Dexamethasone increases αvβ3 integrin expression and affinity through a calcineurin/NFAT pathway

The purpose of this study was to determine how dexamethasone (DEX) regulates the expression and activity of αvβ3 integrin. FACS analysis showed that DEX treatment induced expression of an activated αvβ3 integrin. Its expression remained high as long as DEX was present and continued following DEX removal. FACS analysis showed that the upregulation of αvβ3 integrin was the result of an increase in the expression of the β3 integrin subunit. By real time qPCR, DEX treatment induced a 6.2-fold increase (p < 0.04) in β3 integrin mRNA by day 2 compared to control and remained elevated for 6 days of treatment and then an additional 10 days once the DEX was removed. The increase in β3 integrin mRNA levels required only 1 day of DEX treatment to increase levels for 4 days in the absence of DEX. In contrast, DEX did not alter β1 integrin mRNA or protein levels. The DEX-induced upregulation of β3 integrin mRNA was partly due to an increase in its half-life to 60.7 h from 22.5 h in control cultures (p < 0.05) and could be inhibited by RU486 and cycloheximide, suggesting that DEX-induced de novo protein synthesis of an activation factor was needed. The calcineurin inhibitors cyclosporin A (CsA) and FK506 inhibited the DEX induced increase in β3 integrin mRNA. In summary, the DEX-induced increase in β3 integrin is a secondary glucocorticoid response that results in prolonged expression of αvβ3 integrin and the upregulation of the β3 integrin subunit through the calcineurin/NFAT pathway.     

Molecular modeling studies of atorvastatin analogues as HMGR inhibitors using 3D-QSAR,molecular docking and molecular dynamics simulations

3-Hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR) is generally regarded as targets for the treatment of hypercholesterolemia. HMGR inhibitors (more commonly known as statins) are discovered as plasma cholesterol lowering molecules. In this work, 120 atorvastatin analogues were studied using a combination of molecular modeling techniques including three-dimensional quantitative structure–activity relationship (3D-QSAR), molecular docking and molecular dynamics (MD) simulation. The results show that the best CoMFA (comparative molecular field analysis) model has q² = 0.558 and r² = 0.977, and the best CoMSIA (comparative molecular similarity indices analysis) model has q² = 0.582 and r² = 0.919. Molecular docking and MD simulation explored the binding relationship of the ligand and the receptor protein. The calculation results indicated that the hydrophobic and electrostatic fields play key roles in QSAR model. After MD simulation, we found four vital residues (Lys735, Arg590, Asp690 and Asn686) and three hydrophobic regions in HMGR binding site. The calculation results show that atorvastatin analogues obtained by introduction of F atoms or gem-difluoro groups could obviously improve the inhibitory activity. The new HMGR inhibitor analogues design in this Letter had been submitted which is being currently synthesized by our laboratories.     

Within-day and between-days reliability of quadriceps isometric muscle fatigue using mechanomyography on healthy subjects

This study aimed to examine within-day and between-days intratester reliability of mechanomyography (MMG) in assessing muscle fatigue. An accelerometer was used to detect the MMG signal from rectus femoris. Thirty one healthy subjects (15 males) with no prior knee problems initially performed three maximum voluntary contractions (MVCs) using an ISOCOM dynamometer. After 10 min rest, subjects performed a fatiguing protocol in which they performed three isometric knee extensions at 75% MVC for 40 s. The fatiguing protocol was repeated on two other days, two to four days apart for between-days reliability. MMG activity was determined by overall root mean squared amplitude (RMS), mean power frequency (MPF) and median frequency (MF) during a 40 s contraction. RMS, MPF and MF linear regression slopes were also analysed. Intraclass Correlation Coefficients (ICC); ICC1,1 and ICC1,2 were used to assess within-day reliability and between-days reliability respectively. Standard error of measurement (SEM) and smallest detectable difference (SDD) described the within-subjects variability. MMG fatigue measures using linear regression slopes showed low reliability and large between-days error (ICC1,2 = 0.43–0.46; SDD = 306.0–324.8% for MPF and MF slopes respectively). Overall MPF and MF, on the other hand, were reliable with high ICCs and lower SDDs compared to linear slopes (ICC1,2 = 0.79–0.83; SDD = 21.9–22.8% for MPF and MF respectively). ICC1,2 for overall MMG RMS and linear RMS slopes were 0.81 and 0.66 respectively; however, the SDDs were high (56.4% and 268.8% respectively). The poor between-days reliability found in this study suggests caution in using MMG RMS, MPF and MF and their corresponding slopes in assessing muscle fatigue.     

Effective control of black Sigatoka disease using a microbial fungicide based on Bacillus subtilis EA-CB0015 culture

Black Sigatoka disease caused by the fungus Mycosphaerella fijiensis Morelet is the most devastating disease of bananas worldwide. Its management is reliant on protectant and systemic fungicides despite their environmental concerns. This study evaluated the effect of a microbial fungicide (MF) based on Bacillus subtilis EA-CB0015 and its metabolites for the control of black Sigatoka disease on banana plants in greenhouse and field conditions. The MF applied at 1.5 L/ha and 3.0 L/ha provided control of the disease comparable to the protectant fungicide chlorothalonil in greenhouse. In the field, the MF applied in solution with water at 0.15 L/ha and 1.5 L/ha every 11 days during 10 weeks reduced black Sigatoka disease severity in 20.2% and 28.1% respectively; reductions comparable to those obtained with the protectant fungicides chlorothalonil (1.5 L/ha) and mancozeb (3.8 L/ha). The MF incorporated into different programs with systemic fungicides reduced disease level up to 42.9% with no significant differences with the conventional program. To determine which component of the MF is responsible for the activity against M. fijiensis, greenhouse and in vitro tests were set up to evaluate individually the spores, vegetative cells and secondary metabolites of B. subtilis EA-CB0015. All components reduced the severity of the disease and the germination of ascospores. For both trials the activity of the metabolites was higher and comparable to the activity obtained with the MF, indicating that the efficacy of the MF depends mainly on the metabolites and in lesser extent to B. subtilis EA-CB0015 cells.     

Regulation of isoprenoid/cholesterol biosynthesis in cells from mevalonate kinase-deficient patients

Mevalonic aciduria (MA) and hyper-IgD and periodic fever syndrome (HIDS) are two inherited disorders both caused by depressed mevalonate kinase (MK) activity. MK is the first enzyme to follow the highly regulated 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase (HMGR), which catalyzes the rate-limiting step in the isoprenoid/cholesterol biosynthesis pathway. In fibroblasts of MA patients, but not of HIDS patients, HMGR activity is elevated under normal growth conditions. This activity is down-regulated when cells are supplemented with the isoprenoid precursors geraniol, farnesol, and geranylgeraniol, and a mixture of 25-hydroxycholesterol and cholesterol. This indicates that the regulation of the pathway in these cells is not disturbed. The elevated HMGR activity is probably due to a shortage of non-sterol isoprenoid end products, as indicated by normal HMGR mRNA levels in MA fibroblasts. Furthermore, the HMGR activity in MA cells was more sensitive to geranylgeraniol suppression and less sensitive to sterol suppression than the HMGR activity in low density lipoprotein receptor-deficient cells. HMGR activity in MA cells was down-regulated also by addition of its product mevalonate to the culture medium. Thus, it appears that the elevation of mevalonate levels, which are high in MA patients and moderate in HIDS patients, allows the cells to compensate for the depressed MK activity. Indeed, the isoprenylation of Ras and RhoA protein appeared normal in HIDS and MA fibroblasts under normal conditions but showed increased sensitivity toward inhibition of HMGR by simvastatin. Our results indicate that MK-deficient cells maintain the flux through the isoprenoid/cholesterol biosynthesis pathway by elevating intracellular mevalonate levels.     

Effects of magnetic fields on biomass and glutathione production by the yeast Saccharomyces cerevisiae

The effect of magnetic fields (MF) on glutathione (GSH) production by Saccharomyces cerevisiae ATCC 7754 was studied. For this purpose, a factorial design of experiments was used to determine the influence of the time of exposure (8–16 h) and MF induction (25.0–34.3 mT), in GSH and biomass production. Additionally, control experiments (CE), without the application of MF, were performed. The results indicated the existence of favourable alterations in GSH and biomass concentrations due to the application of MF. In all experiments, the amount of biomass produced was higher than in CE and, with regard to GSH yield, in all the experiments at 24 and 48 h it was higher and in three experiments at 72 h of culture. The highest specific GSH yield (20.9 mg_GSH/g_biomass), GSH yield (340.0 mg/L) and biomass (16.26 g/L) were obtained using a MF induction of 25.0 mT for 16 h. These results were 16.1%, 39.0% and 19.6% higher than in the CE, respectively. Through statistical analysis it was found that the MF induction was a significant factor in GSH yield, and also it was observed that, within the range of the experimental conditions used, the lower MF induction, the higher the GSH yield.     

Matrix metalloproteinase-3 in articular cartilage is upregulated by joint immobilization and suppressed by passive joint motion

Both underloading and overloading of joints can lead to articular cartilage degradation, a process mediated in part by matrix metalloproteinases (MMPs). Here we examine the effects of reduced loading of rat hindlimbs on articular cartilage expression of MMP-3, which not only digests matrix components but also activates other proteolytic enzymes. We show that hindlimb immobilization resulted in elevated MMP-3 mRNA expression at 6 h that was sustained throughout the 21 day immobilization period. MMP-3 upregulation was higher in the medial condyle than the lateral, and was greatest in the superficial cartilage zone, followed by middle and deep zones. These areas also showed decreases in safranin O staining, consistent with reduced cartilage proteoglycan content, as early as 7 days after immobilization. One hour of daily moderate mechanical loading, applied as passive joint motion, reduced the MMP-3 and ADAMTS-5 increases that resulted from immobilization, and also prevented changes in safranin O staining. Intra-articular injections of an MMP-3 inhibitor, N-isobutyl-N-(4-methoxyphenylsulfonyl)-glycylhydroxamic acid (NNGH), dampened the catabolic effects of a 7 day immobilization period, indicating a likely requirement for MMP-3 in the regulation of proteoglycan levels through ADAMTS-5. These results suggest that biomechanical forces have the potential to combat cartilage destruction and can be critical in developing effective therapeutic strategies.     

Effects of eyestalk ablation on carbonic anhydrase activity in the euryhaline blue crab Callinectes sapidus: neuroendocrine control of enzyme expression

Carbonic anhydrase (CA) activity in the gills of the euryhaline blue crab, Callinectes sapidus, was measured in response to acute low-salinity transfer and treatment with eyestalk ablation (ESA) in an attempt to elucidate potential regulatory mechanisms of salinity-mediated CA induction. ESA alone resulted in an approximate doubling of CA activity in the posterior, ion-transporting gills of crabs acclimated to 35 ppt. Transfer of intact crabs to 28 ppt, a salinity at which the blue crab is still an osmotic and ionic conformer, had no effect on CA activity, but treatment with ESA prior to transfer resulted in a 5-fold increase. Hemolymph osmolality was unaffected by ESA. There was a 7-fold induction of CA activity in posterior gills of intact crabs transferred from 35 to 15 ppt, and this was potentiated by about 100% by ESA. Hemolymph osmolality was slightly elevated in the ESA-treated crabs. CA activity in anterior gills did not increase in response to any treatment. Hemolymph concentrations of methyl farnesoate (MF) were measured for all experimental animals. MF concentrations were undetectable in all intact crabs, regardless of salinity. Treatment with ESA resulted in elevated levels of hemolymph MF, but these levels were still relatively low and unrelated to salinity. These results suggest that CA induction is under the control of a regulatory substance located in the eyestalk. This substance appears to be a CA repressor, keeping CA expression at low levels in the gills of crabs acclimated to high salinity. Exposure to low salinity, or treatment with ESA, removes the effects of this putative repressor and allows CA induction to occur.     

Corticosterone may interact with peripubertal development to shape adult resistance to social defeat

Studies of social stress in adult mice have revealed two distinct defeat-responsive behavioral phenotypes; “susceptible” and “resistant,” characterized by social avoidance and social interaction, respectively. Typically, these phenotypes are observed at least 1 day after the last defeat in adults, but may extend up to 30 days later. The current study examined the impact of peripubertal social defeat on immediate (1 day) and adult (30 day) social stress phenotypes and neuroendocrine function in male C57BL/6 mice. Initially, peripubertal (P32) mice were resistant to social defeat. When the same mice were tested for social interaction again as adults (P62), two phenotypes emerged; a group of mice were characterized as susceptible evidenced by significantly lower social interaction, whereas the remaining mice exhibited normal social interaction, characteristic of resistance. A repeated analysis of corticosterone revealed that the adult (P62) resistant mice had elevated corticosterone following the social interaction test as juveniles. This was when all mice, regardless of adult phenotype, displayed equivalent levels of social interaction. Peripubertal corticosterone was positively correlated with adult social interaction levels in defeated mice, suggesting early life stress responsiveness impacts adult social behavior. In addition, adult corticotropin-releasing factor (CRF) mRNA in the paraventricular nucleus of the hypothalamus (PVN) was elevated in all defeated mice, but there were no differences in CRF mRNA expression between the phenotypes. Thus, there is a delayed appearance of social stress-responsive phenotypes suggesting that early life stress exposure, combined with the resultant physiological responses, may interact with pubertal development to influence adult social behavior.     

The regulated in development and DNA damage response 2 (REDD2) gene mediates human monocyte cell death through a reduction in thioredoxin-1 expression

In a previous study, we identified the regulated in development and DNA damage response 2 (REDD2) gene as a highly expressed gene in human atherosclerotic lesions in comparison to normal artery, as well as in cultured human macrophages, and showed its implication in oxidized low-density lipoprotein (LDL)-induced macrophage death sensitivity. In this article, we attempt to identify the mechanism by which REDD2 induces such a phenomenon. Transient transfection of U-937 monocytic cells with a pCI.CMV.REDD2 expression vector increased by approximately twofold the mRNA levels of REDD2 in comparison to control cells transfected with pCI.CMV.GFP. Reactive oxygen species (ROS) production was significantly induced in REDD2-transfected cells compared with control cells (157 ± 48 and 100 ± 8 arbitrary units/mg cell protein, respectively; p < 0.05). Moreover, a significant increase in parameters known to reflect the oxidative modifications of LDL was observed. Among enzymes involved in ROS production or degradation, we found a specific reduction in thioredoxin-1 (Trx-1) mRNA (～ 52 ± 7% decrease, p < 0.01 vs control cells) and protein (～ 60 ± 4% decrease, p < 0.001 vs control cells) levels in cells overexpressing REDD2 in comparison to control cells. In contrast, transfection of U-937 cells with siRNA against REDD2 decreased the mRNA levels of REDD2 by ～ 60% and increased Trx-1 mRNA and protein levels. Moreover, we observed no or a moderate increase in Bax (proapoptotic) and a significant decrease in Bcl2 (antiapoptotic) gene expression in cells that overexpress REDD2 compared to control cells. In addition, we showed that Trx-1 mRNA and protein levels were increased at low H₂O₂ doses and decreased at higher doses. Interestingly, macrophages isolated from human atherosclerotic lesions differentially express REDD2 and Trx-1. Indeed, in certain patients, levels of REDD2 mRNA were low and those of Trx-1 mRNA were high. In contrast, in other patients, levels of REDD2 were high and levels of Trx-1 mRNA were low.     

The effect of isosteviol on hyperglycemia and dyslipidemia induced by lipotoxicity in rats fed with high-fat emulsion

AimsThe aim of present study was to investigate the effects of isosteviol on hyperglycemia and hyperlipidemia in rats fed with high-fat emulsion (HFE).Main methodsHyperglycemia and hyperlipidemia in rats was induced by daily ingestion of HFE for 14 days. Isosteviol (0.2, 1.0, or 5.0 mg/kg/day) was orally administered for 7 days, with rosiglitazone maleate (5.0 mg/kg/day) used as the positive control. The levels of fasting serum glucose (FSG), fasting serum insulin (FSI), total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), and low density lipoprotein (LDL) in serum were assayed. Intravenous glucose tolerance test (IVGTT) was performed with serum glucose and insulin levels monitored. The effect of the supplement of palmitate in HFE on the activity of isosteviol was investigated. Ultrastructural changes in islet β-cells and peroxisome proliferator-activated receptor α (PPARα) mRNA expression profile were determined.Key findingsFSG, FSI, TC and LDL levels and insulin resistance index (IRI) were decreased and HDL level was increased by all doses of isosteviol. During IVGTT, serum glucose levels were decreased by isosteviol and no significant differences were observed in insulin release between isosteviol-treated and control groups. The effects of isosteviol were attenuated by palmitate. Damage to pancreatic islet cells was partially attenuated, and expression profile of hepatic PPARα mRNA was enhanced by isosteviol.SignificanceAntihyperglycemic effects of isosteviol could enhance utilization of glucose in the periphery and reduce β-cell damage induced by dyslipidemia. Modulating-lipidemic effects of isosteviol might be related to the potential enhancement of liver PPARα mRNA expression.     

Evaluation of a method to measure long term cortisol levels

IntroductionElevated levels of cortisol are known to induce various symptoms and diseases, e.g. abdominal obesity, type 2 diabetes, osteoporosis and cardiovascular disease. Measuring serum, saliva and urine cortisol is limited to one time point. Measurement of cortisol in scalp hair is a recently developed method to measure long term cortisol levels. The aim of this study was to investigate whether hair cortisol is a feasible parameter to measure cortisol exposure.ExperimentalWe collected hair samples of 195 healthy individuals, 9 hypercortisolemic and one hypocortisolemic patient and measured hair cortisol levels. Cortisol was extracted from scalp hair using methanol and cortisol levels were measured using a salivary ELISA kit. Measurement of waist and hip circumferences and blood pressure was performed in 46 healthy subjects.ResultsWe found a positive correlation between hair cortisol and both waist circumference (r = 0.392, p = 0.007) and waist-to-hip ratio (WHR) (r = 0.425, p = 0.003). No correlations were found between hair cortisol levels and BMI, blood pressure or age. There was no decline in cortisol levels in six consecutive hair segments. Hair cortisol levels were elevated in patients with known hypercortisolism (p < 0.0001).ConclusionsHair cortisol was positively correlated with WHR, suggesting that hair cortisol reflects cortisol exposure at tissue level, which was also supported by elevated hair cortisol levels in hypercortisolemic patients and concordance between hair cortisol levels and clinical disease course. Cortisol levels in hair are slightly influenced by hair treatment but not by natural hair colour, use of hair products, gender or age.     

Chronic inhibition of nuclear factor kappa B attenuates aldosterone/salt-induced renal injury

AimsRecent studies suggested that nuclear factor kappa B (NF-κB) plays a key role in the pathogenesis of renal injury. This study investigated whether NF-κB inhibition attenuates progressive renal damage in aldosterone/salt-induced renal injury and its mechanisms.Main methodsAdult male rats were uninephrectomized and treated with one of the following for 4 weeks: vehicle (0.5% ethanol, subcutaneously); vehicle/1% NaCl (1% NaCl in drinking solution); aldosterone/1% NaCl (1% NaCl in drinking solution and aldosterone, 0.75 μg/h, subcutaneously); or aldosterone/1%NaCl + pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB (100 mg/kg/day, by gavage). The activity of NF-κB was measured by EMSA and immunohistochemistry, CTGF and ICAM-1 were measured by Western blot and real-time PCR, and TGF-β and CTGF were measured by immunohistochemistry.Key findingsRats that received aldosterone/1% NaCl exhibited hypertension and severe renal injury. Renal cortical mRNA levels of CTGF, TGF-β, ICAM-1 and collagen IV, protein expression of CTGF and ICAM-1, and NF-κB–DNA binding activity were significantly upregulated in rats that received aldosterone/1% NaCl. Treatment with PDTC significantly decreased the percentage of cells positive for CTGF and TGF-β; mRNA levels of CTGF, TGF-β, ICAM-1 and collagen IV, and protein levels of CTGF and ICAM-1 were also inhibited by PDTC.SignificanceThese data suggest that the NF-κB signal pathway plays a role in the progression of aldosterone/salt-induced renal injury.     

Subchronic stress-induced depressive behavior in ovariectomized mice

AimsMood disorders including depression are more common in women than men, particularly in times of lower estradiol levels. In this study, we investigated the effect of estrogen on emotional behavior in mice in a stress environment.Main methodsFemale mice were divided into four groups: two groups were ovariectomized (OVX) and two were sham-operated. One group each of OVX and sham mice was kept in a normal environment and the other groups were assigned to a daily stress (1 h/day) for 7 days from 5 days after operation. On the 14th day after operation, subjects were measured to assess behavioral specificity, locomotor activity, elevated plus-maze (EPM) behavior, passive avoidance (PA) behavior and forced swimming behavior.Key findingsThe OVX plus stress (OVX + S) group showed a significant prolongation of immobility compared with the other groups. In all the groups there were no changes in locomotor activity, EPM behavior or PA behavior. We further examined the effect of estrogen against depressive behavior in the OVX + S group. The vehicle or 17β-estradiol (E2) was administered s.c. to OVX + S mice for 4 days beginning on post-operative day 11. Subchronic E2 treatment decreased the stress response and improved depressive behavior relative to the vehicle group.SignificanceThese data have important implications regarding the prevention of depression in postmenopausal women undergoing estrogen therapy.     

Metabolic profiling and enhanced production of phytosterols by elicitation with methyl jasmonate and silver nitrate in whole plant cultures of Lemna paucicostata

In this study, the effects of methyl jasmonate (MJ) and silver nitrate (SN) treatment on metabolic profiles and yields of phytosterols such as campesterol, stigmasterol, and β-sitosterol in whole plant cultures of Lemna paucicostata were investigated using gas chromatography–mass spectrometry coupled with multivariate statistical analysis. The MJ and SN treatments retarded the growth of L. paucicostata plants, while they enhanced the yields of three phytosterols, compared to control. Higher yields of phytosterols were attained at day 28 compared to day 42. Moreover, stigmasterol yield was the highest at 0.85 mg/g from day 28 plants grown under MJ + SN co-treated culture. Among the various metabolites, the levels of palmitic and stearic acids, which might participate in a defense mechanism, were higher in the MJ + SN condition than in control. To determine the optimal timing of MJ + SN addition, MJ + SN was added on days 21, 28, and 35 after inoculation. The total yield and productivity of phytosterol reached maximum levels when the MJ + SN was added at day 35. The highest productivity of stigmasterol (6.08 mg/L) was also achieved when MJ + SN was added on day 35.     

Interactive effects of elevated temperature and CO2 levels on metabolism and oxidative stress in two common marine bivalves (Crassostrea virginica and Mercenaria mercenaria)

Marine bivalves such as the hard shell clams Mercenaria mercenaria and eastern oysters Crassostrea virginica are affected by multiple stressors, including fluctuations in temperature and CO₂ levels in estuaries, and these stresses are expected to be exacerbated by ongoing global climate change. Hypercapnia (elevated CO₂ levels) and temperature stress can affect survival, growth and development of marine bivalves, but the cellular mechanisms of these effects are not yet fully understood. In this study, we investigated whether oxidative stress is implicated in cellular responses to elevated temperature and CO₂ levels in marine bivalves. We measured the whole-organism standard metabolic rate (SMR), total antioxidant capacity (TAOC), and levels of oxidative stress biomarkers in the muscle tissues of clams and oysters exposed to different temperatures (22 and 27 °C) and CO₂ levels (the present day conditions of ~ 400 ppm CO₂ and 800 ppm CO₂ predicted by a consensus business-as-usual IPCC emission scenario for the year 2100). SMR was significantly higher and the antioxidant capacity was lower in oysters than in clams. Aerobic metabolism was largely temperature-independent in these two species in the studied temperature range (22–27 °C). However, the combined exposure to elevated temperature and hypercapnia led to elevated SMR in clams indicating elevated costs of basal maintenance. No persistent oxidative stress signal (measured by the levels of protein carbonyls, and protein conjugates with malondialdehyde and 4-hydroxynonenal) was observed during the long-term exposure to moderate warming (+ 5 °C) and hypercapnia (~ 800 ppm CO₂). This indicates that long-term exposure to moderately elevated CO₂ and temperature minimally affects the cellular redox status in these bivalve species and that the earlier observed negative physiological effects of elevated CO₂ and temperature must be explained by other cellular mechanisms.     

Elevated serum MMP-9/TIMP-1 ratio in patients with homozygous familial hypercholesterolemia: Effects of LDL-apheresis

ObjectiveMatrix degradation within an atherosclerotic plaque is an important pathogenic factor in atherosclerosis, and is largely modulated by the balance between matrix metalloproteinases (MMPs) and their endogenous inhibitors (i.e., tissue inhibitor of MMPs [TIMPs]). Familial hypercholesterolemia (FH) is a rare inherited disorder associated with premature coronary heart disease. The aim of the present study was to examine MMP-9 and TIMP-1 on plasma and cellular mRNA levels in homozygous FH patients (n = 7) compared with age- and sex-matched heterozygous FH patients (n = 6), and with healthy subjects (n = 7), and to test whether once-weekly LDL-apheresis (three consecutive sessions) of homozygous FH patients show short-term effects on these variables.ResultsThe main findings were that (i) Compared to healthy control subjects, homozygous FH patients have significantly higher serum levels of MMP-9 and lower levels of TIMP-1, and consequently significantly higher MMP-9/TIMP-1 ratio, potentially reflecting higher MMP-9 activity. (ii) Peripheral blood mononuclear cells (PBMC) isolated from FH homozygotes have significantly higher mRNA levels of MMP-9 compared to cells from heterozygotes. (iii) TNFα-stimulated PBMC from FH homozygotes released borderline-significantly more MMP-9 than cells from heterozygotes and healthy controls. (iv) LDL-apheresis (one day before treatment versus fifteen days later, on the day after the weekly treatment) had no significant short-term effect on any of the MMP-9 and TIMP-1 variables measured in serum and cells.ConclusionsThe data may suggest that homozygous FH patients have an enhanced matrix degrading potential as compared with heterozygous FH patients and healthy controls, potentially contributing to the increased cardiovascular risk observed in these patients.