As can be seen in Fig 2B, the replicates of both 2-nitro-1,4-phe

As can be seen in Fig. 2B, the replicates of both 2-nitro-1,4-phenylendiamine GSI-IX clinical trial group closely together in a 2D Sammon projection, indicating a strong robustness and reproducibility of the assay. If triplicate samples

of any one stimulation end up on both sides of the hyperplane, it should be regarded as a sensitizer. Indeed, while the cutoff of a sample being a sensitizer or a non-sensitizer is currently set to zero, this cutoff should and will be evaluated in connection with pre-validation of the assay. Furthermore, a sample being ambiguously classified by the SVM is likely a weak sensitizer, as the absolute value of the decision value may be correlated to the potency of the sensitizer; the further away from the cloud of negative samples a sensitizer is positioned, the higher its potency

as a sensitizer, as discussed in (Johansson et al., 2011). Prediction of a compound’s ability to induce skin sensitization is an important aspect of safety assessment of chemicals, and is currently performed with animal models, such as the murine LLNA. However, a number of factors, such as the REACH legislation and the 7th amendment to the Cosmetics Directive, buy Ibrutinib make animal models unsuitable for assessment of sensitization. Furthermore, these assays are known to not correlate perfectly with clinical experience of human data. Indeed, the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) reported the accuracy of the LLNA to be 72% (Haneke et al., 2001). Genomic allergen rapid detection, GARD, is a novel assay for assessment of sensitization. It is based on a genomic readout,

measuring 200 transcripts in the myeloid cell line MUTZ-3 following compound stimulation. Lepirudin The 200 transcripts, collectively called GARD Prediction Signature, participate in signaling pathways that are involved with recognition of foreign substances. A number of these pathways, such as nuclear factor-erythroid-related factor 2 (NRF2) mediated oxidative response, aryl hydrocarbon receptor (AHR) signaling and Toll-like receptor (TLR) signaling, are known to lead to transcription of cytoprotective enzymes and DC maturation (Johansson et al., 2011) as a response to xenobiotic challenges. Thus, GARD utilizes human MUTZ-3 as an in vitro DC model, taking advantage of its decision-making role in the immune response leading to skin sensitization for predicting sensitizing potency in unknown chemicals. As a consequence of being an assay with a biomarker signature as readout, simultaneously monitoring a number of different cell events, GARD is well suited to detect positive compounds from a wide chemical space. The assay has been shown to be robust and highly reproducible, as well as accurate, with respect to the 38 reference compounds run so far.

Most of these downfalls come from the CFPs inherent top-down appr

Most of these downfalls come from the CFPs inherent top-down approach. The EU has acknowledged the need click here for a regionalization of the CFP, where a greater involvement of stakeholders should be encouraged [21]. The application of collaborative policies, such as co-management, could potentially improve EU fishery policy. The gooseneck barnacle (Pollicipes pollicipes) fishery in the Asturian coast

(North Spain) is currently an important component of the artisanal fleet in this area [27]. In 1994, a co-management system was implemented in the Asturian gooseneck barnacle fishery, which continues to date. According to informal observations, co-management has enabled the sustainability of the system. However, an in-depth study of the system has not been attempted. Here, the implementation and development of this co-management system are explored. Co-management has allowed for an adaptive learning-based approach and a fine-scale management of the fishery (down to 3 m; Fig. 1), thereby endorsing the match of social, biological and management scales. Thus, the co-management system aids in Inhibitor Library the sustainability of the gooseneck barnacle fishery. The illustration of the Asturian gooseneck barnacle system provides insights about the potential for

co-management implementation and its prospects as a management approach in a broader European context. The Asturian co-management

system is located between the Eo estuary (29T 666839 4827388 UTM) and the eastern most part of Cape Peñas (29T 667714 4827400 UTM). It is divided in 7 regions with distinct management, denominated management plans for their Spanish name, which depend on the regional government (Principado de Asturias) and the local fishers׳ associations known as cofradías ( Fig. 1). Currently, the Tapia-Figueras, Viavélez, Ortiguera, Puerto de Vega, Luarca, Cudillero-Oviñana plans are seasonal with a harvest season that starts in October and ends in April, and a total individual daily allowable catch (TAC) per fisher that varies between 6 and 8 kg. However, the Cabo Peñas plan, which comprises the Luanco-Bañugues cofradías, allows harvesting all year with a constant Inositol monophosphatase 1 daily TAC of 8 kg per fisher. The distribution and dimension of the Asturian gooseneck barnacle co-management plans was characterized using the Principado de Asturias Coastal and Marine Geographic Information System. Each co-management plan is subdivided into management zones, which can be separate rocks, groups of rocks, or small coastal strips. Furthermore, information on the commercial quality of each zone was gathered from the Dirección General de Pesca Marítima del Principado de Asturias (DGPM) official records.

Several bands can be viewed in the range of 1700–600 cm−1 The wa

Several bands can be viewed in the range of 1700–600 cm−1. The wavenumber range of 1400–900 cm−1 is characterized by vibrations of several types of bonds, including C–H, C–O, C–N and P–O (Sablinskas et al., 2003 and Wang et al., 2009). Other studies on FTIR analysis of roasted coffees (Briandet et al., 1996 and Kemsley et al., 1995) have reported that carbohydrates exhibit several absorption bands in this region, so it is expected

that this class of compounds will contribute to several of the observed bands. According to Kemsley et al., 1995 and Briandet et al., 1996, and Lyman et al. (2003), chlorogenic acids also present absorption in the region of 1450–1000 cm−1. Chlorogenic acids represent a family of esters formed between quinic acid

and one to four residues ERK inhibitor of certain trans-cinnamic acids, most commonly caffeic, p-coumaric and ferulic ( Clifford, Kirkpatrick, Kuhnert, Roozendaal, & Salgado, 2008). Axial C–O deformation of the quinic acid occurs in the range of 1085–1050 cm−1, and O–H angular deformation occurs between 1420 and 1330 cm−1. The C–O–C ester bond also absorbs in the 1300–1000 cm−1 range ( Silverstein, Webster, & Kiemle, 2005) and therefore the bands located in the range of 1450–1050 cm−1 could be partially due to chlorogenic acids. Hashimoto et al. (2009) studied the influences of coffee varieties, geographical origin and of roasting degree on the mid-infrared spectral characteristics of brewed coffee, and also developed a fast and reliable procedure to determine the Histidine ammonia-lyase caffeine and chlorogenic acid contents in brewed coffee using the ATR-FTIR method. In their method, developed based on the spiking of the coffee brew with different amounts of caffeine, they identified the band at 1242 cm−1

as the most relevant absorption band for characterization of the caffeine content in the brew. In the roasted and ground coffee IR spectra herein obtained for defective and non-defective coffee beans this peak appears shifted to a slightly lower band (1238 cm−1), but it is present in all spectra. Another substance that can be associated to peaks in the 1600–1300 cm−1 range is trigonelline, a pyridine derivative that has been reported to present four bands in this range, due to axial deformation of C C and C N bonds ( Silverstein et al., 2005). A comparison of the average spectra of green and roasted coffees presented in Fig. 2b shows a decrease in the relative absorbance of several bands in the 1700–600 cm−1 region after roasting. Several literature reports confirm that the levels of carbohydrates, trigonelline and chlorogenic acids diminish upon roasting ( Farah et al., 2006 and Franca et al., 2005), so such variations in chemical composition are expected to affect the spectra in the 1700–600 cm−1 range.

, 2005, Rodionova and Panov, 2006 and Janas and Zgrundo, 2007) C

, 2005, Rodionova and Panov, 2006 and Janas and Zgrundo, 2007). Cladocera make up a significant part of the Baltic zooplankton both in numbers and in biomass, especially in summer. Since the early 1990s, the list of cladocerans has been extended by the Ponto-Caspian crustaceans Cercopagis pengoi, Cornigerius maeoticus and Evadne anonyx ( Ojaveer and Lumberg, 1995, Krylov et al., 1999, Panov et al., 1999, Rodionova et al., 2005 and Rodionova

and Panov, 2006). In the Polish coastal zone, and that includes the Gulf of Gdańsk, only C. pengoi has been recorded so far ( Bielecka et al., 2000, Duriš et al., EPZ5676 supplier 2000, Bielecka et al., 2005, Olszewska, 2006 and Bielecka and Mudrak, 2010). Evadne anonyx is an endemic species from the Ponto-Caspian Entinostat mw basin ( Mordukhai-Boltovskoi 1995). Its author classified it among the Caspian Polyphemoidae, which make up the Podonidae group. This marine species, originating from the tertiary period, occurs in shallow water plankton ( Mordukhai-Boltovskoi 1995). The environmental preferences of E. anonyx from the Caspian Sea were described by Aladin (1995), who stated that the salinity and temperature tolerance ranges for E. anonyx were from 4 to as much as 30 PSU and from 11.4 to 26.4° C respectively. That author found that this species, which used to be more

widespread, was forced to abandon the Aral Sea because of increasing salinity, and the Sea of Azov and Black Sea because of growing contamination. The first published report of E. anonyx

in the Baltic Sea, from the Gulf of Finland, related to August 2004 ( Litvinchuk 2005). According to Rodionova & Panov (2006), however, the first specimens of this species were found in the Primorsk oil terminal area in the Gulf of Finland four years earlier. This information was again corrected, this time by Põllupüü et al. (2008), who found that E. anonyx had been observed in the central Gulf of Finland (Tallinn Bay) as from early as 1999. The aim of the present work was to report the first signs of the invasion of the Gulf of Gdańsk by E. anonyx G. O. Sars 1897 and to describe the extent of its range there in 2006. Plankton material was collected in the Gulf of Gdańsk from February to December 2006. The samples were taken from the eastern (Krynica Morska profile – K1–K4, Świbno profile – Sw2–Sw4) and western (Mechelinki station – M2, Sopot profile – So1–So4 and J23) parts of the gulf (Figure 1, Table 1). Hauls were made to a maximum depth of 40 m using a closing Copenhagen plankton net (mesh size 100 μm) from the vessel ‘Oceanograf 2’. The biological material was preserved in 4% formaldehyde solution. The overall zooplankton community analysis was done in the laboratory. All individuals of Evadne anonyx were separated according to the characteristics outlined by Rivier (1998): the number of setae on the exopodites of thoracic limbs I–IV – respectively – and the rounded shape of the cauda.

1), 600 mM KCl, 10 mM MgCl2, 2 mM EGTA, 1 mM EDTA, 1% Triton X-10

1), 600 mM KCl, 10 mM MgCl2, 2 mM EGTA, 1 mM EDTA, 1% Triton X-100 and the protease inhibitors described above. The homogenate was centrifuged at 15,800 × g for 10 min at 4 °C, in an Eppendorf centrifuge, the supernatant discarded and the pellet homogenized with the same volume of the high salt medium. The resuspended homogenate was centrifuged as described and the supernatant

was discarded. The Triton-insoluble IF-enriched pellet, containing NF subunits, Vim and GFAP, was dissolved in 1% SDS and protein concentration was determined. The cytoskeletal fraction was prepared as described above. Equal protein concentrations Ipilimumab price were loaded onto 10% polyacrylamide gels and analyzed by SDS-PAGE. After drying, the gels were exposed to T-MAT films at − 70 °C with intensifying screens and finally the autoradiograph was obtained. Cytoskeletal proteins were quantified by scanning the films with a Hewlett-Packard Scanjet 6100C scanner and determining optical densities with an Optiquant version 02.00 software (Packard Instrument Company, Meriden, CT 06450 USA). Density values were obtained for the studied proteins. Tissues slices were homogenized in 100 μl of a lysis solution containing 2 mM EDTA, 50 mM Tris–HCl, pH 6.8, 4% (w/v) SDS. For electrophoresis analysis, samples were dissolved in 25% (v/v) of solution containing 40% glycerol, 5% mercaptoethanol, 50 mM Tris–HCl, Trichostatin A clinical trial pH 6.8 and boiled for

3 min. Protein homogenate (80 μg) was analyzed by SDS-PAGE and transferred to nitrocellulose membranes (Trans-blot SD semi-dry

transfer cell, BioRad) for 1 h at 15 V in transfer buffer (48 mM Trizma, 39 mM glycine, 20% methanol and 0.25% SDS). The nitrocellulose membranes were washed for 10 min in Tris-buffered saline (TBS; 0.5 M NaCl, 20 mM Trizma, O-methylated flavonoid pH 7.5), followed by 2 h incubation in blocking solution (TBS plus 5% bovine serum albumin and 0.1% Tween 20). After incubation, the blot was washed twice for 5 min with TBS plus 0.05% Tween-20 (T-TBS), and then incubated overnight at 4 °C in blocking solution containing the following antibodies: anti-GFAP (clone G-A-5) diluted 1:500, anti-vimentin (Vim 13–12) diluted 1:400, anti-NF-L (clone NR-4) diluted 1:1000, anti- NF-M (clone clone NN-18) diluted 1:400, anti-NF-H (clone N52) diluted 1:1000, anti-ERK1/2 diluted 1:1000, anti-phosphoERK diluted 1:1000, anti-/JNK diluted 1:1000, anti-phosphoJNK (clone 98F2) diluted 1:1000, anti-p38MAPK (clone A-12) diluted 1:1000, anti-phosphop38MAPK diluted 1:1000, anti-PKAcα diluted 1:1000, anti-PKCaMII diluted 1:500, anti-AKT (clone 2H10) diluted 1:1000, anti-phosphoAKT (clone 244F9) diluted 1:1000, anti-active caspase 3 diluted 1:1000, anti-GSK3β (clone 27C10) diluted 1:1000, anti-phosphoGSK3β, anti-KSP repeats (clone NP1) diluted 1:1000, anti-phoshoNF-LSer55 diluted 1:800, anti-phosphoNF-LSer57 diluted 1:1000 or anti-actin diluted 1:1000.

Cells were washed and resuspended in RPMI 1640 medium supplemente

Cells were washed and resuspended in RPMI 1640 medium supplemented with 20% fetal bovine serum, 2 mM glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin, at 37 °C under 5% CO2. Phytohemagglutinin (4%) was added at the beginning of culture. After 24 h of culture, PBMC were treated with the test substances. The alkaline comet assay was performed as described by Singh et al. (1988) with minor modifications (Hartmann and Speit, 1997), and following the recommendations of the International Workshop on Genotoxicity Test 17-AAG cell line Procedures (Tice et al., 2000). At the end of the treatment, cells

were washed with ice-cold PBS, detached with 100 μL trypsin (0.15%) and resuspended in complete RPMI medium. Next, 20 μL of cell suspension (∼106 cells/mL) were mixed with 100 μL of 0.75% low melting point agarose and immediately spread onto a glass microscope slide precoated with a layer of 1% normal melting point agarose. Agarose was allowed to set at

4 °C for 5 min. The slides were incubated in ice-cold lysis solution (2.5 M NaCl, 10 mM Tris, 100 mM EDTA, 1% Triton X-100 and 10% DMSO, pH 10.0) at 4 °C for a minimum of 1 h to remove cellular proteins, leaving the DNA as ‘‘nucleoids’’. After the lysis procedure, the slides were placed on a horizontal electrophoresis unit. The unit was filled with fresh buffer (300 mM NaOH MAPK inhibitor and 1 mM EDTA, pH > 13.0) to cover the slides for 20 min at 4 °C to allow DNA unwinding and expression of alkali-labile sites. Electrophoresis was carried out for 20 min at 25 V and 300 mA (0.86 V/cm). After

electrophoresis, the slides were neutralized (0.4 M Tris, pH 7.5), stained with ethidium bromide (20 μg/mL) and analyzed using a fluorescence microscope. All the above steps were conducted under yellow light or in the dark to prevent additional DNA damage. Images of 100 randomly selected cells (50 cells from each of two replicate slides) were analyzed for each concentration of test substance. Cells were scored visually and classified PDK4 in 5 grades according to the tail size (from undamaged-0 to maximally damaged-4), and a damage index value was calculated for each sample of cells. Damage index thus ranged from 0 (completely undamaged: 100 cells × 0) to 400 (with maximum damage: 100 cells × 4) (Collins, 2004). The frequency of tailed cells, a DNA damage frequency indicator, was also calculated based on the number of cells with or without tails. In order to determine differences among treatments, data were compared by one-way analysis of variance (ANOVA) followed by the Newman–Keuls test (p < 0.05) using the Graphpad program (Intuitive Software for Science, San Diego, CA). All studies were carried out in triplicate represented by independent biological evaluations. The indirect inhibitory growth effects of α-santonin derivatives (2–4) on HL-60 cells were determined by MTT assay in a previous study (Arantes et al., 2010, 2009).

5 3 1) is reported to be about pH 10 in horse, pH 9 8 in rat and

5.3.1) is reported to be about pH 10 in horse, pH 9.8 in rat and pH 11 in Bacillus brevis. Those working with mammalian systems might favour an assay pH of about 7.2 which is believed to be around the physiological pH within the cell, but clearly this would be unphysiological for gastrointestinal enzymes, such, as pepsin and trypsin, or for lysosomal enzymes. Furthermore, the oxidation of ethanol by liver alcohol dehydrogenase (EC is often followed at higher pH values because the equilibrium of the reaction greatly favours ethanol formation at neutral

pH. Naturally it would be appropriate to use physiological substrates for enzyme assays. However, many studies have used Selleck Cabozantinib unphysiological substrates for ease of manipulation and assay. For

example acetylthiocholine is frequently used to assay acetylcholinesterase (EC because the thiocholine produced can be readily detected by reaction with sulfydryl reagent 5, 5′-dithiobis-2-nitrobenzoate (Nbs2) releasing a yellow coloured compound whose formation can be followed spectrophotometrically at 412 nm (Ellman et al., 1961). Other examples include the use of 4-nitrophenyl phosphate to assay alkaline phosphatase (EC (Schumann et al., 2011). The use of synthetic dyes as electron acceptors in oxidoreductase assays has been common and in some cases the physiological acceptor remains unknown. The demand for higher assay sensitivity and high-throughput procedures has resulted in the development of an increasing number of chromogenic and fluorogenic substrates (Goddard and Reymond, 2004 and Reymond et al., 2009). Clearly, in such cases a considerable amount of work would be necessary to show whether the enzyme behaves identically towards such substrates as it does towards its physiological substrates. It is often recommended that saturating substrate concentrations should be used (i.e. >10Km, for all substrates), as discussed Adenosine triphosphate by Bisswanger

(2014). This, of course, assumes that the Km values have already been determined, at least approximately. Furthermore, this might not always be practicable because of factors such as solubility, the occurrence of high-substrate inhibition or a high absorbance of the assay mixture affecting the behaviour of optical assays ( Dixon et al., 1979 and McDonald and Tipton, 2002). It should also be remembered that any change in the assay conditions (e.g., pH, temperature, ionic strength) may affect the Km values. The buffers and ionic strengths and used in enzyme assays vary widely and are often far from physiological. It might be helpful if it were possible to recommend a simple standard buffer for use in all enzyme assays. Unfortunately, this goal appears to be unobtainable, because at least some enzymes are unhappy in one or other of the common buffers (see e.g., Boyce et al., 2004).

, 2008) and in vitro and in vivo neurotoxicity of Bothrops venoms

, 2008) and in vitro and in vivo neurotoxicity of Bothrops venoms and their PLA2 ( Gallacci and Cavalcante, 2010). Finally, there is the possibility of species differences (bird/rodent vs. human) in the neuromuscular responses to B. b. smargadina venom. Given the arboreal nature of B. b. smargadina and the potency of its venom in avian preparations, this venom may be particularly adapted for dealing with avian prey. The authors thank Dr. Ronaldo Navarro Oviedo (Laboratory of Biological Chemistry,

Academic and Biological School of the National University of San Agustín, Arequipa, Peru) for providing the venom and Gildo B. Leite for technical assistance. This work was supported by Conselho Nacional de Wnt inhibitor Desenvolvimento Científico e Tecnológico (CNPq, Brazil) and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, Brazil). L.R.S. and S.H. are supported by research fellowships from CNPq. “
“The construction of the Itaipu dam complex in the basin of the Alto Paraná river on the border between Brazil and Paraguai submerged the

Seven Falls of Guaira, which were a natural barrier that impeded the dispersion of several species of fishes, including stingrays, to the upper end of the river (Garrone Neto et al., 2007). As a result, Potamotrygon stingrays, whose habitat was originally the basin of the Alto Paraná river, migrated upstream and colonized different regions of its upper reaches. Consequently, the region of Três Lagoas in the Brazilian State of Mato Grosso do Sul, that was once devoid of stingrays, is now overpopulated by Potamotrygon spp. (Potamotrygon falkneri, Potamotrygon motoro and Potamotrygon schuhmacheri) which cause a considerable number of accidents in the riverside population

( Garrone Neto et al., 2007 and Garrone Neto and Haddad, 2009). The local injury caused by these stingrays is due to mechanical penetration of the sting into the tissue and subsequent release of venom leading to the development of local edema, necrosis, intense local pain and cases of secondary infection (Meyer, 1997, Haddad, 2000, Pardal, 2003, Haddad et al., 2004, C-X-C chemokine receptor type 7 (CXCR-7) Barbaro et al., 2007, Garrone Neto and Haddad, 2009 and Dehghani et al., 2010). It is postulated that the local inflammatory reaction and necrosis in freshwater stingray accidents is due to the release into the wound of several proteins with enzymatic activity produced by the protein secretory cells that covers the sting (Barbaro et al., 2007, Pedroso et al., 2007, Magalhães et al., 2008 and Antoniazzi et al., 2011). The protein secretory cells are overlaid by a fin layer of mucus which also covers the entire surface of the stingray and separates the cutaneous tissue from direct contact with the environmental water (Pedroso et al., 2007).

cloacae and C luteola were the most prevalent species amongst is

cloacae and C. luteola were the most prevalent species amongst isolates of Enterobacteriaceae and Pseudomonaceae from both groups. E. cloacae has been frequently identified amongst clinical oral isolates. 11, 12 and 26 However, C. luteola is rarely isolated from the oral cavity, and infections caused by FAK inhibitor this microorganism include sepsis, meningitis, endocarditis, osteomyelitis and peritonitis. 20 and 38 The HIV group showed the greatest diversity of Enterobacteriaceae and Pseudomonas species, many of which can cause opportunistic infections, such as

Escherichia coli infections, gastro-intestinal infections, 39 endocarditis, 40 urinary infections 41 and Klebsiella pneumonia infections that are often involved with aspiration pneumonia. 42 With respect to CD4 cell count, lower counts of enterobacteria and pseudomonas were observed amongst patients in the subgroup with <200 CD4 cells/mm3. This unexpected result warrants further study. The viral load values were distributed equally amongst the subgroups evaluated. Considering that the studies reporting the prevalence of enterobacteria and Pseudomonas in the oral cavities of HIV-positive patients did not correlate their findings with clinical variables, it was not possible to compare our results to the literature. However, increased oral carriage of Gram-negative

bacilli in the HIV group compared to the control group confirms that carriage rates tend to increase in medically compromised individuals, and the presence of such bacteria in the oral cavity as a putative reservoir of infection should be considered. CTLA-4 inhibitor 43 Regarding treatment, it was not possible to have a fixed therapy protocol for all patients because the treatment strategy would change according to the CD4 lymphocyte level and viral load as well as other relevant clinical variables, such as the occurrence of opportunistic diseases and adverse reactions to medication. HAART is correlated

with lower occurrence of oral diseases. It promotes inhibition of viral replication as well as redistribution and restoration of immunity, resulting in an increase in CD4 cell counts. Within the limits of this study, we could not assess the effect of HAART on the oral microflora. The few studies on this subject report that about protease inhibitors may have anti-Candida activity by inhibiting the protease of this microorganism. 1 and 44 Based on the results obtained, it may be concluded that the HIV-positive group showed a higher prevalence of Enterobacteriaceae and Pseudomonadaceae. No difference in staphylococcus counts was found between the studied groups. Funding: Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP (2004/12382-6). Conflict of interest statement: No conflict of interests. Ethical approval: This study was approved by the Local Ethics Committee (protocol number 012-PH/CEP).

, 1990, Azevedo et al , 2002, Leal and Soares, 2004, Falconer and

, 1990, Azevedo et al., 2002, Leal and Soares, 2004, Falconer and Humpage, 2005, Andrinolo et al., 2008 and Funari and Testai, 2008). Independently of the exposure route MCYST-LR preferentially reaches the liver and can also be detect in several organs including lungs (Wang et al., 2008). Recently, our group reported the use of an anti-inflammatory RG7420 research buy drug candidate, LASSBio 596, to treat the pulmonary damage induced by the acute exposure to MCYST-LR. This compound was designed as an agent that modulates TNF-α and inhibits phosphodiesterases (PDEs) (Lima et al., 2002). Briefly, the intraperitoneal administration of LASSBio 596 avoided most of the pulmonary

structural and functional damages, exhibiting a better outcome than dexamethasone. However, both AZD1208 cell line treatments were not effective to avert the liver structural damage (Carvalho et al., 2010). Even though the pharmacokinetics of LASSBio 596 has been described (Rocco et al., 2010), its therapeutic effects on by oral administration are so far unknown. Considering that the intraperitoneal route is not often used in clinical practice and that the treatments did not show effective for liver changes,

an investigation about the therapeutic effects of orally administered LASSBio 596 on pulmonary and hepatic changes seems relevant and could establish the potential of LASSBio 596 as a drug candidate for the treatment of the systemic damage induced by microcystin-LR. Thus, in the present study, we aimed to evaluate the efficacy of LASSBio 596 per os in the treatment of pulmonary and hepatic damage in mice acutely exposed to MCYST-LR. For such purpose pulmonary mechanics, morphology and inflammatory cells influx, as well as the levels of pro-inflammatory mediators both in lungs and liver tissues, were assessed. The present study was approved by the Ethics Committee of the Health Sciences Center, Federal University

of Rio de Janeiro (Protocol IBCCF 012). All animals received humane care according to the “Principles of Laboratory Animal Care” formulated by the National Society for Medical Research and the “Guide for the Care and Use of Laboratory Animals” by the National Academy of Sciences, USA. Twenty-six Swiss mice (35–40 g) were purchased from the animal facilities of the University of Campinas (CMIB/UNICAMP). They were randomly divided HSP90 into 3 groups: In the control group, 40 μl of sterile saline solution (0.9% NaCl, CTRL, n = 8) were intraperitoneally (i.p.) injected, whereas in the other two groups a sub-lethal dose of MCYST-LR (40 μg/kg i.p., purified material kindly provided by Professor Wayne Carmichael, Wright State University, Dayton, OH, USA) was administered. After 6 h, CTRL, TOX (n = 8), and LASS (n = 10) mice received per os 60 μl of a solution composed by 57.5 μl of sterile saline and 2.5 μl of dimethyl sulfoxide (DMSO); in the latter group the solution contained 50 mg/kg of LASSBio 596.