Elevated Matrix Metalloproteinase Concentrations Offer Novel Insight Into Their Role in Pediatric Tuberculous Meningitis.

We collected lumbar and ventricular cerebrospinal fluid and serum from 40 children treated for tuberculous meningitis and measured the concentrations of gelatinases and their inhibitors. The concentrations of matrix metalloproteinase 9 (MMP-9), MMP-2, tissue inhibitor of metalloproteinase 1 (TIMP-1), and TIMP-2 were significantly elevated in the lumbar CSF samples, and we found interesting dynamics for MMP-9 that offer novel insight into its role in pediatric patients with tuberculous meningitis.

The Role of Biomarkers in Acute Kidney Injury.

Several biomarkers have been developed to detect acute kidney injury (AKI) and predict outcomes. Most AKI biomarkers have been shown to be expressed before serum creatinine and to be more sensitive and specific than urine output. Only a few studies have examined how implementation can change clinical outcomes. A second generation of AKI biomarkers have been developed. These markers, including tissue inhibitor of metalloproteinases-2 (TIMP-2) and insulinlike growth factor-binding protein 7 (IGFBP7), have obtained regulatory approval in many countries based on large, rigorous clinical studies and small, single-centered trials and have begun to establish clinical utility.

Plasma matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs and aging and lifelong exercise adaptations in ventricular and arterial stiffness.

The age-associated increase in cardiac and central arterial stiffness is attenuated with lifelong (>25 years) endurance exercise in a dose-dependent manner. Remodelling of the extracellular matrix of cardiovascular structures may underpin these lifelong exercise adaptations in structural stiffness. The primary aim was to examine whether matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) levels are associated with aging and lifelong exercise-related changes in cardiac and central arterial stiffness. Plasma MMPs and TIMPs, left ventricular (LV) (LV stiffness constant) and central arterial stiffness (pulse wave velocity) were examined in healthy adults stratified into five groups based on age and lifelong weekly exercise frequency: (1) young sedentary adults (28-50 years), and older adults (>60 years) who had performed either: (a) sedentary (0-1 sessions/week), (b) casual (2-3 sessions/week), (c) committed (4-5 sessions/week) or (d) athletic (≥6 sessions/week) frequency of exercise. MMP-1 was significantly lower in young compared to older sedentary (p = 0.049). Except for TIMP-2 (p = 0.018 versus committed) and the ratio of MMP-2/TIMP-4 (p = 0.047 versus committed), MMP and TIMP expression was not significantly different in lifelong exercise groups (≥casual) compared to the older sedentary group. MMP-1, -3 had a weak positive relationship with central PWV (r = 0.17-0.25, p ≤ 0.050) but there were no significant relationships between MMPs or TIMPs and LV stiffness constant (p ≥ 0.148). In conclusion, there was not a clear or consistent difference in plasma MMPs and TIMPs with lifelong exercise dose despite exhibiting lower cardiovascular stiffness at the highest exercise levels.

Fusion Peptides CPU1 and CPU2 Inhibit Matrix Metalloproteinases and Protect Mice from Endotoxin Shock Within a Strict Time Window.

Endotoxin shock induction in mice is a commonly used animal model to evaluate the protective effect of biologically active reagents. After an lipopolysaccharides (LPS) stimulus, matrix metalloproteinase-8 (MMP-8) and matrix metalloproteinase-9 (MMP-9) are rapidly degranulated and released by neutrophils, aside other enzymes and effector molecules. MMPs cleave extracellular matrix components and cytokines, and such processes contribute to shock syndrome development. CPU1 and CPU2 are two peptide MMP inhibitors with different in vitro IC50 values to several key enzymes, including MMP-8 and MMP-9. In vivo work confirmed that CPU1 and CPU2 protected mice from endotoxin shock after intravenous and intraperitoneal injections. Furthermore, their minimal effective dose after an intravenous injection and the maximum time interval between intraperitoneal peptide injection (150 mg/kg) and intravenous LPS injection were determined. With the use of an indirect competitive ELISA, plasma CPU1 and CPU2 concentrations in different experimental settings were measured. In addition, the acuteness of MMP-9 release in the mouse circulation after an intravenous LPS injection was confirmed with the zymography technique. Our findings reinforce previous work with other inhibitors about a strict time window within which effective MMP inhibition is needed to obtain significant survival rate improvements and also show that, with strict pharmacokinetic monitoring, potent protease inhibitors may in the future become life-savers in shock conditions.

Tissue inhibitors of matrix metalloproteinases in serum are cardiac biomarkers in Emery-Dreifuss muscular dystrophy.

BACKGROUND: Tissue inhibitors of matrix metalloproteinases (TIMPs) are known to be involved in cardiovascular diseases. Hitherto, they have not been examined in dilated cardiomyopathy in the course of Emery-Dreifuss muscular dystrophy (EDMD). AIM: To define TIMPs in serum because they might help in defining cardiac dysfunction at the early cardiological stages of this disease and detect preclinical stages of cardiomyopathy. METHODS: Twenty-five EDMD patients connected with lamin A/C (AD-EDMD) or emerin (X-EDMD) deficiency and 20 healthy age-matched controls were examined. The serum levels of the tissue inhibitors TIMP-1, -2, -3 were quantified using the ELISA sandwich immunoassay procedure with appropriate antibodies. RESULTS: Serum levels of TIMP-1 were normal in autosomal AD-EDMD and increased in the majority of X-linked EDMD. The level of TIMP-2 was decreased in 25%/21% of AD-EDMD/X-EDMD cases. TIMP-3 serum level was significantly reduced in all the examined patients. Receiver operating curves indicated that in terms of sensitivity and specificity characteristics the performance of TIMP-3 (less that of TIMP-2) makes them the best markers of cardiac involvement among the examined TIMPs. CONCLUSIONS: Evidence shows that the levels of TIMP-3, and in some cases also TIMP-2, are decreased in EDMD. The decrease might be associated with an adverse effect on matrix metalloproteinases and remodelling of the myocardial matrix. The specific decrease of TIMP-3 indicates that this biomarker might help in early detection of cardiac involvement in EDMD. Up-regulation of TIMP-1 in the majority of patients with X-EDMD indicates increased myocardial extracellular matrix turnover, early onset of tissue remodelling, and may contribute to arrhythmia, frequently occurring in this form of the disease.

Plasma and synovial fluid concentration of doxycycline following low-dose, low-frequency administration, and resultant inhibition of matrix metalloproteinase-13 from interleukin-stimulated equine synoviocytes.

REASONS FOR STUDY: To determine whether low-dose, low-frequency doxycycline administration is capable of achieving chondroprotective concentrations within synovial fluid (SF) while remaining below minimum inhibitory concentration 90 (MIC90 ) of most equine pathogens and would be an option in the management of osteoarthritis. OBJECTIVES: To determine whether low-dose, low-frequency oral administration of doxycycline can attain in vivo SF concentrations capable of chondroprotective effects through reduction of matrix metalloproteinase (MMP)-13 activity, while remaining below MIC90 of most equine pathogens. STUDY DESIGN: Descriptive pharmacokinetic study with crossover design. METHODS: Two groups of 6 horses received oral doxycycline. Plasma and SF doxycycline concentrations were measured using high performance liquid chromatography. Group 1 received 5 mg/kg bwt q. 24 h with 21 blood and 8 SF samples collected over 120 h; Group 2 received 5 mg/kg bwt q. 48 h with 27 blood and 11 SF samples collected over 192 h. Cultured synoviocytes were treated with interleukin-1α (1 ng/ml) for 24 h to stimulate MMP synthesis, and then SF was added to the culture medium for 96 h. MMP-13 protein and mRNA were measured in synoviocyte culture medium and synoviocytes, respectively. RESULTS: Mean doxycycline concentration ≥0.043 µg/ml (previously demonstrated to inhibit MMP-13) was achieved in plasma by t = 0.25 h and SF by t = 48 h in Group 1, and in plasma by t = 0.17 h and SF by t = 1 h in Group 2. Synoviocyte culture medium containing doxycycline from Groups 1 and 2 had significantly decreased active MMP-13 protein concentration, and synoviocytes cultured in this medium had significantly decreased MMP-13 gene expression compared to controls. Plasma doxycycline concentration in both groups and SF doxycycline concentration in Group 2 demonstrated a cumulative effect. CONCLUSIONS: Low-dose orally administered doxycycline achieves SF concentrations in vivo capable of diminishing MMP-13 expression. This study supports the use of doxycycline as a disease modifying osteoarthritic drug.

Clinical and immunological aspects of post-kala-azar dermal leishmaniasis in Bangladesh.

We conducted active surveillance for kala-azar and post-kala-azar dermal leishmaniasis (PKDL) in a population of 24,814 individuals. Between 2002 and 2010, 1,002 kala-azar and 185 PKDL cases occurred. Median PKDL patient age was 12 years; 9% had no antecedent kala-azar. Cases per 10,000 person-years peaked at 90 for kala-azar (2005) and 28 for PKDL (2007). Cumulative PKDL incidence among kala-azar patients was 17% by 5 years. Kala-azar patients younger than 15 years were more likely than older patients to develop PKDL; no other risk factors were identified. The most common lesions were hypopigmented macules. Of 98 untreated PKDL patients, 48 (49%) patients had resolution, with median time of 19 months. Kala-azar patients showed elevated interferon-γ (IFNγ), tumor necrosis factor-α (TNFα), and interleukin 10 (IL-10). Matrix metalloproteinase 9 (MMP9) and MMP9/tissue inhibitor of matrix metalloproteinase-1 (TIMP1) ratio were significantly higher in PKDL patients than in other groups. PKDL is frequent in Bangladesh and poses a challenge to the current visceral leishmaniasis elimination initiative in the Indian subcontinent.

Matrix metalloproteinases and the activity of their tissue inhibitors in patients with ST-elevation myocardial infarction treated with primary angioplasty.

BACKGROUND: Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play a role in heart remodelling after acutemyocardial infarction (MI). Their activity is connected with outcome and heart failure development. There is little data on MMP and TIMP activity changes in the setting of ST elevation MI (STEMI) treated with primary percutaneous coronary intervention (pPCI). AIM: To assess the dynamics of activity of MMP-2 and MMP-9 and their endogenous inhibitors TIMP-1 and TIMP-2 in the course of invasive treatment of STEMI. METHODS: The study included 95 patients (age 61.8 ± 12.4 years; 35 women) treated with pPCI with stent implantation due to 100% closure of the target vessel in a setting of STEMI. We measured the activity of MMP-2 and MMP-9 (by zymography,expressed with arbitrary units, AU), CK-MB (U/L), troponin I (ng/mL), TIMP-1, TIMP-2 (ng/mL) concentrations in a peripheral blood before the pPCI, immediately after and 3, 6, 12, 24 and 48 h after the procedure. Left ventricular ejection fraction(LVEF) was estimated at the hospital discharge using the Simpson method. There were two control groups: 15 healthy persons and 15 patients with stable coronary artery disease matched for age and sex with the studied group. RESULTS: The abrupt opening of the target vessel did not produce an early increase in the activity of the MMPs. Their activity was high at the beginning and slowly lowered with time after pPCI so that at 12, 24 and 48 h after pPCI their activity was significantly lower than before and immediately after the pPCI (p < 0.05 for all comparisons). The abrupt opening of the target vessel did not produce significant changes in the TIMP concentration. Only the TIMP-1 showed a slow increase in concentration and achieved a significantly higher level 48 h after the procedure compared to its concentration before and immediately after pPCI (p < 0.05). In 14 patients (15% of the studied group), the post procedure TIMI flow was estimated as lower than 3 (TIMI 1 or 2). There was significantly higher MMP-9 activity in this group before, immediately after and up to 3 h after PCI compared to the group with good angiographic effect (TIMI = 3 after procedure). Patients with lowered LVEF(< 50%) at hospital discharge had higher MMP-9 activity immediately after and 3 h after pPCI compared to patients with preserved LVEF. The same relation was observed for TIMP-2 level, where patients with a higher level before and immediately after pPCI had lowered LVEF at discharge. CONCLUSIONS: 1. The activity level of MMP-2 and MMP-9 is elevated during the STEMI acute phase and falls 12 h after successful pPCI, while TIMP-1 concentration only rises 48 h after the procedure. 2. The abrupt opening of the target vessel in STEMI does not produce acute changes in MMP-2, MMP-9 activity or TIMP-1 and TIMP-2 concentration. 3. The 'no-reflow'phenomenon in STEMI patients occurs more often in those with higher MMP-9 activity before pPCI. 4. Lowered LVEF at hospital discharge is observed in patients with higher periprocedural MMP-9 activity and TIMP-2 level.

Gelatinases and their tissue inhibitors in a group of subjects with metabolic syndrome.

AIM: To evaluate matrix metalloproteases (MMP)-2 and MMP-9 and tissue inhibitor of metalloproteases (TIMP)-1 and TIMP-2 in a group of subjects with metabolic syndrome (MS) subdivided according to the presence or absence of diabetes mellitus. METHODS: We examined in 90 subjects (51 men and 39 women) with MS, defined following the International Diabetes Federation criteria, and subsequently subdivided into diabetic subjects (22 men and 11 women) and nondiabetic subjects s (29 men and 28 women), the plasma concentrations of MMP-2 and MMP-9 and of TIMP-1 and TIMP-2 using specific enzyme-linked immunosorbent assay kits. RESULTS: We found a significant increase in plasma concentrations of MMP-2, MMP-9, TIMP-1, and TIMP-2 in the whole group of MS subjects (P < 0.001) and in both subgroups of MS subjects with diabetes mellitus (P < 0.001) and without diabetes mellitus (P < 0.001) in comparison with healthy controls. We also noted higher concentrations of all the examined parameters in the MS subjects with diabetes mellitus in comparison with the MS subjects without diabetes mellitus. Matrix metalloproteases and TIMPs showed some significant correlations with body mass index and waist circumference and with metabolic parameters in the whole group of MS subjects. CONCLUSION: An altered pattern of MMPs and their inhibitors is demonstrated in MS; the presence of diabetes mellitus strongly influences the concentration of MMP and TIMP, contributing probably to the increased cardiovascular risk of MS subjects.

Serum biomarkers of papillary thyroid cancer.

OBJECTIVE: To identify serum biomarkers of papillary thyroid cancer. METHODS: Prospective analysis was performed of banked tumor and serum specimens from 99 patients with thyroid masses. Enzyme-linked immunosorbent assay (ELISA) was employed to measure levels of five serum proteins previously demonstrated to be up-regulated in papillary thyroid cancer (PTC): angiopoietin-1 (Ang-1), cytokeratin 19 (CK-19), tissue inhibitor of metalloproteinase-1 (TIMP-1), chitinase 3 like-1 (YKL-40), and galectin-3 (GAL-3). Serum levels were compared between patients with PTC and those with benign tumors. RESULTS: A total of 99 patients were enrolled in the study (27 men, 72 women), with a median age of 54 years. Forty-three patients had PTC and 58 cases were benign tumors. There were no statistically significant differences when comparing all five different biomarkers between PTC and other benign thyroid tumors. The p-values were 0.94, 0.48, 0.72, 0.48, and 0.90 for YKL-40, Gal-3, CK19, TIMP-1, and Ang-1, respectively. CONCLUSION: Serum levels of four of the five proteins were elevated in patients with thyroid masses relative to normal values. However, the difference between benign and PTC was not significant. Two of the markers (Gal-3 & TIMP-1) displayed a greater potential difference, which may warrant further investigation. This study suggests that other serum markers should be sought. This is the first study to investigate potential serum biomarkers based on over-expressed proteins in thyroid cancer versus benign pathology.

Altered circulating levels of matrix metalloproteinases 2 and 9 and their inhibitors and effect of progesterone supplementation in women with endometriosis undergoing in vitro fertilization.

OBJECTIVE: To investigate differences in the activity of matrix metalloproteinases (MMPs) 2 and 9 and their respective tissue inhibitors (TIMPs) in follicular fluid of women with endometriosis, to correlate the findings with IVF outcome, and to examine the therapeutic potential of progesterone supplementation in restoring the fine balance between MMPs and TIMPs. DESIGN: Prospective case-control clinical study. SETTING: Infertility clinic and reproductive health research unit. PATIENT(S): A total of 340 infertile women undergoing IVF. INTERVENTION(S): Natural micronized progesterone capsules were administered for luteal support. MAIN OUTCOME MEASURE(S): Association of MMPs 2 and 9 and TIMP-1 with oocyte maturity and embryo development. RESULT(S): An abnormal expression of MMP-2, MMP-9, and TIMP-1 with extensive MMP-9/TIMP-1 imbalance in women with endometriosis undergoing IVF was observed. Transforming growth factor ß1 plays an important role in these women with possible involvement of Smad-2 and -3 proteins. Progesterone supplementation improves the imbalance in MMP-9/TIMP-1 ratio significantly in women with endometriosis who conceive after IVF. CONCLUSION(S): Increase in MMP-2 and -9 and decrease in TIMP-1 expression was associated with poor oocyte and embryo development in women with endometriosis undergoing IVF. MMP-9/TIMP-1 balance was highly affected in these women, and progesterone supplementation appeared to restore this imbalance to a considerable degree.

Circulating biomarkers of pulmonary and extrapulmonary tuberculosis in children.

Tuberculosis (TB) in children is not only more likely to cause more severe disease than that seen in adults, it is also more likely to be extrapulmonary. Moreover, pediatric TB is very difficult to diagnose and suffers from a lack of understanding of host biomarkers for monitoring the progression of disease. Hence, we sought to identify the expression patterns of a variety of biomarkers in the plasma of children with pulmonary TB (PTB) and extrapulmonary TB (ETB), as well as in healthy control (HC) children. Thus, we examined a variety of circulating markers reflecting tissue inflammation, oxidative stress, innate immune activation, fibrosis, and the cytokine response. Children with active TB, compared to HC children, showed markedly elevated plasma levels of matrix metalloproteinases and their endogenous inhibitors. In addition, children with active TB had significantly elevated levels of C-reactive protein, α-2 macroglobulin, and haptoglobin, as well as hemoxygenase 1. Markers of innate immune activation (lipopolysaccharide [LPS] and lipopolysaccharide-binding protein [LBP]) were significantly lower in ETB than in PTB children. Although there were no significant differences between the two groups in their levels of cytokines (type 1 [gamma interferon (IFN-γ), tumor necrosis factor α (TNF-α), interleukin 2 (IL-2), and IL-12], type 2 [IL-4, IL-5, IL-13, and IL-33], and most type 17 [IL-17A, IL-22, IL-1ß, and IL-6] and type 1 interferons [IFN-α and IFN-ß]) or most of the cytokines associated with immune modulation (IL-10 and IL-20), pediatric TB was associated with elevated plasma transforming growth factor ß (TGF-ß), IL-21, and IL-23 levels. Thus, pediatric TB is characterized by elevated levels of a variety of biomarkers at homeostasis, suggesting that these responses may play a crucial role in disease pathogenesis.

Clarithromycin suppresses the periodontal bacteria-accelerated abdominal aortic aneurysms in mice.

BACKGROUND AND OBJECTIVE: Although clarithromycin (CAM) has many biological functions, including regulation of MMPs, little is known about its effect on abdominal aortic aneurysms. Periodontopathic bacteria have been reported to be associated with several kinds of circulatory diseases. The purpose of this study was therefore to clarify the effect of CAM on periodontopathic bacteria-accelerated abdominal aortic aneurysms. MATERIAL AND METHODS: Abdominal aortic aneurysm was produced in mice by the peri-aortic application of 0.25 m CaCl(2). The mice were inoculated once per week with live Porphyromonas gingivalis, which is one of the major periodontopathic bacteria. Test mice (n=8) were given a daily oral dose of CAM, while control mice (n=13) were not. RESULTS: Four weeks after the operation, the P. gingivalis-injected and CAM-treated mice showed a significant decrease in the aortic diameter in comparison with the mice only injected with P. gingivalis. Histopathologically, the samples obtained from the P. gingivalis-injected and CAM-treated mice showed less elastic degradation. Moreover, the plasma MMP-2 concentration of the CAM-treated mice decreased significantly. CONCLUSION: These findings suggest that CAM administration is useful to suppress periodontal bacteria-accelerated abdominal aortic aneurysms via MMP regulation.