6 mmol m−3, the assumption being that these values were constant

6 mmol m−3, the assumption being that these values were constant with depth. No data for the detritus content at the bottom were available, and the instantaneous sinking of detritus was a more arbitrary model assumption. The initial detritus content in the subsurface water layer was prescribed as 100 mgC m−2. However, a constant value of 50 mgC m −3 for pelagic detritus was assumed throughout the water column. For BD and GtD, all the initial values were assumed to

be the same as for the GdD except for the nutrient concentrations, i.e. total inorganic nitrogen – NutrN = 5 mmol m−3 and phosphate – NutrP = 0.5 mmol m−3. The model was validated for GdD (Dzierzbicka-Głowacka et al. 2010a) on the assumption that processes governing POC concentrations CX-5461 solubility dmso in other areas of the Baltic Proper are similar. Thus, the POC concentration and POC dynamics in GtD and BD differ from those in GD owing to differences in nutrient concentration and physical factors. The modelled values of the primary production SP600125 research buy for the 1965–1998 period and POC concentrations for 2010 were compared to the measured values (see Discussion). The most important factors, with an overriding influence on primary production, are PAR, nutrients and temperature. Fourier analysis of

the archived data (34 years) reveals seasonal and annual variations in the sea surface temperature and nutrient concentrations in the past and shows the main trend of increasing temperature and nutrient during more than 40 years in the southern Baltic Sea, mainly in the Gdańsk Deep (GdD). The equation describing long-term variations of hydrological parameters, S=So+A(x−1960)+Bsin(ωx+φ1)+Csin(2ωx+φ2) where A is the average annual

increase of the parameter under investigation, was used by Renk (2000) to analyse the data set from the Sea Fisheries Institute (Gdynia). The tendency for the average temperature in the surface water to increase Immune system by 0.006°C yr−1, and in the upper layer by 0.0117°C yr−1 was evident by the end of the 1965–1998 period ( Renk 2000: Table 4). An increase of 1% of the average annual nutrient value with the exception of summer, when nutrient concentrations are close to zero (i.e. 0.0036 mmolP m−3 and 0.022 mmolN m−3), was recorded in GdD ( Renk 2000: Table 4). This will lead to a nutrient concentration in 2050 higher than in 1965–1998 by ~ 0.18 mmolP m−3 for phosphate and by ~ 1.1 mmolN m−3 for total inorganic nitrogen. For BD and GtD we assumed lower values: 0.0034 mmolP m−3 and 0.021 mmolN m−3. The increase in nutrients includes the inflow of nutrient compounds from the river and atmosphere. This rise in nutrient concentrations in the southern Baltic Sea over a period of many years has enhanced the average annual primary production by about 2 to 3% ( Renk 2000: eq.

Given the volume of oil released by the spill, however, it

Given the volume of oil released by the spill, however, it

is difficult to say that most of the oil was not derived from the spill. There was certainly some weathering of oil which occurred between the seafloor and the surface, and again between the spill site and the shore. This would have been affected by the addition of the dispersant Corexit® at the wellhead and the surface. Local seeps would be the most likely source of additional crude, although the volume of input from seeps would have been negligible in comparison to the spill volume. High concentrations of compounds at the spill site as observed in this study were to be expected, given the volume of the spill. The continental shelf of the northern GOM is known to have hundreds to thousands of small seeps of oil and gas, but the volume of these seeps is negligible compared to the BP/DWH spill volume. In addition, results of the diagnostic Oligomycin A chemical structure ratios of biomarkers were positive, indicating that the source of the oil in our samples Protein Tyrosine Kinase inhibitor was from the BP/DWH spill. Comparing our results with those of other investigators, Aeppli et al. (2012) collected 146 samples in 2010 and 2011 offshore and on the beaches in this region. They focused, however, on the production of oxyhydrocarbons during the weathering process. PAHs were analyzed for a small sub-set of samples

(n = 10); PAHs were not the focus of their analysis. Carmichael et al. (2012) report oiled and non-oiled honeycomb styrofoam material in the GOM surface waters and along the coastal beaches. Naphthalene, fluorene, phenanthrene, and chrysene in the oiled material were

depleted relative to Macondo well oil by 98%, 72%, 43% and 0%, respectively. This highlighted the greater susceptibility of smaller two-ring PAHs to weathering as opposed to the larger multi-ring PAHs. This is consistent with observations made on other oil spills (see Reddy et al., 2011, for data on sub-surface partitioning C-X-C chemokine receptor type 7 (CXCR-7) of n-alkanes and benzene). The distribution of compounds measured in the central region of the northern GOM and in nearby areas are generally consistent with known ocean currents in the region (see Sturges and Lugo-Fernandez, 2005 for a review). The spill site was S–SE of the mouth of the Mississippi River. The river plume is known to be influenced by near-shore coastal currents in the region which split near the mouth of the river, with most of the plume being drawn to the west and the remainder to the east. In addition, the Loop Current is known to produce eddies which impinge on the spill site, potentially carrying petroleum hydrocarbons offshore. Such eddies also break free, potentially carrying such compounds to the west along the edge of the continental shelf. Various impacts extended from June 2010 to at least March 2011. Most samples were collected post-capping (July 15, 2010); thus, geographic patterns of compounds in general represent post-spill distributions.

05) Furthermore, the damage index for AuNps-citrate and AuNps-PA

05). Furthermore, the damage index for AuNps-citrate and AuNps-PAMAM at 1.0 μM did not show a significant effect (p > 0.05) for PBMC. At a concentration of 50.0 μM, the AuNps-PAMAM induced a significant toxic effect (p < 0.05) on PBMC cells, compared to the negative control. ROS and XL184 molecular weight reactive nitrogen species (RNS) are generated during the inflammatory response, especially by phagocytes, and they may contribute to the pathology of many inflammatory conditions (Paino et al., 2011). Furthermore, they represent a disturbance in the balance between pro-oxidant/antioxidant reactions. AuNps cellular uptake were acquired by flow cytometry and appear in Table 4 as a function of side scatter

(SSC), representing the cell granularity, and forward scatter (FSC), representing the cell size. A significant increase

in the SSC values was observed for HepG2 cells only for AuNps-PAMAM treated cells, at a concentration of 50.0 μM. On the other hand, PBMC incubated with citrate- and PAMAM-covered AuNps exhibited an increase (p < 0.05) in the SSC values RG7420 cell line for both concentrations investigated from the negative control, except at 1.0 μM AuNps-citrate. A statistically significant (p < 0.05) measurement of intracellular ROS was observed for both HepG2 and PBMC upon treatment with AuNps, as shown in Fig. 3a and b, respectively. Data from zeta potential analysis, as depicted in Table 1, suggests that cell culture media containing 10% FBS serum influences the nanoparticles stability. Since the medium contains a variety of salts, amino acids and vitamins, its high ionic strength decrease the electrostatic repulsive forces among the nanoparticles, inducing aggregation (Fatisson, 2012). On the other hand, proteins from serum in the medium can adsorb on the nanoparticles creating a protein “corona”, resulting in the stabilization of the colloidal suspensions and preventing aggregation

upon modifying their zeta potential (Fatisson, 2012 and Chithrani et al., 2006), as observed here via DLS or hydrodynamic diameter (Table 1). The interaction between the cells and the nanoparticles could be mediated by nonspecific adsorption of serum proteins onto the gold surface, Succinyl-CoA as proposed by Chithrani et al. (2006). In our case, the AuNp uptake mechanism may occur via receptor-mediated endocytic pathways (clathrin mediated), in agreement to what has been reported by Nativo et al. (2008). Data from literature regarding the cytotoxicity and genotoxicity of citrate or PAMAM-coated AuNps are somehow conflicting (Patra et al., 2007 and Pan et al., 2009). The controversy comes from the variability of parameters, including cell lines used in toxicity assays, concentrations, surface charge and coatings. For example, Connor et al. (2005) demonstrated non-toxic effects of Au nanospheres (diameter from 4 to 18 nm, covered with citrate, cysteine, glucose, biotin, and cetyltrimethylammonium bromide) on human leukemia cell line (K562) cells. On the other hand, Patra et al.

, 2010, Cavallotti et al , 2001, Sandell and Peters, 2002 and San

, 2010, Cavallotti et al., 2001, Sandell and Peters, 2002 and Sandell and Peters, 2003). The areas with greatest neuronal loss are also the regions that exhibit greater changes in microglial phenotype. Whether neuronal loss drives microglial phenotype

changes in ageing, or if changes to the microglia precede and contribute towards neuronal loss, is not known. There are however several mechanisms by which neurons and oligodendrocytes keep microglia in a quiescent state, such as interactions between CD200, fractalkine or CD47 and their cognate receptors on microglia (Gitik et al., 2011, Hoek et al., 2000, Kong et ABT-199 in vivo al., 2007, Koning et al., 2009 and Lyons et al., 2009). Two studies in the healthy adult mouse brain have revealed significant regional variations in the distribution of these molecules. Koning et al. (2009) observed that CD200 expression is greater in grey than white matter, which may contribute to the regional differences in microglial phenotype we report in this study. Fractalkine transcript expression has been reported to be significantly

lower in the cerebellum and other caudal areas such as Idelalisib molecular weight the brainstem than the hippocampus or striatum, which may help to explain the rostral caudal gradient of microglial phenotype changes (Tarozzo et al., 2003). Decreased expression of CD200 in the hippocampus and substantia nigra (Frank et al., 2006 and Wang et al., 2011), and of fractalkine in the hippocampus and forebrain have been demonstrated in aged mice (Lyons et al., 2009 and Wynne et al., 2010). Increased numbers of multinuclear giant cells have also been observed in CD200-/- mice (Hoek et al., 2000), providing a possible explanation for their presence why in the aged brains of our study. A wider assessment of the expression of these immunoregulatory molecules in different regions of the aged brain and how they may correlate with changes in microglial phenotype would be of interest. We anticipated an increase in expression levels of microglia associated molecules after systemic LPS injection, which has previously

been shown to up-regulate FcγRI (Lunnon et al., 2011) and CD11b (Buttini et al., 1996). However, the only molecule we found to be sensitive to systemic LPS injection was FcγRI. CD11b expression was not significantly altered 24 h after systemic LPS challenge. Furthermore, the effect of systemic LPS on FcγRI expression was subtle, region dependent and primarily observed in the white matter regions and the cerebellum of both young and aged mice. A later time point post injection, such as three days, may yield a more robust effect on expression of these molecules (Buttini et al., 1996). Since we had shown that the molecular expression patterns of the microglia in distinct CNS regions were altered with age we used behavioural assays to assess the functional integrity of two regions, the hippocampus and the cerebellum.

The longitudinal changes in these histopathologic end points were

The longitudinal changes in these histopathologic end points were compared against changes in prominent optical parameters as shown in Figure 8, C and D. In the treated group, a major shift in both histology and optical end points was seen, whereas minimal changes were observed across all of these parameters in the control group. In this study, a combination of DRS and AFS was used to investigate cisplatin-induced changes in tumor physiology and morphology across a period of 1 week in a mouse model for

Selleckchem Sotrastaurin hereditary breast cancer. The changes in optical end points were compared against the degree of pathologic response. The results showed that various DRS and AFS parameters in the treated animals significantly changed throughout the course of treatment relative to the untreated animals. These parameters were the Mie-scattering slope (P < .0001), Mie-to-total scattering fraction (P < .001),

tissue oxygenation (P = .035), fat volume fraction (P < .0001), and fluorescence residual (P < .018). ABT-199 solubility dmso Furthermore, the observed changes appeared to be proportional to the degree of vital tumor tissue and the formation of fibrosis. Optical scattering characteristics are dependent on the size and density of cell nuclei and organelles as well as on the composition of the extracellular matrix (e.g., macromolecular aggregates and collagen fibers). In the histopathologic evaluation, considerable alterations in the extracellular matrix (formation of fibrosis) and in the size and the density of (sub) cellular structures were observed in the tumors of the treated animals. These morphologic and structural changes may lead to changes in tissue-scattering

properties that in turn may translate into changes in the Mie-scattering slope and Mie-to-total scattering fraction. Although significant fibrosis and cellular disintegration after treatment with cisplatin may explain these specific changes, further research is needed to provide a better understanding of these relationships. Tumor tissue oxygenation values of untreated animals remained hypoxic over time, whereas tumors of treated animals became progressively more oxygenated. This is consistent with Resveratrol previously reported results where improved oxygenation of tumor tissue was observed due to tumor regression and altered metabolism after treatment with doxorubicin [27], [43] and [44]. For example, Vishwanath et al. performed DRS using a surface probe and showed that mammary-tumor tissue oxygenation in treated mice increased after doxorubicin administration relative to the untreated controls. A particularly interesting finding was the additional fluorescence observed in the treated group. On the basis of two-photon imaging, the extra fluorescence was specifically found in the cellular components of tumor tissue treated with cisplatin. Fluorescence was tumor specific and not observed in liver or muscle tissue of the treated animals.

In fact, Draize testing is the only test formally accepted and va

In fact, Draize testing is the only test formally accepted and validated to assess the full range of irritation severity. Both irreversible and reversible ocular effects can be identified using this test ( Barile, 2010). Eye irritation was traditionally summarized as a “maximum average score” (MAS) which is an average value primarily focused on corneal injury, for individual KU-60019 concentration animals at the time of scoring

( Huhtala et al., 2008). However, many countries had their own scoring systems, which although similar in their approach, led to multiple classifications, labels, and data sheets for the same chemical, dependent upon which country the chemical was been marketed in. In response to this, and as a means of replacing the numerous different classification systems, with a single controlled and unified classification system, the United Nations (UN) developed the current internationally agreed, standard scoring system, known as the Globally Harmonized System (GHS), also known as the “purple book” ( UN, 2013). The GHS utilizes pictograms, signal words, hazard and precautionary statements, and safety data sheets according to standardized levels of physical, health and environmental Androgen Receptor Antagonist research buy hazards. The GHS is based upon averaged single tissue observations which can account for the reversibility of the observed chemical

effects ( Eskes et al., 2005). With regards to eye irritation, there are two primary C-X-C chemokine receptor type 7 (CXCR-7) categories. Substances which cause serious irreversible (up to 21 days) damage/destruction to the cornea, iris and/or conjunctiva are Category 1; substances which cause reversible (within 21 days) irritation including corneal opacity, iritis, redness or chemosis are Category 2. Category 2 chemicals can be split into two subcategories:

2A, irritating to eyes, chemicals which cause reversible irritation to eyes within 21 days; and 2B, mildly irritating to eyes, chemicals which cause reversible irritation to eyes within 7 days. Non irritating chemicals are assigned a GHS No Category classification. The categories are assigned based on calculations of a mean score following observational grading at 24, 48 and 72 h post application of the test chemical. The GHS was adopted in 2002 and published in 2003 ( Silk, 2003). Despite the adoption of the GHS, Draize testing is often criticized due to its subjective and time consuming nature, lack of repeatability, variable estimates, insufficient relevance of test chemical application (Davila et al., 1998), high dosages (Curren and Harbell, 2002) and over-prediction of human responses (Jester et al., 2001), primarily due to interspecies differences. In addition, for most routine and acute toxicity tests, for example skin toxicity tests, there are standardized exposure times and/or delivery methods in place.

The EMG activation was not different from zero in the SC conditio

The EMG activation was not different from zero in the SC condition in middle age group. Mean EMG amplitude between 280 and 300 msec was entered into a group (3) × congruency (3) ANOVA. In this early time window there were no significant congruency effects [F(2,102) = 1.664, p = .1943] or interactions [F(4,102) = .3713, p = .8286] but a group effect approached significance [F(2,51) = 2.48, p = .093]. Mean EMG amplitude between 460 and 480 msec was entered into a group (3) × congruency (3) ANOVA. In the mean amplitude of the 460–480 msec time interval there was a congruency effect [F(2,102) = 7.24, ɛ = .769, p = .0031]. Post hoc Tukey

contrasts on the incorrect hand mean amplitude revealed that the congruent condition had significantly less amplitude than the RC condition (p = .0011, Dasatinib mw .045 vs .07 μV) and SC had significantly less amplitude than RC (p = .0011, .04 vs .07 μV). However there was no difference between congruent and SC in incorrect hand activation. Additionally there was a group × congruency interaction [F(4,102) = 3.06, ɛ = .769, p = .0317]. Tukey post hoc tests showed that in the adolescent group the amplitude in the RC condition (.120 μV) was significantly larger Afatinib chemical structure than the congruent (.06 μV, p = .0198) and SC (.05 μV, p = .0198) conditions. There was no similar difference in the adult and middle

age groups. There was no main effect of group [F(2,51) = 1.014, p = .3698]. Overall in terms of correct hand activity there were no significant group differences however in terms of incorrect hand activity, at the time point between 460 and 480 msec, the adolescent group showed significantly increased incorrect hand activity during the RC condition. This

is in line with our prediction of response level change during adolescence. Following Craik and Bialystoke’s (2006) call to identify the specific nature of age-related change here we systematically tracked neuro-cognitive asymmetries in stimulus and response conflict http://www.selleck.co.jp/products/azd9291.html processing throughout the lifespan within the framework of a single study. We measured ERPs, the LRP, and EMG in an adaptation of the colour word Stroop task that a priori separates stimulus and response level conflict. Behavioural effects, in terms of RT and accuracy, revealed that the congruency manipulations were successful. The RC manipulation yielded the slowest RTs. This replicates previous studies (Houwer, 2003 and Melcher and Gruber, 2009). However, unexpectedly there were no differences between groups in terms of the congruency effects. We predicted that adolescents would be more susceptible to response conflict whereas middle age adults would be sensitive to stimulus conflict however no differences were found behaviourally. At the neural level we found age-related and developmental asymmetries in stimulus and response stages of processing.

Microcontact imprinting method has been used for various proteins

Microcontact imprinting method has been used for various proteins [23], [24], [25], [26] and [27]. BSA (bovine serum albumin) is a protein with the molecular weight of 66.5 kDa and it has many uses in biomedical applications and enzymatic reactions. It is used to prevent adhesion of enzymes during applications [28]. It is a generally used protein reagent in protein assays, like Bradford assay, to measure the concentration of a protein in solution. Furthermore, BSA has a structural homology with HSA (human serum albumin) [29]. Due to this, BSA is frequently studied as a model protein instead of HSA. Moreover, BSA is a commonly used target to analyze when designing new immunochemical

assays. Determination of micro-quantities of BSA is possible with methods like radioimmunoassay (RIA) or enzyme-linked immunosorbent assay Dabrafenib (ELISA) [30]. There are also some determinations like FT-IR spectroscopy, polarographic and fluorimetric measurements used for BSA detection [28], [31] and [32]. Some of the methods require a labelled reagent like a radioisotope or enzyme labeled antibody/antigen [30]. Some of them are really expensive and need time-consuming, complex procedures. Low selectivity

and sensitivity is the another drawback Panobinostat purchase of these methods [33]. Direct, label-free, fast and sensitive measurement of various analytes with biosensors has attracted considerable interest [34]. Highly sensitive biosensor concepts make it possible to assay biomacromolecules at concentrations below the limit of detection of conventional methods [35]. Capacitive biosensors are the electrochemical sensors that measure changes in the dielectric properties second when an analyte interacts with a biorecognition element on the sensor surface, causing a decrease in the capacitance [36], [37], [38], [39], [40] and [41]. Capacitive biosensors have been used for the detection of various analytes like antigens, antibodies,

proteins and heavy metal ions [42], [43], [44], [45], [46] and [47]. These types of biosensors have a lot of advantages like inherent rapidity, high sensitivity, simplicity, low cost, easy manipulation and real-time measurement without labeling. In the study reported here, a capacitive biosensor with an automated-flow injection system was used for BSA detection. BSA is most commonly used model protein in the macromolecular imprinting studies. However, to our knowledge, this is the first microcontact-BSA imprinting study for the detection of BSA with the capacitive biosensor. Microcontact imprinting method was applied for the imprinting of BSA onto the pre-modified gold electrode surface. After modification of the gold electrode surface with poly-tyramine and acryloyl chloride, the protein stamp was brought together with a mixture of monomer and cross-linker in contact with the electrode. Thus, the microcontact BSA imprints were introduced to the electrode surface via UV-polymerization.

A flexible loop-structure protruding from the C-terminal LRR capp

A flexible loop-structure protruding from the C-terminal LRR capping unit of the VLR antibody forms a pocket for the relatively small H-trisaccharide antigen which interacts with residues located in the inner concave surface of the VLR antibody and the C-terminal loop. On the other hand, the C-terminal loop interacts with residues

located in the active site of HEL, an epitope location to which it is notoriously difficult to raise conventional immunoglobulin-based antibodies, which preferentially interact with planar epitopes. We hypothesize that the unique origins and protein architecture of VLR antibodies will render Panobinostat nmr these novel reagents uniquely suited for biomarker discovery. Key to using monoclonal VLR antibodies for this purpose will be their applicability for the capture and purification of protein antigens. Obeticholic Acid Using the monoclonal VLR32 antibody, we demonstrate that lamprey antibodies can be used effectively for immunoprecipitation applications followed by mass spectrometric protein identification. The inability of a monomeric form of the VLR antibody to bind to Jurkat T cells indicates its low affinity, in keeping with recent analyses indicating a Kd of 3.0 × 10− 6 M for monomeric units

of the VLR4 antibody (Kirchdoerfer et al., 2012). However, our data show that a low affinity of the individual antigen-binding unit to the antigen does not impede the use of multimeric VLR antibodies for Non-specific serine/threonine protein kinase protein purification. We observed a weak signal for CD5–GFP fusion proteins in immunoprecipitation experiments using monomeric VLR antibodies, which is likely due to ‘artificial’ multimerization

of these VLR units upon binding to protein G beads. This type of ‘artificial’ multimerization would not occur in flow cytometry assays where we detected no residual binding activity of the recombinant monomeric VLR32 units. CD5 positive human B cells have been described previously in tonsilar tissues (Fischer et al., 1997) and other reports indicate a comparable proportion of peripheral blood B cells (10–25%) expressing the CD5 antigen (Gadol and Ault, 1986 and Ebeling et al., 1993). While tonsilar CD5-positive B cells were readily detected using monoclonal VLR32, we did not detect B cells that bound VLR32 in our initial screen of the VLR library on PBMCs. However, in subsequent experiments we observed a significant inter-person variability of CD5+ cells in blood and found that VLR32 can recognize CD5+ B cells in blood (data not shown), suggesting that the lack of VLR32-binding B cells in our original screen is likely reflective of a donor sample devoid of a substantial CD5+ B cell population. In conclusion, we present monoclonal VLR antibodies as novel reagents for proteomics-based biomarker identification.

Nine independent specimens of human skin were evaluated for the e

Nine independent specimens of human skin were evaluated for the expression of MT-3. All nine specimens displayed immunoreactivity for MT-3 in the epidermis. For each specimen, the immunoreactivity for MT-3 was uniform throughout the epidermis and included staining of the basal keratinocytes ( Fig 1A). Six of the specimens exhibited moderate to strong staining for MT-3 while the other 3 displayed mild to strong intensity of staining ( Table 1). All squamous cell carcinomas (SCC) exhibited staining for MT-3 ( Table 1), five strongly ( Fig 1C), six moderately

( Fig 1E), and one mild to moderate, whereas many of the basal cell carcinomas (BCC) exhibited low staining ( Table 1) with two being totally devoid of MT-3 expression ( Fig 2F), three weakly, and the rest (five samples) were either mild or mild to moderate ( Fig 1D) in MT-3 staining. The BIRB 796 staining of MT-3 was determined

on 9 specimens of nevus. Staining for MT-3 was found for all 9 specimens, exhibited moderate to strong intensity, and was present in over 80% of the cells comprising the nevus ( Table 1, Fig 2A). Staining of MT-3 was also performed on 1 case of dysplastic nevus and 3 cases of in situ melanoma. The staining was similar to that found in the nevus with all specimens displaying moderate to strong staining in over 80% of the cells ( Table 1, Fig 2B). Staining of MT-3 was also performed on 4 cases of superficial spreading melanoma and 5 cases of deeply invasive melanoma. Again, the staining was similar to that found for GSK J4 molecular weight in situ melanoma with moderate to strong staining of MT-3 in over 80% of the melanoma cells ( Table 1, Fig 2C, D, E). Proliferating and confluent cultures of NHEK and HaCaT cells were assessed for their expression of MT-3 mRNA and protein. Real time PCR demonstrated that both resting and dividing NHEK and HaCaT cells had only background levels of MT-3 mRNA expression (Fig 3A, B). Both sets of cells were also shown to express only background levels

of MT-3 protein Inositol monophosphatase 1 (data not shown). Both the NHEK and HaCaT cells were treated with MS-275, a histone deacetylase inhibitor, and 5-aza-2′-deoxycytidine, a DNA methylation inhibitor, to determine if MT-3 expression might be silenced by a mechanism involving histone modification or DNA methylation. The results demonstrated that treatment with MS-275 was effective in restoring MT-3 mRNA expression in both the NHEK and HaCaT cells (Fig 3A,B). While MS-275 treatment was effective in both cell lines, MS-275 increased MT-3 mRNA levels in NHEK cells 10 to 20 fold greater than those of the HaCaT cell line. Treatment of the NHEK and HaCaT cells with 5-aza-2′-deoxycytidine, resulted in a small, but statistically insignificant increase in MT-3 mRNA expression for both cell types (Fig 3A, B).