1b). The method presented in this work uses the following datasets as the basic information necessary for emergency planning, oil spill prevention and oil spill mitigation. Bathymetric data from EMODNET were used in this work (Berthou et al., 2008) (Fig. 1b). The EMODNET Hydrography

data repository stores Digital Terrain Models (DTM) from selected maritime basins in Europe. DTMs used in this study comprise a grid size of 0.25 min. Each grid cell comprises the following data: (a) x, y coordinates, (b) minimum water depth in metres, click here (c) average water depth in metres, (d) maximum water depth in metres, (e) standard deviation of water depth in metres, (f) number of values used for interpolation over the grid cell, (g) number of elementary surfaces used to compute the average grid cell depth, (h) average water depth smoothed by means of a sp line function in metres, and (i) http://www.selleckchem.com/products/DAPT-GSI-IX.html an indicator of the offsets between the average and smoothed water depth as a percentage of water depth. Onshore topography is amongst the principal parameters used in this study to evaluate shoreline susceptibility. Onshore Digital Terrain Models (DTMs) comprise a 3D digital model of the Earth’s surface (McCullagh, 1998 and El-Sheimy et al., 2005). For this work, an onshore digital elevation model was created for Crete through the detailed digitization

of topographic map contours (1:5000 scale maps) from the Hellenic Military Geographical Service (HAGS) (Fig. 3a). The cell size of the digital elevation model was 20 m. Geological data concerning the near-shore structure and the hydrographic network of Crete were included in the database used in this work. Data sources comprise digital geological maps on the 1:50,000 scale (IGME) and local geological maps completed in the period 2005–2013 (Alves and Lourenço, 2010, Kokinou et al., 2012 and Kokinou et al., 2013). Particular care was taken in the identification Glycogen branching enzyme of local structures, bed

dips, rock and soil quality in the regions where shoreline susceptibility was recognised to be high when of the geological mapping of the shoreline. Shoreline susceptibility maps were compiled based on field geological data, later complemented by morphological data acquired from Google Maps©. Our susceptibility maps are based on the application of Adler and Inbar (2007) classification, used in Israel to characterise shorelines according to their susceptibility to oil spills and natural cleaning up capacity (Table 1). The Environmental Susceptibility Index (ESI) proposed by Adler and Inbar (2007) considers a range of values between 1 and 9, with level 1 (ESI 1) representing areas of low susceptibility, impermeable to oil spilt during accidents (Table 1). Conversely, ESI 9 shorelines are highly vulnerable, often coinciding with natural reserves and special protected areas (Table 1). As ESI 9 shorelines coincide with such areas of natural importance, data from the updated NATURA 2000 database (http://cdr.eionet.europa.

4 Gy. As of 2002, all patients were treated with intensity-modulated radiotherapy (IMRT) technique where a five- to seven-field treatment plan was used. EBRT was delivered to the prostate gland and seminal vesicles. The lymph nodes were not incorporated Trichostatin A nmr into the treatment fields. For patients who received neoadjuvant androgen deprivation therapy (ADT; n = 98; 42%), treatment was usually initiated 3 months before the three-dimensional conformal radiotherapy/IMRT and discontinued at the completion of radiotherapy.

The ADT was given to patients with large prostates to achieve pretreatment volume reduction or to high-risk patients, and adjuvant ADT even for high-risk patients was not routinely given. The median duration of ADT used in these patients was 9 months (range, 1–33 months). The median follow-up for the entire cohort of patients was 61.2 months (range, 3–150 Selleck Staurosporine months). Follow-up examinations consisted of an assessment of serum prostate-specific antigen (PSA), patient symptom assessment, and digital rectal examination. New or worsening acute and late GU and GI toxicities were scored according to the National Cancer Institute Common Terminology Criteria for Adverse Events toxicity scale, version 3. Acute toxicity was defined as symptoms experienced by patients during the course of therapy and up to 90 days from the completion of EBRT. The International Prostate

Symptom Score (IPSS) was used to assess urinary functioning (urinary frequency, hesitancy, urgency, intermittence, weak urinary stream, and nocturia) both before and after the treatment. The patient’s status was determined at the time of

analysis in October 2011. The Phoenix definition of biochemical Exoribonuclease failure (absolute nadir plus 2 ng/mL with the corresponding date) was used for this analysis (16). Actuarial likelihood of complication probabilities and disease-specific survival were calculated according to the product-limit estimate (Kaplan–Meier) method. The threshold of statistical significance for differences was set at 0.05. The 7-year PSA relapse-free survival rates were 95% (95% confidence interval [CI], 86.5–100.0%), 90% (95% CI, 84.4–96.9%), and 57% (95% CI, 38.2–80.8%) for low-, intermediate-, and high-risk patients, respectively (Fig. 1). The median follow-up for each risk group was 69 months (range, 11–137 months), 64 months (range, 3–150 months), and 47 months (range, 5–140 months) for low-, intermediate-, and high-risk patients, respectively. In 206 patients who were free of biochemical relapse, 142 patients (69%) were noted to have PSA levels lower than 0.2 ng/mL at the time of last follow-up, and the PSA was undetectable (<0.05 ng/mL) at last follow-up in 85 (36%) of these patients. The 7-year DMs-free survival for low-, intermediate-, and high-risk patients were 95%, 98%, and 83%, respectively.

Measuring oxidized M148 using conventional mass spectrometry methods that utilize spectral counting or extracted ion chromatograms can be lengthy and challenging in large sample sizes. In contrast, MRM is a promising technique that allows multiplexing of several targets and has been successfully applied to quantitate plasma proteins [10] and [14]. Cyclopamine supplier The addition of SIS peptides in MRM allows for absolute quantitation. Since our goal was to develop

an assay to assess the relative ratio of oxidized M148 to the native peptide rather than the absolute concentrations, the SIS peptide for the unmodified M148 was not synthesized. M148(O) SIS peptide was used to correctly identify the peaks of the in vivo M148 peaks, and optimize the transitions. The rationale for not determining the absolute concentration buy Alectinib of M148(O) in plasma was that this concentration can vary because of variations in the ApoA-I concentration. In contrast, monitoring the relative ratio of oxidized M148 to the non-oxidized peptide represents the “quality” of this peptide, is cost-effective and simple with less inherent variability. Thus, this approach

is better suited for comparing M148 oxidation ratios among different patient groups. One advantage of MRM is that different peptide variants can be selected, depending on the goal of the particular project. The M148-containing ApoA-I peptide would not normally

be selected for ApoA-I quantitation because of its susceptibility to methionine oxidation. The “ATEHLSTLSEK” ApoA-I peptide likely gives Protein kinase N1 a better estimate of total ApoA-I concentration. Several limitations of the study deserve mention. First, we have not measured the molar % oxidized of M148, as such measure would require calibration of both forms of the peptide. Second, methionines are susceptible to ex vivo oxidation that can result from inadequate or prolonged freezing, repeated thawing, or centrifugations [15]. Because an anti-oxidant solution was not immediately added before freezing the samples or after HDL isolations, this might have permitted additional oxidation. The ratios of methionine oxidation observed in our study were higher than those reported in an earlier study on diabetes where the samples were preserved in an anti-oxidant solution prior freezing [16]. In this earlier study, however, younger patients with type 1 diabetes were recruited. We have recently demonstrated that immediate freezing of samples at −80 °C without the use of an anti-oxidant solution results in low levels of ApoA-I oxidation that are stable for up to 2 years of storage [17].

Given the potential for synergistic epigenetic modulation between hydralazine and valproic acid, as well as the safety track record for long-term administration in nononcology patients, we conducted this trial to identify a dose appropriate selleck chemicals llc for chronic administration for lung cancer chemoprevention. The results of our trial support further investigation of epigenetic modification as a new therapeutic strategy. The combination of hydralazine and valproic acid is simple, nontoxic, and lends itself to chemoprevention or combination with other treatments. Future studies will need

to be conducted with pharmacodynamic end points, such as the re-expression of defined panels of tumor suppressor genes as a function of therapy. Furthermore, if hydralazine is used, then study patients will need to be stratified by acetylator phenotype, as it is possible that toxicity, and even efficacy, may be determined by such phenotypic expression. Prospective trials will need to assess the role of epigenetic modification through newly discovered epigenetic

mechanisms of action that could be used as biomarkers of efficacy. We acknowledge the efforts of Valerie Parks (RN), Terry Novak (RN), and Mary Pruess (RN) in providing care to the protocol participants as well as in the monitoring of this trial. “
“Breast cancer (BCa) is the most common malignancy among women around the globe, and it is recognized to be the second most common cause of death in women [1]. Its rate is rising rapidly in Asian women and the developing world. According to the Surveillance, Epidemiology, and End Results database, Asian Indian/Pakistani NVP-BKM120 women residing in the United States seem to have a higher frequency of BCa particularly at a younger age (< 40 years) compared to Caucasians

[2]. The data from South Karachi, a pragmatic representative of the population of Bumetanide Pakistan, revealed that BCa accounted for approximately one third of cancers in women [3]. Hormone receptors such as estrogen (ERs) and progesterone receptors (PRs) play a seminal role in determining the treatment strategy and prognosis of patients with BCa. In addition, human epidermal growth factor receptor type 2 (HER2) has been found to be overexpressed in a subset of invasive BCa and is associated with poor prognosis [4] and [5]. According to Surveillance, Epidemiology and End Results database, Asian Indian/Pakistani women residing in the United States had more ER/PR-negative BCa (30.6%) compared to Caucasians (21.8%) [2]. These data are similar to studies undertaken on samples of BCa from women residing in Pakistan that showed that 60% to 65% of the tumors expressed ER/PR [6] and [7]. Furthermore, frequency of HER2 expression has also found to be higher in Pakistani women with BCa (30%-39%) [6], [8] and [9] in contrast to Caucasians (25%-30%) [4] and [5].

3C). The protein expression assessed by western blotting of CRF1, pro-relaxin-3, GAD65 and TPH2 was similar among the naïve, saline injected, true sham PI3K inhibitor (surgery but no infusion) and sham-lesioned (blank saporin infusion) groups (Fig. 4A and B). However, protein levels of CRF1 receptor was considerably reduced in the NI-lesioned rats. There was also a consistent decrease in the levels of pro-relaxin-3 in the NI-lesioned rats as compared to the sham-lesioned rats (Fig. 4A). The same set of samples

was also checked for GAD65 and TPH2 protein levels. A significant decrease in the expression of GAD65 was also observed in the NI-lesioned rats while TPH2 protein levels remained unchanged. Densitometry analysis of the western blot demonstrated a statistically significant reduction in CRF1 (approximately 54%), pro-relaxin-3 (approximately 53%) and GAD65 (approximately

64%) protein expression (Fig. 4B). Further confirmation of the lesion through immunofluorescence labelling verified the loss of CRF1 receptor positive cells in the NI of the NI-lesioned rats as compared to the sham-lesioned rats (Fig. 5A and B). Similarly, relaxin-3 expressing cells SB203580 concentration also decreased considerably in the NI-lesioned rats (Fig. 5D). Examination of the TPH2 expression revealed that TPH2 positive cells lined the midline of the NI and were unaffected by the lesioning procedure (Fig. 5E and F). Positive CRF1 staining of the nearby locus coeruleus (LC) in both sham- and NI-lesioned rats was also detected (Fig. 5G and H). Immunostaining for the glial cell marker, glial fibrillary acidic protein (GFAP), showed up-regulation of glial cells in the NI of both sham-lesioned and NI-lesioned rats at 14 days after surgery (Fig. 6). The levels of pro-relaxin-3 in the MS in naïve, saline, true sham, sham- and NI-lesioned rats were assessed by western blotting. A decrease in relaxin-3 levels was Liothyronine Sodium observed in the MS of NI-lesioned rats (Fig. 7A). Densitometry analysis of the blots showed an approximately 90% decrease in

pro-relaxin-3 levels (Fig. 7B). The NI-lesioned and sham-lesioned rats were tested in a cued fear conditioning paradigm. The rats were first exposed to tone-shock pairing and subsequently freezing behaviour in response to the tone was assessed 24 h later. To determine if the NI-lesioned rats had any locomotor deficits, the total distance travelled by the sham- and NI-lesioned rats were measured during the 15 min habituation phase. There was a slight but insignificant decrease in the distance covered by the NI-lesioned rats (Fig. 8A) possibly due to the increased periods of freezing observed during this phase (Fig. 8B). No significant difference between the percentage of freezing between sham- and NI-lesioned rats was observed during the tone-shock training phase (Fig. 8C), for 2 min before (Fig. 8D) and during the 30 s tone at the 24 h test phase (Fig. 8D).

1; p<0.01; ω2=0.19;), as well as a significant medium effect on total motivation (F(1, 111)=9.9; p<0.05; ω2=0.08;). No significant main effect of school type and of gender

neither on motivation in total nor on any of its subscales was found. The same holds for the rest of the covariates. Within subject effects ( Table 8b): both total motivation and all its subscales showed significant and strong interaction of their temporal development (or time course, TC) with group membership (CC: F(2, 222)=79.1; p<0.01; buy MDV3100 ω2=0.40; SC: F(2, 222)=59.2; p<0.01; ω2=0.34; IM: F(2, 222)=57.1; p<0.01; ω2=0.33; total: F(2, 222)=75.8; p<0.01; ω2=0.39). Significant, but only small up to medium main effects on motivation in total and on two of three subscales were found for the interaction “time course vs. school type” (CC: F(2, 222)=6.1; p<0.05; ω2=0.05; SC: F(2, 222)=11.2; p<0.01; ω2=0.09; total: F(2, 222)=8.1; p<0.05; ω2=0.06). Finally, significant small up to medium size effects were found for the threefold TC×GM×ST interaction as well as the fourfold ZD1839 chemical structure TC×GM×ST×GR interaction on each sub-dimension and motivation in total (TC×GM×ST; CC: F(2, 222)=12.7; p<0.01; ω2=0.09; SC: F(2, 222)=9.8; p<0.01; ω2=0.07; IM: F(2, 222)=3.6; p<0.05; ω2=0.04; total: F(2, 222)=5.9; p<0.05; ω2=0.05; for TC×GM×ST×GR

interaction: see Table 8b). Thus motivation developed differently in time for different treatment groups and school types. in particular, the strongest significant interaction 4��8C characterized by large effects on each subscales and motivation in total was found for the TC×GM interaction. As inspection of the time course (Fig. 2) clearly shows, the TC×GM interaction obviously is due to large differences between treatment and control group in development from before to after intervention (and not a possible

difference afterwards, i.e. from post to follow-up test). Thus, the results of the temporal development of motivation within subjects is consistent with the between subjects main effect of group membership on motivation. The influences of the threefold and the fourfold factor-interaction on each sub-dimension and motivation in total mean, that the positive motivation development of TG compared to CG was different for both school types ST1/2 considered (as clearly visible when comparing Fig. 3 and Fig. 4). This might be due to chance, but a plausible explanation is as follows: in school type 1 (“Realschule”) grade 10, where the instruction took part, is the last year in school; a general drop of motivation towards the very end of the schooling period is a well-known experience of many partitioners supported by the data ( Fig. 3, CG curve). This factor does not exist for school type 2 (“Gymnasium”, Fig. 4), hence the difference found. No influence of gender or of any interaction of gender and other factors neither on motivation in total nor on subscales could be discovered. The holds true for the influence of all other learner covariates considered.

Instead, our data perhaps suggest that improvement in standard nonendoscopic care has led to improved survival, such as the routine administration of intravenous proton pump inhibitor infusions, the routine use

of risk scoring, the implementation of standardized clinical guidelines, and the subsequent local auditing of practice.4, 5 and 30 In conclusion, contrary to previous smaller studies, we have found an encouraging substantial improvement in mortality following hospital admission for upper gastrointestinal hemorrhage. Our study shows that this is partially obscured by changes in age and comorbidity and that the improvements are less marked in the elderly individuals in a manner not explained by comorbidity. We believe that this improvement reflects the effect of changes in the care of gastrointestinal hemorrhage over the last decade, Selleckchem CHIR 99021 but it also suggests the need to focus our ongoing attention on the elderly individuals who may not yet have benefited to the maximum possible extent from these changes. The recent demonstration of under-utilization of endoscopic techniques in the United Kingdom, coupled with the fact that other interventions such as use of proton pump inhibitors are more readily available to the admitting physician worldwide, may suggest areas that could be

further improved.4, Fulvestrant in vitro 5, 31, 32 and 33 The funding bodies had no role in the collection, analysis, or interpretation of the data. “
“Bacillus

coagulans, a non-pathogenic, facultative anaerobic, thermotolerant and acidophilic bacteria, is an important food spoilage microorganism. In the canned vegetable industry where foods are acidified to pH values between 4 and 4.5, this bacterium is frequently found, since spores of B. coagulans are able to grow and germinate at pH values as low as 4 ( De Clerck et al., 2004 and Lucas et al., 2006). Moreover, this bacterium is capable of increasing until the pH of food products to values that may allow for germination of surviving Clostridium botulinum spores ( Viedma et al., 2010). Besides, B. coagulans has caused considerable economic loss for the food industry because of the “flat sour spoilage”, which is a drastic acidification of the food product due to the production of lactic acid without gas formation ( Lucas et al., 2006). For official protocols to validate low acidity foods heat sterilization, C. botulinum spores are the target microorganism and the temperature reference is 121.1 °C. Nevertheless, heat resistant mesophilic spore formers such as Bacillus sporothermodurans ( Periago et al., 2004) and B. coagulans may often determine the stability of foods and safety of industrial processes.

Bothriopsis venoms contain L-amino acid oxidase, esterase, peptidase, phosphodiesterase, phospholipase A2 (PLA2) and proteolytic activities, as well as coagulant, hemorrhagic and myotoxic activities ( Kuch et al., 1996, Porto et al., 2007 and Furtado et al., 2010), in addition to causing

neutrophil migration into the mouse peritoneal cavity ( Porto et al., 2007); other biological activities of these venoms have been poorly studied. In this work, we investigated the neuromuscular activity of venom from the Amazonian forest viper Bothriopsis bilineata smargadina. Male Swiss mice (25–30 g) obtained from the Multidisciplinary Center for Biological Investigation Alectinib molecular weight (CEMIB/UNICAMP) were housed 10/cage at 23 °C on a 12 h light/dark cycle with lights on at 6 check details a.m. Male chicks (4-8 days old) were provided by Granja Ito S/A (Campinas, SP) and housed in metal cages with a sawdust substrate. The mice and chicks had free access to food and water. This study was approved by the institutional Committee for Ethics in Animal Experimentation (CEEA/UNICAMP, protocol no. 2267-1). Bothriopsis

b. smargadina venom was a pool obtained from adult snakes of both sexes captured in the Amazon region. The venom was desiccated and stored at −20 °C until used. When required, the venom was dissolved in 0.9% NaCl prior to use. Chick biventer cervicis nerve-muscle preparations were obtained and mounted (resting tension: 0.5 g) in Krebs solution at 37 °C and allowed to stabilize for 20 min prior to use, as described elsewhere (Borja-Oliveira et al., 2003 and Rodrigues-Simioni et al., 2004). Muscle responses to exogenous acetylcholine (ACh, 110 μM) and KCl (40 mM) were obtained before and after incubation with venom (0.1–30 μg/ml) to screen for postsynaptic

neurotoxicity and myotoxicity (Harvey et al., 1994). Creatine kinase (CK) release was measured for one venom concentration (10 μg/ml) in preparations incubated at 37 °C; activity was assayed using commercial kits ADAMTS5 (CK-NAC, LaborLab, São Paulo, SP, Brazil). The influence of temperature on venom-induced neuromuscular blockade was examined by doing some experiments at 22 °C. Mouse phrenic nerve-diaphragm preparations were mounted in Tyrode solution (composition, in mM: NaCl 137, KCl 2.7, CaCl2 1.8, MgCl2 0.49, NaH2PO4 0.42, NaHCO3 11.9 and glucose 11.1, pH 7.0 at 37 °C after equilibration with 95% O2/5% CO2), as described by Oshima-Franco et al. (2004). After stabilization for 20 min, the preparations were incubated with different venom concentrations (1, 10 and 30 μg/ml, one concentration per preparation) for 120 min and the changes in twitch-tension recorded. To examine the influence of temperature on neuromuscular blockade, some experiments were initially done at 22 °C and the temperature then returned to 37 °C for the rest of the incubation.

This was a 12-month, phase III, multicenter, randomized, double-blind, parallel group, active comparator controlled study in Japanese patients with involutional osteoporosis. Diagnosis of osteoporosis was based on the presence or absence of fragility fracture and BMD measurements specified in the “Guideline for the Diagnosis of Primary Osteoporosis (2000 Revised Version)” established by the Japanese Society for Bone and Mineral Research Enzalutamide molecular weight [20] and [21]. Individuals eligible for this study were ambulatory Japanese male and female subjects aged ≥ 50 years who were diagnosed with osteoporosis, based on the criteria for primary osteoporosis of the Japanese Society for Bone and

Mineral Research [20] and [21]. Primary osteoporosis was defined by the presence of a fragility fracture and BMD < 80% of the ‘young adult mean’ (20 to 44 years of age), or BMD < 70% of the ‘young adult mean’ in the absence of a detectable fragility fracture [21]. In the case of female subjects, ≥ 2 years must have passed since menopause.

The main exclusion criteria were factors which affect selleck screening library efficacy evaluation; secondary osteoporosis and any other disease causing decreased bone mass or affecting lumbar spine BMD (including severe scoliosis of the spine, fracture or severe deformation in any of the L2–L4 lumbar vertebrae, or a spinal X-ray image suggesting severe bone sclerosis [calcification] in any of the L2–L4 lumbar vertebrae); not administration of bisphosphonate within 24 weeks before the first dose of the study

drug; administration of any drug affecting bone metabolism such as SERMs, vitamin D3 and vitamin K2 preparations, and calcitonin analogs, etc. within 8 weeks before the first dose of the study drug. In addition, any subject judged by the attending physician to be unsuitable to participate in the study was also excluded. The study was performed at 60 study sites in Japan between February 2010 and August 2011 in accordance with the ethical principles set out in the Declaration of Helsinki and the ICH Harmonized Tripartite Guideline for Good Clinical Practice, and was approved by the Institutional Review Boards at each study site in line with local regulations. Prior to study registration, all subjects were given a full explanation of the study procedures and provided written informed consent. Subjects fulfilling the inclusion/exclusion criteria were eligible for the study and were randomized (in a ratio of 1:1) to receive risedronate 75 mg once-monthly or risedronate 2.5 mg once-daily. Matching 2.5 mg and 75 mg placebo tablets were administered to maintain double blindness throughout the study. Subjects were instructed to take a single 75 mg risedronate tablet or 75 mg placebo tablet on the same calendar day each month and a single 2.5 mg risedronate tablet or 2.5 mg placebo tablet at the designated time on every day.

, 2009 and Mattsson et al., 2010). Furthermore, systemic bacterial infections are considered to be a risk factor for sporadic AD, connecting infection, inflammation and alterations in amyloid metabolism and leading to cognitive disturbances (Dunn et al., 2005, Honjo et al., 2009 and Eikelenboom et al., 2012). A potential function of the Aβ-peptides in this context may be to opsonize pathogens to support their clearance and/or act as soluble factors with cytokine or chemokine activities.

We have previously reported that monocyte activation by the phagocytosis of polystyrene beads induces APP glycosylation and Aβ-peptide secretion (Spitzer et al., 2010). We observed a relative increase in the release of N-terminally truncated Selleck CAL101 Aβ peptide species from

activated monocytes (Maler et al., 2008 and Spitzer et al., 2010). In the current study, we used mononuclear phagocytes as a model to investigate the impact of various Aβ-peptides on the phagocytosis of polystyrene particles or Escherichiacoli bacteria and on concurrent macrophage polarization. Human peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll (Biochrom, Germany) density gradient centrifugation from buffy coats purchased at Transfusionsmedizin Suhl, Germany. Thrombocytes were removed by under-laying an selleck kinase inhibitor FCS-gradient and centrifugation at 75g for 15 min. Monocytes were isolated from PBMCs by the antibody-mediated removal of non-monocytes using a MACS Monocyte Isolation Kit II (Miltenyi Biotec, Germany) and MACS LS Columns (Miltenyi Biotec, Germany). For the analysis of undifferentiated

monocytes, the cells were resuspended in serum-free AIM-V medium and seeded at a density of 1 × 106 cells/ml in 96-well ultra-low attachment plates (Corning, USA). Differentiated macrophages were obtained by cultivating monocytes for seven days at 8 × 105 cells/ml in RPMI 1640 with 10% FCS in 96-well plates (Biochrom, Germany). For the polarization of macrophages, 40 ng/ml rhGM-CSF (Immunotools, Germany) tuclazepam or 80 ng/ml rhM-CSF (Immunotools, Germany) was added to the culture medium, respectively. After three days, 50% of the medium was exchanged. THP-1 cells were obtained from ATCC and maintained below 1 × 106 cells/ml in RPMI 1640 supplemented with 10% FCS. For differentiation to macrophages, THP-1 cells were cultured at 2 × 106 cells/ml for three days in RPMI 1640 medium supplemented with 10% FCS and 50 ng/ml phorbol 12-myristate 13-acetate (PMA, Sigma–Aldrich, Germany). Primary cultures of porcine microglia were isolated following a protocol adapted from Franke (Franke et al., 2000). Briefly, the secretory areas, cerebellum and meninges were removed from brain hemispheres obtained from a local abattoir. After mincing the tissue, it was incubated for 2 h with 6.5% (v/v) dispase (BD, Germany) at 37 °C. Lipids were removed by mixing 100 mL of the cell suspension with 150 mL of dextran solution (ρ = 1.