Tissue Inhibitors Of Matrix Metalloproteinases Research Articles

Doxycycline-Induced Changes in Circulating MMP or TIMP2 Levels Are Not Associated with Skeletal-Related Event-Free or Overall Survival in Patients with Bone Metastases from Breast Cancer.

Doxycycline is often used as a promoter of inducible gene expression in preclinical models; however, it can also have direct effects on tumor growth and survival. This is due in part to its ability to inhibit cell invasion and regulate matrix metalloproteinase (MMP) expression. Given that doxycycline is also osteotropic, a clinical study to assess its effects on modulation of tumor progression or prevention of skeletal-related events (SRE) in patients with bone metastases from breast cancer (the Achilles trial) was undertaken. Patients received 100 mg of oral doxycycline twice daily for 12 weeks, with serum obtained at baseline and 4, 8 and 12 weeks post-initiation of doxycycline treatment. Exploratory analysis of the effects of doxycycline on circulating levels of MMP or tissue inhibitor of matrix metalloproteinase 2 (TIMP2) was performed in enrolled patients. Statistically significant associations were observed between MMP2, MMP9 and TIMP2 at baseline with significant associations maintained between absolute levels and changes in levels of MMP2 and TIMP2 at weeks 4-12 post initiation of doxycycline. Treatment with doxycycline generally resulted in decreases in MMP2 and MMP9 levels with concurrent upregulation of TIMP2 at 12 weeks post-initiation of doxycycline treatment. Despite this, we observed no association with the levels of any of these factors with either SRE-free or overall survival in this patient cohort. In summary, despite observing hypothesized effects of doxycycline administration on surrogate markers of its anti-tumor activity, measures of circulating levels of these biomarkers were not prognostic in this patient population.

TIMP-2 and IGFBP7 in human kidney biopsies in renal disease.

Tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and insulin-like growth factor binding protein 7 (IGFBP7) are markers of tubular stress and urinary [TIMP-2]*[IGFBP7] is an established biomarker for risk assessment of acute kidney injury. There are no studies of expression profiles or localization of these markers in human renal tissue with confirmed renal disease. We analysed 37 kidney biopsies of patients with renal disease and 10 non-diseased control biopsies for TIMP-2 and IGFBP7 expression using immunohistochemistry. Changes in glomerular morphology were evaluated by a semi-quantitative glomerulosclerosis score (GSI) and tubular interstitial changes were graded by the tubular injury score (TSI) using periodic acid-Schiff-stained paraffin sections. Interstitial fibrosis and tubular atrophy (IF/TA) were graded according to the Banff classification. Urinary [TIMP-2]*[IGFBP7] was collected at the time of biopsy. TIMP-2 and IGFBP7 had significantly greater expression in kidney biopsies from patients with renal disease compared with control tissue, especially in the tubular compartment. Here, IGFBP7 was detected in proximal and distal tubules while TIMP-2 was predominantly localized in the collecting ducts. Renal injury significantly correlated with staining intensity for TIMP-2 and IGFBP7: GSI weakly correlated with glomerular TIMP-2 (r=0.36) and IGFBP7 (r=0.35) and TSI correlated with tubular TIMP-2 (r=0.41) and IGFBP7 (r=0.43). Urinary [TIMP-2]*[IGFBP7] correlated weakly with the histopathological damage score but not with glomerular and tubular expression. Our findings underline the role of TIMP-2/IGFBP7 as an unspecific marker of renal injury that is already in use for early detection of acute kidney injury.

Expression of collagenases (matrix metalloproteinase-1, -8, -13) and tissue inhibitor of metalloproteinase-3 (TIMP-3) in naturally occurring bovine cutaneous fibropapillomas.

Bovine cutaneous fibropapillomas are among the most common skin tumors in cattle; their etiology is associated with infection by bovine papillomavirus (BPV) types-1/-2 which are considered oncogenic. Degradation of the extracellular matrix (ECM), especially collagenolysis, is a key event during a series of relevant physiological processes, including tissue remodeling and repair. Various types of proteins are implicated in the regulation of ECM degradation: among these, matrix metalloproteinases (MMPs), a group of zinc-dependent endoenzymes, and tissue inhibitors of matrix metalloproteinases (TIMPs) are known to play a major role. Previous studies reported that aberrant expression of collagenolytic MMPs (MMP-1/-8/-13) and unbalancing between MMPs and TIMPs represent a critical step in tumor growth and invasion; however, studies regarding this topic in bovine cutaneous fibropapillomas are lacking. The aim of this study was to investigate the expression of the collagenases MMP-1/-8/-13 and TIMP-3 in naturally occurring fibropapillomas harboring BPV-2 DNA and normal skin samples. Here, by immunohistochemistry and western blotting analysis, we demonstrated overexpression of MMP-8/-13 along with a down-regulation of MMP-1, associated with a decrease in TIMP-3 levels in tumor compared with normal skin samples. This is the first study describing MMP-1/-8/-13 and TIMP-3 expression in bovine cutaneous fibropapillomas and our results suggest that an impaired expression of collagenases along with an imbalance between MMPs/TIMPs may contribute to an increased collagenolytic activity, which in turn could be important in ECM changes and tumors development.

Pathomechanism of Liver Fibrosis and Mesenchymal Stem Cells in its Resolution Process

Liver fibrosis is a disease process that, without adequate treatment, can lead to liver failure and can be life-threatening. This disease is reversible and appropriate therapy can prevent further liver damage. Liver transplant therapy is the only treatment for an end-stage liver disease that works, but it has various obstacles and limitations in its implementation. Therefore, nowadays, mesenchymal stem cells (MSCs) have become a hope of therapy for liver fibrosis. Our literature review describes the pathomechanism of liver fibrosis and the steps of its resolution, accompanied by the possible role of MSCs in supporting the process. The activation of several complex pathways regulates liver fibrosis, and its resolution, involving Transforming Growth Factor (TGF)-β, signal transducer and activator of transcription-3, and Wnt/β-catenin signaling is involved in Hepatic Stellate Cells (HSCs) activation, which are precursors of myofibroblasts (MFs) and causes fibrosis. The presence of the High-mobility group box-1 pathway, which also induces the production of proinflammatory cytokines and the role of matrix metalloproteinases (MMPs)/tissue Inhibitors of MMPs s and Syndecan-1, is incorporated into the extracellular matrix (ECM). In repairing liver damage, four steps of liver fibrosis resolution are required, such as preventing further damage, restoring the intrahepatic balance of inflammation, removing and inactivating MFs, and ECM degradation associated with arresting the eight pathways of the fibrosis mechanism. MSCs can help resolve liver fibrosis and speed up wound healing, increase hepatocyte survival, and suppress HSCs activation by blocking fibrosis mechanism pathways such as TGF-β and pro-inflammatory factors such as tumor necrosis factor-alpha, interferon-gamma, IL-6, IL-17, and IL-23, in addition to an elevated level of an anti-inflammatory factor like IL-10.

TIMPs Expression as A Maternal Cell Free Plasma Biomarker of Severe Preeclampsia: A Case-Control Study.

Preeclampsia (PE) is a pregnancy related disorder with prevalence of 6-7%. Insufficient trophoblastic invasion leads to incomplete remodeling of spiral arteries and consequent decrease in feto-placental perfusion. Altered placental expression of tissue inhibitors of matrix metalloproteinase (TIMPs) is considered to be involved in this process while the balance between matrix metalloproteinases (MMPs) and TIMPs contributes to remodeling of the placenta and uterine arteries by degradation and refurbishing of extracellular matrix (ECM). Therefore, TIMPs, fetal expression pattern was evaluated with the aim of its potential to be used as a determinant for the (early) detection of PE. In this case-control study, cell free fetal RNA (cffRNA) released by placenta into the maternal blood was used to determine expression patterns of TIMP1, 2, 3 and 4 in the severe preeclamptic women in comparison with the normal pregnant women. Whole blood from 20 preeclamptic and 20 normal pregnant women in their 28-32 weeks of gestational age was collected. The second control group consisted of 20 normal pregnant women in either 14 or 28 weeks of gestation (each 10). cffRNA was extracted from plasma and real-time polymerase chain reaction (PCR) was done to determine the expression levels of TIMP1, 2, 3 and 4 genes. Statistical analysis of the results showed significant higher expression of TIMP1-4 in the preeclamptic women in comparison with the control group (P=0.029, 0.037, 0.037 and 0.049, respectively). Also, an increased level of TIMPs expression was observed by comparing 14 to 28 weeks of gestational age in the normal pregnant women in the second control group. An increased cffRNA expression level of TIMPs may be correlated with the intensity of placental vascular defect and may be used as a determinant of complicated pregnancies with severe preeclampsia.

Ratio of Serum Matrix Metalloproteinases and Their Inhibitors in Chronic Insomnia Patients Running Title: MMP/TIMP Balance in Chronic Insomnia

Objective: Sleep is an essential part of a healthy life. Many people experience insomnia due to their living conditions. Matrix metalloproteinases (MMPs) play an essential role in remodeling the microenvironment. Tissue inhibitors of MMPs (TIMPs) maintain a balance with MMPs. Disruption of the balance causes various pathologies. The aim of the study is to elucidate the effects of sleep disturbance on matrix metalloproteinases and inhibitors by comparing MMP-2/TIMP-2 and MMP-9/TIMP-1 ratio in the serum of patients with chronic insomnia to healthy controls. Material and Methods: This study included 40 adult males diagnosed with chronic insomnia and 40 healthy individuals as a control group. Blood samples were obtained from the brachial vein and subsequently centrifuged at 2000 rpm for 15 minutes to collect serum samples. MMP-2, MMP-9, TIMP-1, and TIMP-2 levels were analyzed using commercial Enzyme-linked immunosorbent assay (ELISA) kit protocols with the obtained serum samples. Results: Consequently, in this study it was demonstrated a higher MMP-2/TIMP-2 ratio in chronic insomnia patients when compared to healthy controls, whereas the MMP-9/TIMP-1 ratio remained unchanged. Conclusion: These results suggest that higher MMP2/TIMP2 ratio may potentially contribute to the pathogenesis of diseases associated with sleep deprivation.

Restoration of Motor Function through Delayed Intraspinal Delivery of Human IL-10-Encoding Nucleoside-Modified mRNA after Spinal Cord Injury.

Efficient invivo delivery of anti-inflammatory proteins to modulate the microenvironment of an injured spinal cord and promote neuroprotection and functional recovery is a great challenge. Nucleoside-modified messenger RNA (mRNA) has become a promising new modality that can be utilized for the safe and efficient delivery of therapeutic proteins. Here, we used lipid nanoparticle (LNP)-encapsulated human interleukin-10 (hIL-10)-encoding nucleoside-modified mRNA to induce neuroprotection and functional recovery following rat spinal cord contusion injury. Intralesional administration of hIL-10 mRNA-LNP to rats led to a remarkable reduction of the microglia/macrophage reaction in the injured spinal segment and induced significant functional recovery compared to controls. Furthermore, hIL-10 mRNA treatment induced increased expression in tissue inhibitor of matrix metalloproteinase 1 and ciliary neurotrophic factor levels in the affected spinal segment indicating a time-delayed secondary effect of IL-10 5 d after injection. Our results suggest that treatment with nucleoside-modified mRNAs encoding neuroprotective factors is an effective strategy for spinal cord injury repair.

Assessment of YME1L and mitofusin2 as a possible diagnostic and/or therapeutic target in hepatocellular carcinoma

Mitochondrial dynamics are a contraversal issue in hepatocellular carcinoma. The present study tries to illustrate the role of mitochondrial dynamics proteins (mitofusin-2 (Mfn2) and YME1L) in hepatocarcinogenesis. Five groups were used: the control group and three HCC groups (after 8, 16, and 24 weeks from DENA induction). The last group was treated with Sorafenib (SP) (10 mg/kg), via oral gavage for 4 weeks after cancer induction. This study revealed that Mfn-2 was downregulated and YME1l was overexpressed in different HCC groups. This dysregulation of mitochondrial dynamics proteins was associated with high hepatic levels of cyclin D1, MMP-9, and MDA and overexpression of ki67 as well as decreasing the hepatic expression of tissue inhibitor of matrix metalloproteinase-3 (Timp-3) and Bax. To confirm the possible role of Mfn2 and YME1L in HCC, we assessed the effect of sorafenib on these parameters and its related HCC characteristics. Sorafenib corrected the level of Mfn2 and YME1L and decreased tumor cell proliferation as well. We also elucidated that mitochondrial dynamics proteins (Mfn2 and YME1L) could be a good therapeutic target for HCC.

The Biology and Function of Tissue Inhibitor of Metalloproteinase 2 in the Lungs.

Tissue inhibitors of matrix metalloproteinases (TIMP) are a family of four endogenous proteins that primarily function to inhibit the activities of proteases such as the matrix metalloproteinases (MMP). Altered MMP/TIMP ratios are frequently observed in several human diseases. During aging and disease progression, the extracellular matrix (ECM) undergoes structural changes in which elastin and collagens serve an essential role. MMPs and TIMPs significantly influence the ECM. Classically, elevated levels of TIMPs are suggested to result in ECM accumulation leading to fibrosis, whereas loss of TIMP responses leads to enhanced matrix proteolysis. Here, we outline the known roles of the most abundant TIMP, TIMP2, in pulmonary diseases but also discuss future perspectives in TIMP2 research that could impact the lungs. TIMP2 directly inhibits MMPs, in particular MMP2, but TIMP2 is also required for the activation of MMP2 through its interaction with MMP14. The protease and antiprotease imbalance of MMPs and TIMPs are extensively studied in diseases but recent discoveries suggest that TIMPs, specifically, TIMP2 could play other roles in aging and inflammation processes.

Biomarkers for Predicting the Occurrence and Progression of Atrial Fibrillation: Soluble Suppression of Tumorigenicity 2 Protein and Tissue Inhibitor of Matrix Metalloproteinase-1.

Soluble suppression of tumorigenicity 2 protein (sST2) and tissue inhibitor of matrix metalloproteinase (TIMP)-1 are involved in multiple pathogenic pathways, including cardiac remodeling, which is the main pathology of atrial fibrillation (AF). This study aims to investigate the previously unexplored relationship between the serum levels of sST2, TIMP-1, and AF. This was a prospective cross-sectional study conducted at the Capital Medical University Affiliated Beijing Anzhen Hospital between June 2019 and July 2020, with a total of 359 participants. The clinical characteristics and laboratory results of the patients were compared, and multivariable ordinal logistic regression was used to evaluate the relationship between serum sST2, TIMP-1, and AF. The participants included 110 patients with sinus rhythm (SR), 113 with paroxysmal AF (the paroxysmal AF group), and 136 with persistent AF (the persistent AF group). It was found that the sST2 levels gradually increased in these three groups, from 9.1 (6.7-12.4 pg/ml) in the SR group to 14.0 (10.4-20.8 pg/ml) in the paroxysmal AF group and to 19.0 (13.1-27.8) pg/ml) in the persistent AF group (p < 0.001). The multivariable ordinal logistic regression model for sST2 and TIMP-1 demonstrated that sST2 had an area under the receiver operating characteristic (ROC) curve (AUC) of 0.797 (95% confidence interval (CI) 0.749-0.846, p < 0.001) and TIMP-1 had an AUC of 0.795 (95% CI 0.750-0.841, p=0.000). The multivariable ordinal logistic regression model for sST2 and TIMP-1 showed good discrimination between SR and AF, with an AUC of 0.846, and the addition of clinical factors, such as brain natriuretic peptide (BNP), left atrial diameter, age, and gender, to the biomarker model improved the detection of SR and AF (AUC 0.901). In this cohort study, sST2 and TIMP-1 were associated with AF progression, independent of clinical characteristics and biomarkers. Soluble ST2 and TIMP-1 combined with age, elevated N-terminal-pro hormone BNP(NT-BNP), and an enlarged left atrium were able to demonstrate the progression of AF reliably.

Early Phase Increase in Serum TIMP-1 in Patients with Acute Encephalopathy with Biphasic Seizures and Late Reduced Diffusion.

Acute encephalopathy with biphasic seizures and late reduced diffusion (AESD) is the most frequent subtype of acute encephalopathy syndrome among Japanese children. Exanthem subitum is the most common causative infectious disease of AESD. We herein retrospectively analyzed serum and cerebrospinal fluid (CSF) concentrations of matrix metalloproteinase-9 (MMP-9), tissue inhibitor matrix metalloproteinase-1 (TIMP-1), and seven cytokines in patients with AESD or prolonged febrile seizure (FS) to assess the pathophysiology of AESD and detect biomarkers for diagnosing AESD in the early phase. Serum and CSF samples were obtained from 17 patients with AESD (1st seizure phase group, n = 7; 2nd seizure phase group, n = 10) and 8 with FS. The concentrations of MMP-9, TIMP-1, and seven cytokines were measured by enzyme-linked immunosorbent assays or cytometric bead arrays. Serum concentrations of TIMP-1 were significantly higher in the 1st seizure phase group than in the 2nd seizure phase group. No significant differences were observed in serum concentrations of MMP-9 or the MMP-9/TIMP-1 ratio. The MMP-9-independent increase in circulating TIMP-1 concentrations observed in the present study may be associated with the pathophysiology of AESD in the 1st seizure phase.

Kidney Cell Cycle Arrest and Cardiac Biomarkers and Acute Kidney Injury Following Angiography: The Prevention of Serious Adverse Events Following Angiography (PRESERVE) Study

Recent studies in patients with chronic kidney disease (CKD) indicate that most cases of contrast-associated acute kidney injury (CA-AKI) are mild and are not associated with elevation in kidney injury biomarkers. We used highly sensitive kidney cell cycle arrest and cardiac biomarkers to assess the risk of CA-AKI and major adverse kidney events in patients with CKD undergoing angiography. A retrospective study. A subset of 922 participants from the Prevention of Serious Adverse Events following Angiography trial. Pre- and postangiography urinary tissue inhibitor of matrix metalloproteinase [TIMP]-2 and insulin growth factor binding protein [IGFBP]-7 were measured in 742 subjects, and plasma β natriuretic peptide (BNP) and high-sensitivity C-reactive protein (hs-CRP), and serum troponin (Tn) in 854 participants using samples obtained 1-2 hours before and 2-4 hours after angiography. CA-AKI and major adverse kidney events. We fitted logistic regression to examine association and area under the receiver operating characteristic curves for risk prediction. There were no differences in postangiography urinary [TIMP-2]•[IGFBP7], plasma BNP, serum Tn, and hs-CRP concentrations among patients with and without CA-AKI and major adverse kidney events. However, higher pre- and postangiography median plasma BNP (pre: 200.0 vs 71.5, pg/mL, P=0.05; post: 165.0 vs 81pg/mL, P=0.02); serum Tn (pre: 0.03 vs 0.01, ng/mL, P<0.001; post, 0.04 vs 0.02, ng/mL, P=0.01); and hs-CRP (pre: 9.55 vs 3.40mg/L, P=0.01; post: 9.90 vs 3.20mg/L, P=0.002) concentrations were associated with major adverse kidney events, although their discriminatory capacity was only modest (area under the receiver operating characteristic curves<0.7). Most participants were men. Most mild CA-AKI cases are not associated with urinary cell cycle arrest biomarker elevation. Significant elevation in preangiography cardiac biomarkers may reflect patients with more significant cardiovascular disease that may predispose to poor long-term outcomes independent of CA-AKI status.

Structural and functional features of the heart and the dynamics of biomarkers in the progression of chronic heart failure with a moderately reduced ejection fraction of the left ventricle in patients with diabetes mellitus

Aim. To study the relationship between serum levels of aldosterone, tissue inhibitor of matrix metalloproteinases-1 (TIMP-1), tumor necrosis factor-alpha (TNF-) and structural and functional changes in the heart during the progression of chronic heart failure (CHF) with a moderately reduced ejection fraction (CHFunEF) in patients with type 2 diabetes mellitus (DM).&#x0D; Material and methods. 58 patients were examined 30 (51.7%) men and 28 (48.3%) women diagnosed with DM and CHFunEF (EF=4149%), with functional class (FC) I, II and III of CHF according to New York Heart Association (NYHA). The control group consisted of 16 people. Patients underwent echocardiography, determination of serum levels of aldosterone, TIMP-1 and TNF- by ELISA. The patients were divided into 2 groups: 1st group patients with DM and CHFunEF who had a myocardial infarction (MI) 27 people (46.6%), 2nd group patients with DM and CHFunEF 31 people (53.4%). Statistica v. 10.0 software (USA) was used for statistical analysis. Differences were considered statistically significant at р0.05.&#x0D; Results. Serum levels of aldosterone, TIMP-1 and TNF- were significantly higher than those of the control group and increased with an increase in CHF FC. In patients of the 1st group, the volume and index values of the left parts of the heart and serum levels of biomarkers significantly exceeded those of the 2nd group.&#x0D; Conclusion. Activation of the aldosterone system, increased profibrotic and immuno-inflammatory processes with the progression of CHFunEF in patients with DM lead to an increase in serum levels of aldosterone, TIMP-1 and TNF- with an increase in CHF FC. Patients with DM and CHFunEF who have undergone MI have more pronounced structural and functional changes in the heart and greater values of volumetric and index parameters of the left heart than patients with DM and CHFunEF. Serum levels of aldosterone, TIMP-1 and TNF- in patients with DM and CHFunEF who have undergone MI exceed those in patients with DM and CHFunEF, these biomarkers can be used to diagnose the progression of CHFunEF in patients with DM.

Alternanthera brasiliana L. extract alleviates carbon tetrachloride-induced liver injury and fibrotic changes in mice: Role of matrix metalloproteinases and TGF-β/Smad axis

Ethnopharmacological relevanceAlternanthera brasiliana L. is a flowering plant belonging to the family Amaranthaceae and is popularly known as “penicillin”. It is used in folk medicine to treat infections, coughs, wound healing, and inflammatory diseases.Aim of the study: We investigated the effect of Alternanthera brasiliana L. leaves hydroalcoholic extract (AB) against oxidative stress, inflammation, and fibrotic changes in an experimental model of carbon tetrachloride (CCl4)-induced liver injury and fibrosis in mice. Materials and methodsThirty-six male Balb/C mice were randomized into five groups: normal control, AB control, CCl4 control, CCl4 + AB-200 mg/kg, and CCl4 + AB-400 mg/kg. In mice, liver injury was induced by intraperitoneal injection of CCl4 (20% in corn oil, 5 ml/kg body weight) thrice a week for six consecutive weeks. AB extract at two doses (200 mg/kg and 400 mg/kg body weight) was administered orally for six consecutive weeks. Liver injury-related serum markers (ALT, AST, ALP), antioxidants (GSH, GST, SOD, and vitamin C), pro-inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-18, ultrasonographic and histological alterations, proteins of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinase-1 (TIMP-1), nuclear factor-κB (p65) (NF-κB), nod-like receptor protein 3 (NLRP3), and TGF-β/Smad signaling were accessed. LC-Q-TOF-MS/MS analysis of AB was performed. ResultsAB treatment significantly decreased the CCl4-induced rise in serum ALT, AST, and ALP activities and improved the histological alterations. Compared with the CCl4-treated group, treatment with AB significantly restored the hepatic antioxidants and reduced the pro-inflammatory cytokines in the liver. The antioxidant activity of AB may be attributed to its terpenoid constituents, which was confirmed by LC-Q-TOF-MS/MS analysis. The CCl4-induced rise in expression of MMP-2 and MMP-9 and decrease in TIMP-1 were markedly restored in the AB-treated groups. Further findings revealed a significant reduction in the protein levels of phospho–NF–κB (p65), NLRP3, TGF-β, pSmad2/3, collagen I, and α-smooth muscle actin (α-SMA) in the AB treatment groups. ConclusionsThe hepatoprotective effect of AB may be attributed to the high content of terpenoid compounds and alleviates liver injury and associated fibrotic changes through modulating MMPs, NF-κB (p65), and the TGF-β/Smad axis.

Expression of matrix metalloproteinases (MMPs)-2/-7/-9/-14 and tissue inhibitors of MMPs (TIMPs)-1/-2 in bovine cutaneous fibropapillomas associated with BPV-2 infection.

Bovine papillomaviruses -1/-2 (BPVs) are small non-enveloped double-stranded DNA viruses able to infect the skin of bovids and equids, causing development of neoplastic lesions such as bovine cutaneous fibropapillomas and equine sarcoid. Matrix metalloproteinases (MMPs) are a group of zinc-dependent endopeptidases that degrade basal membrane and extracellular matrix, whose function is essential in physiological processes such as tissue remodeling and wound healing. MMPs activity is finely regulated by a balancing with expression of tissue inhibitors of MMPs (TIMPs), a process that is impaired during tumour development. BPV infection is associated with upregulation of MMPs and /or their unbalancing with TIMPs, contributing to local invasion and impairment of extracellular matrix remodeling in equine sarcoid; however, studies regarding this topic in bovine fibropapillomas are lacking. The aim of this study was to perform an immunohistochemical and biochemical analysis on a panel of MMPs and TIMPs in BPV-2 positive bovine cutaneous fibropapillomas vs. normal skin samples. Immunohistochemistry revealed a cytoplasmic expression of MMP-2 (15/19), a cytoplasmic and perinuclear immunoreactivity of MMP-7 (19/19) and MMP-9 (19/19), along with a cytoplasmic and nuclear pattern of MMP-14 (16/19), accompanied by a cytoplasmic expression of TIMP-1 (14/19) and TIMP-2 (18/19) in tumour samples; western blotting revealed an overexpression of MMP-2 (8/9), MMP-7 (9/9) and MMP-9 (9/9), and a decreased level of MMP-14 (9/9), TIMP-1 (9/9) and TIMP-2 (9/9) in tumour versus normal skin samples. Moreover, gelatine zymography confirmed the expression of pro-active MMP-2 (9/9) and MMP-9 (9/9) and, most importantly, indicated the presence and increased activity of their active forms (82 and 62 kDa, respectively) in tumour samples. This is the first study describing MMPs and TIMPs in bovine cutaneous fibropapillomas and our results suggest that their unbalanced expression in presence of BPV-2 may play a significant role in tumour development. A further analysis of supplementary MMPs and TIMPs could bring new important insights into the papillomavirus induced tumours.

Matrix Metalloproteinases 2 and 9 and Their Tissue Inhibitors in the Diagnostics of Medullary Thyroid Carcinoma.

Medullary Thyroid Carcinoma (MTC) is a tumor of the neuroendocrine system. In recent years, the need to assess the MTC diagnostic-related parameters has emerged with the aim to elucidate the mechanisms involved in this pathology. The objective of this study was to evaluate the role of Matrix Metalloproteinases (MMPs) 2 and 9, their tissue inhibitors of matrix metalloproteinases (TIMPs), S100 protein, and amyloid in the diagnostic of MTC. Thirty-two samples with MTC (72% women) were included in this cross-sectional study and divided by groups: T category 1 (T1)≤20mm and T category 2 (T2) 20 to 40mm of tumor size. MMPs 2 and 9, TIMPs 2 and 1, S100 protein, and calcitonin in tissues were obtained by immunohistochemical techniques. The presence of amyloid in tissue sections was detected on Thioflavin T-stained slides under fluorescent microscope. Percentage of positive cells (P) observed for MMP-2 was higher in those samples presenting T2 MTC with respect to those with T1 MTC ( P <0.05). Moreover, P-MMP-2 showed a direct correlation with higher T category of MTC (Rho=0.439, P < 0.001), whereas P-MPP-9 was directly correlated with S100 protein and the intensity of calcitonin in tissues (Rho=0.419, P =0.017; Rho=0.422, P =0.016, respectively. Therefore, MMPs were directly correlated with some traditional biomarkers of MTC. In this regard, P-MMP-2 was more expressed in type 2 MTC. Combining the analysis of traditional and other useful biomarkers of MTC as MMPs 2 and 9 could be a useful strategy in the diagnostic of MTC.

Arsenic trioxide sensitizes pancreatic cancer cells to gemcitabine through downregulation of the TIMP1/PI3K/AKT/mTOR axis

Gemcitabine (GEM) is the first-line medication for pancreatic ductal adenocarcinoma (PDAC). However, over some treatment cycles, GEM sensitivity declines and chemotherapeutic resistance develops, resulting in tumor recurrence and metastasis. Therefore, it is critical to elucidate the mechanism of GEM chemoresistance. And a specific drug that is closely related to the mechanism is urgently required to sensitize GEM. Here, tissue inhibitor of matrix metalloproteinases 1 (TIMP1) and phosphorylated mammalian target of rapamycin (p-mTOR) were found to be substantially elevated in PDAC patients and were associated with worse overall survival. The TIMP1/PI3K/AKT/mTOR pathway was found in GEM-resistant PDAC cells and was revealed to be involved in epithelial-mesenchymal transition (EMT) and apoptosis. Furthermore, arsenic trioxide (ATO), a basic therapeutic drug for acute promyelocytic leukemia, mediated TIMP1 reduction by inducing reactive oxygen species generation and hampered the subsequent PI3K/AKT/mTOR axis. Moreover, the combination of ATO and GEM cooperatively suppressed the TIMP1/PI3K/AKT/mTOR pathway, synergistically inhibited EMT and promoted apoptosis. In vitro and in vivo, ATO combined with GEM has a collaborative anticancer effect, inhibiting cancer cell proliferation, migration, invasion, and suppressing tumor growth both in PDAC parental and GEM-resistant cells. Overall, the TIMP1/PI3K/AKT/mTOR pathway is present in PDAC and linked to GEM resistance. ATO suppresses the axis to sensitize GEM and reverse GEM resistance, suggesting a promising treatment for the disease.

Abstract 10409: Performance of the Hart Cve Panel for Prediction of Cardiovascular Events in Patients With Chronic Kidney Disease

Background: Patients with chronic kidney disease (CKD) are at increased risk for cardiovascular events (CVE: incident myocardial infarction [MI], stroke, or cardiovascular death) following an angiographic procedure. Methods: Among 446 patients undergoing coronary or peripheral angiography, we applied a previously-developed biomarker panel to predict incident cardiovascular events (HART CVE, Prevencio, Kirkland WA); the panel consists of 4 biomarkers (N-terminal pro-B type natriuretic peptide, kidney injury molecule-1, osteopontin, and tissue inhibitor of matrix metalloproteinase-1) with results expressed in 2 risk levels (CVE high risk and low risk). Patients were followed for 2 years for incident cardiovascular events. Results: During the 2-year follow-up, 74 CVE occurred. Participants were categorized into four groups based on CKD stages (Stage 1-2 vs Stage 3-5) and CVE risk level (high risk and low risk). In our multivariable model, considering patients at CKD stage 1-2/CVE low-risk as a reference, the hazard ratio (95 % confidence interval) of cardiovascular events was 2.28 (1.53-5.22) for CKD stage 1-2/CVE high-risk, 0.50 (0.07-3.73) for CKD stage 3-5/CVE low-risk and 4.56 (2.54-8.18) for CKD stage 3-5/CVE high-risk ( Table ). The C-statistic for predicting 2-years cardiovascular events in all 446 patients was 0.77 (0.72, 0.82). Conclusion: Patients with CKD are at high risk for incident MI, stroke, and CV death. Utilizing a multiple-protein biomarker panel can assist clinicians in identifying patients at higher risk of CVE, particularly among those at high stages of CKD.

Association between transforming growth factors-β and matrix metalloproteinases in the aqueous humor and plasma in myopic patients

To investigate the relationship between levels of transforming growth factor beta (TGF-beta), matrix metalloproteinase (MMP) and tissue inhibitors of MMP (TIMP) in the aqueous humor (AH) and plasma (PL) of myopic patients. Aqueous humor and plasma were collected intraoperatively from 37 myopic patients with various axial lengths (AL) during ICL surgery. The concentrations of TGF-β1, TGF-β2, TGF-β3, MMP-1, MMP-2, MMP-7, MMP-9, MMP-10, TIMP-1, TIMP-2, TIMP-3 and TIMP-4 were measured using Luminex xMAP Technology kits (Milliplex xMAP kits). The association between TGFβ and MMP/TIMP levels were analyzed based on the Spearman's correlation test or Pearson's correlation test. There was a negative correlation between TGF-β1 and MMP-1, TIMP-2, TIMP-3 and TIMP-4 in the AH, and a positive correlation between TGF-β1 and MMP-1 in the PL. In the AH, levels of TGF-β2 were positively correlated with levels of TIMP-3 (r=0.441; P<0.001). In the PL, levels of TGF-β2 also correlated positively with levels of TIMP-3 (r=0.427; P<0.001)). The level of TGF-β2 was the highest among TGF-βs in the AH. A consistent positive correlation between TGF-β2 and TIMP-3 was found both in the AH and PL, indicating that systemic levels of TGF-β2 and MMPs/TIMPs may also play a role in myopic progression.

Utilizing the Knowledge of Role of Matrix Matrixmetalloproteinases Along with Tissue Inhibitors of Matrix Metalloproteinasesin Various Gynaecological Disorders with Great Emphasis on Preeclampsia, PCOs Along with Gynaecological Malignancies-A Systematic Review

Our insight has escalated over the past decade regarding the understanding of Molecular mechanisms underlying the generation of preeclampsia, with us reviewing the role of epigenetics in the development of preeclampsia along with intra uterine growth restriction (IUGR) generation. Matrix Metalloproteinases (MM’s) MMP’s represent calcium-based zinc-endopeptidases that belong to the metzincin superfamily possessing a lot of functions like a physiological part in tissue morphogenesis, cell migration along with angiogenesis as well as in pregnancy in invasion along with migration of trophoblast besides spiral artery remodeling. Matrix Metalloproteinases (MMP), cytokines, growth factors have a role in physiological events like embryo implantation, trophoblast invasion as well as migration along with decidualization with studies documented that blastocysts are direct generators of MMP2. Thus here we decided to conduct a systematic review on the role of Matrix Metalloproteinases (MMP’s) along with their inhibitors,namely the, Tissue inhibitors of Matrix Metalloproteinases (TIMPs) in gynaecological disorders specifically in generation of preeclampsia along with IUGR; in women with Polycystic Ovarian Syndrome(PCOS); besides in spontaneous abortion along with generation of various cancers of the whole body but main emphasis on gynaecological cancers .Hence utilizing the search engine Thus we utilized Pubmed, MEDLINE, Google Scholar, Scopus, Embase, Web of Sciences, Cochrane library search engines where we utilized the MeSH terms like MMP’s in pregnancy physiological roles; pathological in preeclampsia, IUGR; miRs in these disorders; spontaneous abortion; gynaecological disorders like PCOS; gynaecological cancer along with association of certain MIR’s abnormal in IUGR that were linked to certain gliomas along with MMP’S and IUGR and further utilization of MMP inhibitors clinically, besides that since no efficacious therapy available other therapies like role of extracellular vesicles (ECV’s) that target combined miR’S that are altered and associated with MMP’s in various pathologies and effectively we can treat those disorders from 1970 till January 2021 till date. We found a total of 500 articles out of which we selected 145 articles for this review following evaluation of cross references. No meta-analysis was done. Here we provide an insight on how this knowledge on molecular mechanisms interactions of MMP’s, TIMP’s micro RNA’S might get utilized in generating therapies to prevent or treat preeclampsia, IUGR, various gynaecological malignancies developing extracellular vesicles carrying miR;s targeting specific MMP’s in particular disease or malignancy.