PTMs often occur at low stoichiometry and thus efficient enrichment techniques are key for their successful and comprehensive identification. In general different chemical affinities between the modified and unmodified species are utilized for differential binding to a resin or chromatographic media yielding

positive or negative selectivity and enrichment. All approaches share the common hurdle of unspecific carryover and loss following binding to surfaces. A great advance for the enrichment of peptides bearing PTMs is the replacement of resins by soluble hyper branched polyglycerol polymers leading to massively decreased nonspecific binding while increasing binding capacity Y-27632 mouse [6•]. Upon successful peptide enrichment mass spectrometry is used for peptide and PTM identification. Unlike identification

of the entire protein by multiple peptides in one shotgun experiment, identification of a specific modification and often the protein bearing the PTM, is based on the observation of one single peptide only. For proteins having two or more such modifications, protein identification can often be made by two or more different and unique peptides. However, for single peptides bearing a PTM, such as phosphopeptides, unambiguous protein identification is problematic. For Romidepsin chemical structure the identification of protein termini we and others introduced high confidence protein identification from single peptide identifications based on multiple peptide variants [7 and 8]. In the past ten years since its introduction [5] degradomics and its subfield, terminomics, have developed from a small field covered by only a few publications a year to a vibrant community publishing over 40 papers in 2011 (Figure 1). For in depth comparison of available mass spectrometry based methods for the proteome-wide

analysis of limited proteolysis and their subsequent modification we refer to a recent review by Huesgen and Overall [9••] and by the accompanying paper in this issue from 4-Aminobutyrate aminotransferase the Gaevert laboratory [10]. Since the function of a protease is inherently linked to the effect of proteolysis on its substrates, and since more than half of all proteases have no annotated substrates in MEROPS, the protease database (http://merops.sanger.ac.uk), since 2000 a major focus has been in the identification of protease substrates [11]. These include matrix metalloproteinases (MMPs) 2, 9, 14, 25 [6•, 12, 13, 14, 15 and 16], cathepsins D and E [8 and 17] and caspases 2, 3, 7 [18], meprins, astacins, and the methionine aminopeptidase-2 [19]. In vivo the cleavage rate differs greatly between individual substrates by the same protease [ 20•]. The cleavage site specificity of proteases has been investigated in depth using standard and specifically tailored degradomics approaches using database searchable, proteome-derived peptide libraries in a procedure called PICS [ 21].

Furthermore, this electrode has multiple layers on top permitting repeated uses after washing, these layers also provide significant durability and resistances against interferential substances in the solutions as described in previous studies [6] and [7]. Fig. 4(a) represents the comparisons between the amperometric responses on the first day of measurements and those after 30 days with the same chips. The chips were stored in a fridge when not being used. Compared with our previous study using FGO-Au-PCB chips without multiple layers [13], the overall level of measured current increased by 20 times as well as the long term stability

was increased up to 5.6%. It was demonstrated that current generated by the multiple layer-Au-PCB find more drops

to overall 8.7% of its initial value within 30 days. The resistant ability of the Au-PCB electrode modified with multiple layers was investigated under additions of different interferential substances, such as ascorbic acid, uric acid, acetaminophen, click here creatinine and all these substances mixed together (Fig. 4(b)). The Au-PCB chip exhibited no variations with the increases of the added interferential substances, indicating that the layers on top of the electrode efficiently restrict those substances from penetrating them to reach the electrode which explains the increase of current level as well as long term stability of our fabricated chips. In addition, no changes were also observed when the inferential substances were added both in time and concentration dependent manners. The amperometric response in urine was measured from the patients (n = 30) with hyperglycemia and their patterns of responses were compared with the concentration of glucose in blood measured with a commercially available glucose meter. As can be seen in Fig. 5(a), the amperometric

responses from a single chip, which are represented by black solid circle and left Y axis, have a similar pattern to the measured blood glucose (red solid square and right Y axis) suggesting that our oxyclozanide system is able to measure the level of glucose in an accurate manner as well as being stable during multiple uses in real samples. Fig. 5(b) shows the high correlation between blood glucose and glucose in urine with squared R of 0.91, which means the amount of glucose in blood is likely to be linearly correlated with the concentration of glucose in urine. In summary, we fabricated functionalized graphene oxide, which is an integration of metalloid polymer hybrids with oxidized graphene oxide nanosheets. Functionalized graphene oxide was then adsorbed on gold electrodes to form a FGO-Au-electrode. The FGO-Au-electrode chips with multiple layers were prepared by spin coating to form a multilayer-FGO-Au-electrode and then each of them was implemented on the PCBs.

Previously, DEK expression was reported to be 10-fold lower in mature hematopoietic cells as compared to immature CD34 positive cells [6]. Since four studies analyzing DEK expression in leukemia were inconclusive the aim of this study was to characterize DEK expression in a large multi-center cohort of AML cases. As an initial reference, DEK expression was profiled during normal hematopoietic differentiation of the myeloid lineage in both human

and mouse using the Hemaexplorer database [31]. Analysis of DEK expression in primary AML samples was compared to normal bone marrow using both the Microarray Innovations in Leukemia (MILE) study [32] and acute myeloid leukemia dataset (LAML) from Ley et al [33] and mapped back to the normal hematopoietic expression. This was validated and confirmed in independent cohorts of primary AML patient samples at the RNA level by

click here qRT-PCR and at the protein level by immunohistochemistry using a newly assembled AML-specific tissue microarray (TMA). Finally, DEK expression was evaluated in relation to overall survival of AML patients and prognostic relevance using the LAML dataset [33]. DEK expression during normal hematopoiesis in both human and murine models was assessed RO4929097 cost using the publicly available Hemaexplorer database (http://servers.binf.ku.dk/hemaexplorer) [31], which enabled DEK gene expression levels to be profiled in hematopoietic cells during different maturation stages based on curated microarray data. The data was analyzed using the Partek Genomics Suite v 6.6 (Partek Inc., Missouri, USA) Bay 11-7085 and GraphPad Prism 5 (GraphPad, California, USA).

All data was normalized and batch corrected. DEK expression levels in AML compared to normal bone marrow (NBM) were determined using the Affymetrix CEL files generated for the MILE study database GSE13204 [32] and the LAML dataset [33], and analyzed using Partek Genomics Suite v 6.6. ANOVA was carried out on microarray results by comparing DEK expression in NBM controls to leukemia in addition to comparison tests between NBM and specific AML subtypes. Overall patient survival associated with DEK expression was analyzed using the alternative microarray dataset LAML generated as part of The Cancer Genome Atlas (TCGA; [33]). RNA was extracted and purified from samples of 30 patients with AML (OREC 08/NIR01/9). Synthesis of cDNA was performed using the High-Capacity cDNA reverse transcription (RT) kit according to the manufacturer’s protocol (Applied Biosystems, California, USA). RT was performed using the Veriti Thermal Cycler (Applied Biosystems) at the following conditions: 25 °C for 10 min, 37 °C for 2 h, 85 °C for 5 min and a 4 °C hold period. All qRT-PCR was executed using the SYBR green mastermix (Roche) on the 7900HT Fast Real-time PCR platform (Applied Biosystems) with standard cycling conditions (95 °C for 10 min followed by 40 cycles of 95 °C for 10 s and 60 °C for 30 s).

A AGV esteve independentemente correlacionada com

níveis de 25(OH)D3, mesmo após ajustes para fatores confundidores. Em adição, os níveis séricos de 25(OH)D3 pareciam diminuir em 0,26 ng/ml por cada aumento de 10 cm2 na AGV.16 Com o aumento da longevidade mundial, tem‐se observado a emergência Natural Product Library de verdadeiras “epidemias”, tais como a síndrome metabólica, a obesidade, o DM2 e a deficiência de vitamina D. Não obstante, tem‐se tentado estabelecer relação de causalidade ou consequência entre elas. Dados nacionais provenientes de estudos em mulheres na pós‐menopausa já comprovaram ser o Brasil um país com deficiência de vitamina D, apesar de sua localização geográfica privilegiada. A frequência de excesso de peso na população superou buy Ivacaftor oito vezes

o déficit de peso entre as mulheres e em 15 vezes o da população masculina, segundo dados da 2 a etapa da Pesquisa de Orçamentos Familiares 2002‐2003 do IBGE. Constatou‐se que a obesidade entre elas cresceu 50% de 1974 a 1989 e manteve‐se estável entre 1989 e 2003. As consequências da obesidade incluem uma das mais comuns doenças crônicas em nossa sociedade, o DM2. Esse, por sua vez, é fator de risco para DCV, hipertensão arterial, doenças vasculares cerebrais, hiperlipidemia, osteoartrite e apneia obstrutiva do sono. Tais comorbidades estão presentes em muitas mulheres brasileiras na pós‐menopausa. A despeito do grande número de estudos transversais e do número limitado de trials clínicos que avaliaram as concentrações de 25(OH)D como um potencial determinante

de DCV e DM2, permanece ainda incerto se a melhoria do seu status poderia reduzir o risco dessas condições. Mediante o exposto, torna‐se necessário o conhecimento da aplicabilidade do uso de vitamina D em mulheres na pós‐menopausa e diabéticas no que diz respeito ao controle metabólico e à redução dos riscos cardiovasculares. Os autores declaram não haver conflitos de interesse. “
“Rape is an underreported heinous crime that affects women and men around the world with physical and psychological Protirelin harm, at risk of contracting infectious diseases and which may result in an unwanted pregnancy. Numerous pregnant women for rape decide late to seek their right to legal abortion up to 22 weeks of gestation.1 Besides representing a serious public health problem in developing countries, discuss the termination of unwanted pregnancy, whether as a result of rape or not, involves rethinking the legal, moral, religious, social and cultural aspects that are linked to it.2 Even nearly 7% of rape cases in Brazil result in pregnancy. Under Brazilian law, the victim of such violence has the right to abort, but 67.4% of women who have gone through this suffering had no access to legal abortion services in the public health and have just tried abortion unsafely or sought the service late.

0 ± 0.02 μL of distilled water was added. The pan was hermetically sealed and equilibrated at room temperature for 24h, then heated at the rate of 10 °C/min from 15 to 110 °C with an empty sealed pan as a reference. Parameters including onset (T0), peak (Tp), conclusion (Tc) and enthalpy (δH) were determined. Temperature at which storage modulus increased, storage modulus at the end of

heating (G′h) and storage modulus at the end of cooling (G′c) were measured with a Paar Physica Controlled Stress Rheometer (MCR 300, Gaz, Austria), buy Epacadostat equipped with parallel plate geometry. Measurements were made in the linear viscoelastic region determined in tests of constant frequency and variable amplitude. Strain and frequency were set at 0.01% and 1 Hz, respectively. The temperature of the bottom plate was controlled with a Peltier system (Viscotherm VT2, Paar Physica, Gaz, Austria), and liquid paraffin was applied to the sample’s exposed surface to prevent water evaporation. Native and extruded amaranth flour aqueous suspension selleck inhibitor (0.20 g wt) were heated from 20 °C to 90 °C at a rate of 10 °C/min, kept at 90 °C for 10 min (sufficient time to allow the storage modulus equilibrium), then cooled to 20 °C at

10 °C/min and held for 10 min at this temperature. All analyses were carried out in at least duplicate and data expressed as mean ± standard deviation employing the Statistica version7.1 software (Statsoft Inc., Tulsa, OK, USA). The proximate composition, on a dry

basis, of the native and extruded amaranth flours are depicted in Table 1. The results obtained for native flours are in agreement with those reported by previous studies on the same amaranth variety: protein at around 15 g/100 g (the nitrogen factor used was 5.85 according Selleck Gemcitabine to Berghofer & Schoenlechner, 2002), and lipid of around 7 g/100 g (Capriles, Coelho, Matias, & Arêas, 2006). Starch, fiber and ash amounts were also in accordance with Capriles et al. (2006) and Mendonça et al. (2009). The extruded flour compositions were similar to those of native flour. Thus, both mild and severe extrusion process did not significantly affect the composition of the flours. Although vitamin and mineral amounts were not determined in the present study, according to Cheftel (1986), the thermoplastic extrusion process did not reduce these nutrients. Hunter color values (L∗, a∗, b∗) of flours are shown in Table 2. Many reactions take place during extrusion cooking that may affect color. The color observed in extruded products might be due to caramelization or the Maillard reaction (Cheftel, 1986). Lysine and other amino acids present in the raw material probably react with the reducing sugars, favored by the processing conditions, which lead to darkening of the extruded products (Gutkoski & El-Dash, 1999). Luminosity (L∗ value) was decreased by the extrusion process whereas a∗ and b∗ values were increased, findings which are consistent with those of Ilo, Liu, and Berghofer (1999).

Results suggest that face-to-face administration of the TAND Checklist led to increased clarity, providing good support for the face-to-face approach when using the TAND Checklist. Examination GSI-IX mw of internal consistency suggested that the TAND Checklist has acceptable to excellent internal consistency within the domains and subdomains measured. The items from the psycho-social domain did not appear to have good internal consistency. On closer inspection, the three elements of this item include intra- and interpersonal

factors (self-esteem, family stress and parental relationship stress), where high internal consistency may not be expected. We suggest that the psycho-social domain should therefore be used simply as an introduction

to a conversation about this important level of investigation. One of the main objectives of the study was to investigate external validity of the TAND Checklist domain and subdomains. The behavioural domain items of the TAND Checklist correlated very strongly with the total difficulties score of the SDQ, suggesting that the TAND Behaviour Question may be helpful at identifying a range of behavioural difficulties that may underlie a range of psychopathologies as screened for using the SDQ. Results within the subdomain of hyperactivity also showed strong correlation between items associated with hyperactivity in the TAND Checklist and GSK126 the total hyperactivity/inattention score produced by the SDQ assessment tool, suggesting that endorsement of the hyperactivity items on the TAND Checklist should raise the clinical suspicion of ADHD or an attention-related disorder. The TAND Checklist social communication subdomain constructs

over correlated strongly with items from the SCQ, highlighting behaviours associated with autism spectrum disorders. Findings suggested that these items may be very useful markers of risk for ASD which is known to have a very high prevalence in TSC. Overall, results from the behavioural domain suggested that ADHD-related and ASD-related behaviours, two key developmental challenges in TSC, may usefully be identified through the TAND Checklist. There was a moderate correlation between the level of intellectual ability as perceived by parents and researcher judgement based on the Wessex scale. Results suggest that parental perception of intellectual development is generally reasonably accurate. Given the multi-componential nature of intelligence, all individuals with TSC are recommended to have a formal assessment of their intellectual strengths and weaknesses at key developmental timepoints.9 At the neuropsychological level, the TAND Checklist showed very strong correlation with the BRIEF.

Nas situações em que existe dominância do VHD, verifica-se a presença de AcHBe e baixa carga viral VHB, quadro este similar ao do doente apresentado, em que de facto, os dados confirmaram que o VHD tinha um papel preponderante na atividade necroinflamatória da doença. Pode no entanto existir replicação do VHB (AgHBe positivo e elevados

níveis de ADN – VHB), devendo a terapêutica nestes casos ser dirigida ao VHB2, 4, 7 and 8. Tendo em conta estes dados, e como apenas numa minoria de doentes a infecção VHD crónica se resolve espontaneamente, acompanhada BMS-907351 de perda do AgHBs (0,94%/ ano) e formação de Ac Hbs (0,27%/ano)1, as recomendações atuais aconselham o início de terapêutica dirigida ao vírus dominante em todos os doentes com doença hepática crónica VHD-VHB compensada7 and 8. Antes do início do tratamento, devem avaliar-se as características da

infecção VHB e excluir infecções concomitantes (nomeadamente, VHC e VIH). O correto estadiamento buy Alectinib da doença hepática com biopsia hepática é essencial, uma vez que não existem estudos que validem os métodos não invasivos como a elastografia hepática na avaliação do grau de fibrose nos doentes com infecção VHD4. Nos doentes com infecção crónica, em que o VHD é o vírus dominante, apenas um fármaco está atualmente recomendado: o interferão-alfa (clássico ou peguilado)4, 7 and 8. Vários estudos têm sido desenvolvidos nos últimos anos, utilizando outros fármacos nomeadamente a famciclovir, a ribavirina, o adefovir, a lamivudina e a clevudina, que não evidenciaram qualquer vantagem4. Num

estudo efectuado em Itália por Farci et al., a terapêutica com INFα na dose de 9 MUI, 3x/semana, por um período de 1 ano, foi mais eficaz que o placebo e que o INFα na dose de 3 MUI, 3x/semana, na normalização das aminotransférases. No entanto, o tratamento mostrou-se ineficaz na manutenção da resposta virológica sustentada (RVS)2 and 4. Apesar disso, no seguimento destes doentes, 10 anos depois, os autores observaram melhoria histológica no primeiro grupo de doente (INFα – 9 MUI), tendo inclusive ocorrido casos de Microtubule Associated inhibitor completa regressão da fibrose e da cirrose hepática10. Posteriormente, 2 estudos publicados em 2006 avaliaram a eficácia do PegINFα- 2b no tratamento da hepatite crónica VHD. A terapêutica mostrou RVS de 17 e 43%. Em 2007, Wedmeyer et al., estudaram a eficácia do PegINFα-2a tendo demonstrado uma taxa de resposta sustentada de 23% após 48 semanas de tratamento2 and 4. As orientações internacionais são consensuais, no que diz respeito ao único tratamento aprovado para a infecção crónica VHD: interferão clássico ou peguilado. A sociedade europeia recomenda o doseamento do ARN-VHD às 24 semanas para avaliação da resposta ao tratamento7 and 8. Ambas as sociedades referem que a duração da terapêutica deverá ser de 1 ano.

3) [6]. Several studies were reported on ultrasound perfusion imaging in healthy volunteers using perfusion weighted

MRI as reference for ultrasound perfusion imaging (Contrast Burst and Time Variance Imaging as well as high MI harmonic imaging) [5] and [10]. In these studies the time to peak intensity and in one study [5] the area under the time–intensity curve of ultrasound perfusion imaging showed a good correlation to the time to peak intensity as measured in perfusion weighted MRI. In most clinical studies on ischemic stroke patients contrast bolus kinetics was analyzed using different high MI harmonic imaging modalities (harmonic imaging, power modulation, and pulse inversion imaging). Levovist™, Optison™, and SonoVue™ were used AZD2281 research buy as contrast agents [12], [13], [14], [15] and [16]. With new, more sensitive multi-pulse ultrasound technologies it is possible to analyze brain perfusion not only in the ipsilateral but also in the contralateral hemisphere within one Cyclopamine investigation improving the geometry of the insonation plane and overcoming near-field artifacts [16]. When using this approach, additional artifacts (calcification of pineal gland and choroid plexus of lateral ventricles causing shadowing artifacts) have to be considered. In recent low MI real time refill kinetics studies [17] and [18] perfusion deficits in acute ischemic stroke patients could

be visualized qualitatively with high sensitivity in the ipsilateral hemisphere. The maximal area without detectable contrast signal correlates with the severity of stroke symptoms [17]. Besides this, quantitative thresholds for the occurrence of ischemia were calculated (β < 0.76 and A × β < 1.91 [18]). Different parameters of the bolus kinetics curve acquired from ischemic brain regions in the acute phase of stroke were compared with follow-up CT to visualize the infarction. A combination Hydroxychloroquine in vitro of the peak intensity increase (PI) and time-to-peak (TTP) proved to be most helpful in detecting the area of infarction, with a sensitivity between 75% and 86% as well

as a specificity between 96% and 100% [13] and [15]. In more recent studies color-coded parametric images were evaluated [12] and [19]. They provide information on the time–intensity data in all pixels under evaluation, thus facilitating the visualization of the perfusion state [19]. Although the supplying artery was found patent by color-coded duplex, in 13–14% of acute ischemic stroke patients a perfusion deficit in the middle cerebral artery territory could be identified with parametric perfusion imaging [13] and [19]. The areas of disturbed perfusion in the parametric images (especially the PPI image) correlate with the area of infarction in follow-up CT and the severity of stroke symptoms in the early phase as well as after four months [16].

, 2008).

In the present study, the VITROCELL® 24 air–liquid exposure system (VITROCELL® Systems GmbH, Waldkirch, Germany) was investigated in combination with the comet assay to assess DNA damage in 2 human lung cell lines, human lung adenocarcinoma cells (A549) and human bronchial epithelial cells (BEAS-2B), after exposure to WS. While similar WS exposure systems have been successfully used with human bronchial epithelial cell lines (Fukano et al., 2006, Massey et al., 1998 and Wolz et al., 2002) the VITROCELL® 24 has the added advantage of enabling exposure to multiple doses of WS within the same plate in a single run because it uses 24-well plates. Results showed a repeatable and reproducible dose–response relationship between DNA learn more damage and increased WS dose in both cell lines, demonstrating that the combination of the comet assay with the VITROCELL® 24 is a valuable new in vitro test system to screen and assess DNA damage in human lung cells exposed to cigarette smoke. Human lung cell lines A549 and BEAS-2B were exposed to diluted WS from the Reference Cigarette 3R4F in the VITROCELL® 24 and DNA damage was evaluated using the comet assay. Five

independent biological assay replicates were performed per cell line: 3 assays on the same day and 2 assays on 2 different days. The cells from 4 wells per dilution, per plate, were run in triplicate (cells split on 3 slides) for each independent assay and both intra-day and inter-day variability were assessed. An overview of the study design is presented in Fig. 1. A549. Lapatinib concentration cells and BEAS-2B cells (American Type Culture Collection, Manassas, VA, USA; number CCL-185™ and CRL-9609™, respectively) were cultured in Dulbecco’s modified Eagle medium containing 10% fetal bovine serum at 37 °C in a humidified incubator with 5% CO2 in air and 85% relative humidity. Cultures were screened for the presence of mycoplasma contamination using the Myco Alert Mycoplasma Detection Kit

(Lonza, Rockland, ME, USA). Cigarettes (University of Kentucky Reference Cigarette 3R4F; total particulate matter yield approximately 10 mg/cig) were smoked on the VC 10 smoking robot (Fig. 2A) in conformity with the International Organization for Standardization 5-Fluoracil concentration (ISO) smoking regimen (ISO, 2000). Two subsequent runs of 5 cigarettes were performed for each exposure, the smoking run was stopped after 7 puffs, and the overall exposure time was 14 min. Fresh WS (5 puffs per minute × 35 ml = 175 ml/min) from 5 cigarettes was passed puff-wise through the dilution system (Fig. 2B) and diluted with at least 5 different velocities of humidified synthetic air (SA; 85% nitrogen and 15% oxygen; Praxair, Düsseldorf, Germany). Dilution velocities ranged from 4 l/min to 0.2 l/min (from low to high smoke concentrations; Table 1).

pylori urease activates platelets through a lipoxygenase-mediated pathway, leading to ADP exocytosis and, therefore, platelet aggregation ( Wassermann et al., 2010). In this study we aimed to evaluate the participation of H. pylori urease in the acute inflammatory process promoted by CAL-101 molecular weight this bacterium. For that purpose we worked with a purified recombinant HPU (rHPU) produced in Escherichia coli. Our results showed that rHPU induces: (i) mouse paw edema; (ii) human neutrophil migration; (iii) protection of human neutrophils against apoptosis; (iv) production of

reactive oxygen species by neutrophils, and (v) induction of expression of lipoxygenase(s) in human neutrophils. H. pylori recombinant urease (rHPU) was produced by heterologous expression ( McGee et al., 1999) in E. coli SE5000 transformed with plasmid pHP8080, kindly provided by Dr. Harry T. Mobley, University of Michigan Medical School. rHPU was purified from bacterial extracts as previously described ( Wassermann et al., 2010). For the experiments, the purified protein was concentrated using Centriprep cartridges (30 kDa cut-off) to give a 0.5 mg protein/mL solution ( Suppl. Fig. 1) and dialyzed against 20 mM sodium phosphate,

pH 7.5, 1 mM EDTA, 5 mM 2-mercaptoethanol. The buffer from the last dialysis change was used as a negative control in all bioassays. Protein content of samples was determined by their absorbance at 280 nm, or by the APO866 purchase Coomassie

dye binding method (Bradford, 1976). Urease activity was measured colorimetrically by the alkaline nitroprussiate method (Weatherburn, 1967). For studies of urease inhibition, protein solutions were incubated overnight at 4 °C with 1 mM p-hydroxy-mercurybenzoate followed by extensive Tideglusib dialysis to remove excess of inhibitor (Follmer et al., 2001). Neutrophils were isolated from EDTA (0.5%)-treated peripheral venous blood of healthy human volunteers by Percoll gradient (Coelho et al., 2004) and suspended in RPMI medium (97% of viable cells, as assessed by trypan blue exclusion). Residual erythrocytes were removed by hypotonic lysis. Chemotaxis was assayed in 48-well microchemotaxis chambers (NeuroProbe, Gaithersburg, MD) using 5-μm polycarbonate filter (Coelho et al., 2004). Neutrophils (106 cells/mL in RPMI, 0.01% bovine serum albumin, BSA) were allowed to migrate toward formyl-methionyl-leucyl-phenylalanine (fMLP, 100 nM), rHPU (10 nM, 30 nM, 100 nM) or medium (random migration; 37 °C, 5% CO2). After 1 h, filters were removed, fixed, stained, and neutrophils that migrated through the membrane were counted under a light microscope on at least 5 randomly selected fields (Coelho et al., 2004). To evaluate the participation of 5-lipoxygenase products, cells were treated with AA861 (10 μM) for 15 min prior to stimulation with rHPU. Each treatment was assayed in triplicate. Results are expressed as mean ± S.E.M.