=0.003) and WT MI (p=0.02) mice after AMI . Compared to WT, BMP9-/- mice exhibited an increased ratio of circulating markers of collagen degradation and synthesis (ICTP:PINP) after AMI (p=0.02). In H-Atr-Fb, siRNA mediated knockdown of BMP9 increased protein levels of Type I collagen (p<0.05) and MMP9 (p=0.04) and increased MMP9 activity (p=0.003). <h3>Conclusion</h3> We report that BMP9 is necessary for survival after AMI and identify a novel functional role of BMP9 as a negative regulator of MMP9 activity and collagen synthesis in the LV during AMI. These findings suggest that BMP9 may be an important novel target of therapy to improve myocardial recovery and survival after AMI.

Heat-Inactivated Akkermansia muciniphila Improves Gut Permeability but Does Not Prevent Development of Non-Alcoholic Steatohepatitis in Diet-Induced Obese Ldlr-/-.Leiden Mice.

The development of non-alcoholic steatohepatitis (NASH) has been associated with alterations in gut microbiota composition and reduced gut barrier function. Akkermansia muciniphila is a gut microbe that is thought to have health-promoting properties, including the ability to improve gut barrier function and host metabolism, both when administered live and after heat-inactivation. We questioned whether heat-inactivated A. muciniphila may reduce NASH development. Ldlr−/−.Leiden mice, a translational, diet-induced model for NASH, were fed a NASH-inducing high-fat diet (HFD) supplemented with heat-inactivated A. muciniphila. After 28 weeks, effects of the treatment on obesity and associated metabolic dysfunction in the gut (microbiota composition and permeability), adipose tissue, and liver were studied relative to an untreated HFD control. Treatment with heat-inactivated A. muciniphila did not affect body weight or adiposity and had no effect on plasma lipids, blood glucose, or plasma insulin. Heat-inactivated A. muciniphila had some minor effects on mucosal microbiota composition in ileum and colon and improved gut barrier function, as assessed by an in vivo functional gut permeability test. Epidydimal white adipose tissue (WAT) hypertrophy and inflammation were not affected, but heat-inactivated A. muciniphila did reduce hypertrophy in the mesenteric WAT which is in close proximity to the intestine. Heat-inactivated A. muciniphila did not affect the development of NASH or associated fibrosis in the liver and did not affect circulating bile acids or markers of liver fibrosis, but did reduce PRO-C4, a type IV collagen synthesis marker, which may be associated with gut integrity. In conclusion, despite beneficial effects in the gut and mesenteric adipose tissue, heat-inactivated A. muciniphila did not affect the development of NASH and fibrosis in a chronic disease setting that mimics clinically relevant disease stages.

Dynamics of fibrotic and vascular endothelial dysfunction markers in elderly hypertensive patients after ischemic stroke receiving beta-blockers

Aim. To compare the effect of beta-blocker therapy (bisoprolol and nebivolol) on the dynamics of fibrotic and vascular endothelial dysfunction markers in elderly hypertensive patients after ischemic stroke (IS).Material and methods. This prospective cohort study included 75 hypertensive patients who were admitted to the hospital due to IS. The mean age of patients was 67±6 years. The average National Institutes of Health Stroke Scale (NIHSS) score was 7±3. The followup period was 6 months. The control group consisted of 20 elderly people with hypertension without prior myocardial infarction. The patients were divided into groups based on received therapy: group 1 (n=38) — bisoprolol; group 2 (n=37) — nebivolol. The level of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) was determined by enzyme-linked immunosorbent assay (ELISAKit, USA). Vascular ultrasound was carried out using a LOGIQP9 (GE) system according to the Celermajer method.Results. After 6-month nebivolol, we revealed a decrease in the level of MMP-9 by 30,2% (p&lt;0,01), TIMP-1 by 15,6% (p&lt;0,05). After 6-month bisoprolol therapy, the level of MMP-9 decreased by 14,5% (p&lt;0,05), while TIMP-1 did not change. Intergroup comparison found that when using nebivolol, there was a higher decrease in the level of MMP-9 by 15,7% (p&lt;0,05), TIMP-1 by 9,7% (p&lt;0,05), MMP-9/TIMP-1 by 7,8% (p&lt;0,05) than with bisoprolol therapy. After 6-month bisoprolol therapy, there was a decrease in the proportion of patients with severe endothelial dysfunction (ED) by 7,9% (p&lt;0,05). Two patients from the nebivolol group moved into mild ED category. The number of patients with moderate ED increased by 19% (p&lt;0,01), while prevalence of severe ED decreased by 24,4% (p&lt;0,01).Conclusion. The results obtained indicate that the beta-blocker nebivolol at an average dose of 8,55+1,75 mg/day significantly reduces the vascular fibrosis, normalizes the ratio of collagen synthesis and degradation markers, improves the vasodilation brachial artery properties in comparison with bisoprolol in elderly hypertensive patients after IS.

Systemic vascular basement membrane markers linked to synovial vascular remodeling are biomarkers of hemarthrosis in patients with hemophilia

IntroductionInterstitial, cartilage, and bone collagens have been proposed as biomarkers of joint deterioration in hemophilic arthropathy. The role of basement membrane (type IV and VIII) collagens as biomarkers of endothelial turnover in relation to acute joint bleeding is not understood. MethodsThirty‐one adult patients with hemophilia were studied prospectively for 3 years with musculoskeletal ultrasound/power Doppler (MSKUS/PD) during pain‐free intervals and painful events for joint bleed status, synovial vascular flow, and 10 plasma markers of collagen turnover. Joint health was determined using Hemophilia Joint Health Scores and Pettersson scores. In animal studies, bleeding was induced in factor VIII−/− mice by knee joint injury. Synovial vascular remodeling was assessed using MSKUS/PD and histology. Murine plasma samples were analyzed for type IV collagen turnover markers. ResultsNinety‐one patient visits were compiled. Twenty‐five were due to acute painful episodes, with 16 confirmed hemarthroses. Type IV collagen turnover markers (PRO‐C4 and C4M), and a type VIII collagen synthesis marker (PRO‐C8), were transiently elevated during acute hemarthrosis. Hemarthrosis was accompanied by increased synovial microvascular flow (MSKUS/PD), and levels of type IV collagen markers correlated with PD signals in the joint. In factor VIII‐deficient mice, plasma levels of type IV collagen turnover markers correlated negatively with synovial αSMA staining, indicating that reduced type IV collagen turnover was associated with thicker vessels. ConclusionsOur findings suggest that basement membrane turnover markers, closely linked to synovial vascular remodeling, may be systemic biomarkers of acute hemarthrosis. Vascular instability during neovascularization may be involved in the dynamics of hemarthrosis.

Metformin prevents PFKFB3-related aerobic glycolysis from enhancing collagen synthesis in lung fibroblasts by regulating AMPK/mTOR pathway.

Aerobic glycolysis has been shown to contribute to the abnormal activation of lung fibroblasts with excessive collagen deposition in lipopolysaccharide (LPS)-induced pulmonary fibrosis. Targeting aerobic glycolysis in lung fibroblasts might therefore be considered as a promising therapeutic approach for LPS-induced pulmonary fibrosis. In the present study, the aim was to investigate whether metformin, a widely used agent for treating type 2 diabetes, could alleviate LPS-induced lung fibroblast collagen synthesis and its potential underlying mechanisms. Different concentrations of metformin were used to treat the human lung fibroblast MRC-5 cells after LPS challenge. Indicators of aerobic glycolysis in MRC-5 cells were detected by measuring glucose consumption and lactate levels in culture medium in addition to lactate dehydrogenase activity in cellular lysates. The glucose consumption, lactate levels and the lactate dehydrogenase activity were measured respectively using colorimetric/fluorometric and ELISA kits. The effects of metformin in AMP-activated protein kinase (AMPK) activation was assessed by mitochondrial complex I activity kits. Collagen I, α-smooth muscle actin (α-SMA) and collagen III were used as markers of collagen synthesis, which was measured using western blotting, whereas phosphorylated (p-) AMPK, AMPK, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) and mTOR were detected by western blotting. Metformin significantly decreased mitochondrial complex I activity and upregulated the expression of p-AMPK/AMPK protein in a concentration-dependent manner. Furthermore, the aerobic glycolysis mediated by PFKFB3 and collagen synthesis in LPS-treated MRC-5 cells was gradually inhibited with increasing concentrations of metformin. However, this inhibitory role of metformin on PFKFB3-meditaed aerobic glycolysis and collagen synthesis was prevented by treatments with 3BDO and compound C, which are specific mTOR activator and AMPK inhibitor, respectively. Taken together, the findings from this study suggested that metformin may prevent PFKFB3-associated aerobic glycolysis from enhancing collagen synthesis in lung fibroblasts via regulating the AMPK/mTOR pathway.

Relationships between circulating galectin-3, extracellular matrix fibrosis and outcomes in dilated cardiomyopathy.

Galectin-3 is an emerging biomarker in cardiovascular disease. Myocardial galectin-3 is involved in the pathology of cardiac fibrosis; however, the role of circulating galectin-3 is not yet established. To assess the relationships between circulating galectin-3, fibrosis and outcomes in dilated cardiomyopathy (DCM). We included 70 patients (age: 48 ±12.1 years, ejection fraction (EF) 24.4 ±7.4%) with new-onset DCM (n = 35, ≤6 months). Galectin-3 and procollagen type I and III (PICP, PINP, PIIICP, and PIIINP), transforming growth factor β (TGF-β), connective tissue growth factor (CTGF), osteopontin (OPN), matrix metalloproteinases (MMP-2 and -9), and tissue inhibitor (TIMP-1) were determined in serum at baseline and after 3 and 12 months. Patients underwent endomyocardial biopsy. The endpoint was a combination of death and urgent hospitalization at 12 months. Galectin-3 did not correlate with biopsy-determined fibrosis. Baseline galectin-3 correlated with OPN,, TIMP-1, PIIICP, and MMP-2. In new-onset DCM, galectin-3 levels at baseline were higher than at 3 and 12 months, whereas in chronic DCM there was no difference. Galectin-3 was a predictor of the endpoint (hazard ratio (HR) = 1.115; 95% confidence interval (95% CI) = 1.009-1.231; p < 0.05). The best cut-off value was 14.54 ng/mL (area under the curve (AUC) = 0.67). Patients with galectin-3 ≥14.54 ng/mL had an increased risk of events (HR = 2.569; 95% CI = 1.098-6.009; p < 0.05). Circulating galectin-3 is unrelated to fibrosis. Serial measurements of galectin-3 correlated with markers of fibrosis, including markers of collagen synthesis and OPN. Circulating galectin-3 was independently associated with cardiovascular (CV) outcomes in DCM.

Integrative characterization of asymptomatic vs. symptomatic patients with moderate-severe primary mitral regurgitation

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): This work was supported by NHLBI and Specialized Centers of Clinically Oriented Research grant in cardiac dysfunction, by Department of Veteran Affairs for Merit Review and NIH Grant P01. Background Optimal timing for surgery in primary mitral regurgitation (PMR) is a subject of debate. Purpose Use a combination of imaging, circulating biomarkers and histology to define the transition from asymptomatic (asx) to symptomatic (sx) stage in moderate-severe PMR. Methods 57 controls, 82 sx and 47 asx patients with moderate to severe PMR underwent cardiac MR(CMR), plasma BNP, levels of xanthine oxidase (XO) activity, and circulating markers of collagen (Col) synthesis (propeptide of procollagen type I, PICP) and degradation (Col type I fibers, ICTP). Results A progressive worsening in the severity of adverse LV remodeling was noted based on CMR indices listed in Table 1. The 3D mid-wall LV endocardial curvature to wall thickness was lower in normal and similar in asx and sx patients.Plasma BNP was progressively higher in normal vs. asx vs sx (Figure 1A). The levels of XO activity were higher in sx vs. asx (Figure 1B,1C). Levels of PICP were similar among the three; levels of ICTP were significantly higher in the sx vs the asx (Figure 1D,1E). LV biopsies revealed large areas of patchy replacement fibrosis and areas devoid of interstitial Col (Figure 1F). From the subgroup of 56 patients who underwent CMR 6 months after surgery, 27% had LVEF &amp;lt; 50%. The preoperative LVEF, LVESV, LVESD and LA emptying fraction were predictors of postoperative drop in LVEF. Conclusion Increased COl breakdown coupled with markers of oxidative stress and CMR-based LV remodeling characterize the transition from asx to sx PMR. Longitudinal studies are needed to define cutoff"s for patient selection and optimal timing for MV surgery. CMR indices in normal, asx and sx PMR. Normal (57) Moderate/Severe MR Asymptomatic (56) Pre-Surgery MR Symptomatic (81) P-value Age 48 (34, 55) 54 (46, 62) * 57 (50, 67) * &amp;lt;0.0001 Female/Male 30(53%)/27(47%) 35(63%)/21(38%) 25(31%)/56(69%) LVEF (%) 64 (61, 67) 62 (59, 66) 63 (59, 67) 0.3482 LV EDV (mL/m2) 71 (61, 76) 89 (73, 103) * 100 (83, 123) *# &amp;lt;0.0001 LV ESV (mL/m2) 24 (21, 29) 32 (26, 38) * 36 (28, 47) * &amp;lt;0.0001 LVSV (mL/m2) 43 (37, 50) 54 (43, 66) * 60 (54, 77) *# &amp;lt;0.0001 LVED Mass/Volume 0.7 (0.6, 0.8) 0.6 (0.5, 0.7) * 0.6 (0.6, 0.7) * 0.0012 LVED Mass/Vol x SI 1.29 (1.05, 1.50) 0.97 (0.77, 1.15) * 0.94 (0.77, 1.14) * &amp;lt;0.0001 Regurgitant Volume (mL) ― 38 (26, 54) 57 (38, 83) # 0.001 Regurgitant Fraction (%) ― 39 (28, 50) 46 (36, 56) # 0.042 Table 1 shows the CMR indices of LV remodeling in normal, asymptomatic and symptomatic patients with moderate-severe PMR. p-values *vs. Normal; # vs. Asymptomatic. Kruskal-Wallis test or unpaired t-test Abstract Figure 1.

Pathogenetic substantiation of preterm births prevention methods

The study of the pathogenesis of premature birth is one of the important issues of modern obstetrics. This could contribute developing a scientifically sound program for predicting and preventing this complication of pregnancy. The aim of the study was to investigate the blood variables of the immune system and connective tissue metabolism in women with preterm birth at 23-36 weeks of gestation. Materials and methods. A total of 227 pregnant women were examined, 190 of whom had clinical signs of threatened preterm birth. Group I included 48 women with a threatened preterm delivery resulted in childbirth between 23 and 27 weeks. In group II, 142 women at risk of preterm birth were observed from 28 to 36 weeks. Group III included 37 women with a healthy pregnancy, completed at 38-41 weeks. Result. The number of lymphocytes and their subpopulations, the content of connective tissue metabolites in the blood of the examined patients were studied. In women of group, I relatively to women of group III there was an evident (p &lt; 0.001) decrease in blood level of CD3 + -marker of T-lymphocytes by an average of 23 %, which suggests the presence of immunodeficiency and considered as a stress response. In women with preterm birth at 28-36 weeks of gestation (group II), the relative level of CD3 + in the blood, on the contrary, slightly but probably (p = 0.014) increased compared to women with healthy pregnancy by an average of 13%. A significant decrease in the immunoregulatory index in women of group I is associated with the suppression of the T-helper component and emphasizes the presence of T-cell immunosuppression. The decrease in the index in women of group II occurred due to cytotoxic T-lymphocytes. Сonclusions. Thus, the work confirms the opinion of R. Romero on the role of immune "rejection" in the third trimester, associated with the HLA-incompatibility of mother and fetus. In women of group I, compared with women of group III, there was a more pronounced increase in serum free oxyproline - a marker of collagen breakdown than bound oxyproline - a marker of collagen synthesis. Group II women also had a simultaneous increase in the level of the two oxyproline fractions, but more pronounced for the protein-bound fraction. This suggests that the violation of the metabolism of connective tissue leads to the destruction of the collagen matrix of the cervix and its shortening. Studies suggest that cerclage or insertion of pessary should be used to prevent cervical incompetence in the second trimester due to metabolic disorders in the connective tissue. The use of progesterone is appropriate for prevention of "short cervix" syndrome, as well as the possible immunosuppressive effect of the latter. The established signs of autoimmune processes require screening for antiphospholipid syndrome and the appointment, if confirmed, of anticoagulants.