Ratio Of MMP-9 Research Articles

Expression of MMP-2 and TIMP-1 during rapid maxillary expansion in rats

ObjectiveThe aim of this study was to investigate the expression of MMP-2 and TIMP-1 during midpalatal suture expansion in rats. Design72 male Wistar rats were randomly divided into 2 groups: the experimental group and the control group. In the experimental group, opening loops were applied across the midpalatal suture with an initial force of 50g, whereas in the control group, rats were subjected to sham installation of opening loops without activation. On day 1, 4, 7 and 14, nine rats from each group were sacrificed, and the maxillae were dissected and prepared for Immunohistochemistry (IHC) and RT- PCR examination of MMP-2 and TIMP-1 expression. ResultsThe results of IHC and Real Time PCR revealed that both protein and mRNA expression of MMP-2 and TIMP-1 were significantly increased after midpalatal expansion, and the ratio of MMP-2/TIMP-1 was also significantly enhanced. ConclusionsThe data suggested that MMP-2 and TIMP-1 might play an important role during the mid-palatal suture remodeling process of maxillary expansion.

Priming anoestrous Corriedale ewes with progesterone and gonadotrophin-releasing hormone causes cervical tissue remodelling due to metalloproteinase-2 (MMP-2) activity

The aim was to obtain experimental evidence of cervical collagen degradation in anoestrous Corriedale adult ewes induced to ovulate with progesterone (P) and gonadotrophin-releasing hormone (GnRH), at the expected time of induced ovulation and early luteal phase. In Experiment 1, anoestrous ewes were treated with P for 10 days (P, n = 4), with nine micro-doses of GnRH followed by a GnRH bolus injection (GnRH, n = 4) or with P plus GnRH treatments (P+GnRH, n = 3), and cervices were obtained either without treatment (A, n = 4), when P was removed, or 24 h after the GnRH bolus injection. In Experiment 2, cervices were obtained 1 (group P+GnRH, n = 5) or 5 (P+GnRH 5, n = 6) days after the GnRH bolus injection with P pretreatment. MMP-2 activity was detected in all samples; however, MMP-9 activity was only detected in 15% of the samples. The activity of the latent (L) form of MMP-2 in the cranial zone of group A was higher than in the cranial zone of groups P, GnRH and P+GnRH, and was also higher than that in the caudal zone of the same group (P < 0.05). The collagen concentration was lower in group P+GnRH 1 than in group P+GnRH 5 (P < 0.04). The activity of the activated (A) form of MMP-2 and the A/L MMP-2 ratio were higher in group P+GnRH 1 than in group P+GnRH 5 (P < 0.05). Data suggest that the L form of MMP-2 was expressed mainly in a constitutive form in the cervix of anoestrous ewes and that an oestrogen-dependent activation mechanisms due to the GnRH treatment may be responsible for the lowest collagen content at the moment of the induced ovulation. This work provides evidence about cervical collagen remodelling in anoestrous ewes treated with P + GnRH.

MiR-29a Assists in Preventing the Activation of Human Stellate Cells and Promotes Recovery From Liver Fibrosis in Mice.

The microRNA-29 (miR-29) family is known to suppress the activation of hepatic stellate cells (HSCs) and reversibly control liver fibrosis; however, the mechanism of how miR-29a controls liver fibrosis remains largely unknown. This study was conducted to clarify the mechanism of anti-fibrotic effect of miR-29a and to explore if miR-29a is a promising candidate for nucleic acid medicine against liver fibrosis. Two liver fibrosis murine models (carbon tetrachloride or thioacetamide) were used. MiR-29a mixed with atelocollagen was systemically administered. Hepatic fibrosis was evaluated by histological analysis and the expression levels of fibrosis-related genes. We observed that miR-29a treatment dramatically accelerated the reversion of liver fibrosis in vivo. Additionally, miR-29a regulated the mRNA expression of collagen type I alpha 1 (COL1A1) and platelet-derived growth factor C (PDGFC). We also noted that miR-29a significantly suppressed COL1A1 mRNA expression and cell viability and significantly increased caspase-9 activity (P < 0.05) in LX-2 cells. Pretreatment of miR-29a inhibited activation of LX-2 cell by transforming growth factor beta treatment. MiR-29a exhibited anti-fibrotic effect without cell toxicity in vivo and directly suppressed the expression of PDGF-related genes as well as COL1A1 and induced apoptosis of LX-2 cells. MiR-29a is a promising nucleic acid inhibitor to target liver fibrosis.

PS 02-49 ASSOCIATION OF MATRIX METALLOPROTEINASES WITH THE RENAL BLOOD FLOW AND RENAL FUNCTION IN PATIENTS WITH RESISTANT HYPERTENSION AND TYPE 2 DIABETES MELLITUS

Objective: We wished to assess the relationship between the plasma concentrations of matrix metalloproteinases (MMP) and their tissue inhibitors with intrarenal vascular resistance level, renal function and development of microalbuminuria in patients with resistant hypertension (RH) and type 2 diabetes mellitus (DM). Design and Method: A total of 18 patients with RH and type 2 DM were included in the study (mean aged 58.6 ± 7.5 years, 6 male, office blood pressure (BP) 174.4 ± 20.2/94.1 ± 15.2 mmHg, basal glycaemia 8.8 ± 2.3 mmol/L, HbA1c 6.8 ± 0.7%, an estimated glomerular filtration rate (eGFR) 74.2 ± 27.9 mL/min/1.73m 2), which submitted to office BP measurements, renal Doppler ultrasound and laboratory tests to assess microalbuminuria (defined as 30–300 mg/24 h), eGFR (MDRD formula), HbA1c and basal glycaemia levels, plasma concentrations of MMP-9, MMP-2, tissue inhibitor of MMP type 1 (TIMP-1). Results: Decreased MMP-9 concentration was linearly associated with increased resistive index (RI) in renal arteries (in the main renal arteries: R = −0.60, p = 0.009 (right), R = 0.−60, p = 0.008 (left); in segmental arteries: R = −0.49, P = 0.038 (right) and R = −0.59, P = 0.012 (left)), while growth of TIMP-1/MMP-9 ratio was positively correlated with RI at segmental arteries (R = 0.51, R = 0.028- on the right, R = 0.46, P = 0.04- on the left). Additionally, decreased MMP-9 concentration was linearly associated with decreased eGFR (R = 0.53, P = 0.029). No correlation was observed between MMP-2, MMP-9, TIMP-1, TIMP-1/MMP-9, TIMP-1/MMP-2 ratios and microalbuminuria, however, microalbuminuric patients had a higher TIMP-1/MMP-2 ratio than normoalbuminuric patients (2.97(2.15–3.80) and 1.58(1.39–1.99)ng/mL, accordingly, P = 0.026). Besides, microalbuminuric patients had a higher TIMP-1/MMP-2 ratio than normoalbuminuric patients [2.97 (2.15–3.80) and 1.58 (1.39–1.99), accordingly, P = 0.026)]. Conclusions: In patients with RH and type 2 DM the decreased MMP-9 activity and increased TIMP-1/MMP-9 ratio are associated with growth of intrarenal vascular resistance and decline of renal filtration function. Furthermore, increased TIMP-1/MMP-2 ratio in these patients was due primarily to increase in basal glycaemia level and linked to development of microalbuminuria.

Differential Diagnosis of Autoimmune Pancreatitis From Pancreatic Cancer by Analysis of Serum Gelatinase Levels

The aim of this study was to analyze serum gelatinases as part of the clinical strategy for the preoperative differentiation between autoimmune pancreatitis (AIP) and pancreatic ductal adenocarcinoma (PDAC). The finding of differential markers will prevent unnecessary surgical resection and allow optimal treatment of these diseases. Quantitative gelatin zymography was applied to analyze all individual gelatinase forms in serum and to define proteinase alterations associated with AIP and PDAC. For this purpose, sera of 130 patients, being 29 with AIP, 33 with chronic pancreatitis, 32 with PDAC, and 36 healthy controls, were first assayed for gelatinase levels by quantitative zymography before further validation by the analysis with commercial sandwich enzyme linked immunosorbent assays. Serum profiling data obtained by zymography analysis revealed that gelatinase B/matrix metalloproteinase 9 (MMP-9), the neutrophil gelatinase B-associated lipocalin/MMP-9 complex, and gelatinase A/MMP-2 levels were significantly increased in patients with AIP. These proteins are promising markers to discriminate between AIP and PDAC. The best composite parameter, being the ratio of total MMP-9 over MMP-2 levels, can predict 93% of the AIP and 75% of the PDAC correctly. With enzyme linked immunosorbent assay, we confirmed the zymography results. Differential gelatinase serum profiles as AIP markers, together with other clinical tests, help to assure the diagnosis of PDAC or AIP.

Tear Mediators in Corneal Ectatic Disorders.

PurposeTo compare the concentrations of 11 tear mediators in order to reveal the biochemical difference between pellucid marginal degeneration (PMD) and keratoconus (KC).MethodsWe have designed a cross-sectional study in which patients with corneal ectasia based on slit-lamp biomicroscopy and Pentacam HR (keratometry values (K1, K2, Kmax), astigmatism, minimal radius of curvature (Rmin), corneal thickness (Apex and Min), indices (surface variation, vertical asymmetry, keratoconus, central keratoconus, height asymmetry and decentration)) were enrolled. Eyes of keratoconic patients were similar to the PMD patients in age and severity (K2, Kmax and Rmin). Non-stimulated tear samples were collected from nine eyes of seven PMD patients, 55 eyes of 55 KC patients and 24 eyes of 24 healthy controls. The mediators’ (interleukin -6, -10, chemokine ligand 5, -8, -10, matrix metalloproteinase (MMP) -9, -13, tissue inhibitor of metalloproteinases (TIMP)-1, tissue plasminogen activator, plasminogen activator inhibitor, nerve growth factor) concentrations were measured using Cytometric Bead Array.ResultsMMP-9 was the only mediator which presented relevant variances between the two patient groups (p = 0.005). The ratios of MMP-9 and TIMP-1 were 2.45, 0.40 and 0.23 in PMD, KC and the controls, respectively.ConclusionAs far as we are aware, this is the first study that aims to reveal the biochemical differences between PMD and KC. Further studies of biomarkers to investigate the precise role of these mediators need to be defined, and it is important to confirm the observed changes in a larger study to gain further insights into the molecular alterations in PMD.

Identification of Biomarkers for Footpad Dermatitis Development and Wound Healing.

Footpad dermatitis (FPD) is a type of skin inflammation that causes necrotic lesions on the plantar surface of the footpads in commercial poultry, with significant animal welfare, and economic implications. To identify biomarkers for FPD development and wound healing, a battery cage trial was conducted in which a paper sheet was put on the bottom of cages to hold feces to induce FPD of broilers. Day-of-hatch Ross 308 male broiler chicks were fed a corn–soybean meal diet and assigned to 3 treatments with 8 cages per treatment and 11 birds per cage. Cages without paper sheets were used as a negative control (NEG). Cages with paper sheets during the entire growth period (d 0–30) were used as a positive control (POS) to continually induce FPD. Cages with paper sheets during d 0–13 and without paper sheets during d 14–30 were used to examine the dynamic of FPD development and lesion wound healing (LWH). Footpad lesions were scored to grade (G) 1–5 with no lesion in G1 and most severe lesion in G5. Covering with paper sheets in POS and LWH induced 99% incidence of G3 footpads on d 13. Removing paper sheets from LWH healed footpad lesions by d 30. One representative bird, with lesions most close to pen average lesion score, was chosen to collect footpad skin samples for biomarker analysis. Total collagen protein and mRNA levels of tenascin X (TNX), type I α1 collagen (COL1A1), type III α1 collagen (COL3A1), tissue inhibitor of metalloproteinase 3 (TIMP3), and integrin α1 (ITGA1) mRNA levels were decreased (P < 0.05), while mRNA levels of tenascin C (TNC), tumor necrosis factor (TNF) α, Toll-like receptor (TLR) 4 and vascular endothelial growth factor (VEGF), IL-1β, and the ratio of MMP2 to all TIMP were increased (P < 0.03) in G3 footpads in POS and LWH compared to G1 footpads in NEG on d 14. These parameters continued to worsen with development of more severe lesions in POS. After paper sheets were removed (i.e., LWH), levels of these parameters gradually or rapidly returned to levels measured in NEG. Regression analysis indicated significant quadratic changes of these parameters to footpad lesion scores. In summary, these biomarkers were interrelated with dynamic changes of footpad lesion scores, suggesting they may be used as potential biomarkers for footpad lesion development and wound healing process.

Reference ranges of matrix metalloproteinase-1, -2, -9 and tissue inhibitor of matrix metalloproteinases-1 concentrations in amniotic fluid in physiological pregnancy

The aim of this study was to determine reference values of matrix metalloproteinase-1 (MMP-1), MMP-2, MMP-9 and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) in the amniotic fluid at the first stage of labor in physiological pregnancy. 89 women at the first stage of term labor have been examined. Samples of amniotic fluid were taken at the first period of labor by vaginal amniotomy. Concentrations ofMMP-1, MMP-2, MMP-9, and TIMP-1 were investigated in amniotic fluid by ELISA kits. We have determined normal concentration ranges for MMP-1, MMP-2, MMP-9, TIMP-1, and ratios of concentrations of MMPs and TIMP-1 (MMP-1/TIMP-1, MMP-2/TIMP-1, MMP-9/TIMP-1) in the amniotic fluid at the first period of labor in physiological pregnancy. These included: MMP-1--5.1-16.8 pg/mg of protein, MMP-2--238.3-374.1 pg/mg of protein, MMP-9--66.1-113.3 pg/mg of protein, TIMP-1--4.7-13.6 pg/mg of protein, ratio of MMP-1/TIMP-1--0.1-2.2, ratio of MMP-2/TIMP-1--19.9-55.7, ratio of MMP-9/TIMP-1--4.2-17.2.

Myocardial Collagen Cross-Linking Is Associated With Heart Failure Hospitalization in Patients With Hypertensive Heart Failure

BackgroundExcessive myocardial collagen cross-linking (CCL) determines myocardial collagen’s resistance to degradation by matrix metalloproteinase (MMP)-1 and interstitial accumulation of collagen fibers with impairment of cardiac function. ObjectivesThis study sought to investigate whether CCL and a newly identified biomarker of this alteration are associated with hospitalization for heart failure (HHF) or cardiovascular death in patients with HF and arterial hypertension in whom other comorbidities were excluded. MethodsEndomyocardial biopsies and blood samples from 38 patients (invasive study), and blood samples from 203 patients (noninvasive study) were analyzed. Mean follow-ups were 7.74 ± 0.58 years and 4.72 ± 0.11 years, respectively. Myocardial CCL was calculated as the ratio between insoluble and soluble collagen. The ratio between the C-terminal telopeptide of collagen type I (CITP) and matrix metalloproteinase-1 (CITP:MMP-1) was determined in blood samples. ResultsInvasive study: CCL was increased (p < 0.001) in patients compared with controls. Patients were categorized according to normal or high CCL values. Patients with high CCL exhibited higher risk for subsequent HHF (log-rank test p = 0.022), but not for cardiovascular death. CITP:MMP-1 was inversely associated with CCL (r = −0.460; p = 0.005) in all patients. Receiver operating characteristic curves rendered a CITP:MMP-1 cutoff ≤1.968 (80% sensitivity and 76% specificity) for predicting high CCL. Noninvasive study: Patients were categorized according to CITP:MMP-1 ratio values as normal ratio (>1.968) or low ratio (≤1.968). Patients with a low ratio exhibited higher risk for HHF (log-rank test p = 0.014), which remained significant after adjustment for relevant covariables (adjusted hazard ratio: 2.22; 95% CI: 1.37 to 3.59, p = 0.001). In addition, CITP:MMP-1–based categorization yielded significant integrated discrimination and net reclassification improvements (p = 0.003 and p = 0.009, respectively) for HHF over relevant risk factors. CITP:MMP-1 was not associated with the risk of cardiovascular death. ConclusionsExcessive myocardial CCL is associated with HHF in hypertensive patients with HF. In this population, the serum CITP:MMP-1 ratio identifies patients with increased CCL and high risk of HHF.

Forensic Potential of MMPs and CC Chemokines for Wound Age Determination.

In this study, we investigated time-dependent expression of matrix metalloproteases (MMP)-2, MMP-9, chemokine CC motif ligand (CCL)-2, CCL-3, CCL-5, IL-1β, IL-6, and TNF-α mRNA at the skin injury site and sought their forensic potentials during the skin wound repair process. The tested wound ages in 42 mouse skin wounds were distributed at 0d, 1d, 3d, 5d, 7d, 10d, and 14d, respectively and then followed by real-time polymerase chain reaction. Ultimately, MMP-2 played an important role in the inflammation phase. On the contrary, MMP-9 became involved at a later phase during wound healing. Meanwhile, CCL-2 and CCL-3 were active throughout almost all of the process. However, CCL-5 mRNA had no significance. Collectively, an MMP-9/MMP-2 ratio of over 0.84 indicated that skin wound healing age was strongly 5days or less. So elevated gene expressions of cytokines and chemokines in different phases of wound ages implied that combined exploration could make wound age determination more accurate and objective.

Alterations in serum matrix metalloproteinases during different reproductive stages of Murrah buffaloes

A comparative study on serum gelatinase was carried out in Murrah buffaloes. Healthy buffaloes (24), 3-6 year old were selected and divided into 4 groups, each comprising 6 animals (group 1: pregnant; group 2: estrus; group 3: anestrus; and group 4: regular cyclic). Blood samples were collected in heparinised vacutainer and immediately transported to the laboratory. These samples were centrifuged at 3,000 rpm for 15 min and serum was preserved at -20°C in deep freezer. The serum samples were subjected to gelatin zymography. In group 1, major bands at 220, 92 and 72kDa bands were observed. The 92kDa MMP-9 band was very prominent and its activity was 3 times higher than that of MMP-2 and also 72KDa of MMP-2 band was very prominent as compared to the human marker. The 220 kDa band appeared as doublet of pro and active forms and above the level of 92kDa band. Minor catalytic breakdown products of 135 kDa were also observed. Below the 92 kDa band, 82kDa active form of MMP-9 was also observed as a band. In group 2, bands were observed at 220, 92 and 72kDa. The activity of MMP-9 band was about half a time lesser than that of human marker MMP-9 (92kDa). All the 3 enzymes were found only in their proform. The level of expression of MMP2 (72kDa) was equal to that of human marker. The ratio of MMP-9 and MMP- 2 was about 2.5. In group 3, there is a very prominent band of 72kDa MMP-2 and a fainter band of 92kDa MMP-9 and the faintest band of 220kDa MMP-9 were observed. Among the 3 bands, the MMP-2 was very prominent and the ratio of MMP-9 and MMP-2 was 0.25. Finally, in group 4, three bands were observed at 220, 92 and at 72 kDa. The level of expression of MMP-9 was greater than MMP-2 and the ratio MMP-9/MMP-2 was about 1.25. It is concluded that MMP2 and MMP 9 was demonstrated in the serum of all the 4 groups. The level of MMP-9 expression was greater in estrus buffaloes as compared to that of regular cyclic buffaloes and it was lowest in anestrus buffaloes.

Matrix metalloproteinase activation by free neutrophil elastase contributes to bronchiectasis progression in early cystic fibrosis.

Neutrophil elastase is the most significant predictor of bronchiectasis in early-life cystic fibrosis; however, the causal link between neutrophil elastase and airway damage is not well understood. Matrix metalloproteinases (MMPs) play a crucial role in extracellular matrix modelling and are activated by neutrophil elastase. The aim of this study was to assess if MMP activation positively correlates with neutrophil elastase activity, disease severity and bronchiectasis in young children with cystic fibrosis.Total MMP-1, MMP-2, MMP-7, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-2 and TIMP-1 levels were measured in bronchoalveolar lavage fluid collected from young children with cystic fibrosis during annual clinical assessment. Active/pro-enzyme ratio of MMP-9 was determined by gelatin zymography. Annual chest computed tomography imaging was scored for bronchiectasis.A higher MMP-9/TIMP-1 ratio was associated with free neutrophil elastase activity. In contrast, MMP-2/TIMP-2 ratio decreased and MMP-1 and MMP-7 were not detected in the majority of samples. Ratio of active/pro-enzyme MMP-9 was also higher in the presence of free neutrophil elastase activity, but not infection. Across the study cohort, both MMP-9/TIMP-1 and active MMP-9 were associated with progression of bronchiectasis.Both MMP-9/TIMP-1 and active MMP-9 increased with free neutrophil elastase and were associated with bronchiectasis, further demonstrating that free neutrophil elastase activity should be considered an important precursor to cystic fibrosis structural disease.

Effect of G-CSF and TPO on HIBD in neonatal rats.

To observe effect of granulocyte colony-stimulating factor (G-CSF) and restructure human thrombopoietin on hypoxic-ischemic brain damage (HIBD) in new born rats. A total of 60 neonatal SD rats were selected and divided into 4 groups, with 15 in each group. Group A served as control group. Rats of Groups B-D were prepared for HIBD model by ligation of left common carotid artery combined with hypoxia method. Rats of Group A were only completed with free left common carotid artery without ligation and hypoxia operation. After HIBD model preparation, Group B was administrated with subcutaneous injection of normal saline for placebo treatment; Group C was administrated with cervical subcutaneous injection of 0.5 μg/10 g granulocyte colony stimulating factor (G-CSF) for 5 d (Once a day); Group D was administrated with intraperitoneal injection of 15 U/10 g recombinant human thromobopoietin (rhTPO) for treatment. After modeling for 7, 14 and 21 d, 5 rats were sacrificed in each group, respectively. Brain quality damage (%) conditions of experimental animals in each group were compared in different time points, and cerebral histopathological changes of each group were observed. Expression of nestin in rats of each group was detected by immunohistochemical method. After modeling for 7, 14 and 21 d, brain quality damages (%) of Groups B, C and D were significant higher than that of in Group A (P<0.05), while brain quality damage (%) degree of Group B was the highest in different time points, followed by Groups D and C, respectively. It was significant different compared among groups (P<0.05). The histopathological observation showed that degrees of brain damages in Groups C and D were significant lower than that of in Group B. After modeling for 7, 14 and 21 d, nestin positive cell populations in Groups B, C, and D were significant higher than Group A (P<0.05). Nestin cell populations of Group C in different time points were significant higher than Groups B and D (P<0.05). There was no significant difference in nestin positive cell populations in the above time points between Groups B and D (P>0.05). Both G-CSF and TPO can protect the nervous system of HIBD neonatal rats. G-CSF can promote the proliferation and differentiation of neural precursor cells to decrease the degeneration and necrosis of nerve cell. TPO can obviously ameliorate morphology index of HIBD rats. Through regulating ratio of TIMP-1 and MMP-9, TPO can maintain the integrity of blood brain barrier to relieve the occurrence of brain damage.

Abstract W P79: Hyperhomocysteinemia Is Associated With Vulnerability of the Cerebral Aneurysmal Wall to Rupture in Ovariectomized Rats

Introduction: The pathogenesis of cerebral aneurysms is multifactorial. Hyperhomocysteinemia (HHcy) leads to endothelial and platelet deterioration and may be associated with the formation of cerebral aneurysms. We tested the hypothesis that HHcy plays a role in the growth and/or rupture of cerebral aneurysms. Methods: We exposed 13-week-old female Sprague-Dawley rats fed a high-salt diet to estrogen deficiency, hemodynamic stress, and induced hypertension. To induce HHcy we added methionine to their drinking water. Results: In methionine induced rats the aneurysms the total rupture rate was higher than in rats not treated with methionine. One fourth of the aneurysms arising at the anterior communicating (Acom) artery, and half of the aneurysms at the posterior half of the circle of Willis, but no aneurysms at the anterior cerebral artery-olfactory artery (ACA-OA) bifurcation ruptured. Immunohistochemically, the infiltration of M1 macrophages was increased at the Acom artery in methionine induced rats and the messenger ribonucleic acid level of matrix metalloproteinase (MMP)-9, the ratio of MMP-9 to tissue inhibitor of metalloprotease (TIMP)2, and the interleukin-6 level were higher at the Acom artery than the ACA-OA bifurcation. Treatment with folic acid abolished the exacerbated effects due to HHcy, suggesting that the propensity of cerebral aneurysms at the Acom artery to rupture is attributable to disequilibrium between the degradation molecules and their inhibitors and to macrophage infiltration. Conclusion: HHcy may be associated with vulnerability of the cerebral aneurysmal wall to rupture in hypertensive ovariectomized rats.

Evaluation of matrix metalloproteinase-9 and tissue inhibitor metalloproteinase-1 levels in bronchoalveolar lavage of apparently healthy smokers

Cigarette smoking has a destructive effect on the extracellular matrix (ECM), so it is considered the most important risk factor for the development of emphysema as it leads to recruitment of activated macrophages and neutrophils in the lung leading to release of proteolytic enzymes as matrix metalloproteinases (MMPs) or an imbalance between MMPs and the tissue inhibitors of MMP (TIMPs), playing a role in the pathogenesis of emphysema. Aim of the workThe aim of this work was to evaluate the level of MMP-9 and tissue TIMP-1 in bronchoalveolar lavage of apparently healthy smokers. Methods and resultsMMP-9 and TIMP-1 levels were measured by ELISA in BAL in 10 healthy controls, 15 apparently healthy smokers and 15 emphysematous patients. Smokers and COPD patients had a higher concentration of MMP-9 and TIMP-1 compared with controls. The ratio of MMP-9 to TIMP-1 was significantly higher in the emphysema group than the two other groups. Also the smoker group was higher than the control group but without statistic significant difference. ConclusionsHealthy smokers had a higher concentration of total MMP-9 and that concentration was correlated with their exposure to tobacco smoke. Maintenance of normal MMP-9/TIMP-1 ratio in healthy smokers may explain the absence of progressive airway obstruction. Measurement of active MMP-9 concentration could be useful for assessment of airway remodeling.

Serum Matrix Metalloproteinase-9 and Tissue Inhibitor of Metalloproteinase-1 Expression in Patients with Non-alcoholic Fatty Liver Disease

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in developed countries. NAFLD may progress to non-alcoholic steatohepatitis (NASH) and cirrhosis. Emerging evidence suggests that NAFLD is the hepatic manifestation of metabolic syndrome (MetS). NAFLD is closely linked to MetS, with a significant increase in cardiovascular risk. Several matrix metalloproteinases (MMPs) and tissue inhibitors ofMMPs (TIMPs) play important roles in the pathophysiology of atherosclerosis and liver fibrosis. In this study, we investigated the usefulness of serum metalloproteinases as noninvasive markers of NAFLD. 46 patients with NAFLD and twenty-six healthy controls were enrolled into the study, in Gulhane Military Medical Academy, Haydarpasa Training Hospital. Liver biopsies were performed on all patients with NAFLD and histopathological evaluations were made by an experienced pathologist. All NAFLD patients were divided into 2 subgroups according to MetS status using ATP III criteria. MMP-9 and TIMP-1 were studied in serum samples of all groups. Results were compared between both groups and subgroups. In this study, the NAFLD and control groups did not differ significantly on MMP-9, TIMP-1, and TIMP-1/MMP-9 ratio (p>0.05). However, we found a significant relationship between the HOMA and TIMP-1 (p<0.05). Moreover, MMP-9 and TIMP-1/MMP-9 levels were significantly correlated with waist circumference (p<0.05). Our findings are not sufficient to suggest that MMP-9, TIMP-1, and TIMP-1/MMP-9 ratio might be used as noninvasive biochemical diagnostic tests among NAFLD patients.

Changes of Proteases, Antiproteases, and Pathogens in Cystic Fibrosis Patients' Upper and Lower Airways after IV-Antibiotic Therapy.

Background. In cystic fibrosis (CF) the upper (UAW) and lower airways (LAW) are reservoirs for pathogens like Pseudomonas aeruginosa. The consecutive hosts' release of proteolytic enzymes contributes to inflammation and progressive pulmonary destruction. Objectives were to assess dynamics of protease : antiprotease ratios and pathogens in CF-UAW and LAW sampled by nasal lavage (NL) and sputum before and after intravenous- (IV-) antibiotic therapy. Methods. From 19 IV-antibiotic courses of 17 CF patients NL (10 mL/nostril) and sputum were collected before and after treatment. Microbiological colonization and concentrations of NE/SLPI/CTSS (ELISA) and MMP-9/TIMP-1 (multiplex bead array) were determined. Additionally, changes of sinonasal symptoms were assessed (SNOT-20). Results. IV-antibiotic treatment had more pronounced effects on inflammatory markers in LAW, whereas trends to decrease were also found in UAW. Ratios of MMP-9/TIMP-1 were higher in sputum, and ratios of NE/SLPI were higher in NL. Remarkably, NE/SLPI ratio was 10-fold higher in NL compared to healthy controls. SNOT-20 scores decreased significantly during therapy (P = 0.001). Conclusion. For the first time, changes in microbiological patterns in UAW and LAW after IV-antibiotic treatments were assessed, together with changes of protease/antiprotease imbalances. Delayed responses of proteases and antiproteases to IV-antibiotic therapy were found in UAW compared to LAW.

Biomarkers in Trypanosoma cruzi-infected and uninfected individuals with varying severity of cardiomyopathy in Santa Cruz, Bolivia.

BackgroundTwenty to thirty percent of persons with Trypanosoma cruzi infection eventually develop cardiomyopathy. If an early indicator were to be identified and validated in longitudinal studies, this could enable treatment to be prioritized for those at highest risk. We evaluated cardiac and extracellular matrix remodeling markers across cardiac stages in T. cruzi infected (Tc+) and uninfected (Tc−) individuals.MethodsParticipants were recruited in a public hospital in Santa Cruz, Bolivia and assigned cardiac severity stages by electrocardiogram and echocardiogram. BNP, NTproBNP, CKMB, troponin I, MMP-2, MMP-9, TIMP-1, TIMP-2, TGFb1, and TGFb2 were measured in specimens from 265 individuals using multiplex bead systems. Biomarker levels were compared between Tc+ and Tc− groups, and across cardiac stages. Receivers operating characteristic (ROC) curves were created; for markers with area under curve>0.60, logistic regression was performed.ResultsAnalyses stratified by cardiac stage showed no significant differences in biomarker levels by Tc infection status. Among Tc+ individuals, those with cardiac insufficiency had higher levels of BNP, NTproBNP, troponin I, MMP-2, TIMP-1, and TIMP-2 than those with normal ejection fraction and left ventricular diameter. No individual marker distinguished between the two earliest Tc+ stages, but in ROC-based analyses, MMP-2/MMP-9 ratio was significantly higher in those with than those without ECG abnormalities.ConclusionsBNP, NTproBNP, troponin I, MMP-2, TIMP-1, and TIMP-2 levels rose with increasing severity stage but did not distinguish between Chagas cardiomyopathy and other cardiomyopathies. Among Tc+ individuals without cardiac insufficiency, only the MMP-2/MMP-9 ratio differed between those with and without ECG changes.

The Correlation of Matrix Metalloproteinase 9–to-Albumin Ratio in Wound Fluid with Postsurgical Complications after Body Contouring

The authors' earlier retrospective report of surgical complications after abdominal contouring surgery provided evidence that post-bariatric surgery patients are at increased risk of developing wound complications compared with a normal population. This prospective pilot study was designed as a comparative analysis of both surgical and wound healing characteristics between massive weight loss and normal patients who present for abdominal contouring surgery. Excisional wounds were created and polytetrafluoroethylene tubing was inserted during the preoperative period for later harvesting in patients undergoing abdominal contouring following Roux-en-Y gastric bypass for weight loss (n = 16) or abdominoplasty (n = 17). Wound fluids were sequentially collected from drains and subjected to matrix metalloproteinase (MMP) analysis. Standard postsurgical complications were documented. Surgical complications were more common in weight loss patients (47 percent) than in control patients (25 percent). MMP analyses showed that MMP-9 levels remained significantly elevated at postoperative day 4 in patients who subsequently experienced complications in either the weight loss group (p = 0.02) or the control group (p = 0.03). Other parameters showed no significant differences between massive weight loss patients and controls. Although many markers were examined, the ratio of MMP-9 to albumin was the only predictor of postsurgical complications in any group. This lends further support to growing evidence that MMP-9 may be a useful biomarker of postsurgical complications. This pilot work showed no causal factors that explain the higher rates of postsurgical complications in the post-bariatric surgery patient population. Risk, II.

Changes of TGF-β2, MMP-2, and TIMP-2 levels in the vitreous of patients with high myopia.

To investigate the concentrations of transforming growth factor (TGF)-β2, matrix metalloproteinase (MMP)-2, and tissue inhibitor of metalloproteinase (TIMP)-2 in the vitreous of patients with high myopia. Twenty-six patients with high myopia (HM) who received vitrectomy for macular retinoschisis or macular hole were enrolled in this prospective study. Twenty-six patients with idiopathic macular hole or macular epiretinal membrane were chosen as a control group. Vitreous samples were obtained during the vitrectomy surgery. The levels of TGF-β2、MMP-2、TIMP-2 in the vitreous samples were measured by enzyme-linked immunosorbent assay. The MMP activity was determined by a fluorometric assay. There was no significant difference in the vitreous level of TGF-β2 between HM (1.64 ± 0.38 ng/ml) and the control group (1.56 ± 0.32 ng/ml, p = 0.56). The vitreous levels of MMP-2 in HM (32.40 ± 14.90 ng/ml) were significantly higher than in the control group (21.42 ± 6.74 ng/ml, p < 0.01). The ratio of MMP-2/TIMP-2 was significantly elevated in the vitreous samples from HM (0.61 ± 0.19), compared to the control group (0.48 ± 0.11, p < 0.05). The MMP activity was also significantly elevated in the vitreous samples from HM (4,030.8 ± 1,257.3 FIU), compared to the control group (3,245.8 ± 835.6 FIU, p < 0.05). The elevated MMP/TIMP ratio and MMP activity may play a role in the pathogenesis of human high myopia. Large prospective studies are needed to further investigate the effect of MMPs in the pathogenesis of human high myopia.