Obtaining knowledge to identify proteins associated with a partic

Obtaining knowledge to identify proteins associated with a particular physiological or pathological state, has a great significance in understanding disease states and to develop new diagnostic and prognostic assays [19] and [20]. Neuroproteomics include comparative analysis of protein expression in normal and diseased states to study the dynamic

properties associated with neuropeptide processing in biological system of diseases [21]. This review will discuss several key neuroproteomic areas that not only address CNS injury research but also will address the translational potential from animal studies to clinical practice. We will cover three major neuroproteomic platforms: differential neuroproteomics, quantitative proteomics, and imaging mass spectrometry (IMS) approach.

Differential CYC202 chemical structure proteomic approach is ideally suited to discover protein biomarkers that might be differentially expressed or altered by contrasting two or more biological samples (Fig. 1). The complexity, immense size, variability of the neuroproteome, extensive protein–protein and protein–lipid interactions, proteins in the CNS tissues are extraordinarily resistant to isolation selleckchem [10] and [22]. Therefore, high resolving protein/peptide separation methods are essential for the separation and identification. The development of modern separation techniques coupled online with accurate and high resolving mass spectrometric tools have emerged as preferred components for diagnostic,

prognostic and therapeutic protein biomarkers discovery that expands the scope of protein identification, quantitation Resveratrol and characterization. Proteomics has two major approaches. The bottom-up (or shotgun) approach involves direct digestion of a biological sample using a proteolytic enzyme (such as trypsin) that cleaves at well-defined sites to create a complex peptide mixture. The digested samples can then be analyzed by liquid chromatography (single or multi-dimensional) prior to tandem mass spectrometry (LC–MS/MS) [23]. The second approach is top-down that involves separating intact proteins from complex biological samples using separation techniques such as liquid chromatography or 2-D gel electrophoresis (isoelectofocusing + SDS-gel electrophoresis – separation by relative molecular weight) followed by differential expression analysis using spectrum analysis or gel imaging platforms. This is sometimes assisted by differential dye-labeling of two samples (e.g. with Cy-3, Cy-5 dye) and equal amount of the labeled samples are mixed and resolved by 2-D gel, creating a differential gel map or differential gel electrophoresis (DIGE) where differentially expressed proteins (up- or down-regulated proteins) can be identified by fluorescence scanning and band cut out for protein identification [24].

It is not unusual for an organism not to be identified on the ple

It is not unusual for an organism not to be identified on the pleural fluid culture and therefore broad-spectrum antibiotic coverage should be instituted when the diagnosis of empyema is made. This can be modified if the culture data identifies selleck kinase inhibitor an organism. Early VATS combined with early rehabilitation offers excellent results, radically improving the outcome in both the fibrinopurulent, as well as in organizing stages of PE in children, but surgeon should be experienced in the less invasive technique. The method seems to be successful even in very neglected cases, if not patient could benefit

from fibrinolytic therapy. According to order. None declared. “
“Vaccines are among the greatest and most effective public health interventions in

preventing morbidity, mortality and public health costs caused by infectious diseases [1]. Today, incidence rates of vaccine Everolimus manufacturer preventable disease (VPDs) in the U.S. have declined to an all time low [2]. Despite the undoubted success, the nearly forgotten VPDs in the U.S. are back. From 2001 – 2008, a median of 56 (range: 37–140) measles cases were reported to the CDC annually. During the first 19 weeks of 2011, 118 cases of measles were reported, the highest number reported for this period since 1996. Of the cases, 105 (89%) were imported from other countries and unvaccinated persons accounted for 105 (89%) [3]. There were outbreaks of mumps [4], an invasive HiB disease [5]. The CDC reports on its website that in 2010, 9 143 cases of pertussis were reported in California, the most cases reported since 63 years. Among them were 10 infants who died from the disease. There were outbreaks in Michigan, Ohio and other states [6]. The USA is on the verge of becoming a victim of this

success, because increasing numbers of parents, who apparently love their children, refuse to vaccinate them. Why does it happen? The answer to this question is not easy and straightforward. Robert Chen Osimertinib molecular weight tried to answer this question showing the graph dubbed – “Natural history of an immunization program” (Fig. 1) [7]. In the first, pre-vaccine period, people feel threatened by the disease, especially if the disease is communicable and hard to treat. They often know victims of the disease, who either died or suffered from the complications. When a vaccine becomes available, people widely and enthusiastically accept it, even despite side effects the vaccine can cause. The best example of this is a national enthusiasm in the USA after developing the polio vaccine in the 1950s. In the second period, when a vaccine causes a massive decrease in VPD cases and deaths, people start to forget the threat, a memory of the victims and social disruptions of the disease fades. With the increased use of a vaccine, the focus is on real and imaginary side effects of vaccination.

xylosus, S saprophyticus and S hominis have been reported in pr

xylosus, S. saprophyticus and S. hominis have been reported in previous studies.

26, 27, 29, 30, 31 and 32 In our results, S. sciuri, S. simulans and S. chromogenes were identified. These species were not found in previous studies of oral samples. Some of these species, although isolated infrequently, may cause infections in humans, such as urinary tract infections, LBH589 nmr bacteremia, endocarditis, osteomyelitis, cellulitis and cerebral empyema. 33 and 34 Counts of staphylococci were lower in the oral cavities of patients with low viral load (<400 copies/ml), but no difference was observed in relation to CD4 cells. No previous studies were found with which to compare these data. Enterobacteria and pseudomonas were identified in the oral cavities of 77.7% of the HIV-positive group. The control group showed a lower isolation frequency (44.4%) in the oral cavity. The increased oral prevalence of these microorganisms seems to be associated with systemic and local factors. However, data in the literature are still controversial. Jobbins et al.3 reported isolation of coliforms from 49% of patients with malignancy. A low prevalence of these microorganisms in the elderly and mentally disabled patients was reported.17 and 18 Senpuku et al.14 isolated Enterobacteriaceae

from 16% of elderly patients and from 6% of controls. A higher prevalence of enterobacteria in the oral cavity was observed by Santos and Jorge 11 in healthy Brazilian individuals (51%). Hägg et al. 35 reported a significant increase in the prevalence of enterobacteria Everolimus manufacturer after insertion of fixed orthodontic appliances. Zhu et al., 36 studying

stroke selleck chemicals llc patients at three different stages (acute phase, upon discharge from the hospital and 6 months later), observed that the oral carriage rate of coliforms was significantly lower at 6 months after hospital discharge, but found no significant relationship between the presence of coliforms and other variables studied (age, gender, plaque index, bleeding index, DMFT, denture wearing, dysphagia, smoking, diabetes and tooth brushing difficulty). Other studies with HIV-positive patients in different countries found a low prevalence of enterobacteria and/or pseudomonas in the oral cavity. Schmidt-Westhausen et al.37 obtained an enterobacteria prevalence of 22%. Tsang and Samaranayake15 reported the isolation of Enterobacteriaceae (26.3%) and P. aeruginosa (15.1%). The only Brazilian study regarding these microorganisms in the oral cavities of HIV-positive patients was conducted by Figueirêdo et al., 16 who found Enterobaceriaceae in 96.4% of isolates and P. aeruginosa, the only Pseudomonas identified, in 3.6% of isolates. According to Santos and Jorge, 11 some authors have noted discrepancies in the prevalence of these microorganisms in the oral cavities of individuals from developed and developing countries, hypothesizing that the incidence of these microorganisms in the oral cavity may be related to high numbers of coliforms in drinking water and foods.

The elevated TSS levels alter natural sedimentation processes in

The elevated TSS levels alter natural sedimentation processes in watercourses and can result in increased turbidity, depletion of dissolved oxygen, inhibition of benthic aerobic microorganisms and impairment of photosynthesis (Marsalek et al., 2005 and Sujkova et al., 2012). Chloride ions are natural components of surface waters, but the continuous discharge of wastes with high chloride ion concentrations can increase the total water salinity. Both aquatic and terrestrial ecosystems can be affected by exposure to high chloride ion concentrations (Perera et al. 2013). Secondary salinisation of rivers is a growing threat (Cañedo-Argüelles et al. 2013): elevated chloride levels

render surface waters unsuitable as an environment for many freshwater limnetic organisms and as a potable water supply. Angiogenesis inhibitor Moreover, chloride ions can alter the equilibrium between adsorbed and dissolved metals in snowmelt (Bäckström et al. 2004), thus leading to increased releases of the dissolved metals to watercourses. The overall mean concentrations of ammonium and phosphate ions in the snowmelt runoff exceeded MPCs 2.3 and 13.3 times respectively. The discharge of effluents with elevated levels of nutrients Staurosporine cell line (e.g. ammonium and phosphate)

can improve the survival and growth of aquatic plant organisms, but can also contribute to the eutrophication of the receiving waters (Bartlett et al. 2012). Long-term observation data indicate that the water in the River Mukhavets is constantly contaminated by phosphate, nitrite and ammonium ions; hence, surface runoff contributes to the total pollution by Methocarbamol components of prime concern (Loginov, 2009, Loginov, 2010, Loginov, 2011 and Loginov, 2012). The concentrations of most of the tested contaminants vary in a similar way, increasing from snow to snowmelt runoff samples (Table 1 and Table 2, Figure 2b). It is obvious that these impurities did not originate only from atmospheric precipitation. They became accumulated in the snow layer during its formation and contribute to their excessive outflow

in the snowmelt surface runoff. The concentrations of several HMs exceeded MPC levels. The concentration of Zn exceeded MPC in all the samples of snow and snowmelt runoff, and Cu and Mn concentrations also exceeded MPCs in all the tested runoff samples (the overall mean concentration of Zn in snowmelt runoff exceeded MPC 3.2 times, the overall mean concentrations of Cu and Mn exceeded MPCs 4 and 3.1 times respectively). The small decrease in the mean concentration of Cu and Zn in the runoff compared to snow at site 2 is explained by the fact that we were not able to completely avoid the influence of traffic emissions when sampling the snow, and snowmelt runoff was most probably diluted by effluent from another part of the site with a lower concentration of these metals.

16, 95% CI: 0 10–0 26, p < 0 0001; EURTAC: 9 7 vs 5 2 months, re

16, 95% CI: 0.10–0.26, p < 0.0001; EURTAC: 9.7 vs. 5.2 months, respectively, HR = 0.37, 95% CI: 0.25–0.54, p < 0.0001). Until now, erlotinib has not been prospectively evaluated in Japanese

patients with EGFR mutation-positive NSCLC. This prospective, phase II, open-label study (JO22903) was initiated to obtain confirmatory efficacy and safety data in the first-line setting for Japanese patients with EGFR mutation-positive NSCLC, in order to corroborate data from Chinese and Caucasian populations. JO22903 was a phase II, multicenter, open-label, non-randomized study conducted at 25 centers in Japan. Eligible patients were aged ≥20 years with advanced, untreated, metastatic (stage IIIB/IV), E7080 or relapsed NSCLC, with an Eastern Cooperative Oncology Group performance status (ECOG PS) of 0 or 1 and tumors harboring confirmed activating mutations of EGFR (exon 19 deletion or L858R point mutation in exon 21), with at least 1 measurable lesion according

to Response Evaluation Criteria in Solid Tumors (RECIST) version 1.0. Staging was assessed by TNM classification (7th edition). The study was carried out in accordance with the Declaration of Helsinki and Japanese Good Clinical Practice guidelines. The protocol was approved by ethics committees and all patients gave informed consent for study participation. Eligible patients received oral erlotinib 150 mg/day until disease progression (PD) or unacceptable toxicity (Fig. 1). Dose reductions (in 50-mg decrements) and/or interruptions Gefitinib order (of up to 2 weeks) were permitted to manage adverse events (AEs) related to erlotinib treatment. Treatment was interrupted if interstitial lung disease (ILD) was suspected; for patients with confirmed ILD diagnosis, erlotinib was discontinued immediately. In cases of gastrointestinal perforation or any grade 4 AE, erlotinib was discontinued. Patients were screened for EGFR mutations in a local or central laboratory. In the central laboratory, EGFR mutation status was determined using Scorpion ARMS [5].

For exploratory analyses, tumor samples were obtained from hospital archives for patients who were screened in their local laboratory to confirm the concordance between several local methods and Scorpion ARMS. In addition, serum samples were collected at screening from all patients who provided informed consent to participate Pazopanib order in the exploratory research (n = 95). DNA was isolated from serum with the QIAmp MinElute Virus Spin kit (Qiagen, Hilden, Germany). Scorpion ARMS was used for EGFR mutation testing for circulating DNA in the serum. Tumor response was assessed by an independent review committee (IRC) using RECIST version 1.0. Tumor response evaluation was scheduled every 6 weeks. AEs were graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI-CTC AE) version 4.0. At baseline mandatory lung and abdominal scans (CT/MRI), brain scans (CT/MRI) and bone scans (bone scintigraphy, PET, CT and MRI) were performed.

This study examined the influence of semantic information on read

This study examined the influence of semantic information on reading aloud, and whether individual differences in the use of this information were related to anatomical differences in relevant parts of the neural circuits for reading. Effects of imageability on RT ranged widely (Fig. 1B), suggesting that skilled readers differ in the extent to which they use semantic information in reading aloud. This variation was associated with the

volume of white matter tracts passing through both the ITS, an area that supports lexical semantic processing, and the pMTG, an area implicated in phonological processing. A similar effect was found for the volume of tracts passing through both the AG, an area associated with semantic processing, and the pSTG, an area associated with phonological processing. Variability in how words are read is often attributed to use of different strategies

or styles; our results show that one type of individual difference, see more ABT-888 supplier in the use of semantics in reading aloud, is associated with neuroanatomical differences. Further research will be needed to determine the origins of these individual differences. There may be differences in brain development and structure that cause individuals to vary in how they read aloud. Alternatively, the neuroanatomical differences could result, wholly or in part, from experiential factors including the nature of early language and reading experience, and how reading is taught. The latter alternative is suggested by a study showing white matter changes associated with interventions for reading problems (Keller & Just, 2009). Further studies of this type using other methods in which participants acquire new reading skills (Bailey et al., 2004, Carreiras et al., 2009 and Dehaene et al.,

2010) are necessary, however. It may also be possible to track the development of these pathways in longitudinal studies of children who transition from pre-readers to reading (for an example focused on the pOTS see Ben-Shachar, Dougherty, Deutsch, & Wandell, 2011). The analyses we conducted were hypothesis-driven, testing whether individual differences in reading aloud would be related to neuroanatomical differences in connectivity between areas thought to be involved check details in mappings between semantics and phonology, as indicated by other findings. However, the results are novel and require both replication (e.g., with additional subject populations, such as younger readers and adults who vary widely in reading skill) and extension (e.g., addressing individual differences involving other types of information and tasks, and in English and other writing systems). The main result concerning relations between behavioral and neuroanatomical differences is correlational, and the functions of the two semantic-phonological pathways are underdetermined. These are important directions for future research stimulated by interesting results in a promising new area.

, 2010) In both situations, the proteinuria can represent

, 2010). In both situations, the proteinuria can represent MK0683 clinical trial a clearance effect of lipoic acid, regarding intense proteolysis in the case of Bothrops venom ( Gonçalves et al., 2008), and eventual myolysis in the case of Crotalus venom ( Monteiro et al., 2001), but this proteinuria can also be only a consequence of the ability of lipoic acid in promoting the solubilization and/or remotion of proteins bounded to membranes ( Alegre et al., 2010). The simvastatin is prominent to mitigate the decrease of plasma urea, urinary hyperosmolality, hypercreatinuria and the decrease of APN in the soluble and membrane fractions

of the renal cortex of envenomed mice, and besides its inefficacy to restore the creatinemia, the unique possible deleterious effect of treatment of envenomed mice with simvastatin seems to be the decrease of DPPIV activity in the membrane fraction of renal cortex and medulla, which also occurs with lipoic acid. Therefore, considering the comparison between deleterious and favorable effects and that the antioxidant effect of both drugs is the primordial factor to reduce damage to renal tissue caused by the venom of B. jararaca, it seems unjustified the combined administration of both drugs to treat this envenomation, but it seems better the administration of simvastatin alone. Also considering that several important effects of B. jararaca venom (hypoproteinemia,

decrease of PIP activity in the membrane and soluble fractions of renal cortex and decrease of protein content in the membrane Selleck MAPK Inhibitor Library fractions of the

mafosfamide renal medulla, decrease of PIP and APN activities in the soluble fraction of the renal medulla, and decrease of PIP, CAP and PAP in the membrane fraction of the renal medulla) were not attenuated by treatment with these drugs, other antioxidant and nephroprotector agents should be further investigated to treat the snake bite accidents caused by the genus Bothrops. These data permit to distinguish the AKI induced by B. jararaca venom as characterized by hyperuricemia, hypercreatinemia, urinary hyperosmolality, decreased hematocrit, and decreased protein content in plasma and in the membrane fraction of the renal cortex and medulla. Also, alterations on several renal aminopeptidases activities are revealed among the mechanisms and consequences of the nephrotoxic effects of this venom. Overall, this investigation shows that lipoic acid and simvastatin exhibit preponderant beneficial effects on important parameters affected by B. jararaca venom, especially on hematocrit, creatinemia, uricemia and renal redox status, which recommend a clinical investigation, primordially of simvastatin (with fewer undesirable effects than lipoic acid), as coadjuvant in the serotherapy of this snake bite. This investigation was supported by a Research Grant 06/06926-9 from FAPESP (Fundação de Amparo àPesquisa do Estado de São Paulo, Brazil). P.F.S.

Larvae removed from seeds of V unguiculata were transferred to a

Larvae removed from seeds of V. unguiculata were transferred to a cavity produced in the compacted mass of flour in one half of the gelatin capsule, at a ratio of three larvae per capsule. Following this, the two halves of the capsules were carefully joined together in order to permit the feeding ABT 199 movements of the larvae and maintained in the dark. Controls were used in which only FITC was mixed with seed flour at the concentration of 2.0% (w/w) in order to assess the level of FITC absorption. Capsules containing only cowpea flour

were used as controls to evaluate auto-fluorescence of the internal organs. Larvae were left to complete their metamorphosis until emergence of adults. In order to visualize and document the presence of labelled vicilins by microscopy from gonads and eggs, fresh portions were mounted on glass slides and visualized using a laser Confocal microscope (Leica DMI6000 B Microscope). Vicilin–FITC fed and mated females 3-days after emergence were transferred to glass vials and maintained during 24 h inside an incubator at 28 °C and 70% RH and without access to males. After this time, the eggs laid on cowpea seeds were removed with a fine needle, placed in a 1.5 mL tube and homogenized (50 eggs/150 μL) in 250 mM NaCl at 4 °C. The homogenate was centrifuged at

15,000 × g for 15 min at 4 °C and the proteins in the supernatant were fractionated by SDS–PAGE as previously described. Virgin vicilin–FITC fed males and control females

check details that copulated with some of those males were dissected and their genitalia and fat bodies were collected. Following collection, some genital tracts were freshly prepared for confocal microscopy and pooled genitalia were homogenized in water using a hand-held Potter–Elvehjem homogenizer immersed in ice. Tissue homogenates were centrifuged at 15,000 × g for 30 min at 4 °C and the supernatants were used for protein determination and fractionation by SDS–PAGE as previously described. Preparative gel electrophoresis (SDS–PAGE) comprising ca 30 μg of proteins from Phosphatidylinositol diacylglycerol-lyase C. maculatus whole egg homogenates and 50 μg of protein from genitalia of both males and females were run as above and stained with Coomassie Blue. Protein bands with Rf similar to peptides recognized by the anti-vicilin antibody (see Souza et al., 2010) were then located on the preparatory gels and excised manually. Gel slices were distained (0.1 M ammonium bicarbonate and 40% acetonitrile), dehydrated (100% acetonitrile) and dried in a speed-vac. Protein digestion was performed as described by Demartini et al. (2011). The tryptic peptides collected after digestion were analyzed by reversed-phase HPLC coupled with tandem mass spectrometry (LC–MS/MS) performed in an electrospray ionization quadrupole time-of-flight mass spectrometer (Q-TOF Micro™, Micromass, Waters, Milford, United States).

At some corner compartments, Re~1000Re~1000

for the squar

At some corner compartments, Re~1000Re~1000

for the square tank, Re~600Re~600 for the ‘J’-type tank. At the start of each experiment, the tank was filled with clear water, and then a dilute methylene blue dye solution (concentration of 0.1 mg/l) was pumped into the tank via the inlet. Images were taken at a rate of 7.5 frames per second by learn more an Allied Vision Dolphin machine vision and saved as a BMP file every 100 frames. Matlab Image Processing Toolbox was employed to analyse these images. The experiments involved measuring the fraction of initial water in each compartment that is flushed out when water is injected into the tank. With the help of the inclined mirror, the camera captured a plan view of the tank. Dye water was injected into the tank (see Kamada et al., 2004). An optical method was used to assess

the mass of dye within BMS-354825 ic50 each compartment based on the classical absorption theory of Lambert–Beer (see Cenedese and Dalziel, 1998, Rahim et al., 2010, Zeng et al., 2010 and Suhling et al., 2001). The image processing was based on the principle that the depth integrated dye concentration in water can be related to the intensity of light passing through the water and the distance travelled by the light in the water. The dye concentration in the water at point (x  ,y  ) can then be related to the change in light intensity through equation(17) CI(x,y)=∫0lC(x,y,z)dz=f(logI0(x,y)I(x,y)),where l   is the distance in the z  -direction that the light travels in the water, I  0 is the light intensity after the light travels through clear water, and I   is the light intensity after the light travels through dye water. The function f  (x  ) is determined by a series of calibration tests for fixed l  . The volume averaged flushed fraction in compartment [i  ][j  ] is equation(18) C[i][j](T)=∫A[i][j]CIdA∫A[i][j]CI,ηdA,where CI,ηCI,η is the depth integrated 5-Fluoracil dye concentration when compartment [i][j] is completely filled with dye water, calculated from (17), and A[i][j] is the base area of compartment [i][j]. The main point was to determine

the fraction of initial fluid in each compartment that is removed, as a function of time. The diagnostic tools defined in Section 2.2 to analyse the model predictions were applied to analyse the experimental data. For images captured from the experiments, each compartment from the plan view was individually masked so that its time series (18) could be evaluated. We estimated T1/2,[i][j]T1/2,[i][j] by interpolating C[i][j]C[i][j] to determine when C[i][j]=0.5C[i][j]=0.5. We estimated α1/2,[i][j]α1/2,[i][j] by linearly regressing C[i][j]C[i][j] with T   over the interval |C[i][j]−0.5|≤0.1|C[i][j]−0.5|≤0.1 and identified α1/2,[i][j]α1/2,[i][j] proportional to the slope of the curve. The major experimental measurement errors are caused by masking and calibration.

) After cultivation of the following 24 h, the GFP expression wa

). After cultivation of the following 24 h, the GFP expression was analyzed using Olympus CKX41 fluorescent microscope and ELISA reader (BioTek

synergy HT). Cells with GFP expression indicated it was successful in construction of target gene reporter plasmid. Cells with an apparent absence of green fluorescence indicated gene silencing. Cell viability assay was performed right learn more after the fluorescent analysis. The protocol of transfection of reporter plasmid was according to the manufacturer’s instruction (Clontech). The experiment of knockdown endogenous MMP1 gene was performed in MeWo cells. MeWo cell is human melanoma cell and the morphology is fibroblast, therefore, it can express the MMP1 protein. Exogenous delivery of siRNA duplexes to mammalian cells was carried out with the Xfect™ siRNA Transfection Reagent (Clontech Laboratories, Inc.) in a

24 well plate, which was developed for the delivery of siRNA. Absence of transfection reagents, siRNA duplexes were not taken up by cells. The protocol was according to the manufacturer’s instruction (Clontech). After transfection with 859 siRNA and further 24 h incubation, cells were lysed in a mammalian cell lysis buffer (Clontech Laboratories, Inc.). Western blot analysis was then performed using conventional protocols. In brief, protein concentration was determined with Bradford assay (Bio-Rad) with Epigenetic inhibitor bovine serum albumin as a standard (Sigma). Equal amounts of total protein were then separated on 12% polyacrylamide gels by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride (PVDF) membrane. Antibodies and dilutions used in this study included anti-MMP1 (1:1000 dilution, Millipore, Billerica, MA, USA) and anti-GAPDH (1:2000 dilution, Millipore, Billerica, MA,

USA). After being washed extensively, the membranes were incubated with goat anti-rabbit IgG peroxidase conjugate antibody (1:10000 dilution) for 1 h at room temperature and developed with chemiluminescent HRP substrate (Millipore, Billerica, MA, USA). Membranes probed for hMMP1 were re-probed for GAPDH to normalize for loading and/or quantification errors and to allow comparisons of target protein expression Teicoplanin or inhabitation to be made. Band density was measured by photoimage (Fusion-SL2-3500WL, Vilber Lourmat, France, www.vilber.com). To detect the potential toxicity to the cell during the experiments, the cell viability was determined in 24 well plates. After specified periods of cell incubation (48 h post-transfection), 0.5 mL of MTT solution (1.5 mg/mL) was added to each well and incubated at 37 °C for 4 h. After removal of media, 0.5 mL of DMSO was added and the absorbance at 540 nm was measured. The viabilities were normalized to the absorbance of non-treated cells. The expression of MMP1 mRNA was analyzed by real time-PCR assay.