We also reviewed the molecular basis of Fas-mediated apoptosis in malignant gliomas. Glioblastoma specimens from 97 patients who had not been previously treated were retrieved from the archives of the Departments of Pathology at São Paulo Federal University (n = 60) and Ribeirão Preto Medicine Faculty

at São Paulo University (n = 37). The tumor specimens were re-examined and confirmed to be glioblastomas according to the criteria of the most recent WHO Classification of Central Nervous System Tumors [22]. All of the patients had undergone surgery during the 15-year period from 1992 through 2006. This study was approved by the Ethics Committees of both institutions (Resolution No. 196 of Brazilian National Health Council). Histological sections (4 μm) were cut from each tissue block, find more stained by hematoxylin–eosin, and carefully reviewed by 3 independent pathologists. The areas most representative of each tumor were selected

for analysis. Cylindrical cores were removed and used in the construction of tissue microarray (TMA) blocks. Five TMA blocks were constructed using a Beecher tissue array instrument™ (Beecher Instruments, Silver Spring, MD, USA), according to the manufacturer’s instructions, in the following stages: (1) Two different areas of the tumor were marked in the original donor block for sampling (necrotic zones and perinecrotic palisading cells were not included in the samples), (2) cylindrical holes were created in the receptor block using the TMA platform. Positions were created in the receptor CCI-779 clinical trial blocks and were separated by approximately 500 μm such that a matrix of holes for the tissue samples was created, (3) 1-mm diameter cylinders of tissue were extracted from the areas of interest in the donor blocks using a 1-mm-diameter needle (TMArrayer Punch Beecher Instruments™), Roflumilast (4) the cylindrical tissues obtained from the donor blocks were

transferred to the holes in the receptor blocks, and (5) finally, the quality of the blocks (representativeness of the tumor samples) was assessed before storage. Twenty-five control cores obtained from normal brains harvested from 25 autopsied patients (6–12 h postmortem) were included as controls. The immunohistochemical procedures were performed on 4-μm-thick sections that were obtained from the TMA blocks and mounted on slides pretreated with 3-minopropyl-triethoxysilane (Sigma). To aid in the adhesion of the slices from the TMA blocks to the silane-treated slides, an adhesive tape system (Instrumedics Inc., Hackensak, NJ, USA) was also used. Briefly, for immunostaining, the slides were deparaffinized, and rehydrated through a graded ethanol series. For antigen retrieval, slides were placed in a 0.01 M citrate buffer (pH 6.0), heated in a steam bath for 3 min, and allowed to cool at room temperature for 30 min. Endogenous peroxidase activity was blocked using 3% hydrogen peroxide for 15 min, followed by washing in 0.05 M Tris buffer (pH 9.5).

Green et al. verificaram que a maioria dos participantes do seu estudo tinham sido diagnosticados entre a quarta e a sexta décadas de vida11. Neste estudo, a mediana da idade de diagnóstico é inferior ao reportado na literatura, o que poder-se-á dever ao método utilizado para a seleção da amostra. O

método de referência para efetuar o diagnóstico de DC continua a ser selleck screening library a avaliação histológica com biopsia intestinal5. Quando questionados acerca da realização deste exame, apenas 79% dos inquiridos afirmaram tê-lo feito, valor semelhante aos 75% encontrados num estudo realizado nos Estados Unidos da América11, mas bem inferior ao valor encontrado num estudo canadiano33. Uma limitação do presente estudo prende-se com o facto de não se ter questionado os participantes que não realizaram avaliação histológica com biopsia intestinal sobre quais os critérios ou testes realizados que estiveram na origem do seu diagnóstico. A partir do momento em que são diagnosticados com DC os indivíduos devem iniciar DIG, que deve ser mantida para toda a vida1, 6 and 35. Neste trabalho, verificou-se que apesar de 97,4% dos inquiridos tentar cumprir a DIG na sua alimentação diária, 47,7% reportaram ingerir glúten com frequência variável. Já Lamontagne et al. verificaram que, embora 90%

dos participantes evitasse tanto quanto possível a ingestão de glúten, 72% admitia consumir alimentos com o agente tóxico20. A percentagem de inquiridos selleckchem que afirmaram consumir alimentos com glúten por escolha própria neste estudo foi semelhante à observada por Lamontagne et al.: 35,4 e 36%, respetivamente20. Numa revisão sistemática recente apontava-se que as proporções de adesão estrita à DIG auto reportadas variavam

de 42-91%, sendo que fatores como a disponibilidade e o preço dos AESG, o saber interpretar a rotulagem alimentar, ter a capacidade de manter a DIG aquando de viagens, no trabalho e durante eventos sociais, contribuíam de forma positiva para o cumprimento da dieta19. Cerca de 54% dos inquiridos neste estudo referiram ter diminuído a frequência de consumo de refeições fora de casa após o diagnóstico, percentagem superior à encontrada num estudo realizado no Reino Unido – 44,2%36. Já no estudo de Lee e Newman, 86% dos participantes afirmaram que a DIG prejudicava a realização de refeições fora de casa37. Lepirudin A diminuição da frequência de refeições fora de casa terá, certamente, a ver com o facto dos inquiridos não se sentirem seguros nas escolhas alimentares, dada a natureza restritiva da DIG. É facilmente percetível que, quer pela sintomatologia associada à ingestão inadvertida de glúten quer pela dificuldade em seguir uma DIG pelas mais diversas razões, a DC possa afetar a auto perceção do estado de saúde e da qualidade de vida dos doentes celíacos. Para avaliar estes domínios usou-se a escala SF-36, comummente utilizada para relacionar qualidade de vida com desfechos de saúde.

The sponsors were not provided with a copy of this manuscript for review prior to submission, nor allowed input in the conduct or reporting of the work. The statements

in this paper are the authors’ and not those of their employers or the sponsors. The authors are or were employed by Exponent, a scientific and engineering selleck chemicals llc research and consulting firm, and have provided these services for private and government clients, including on projects involving arsenic. JST has presented on arsenic risk assessment issues in public comments to EPA and the NAS on behalf of industry and trade associations with interests in arsenic. JST has been retained in defense and plaintiff litigation cases related to arsenic. Transparency Document. The authors thank Betty Dowd, Mary Becker, Eileen McAuliffe, and Christine Shirley for graphics, editorial, and technical assistance. “
“En el artículo «Anemia ferropénica y uso de hierro endovenoso en patología digestiva» (Gastroenterol Hepatol.

2010;33[8]:605-613) de Fermín Mearin et al., se ha detectado un error en el nombre de uno de los autores. El learn more nombre correcto es: Javier P. Gisbert. “
“Peripheral neuropathy is a common adverse effect of several classes of anti-cancer drugs, including vincristine, paclitaxel, oxaliplatin, cisplatin and bortezomib (Wolf et al., 2008). These agents exert direct and indirect effects on sensory nerves to reduce the amplitude of action potential, slow conduction velocity and induce pain in patients, especially those who experience

nociceptive sensory loss during treatment. Cancer chemotherapy-induced peripheral neuropathy (CIPN), which can Vildagliptin be extremely painful, results in patient suffering and also limits the treatment with potentially useful anticancer drugs. The incidence of CIPN varies depending on the conditions with severe neuropathy (3–7%) with single agent, but can rise up to 38% with combination regimens (Connelly et al., 1996). Clinically, paclitaxel-induced neurotoxicity typically presents as a sensory neuropathy with the most common complaints being numbness, tingling and burning pain. The subjects mainly experience tingling and allodynia that often occur in a “glove and stocking” distribution. Sensory symptoms usually start symmetrically in the feet, but also appear simultaneously in both hands and feet (Dougherty et al., 2004). The most cases resolve within months after paclitaxel treatment is discontinued, but the sensory abnormalities and pain sometimes become a chronic problem (Rowinsky et al., 1993). The occurrence and severity of the neuropathy is dependent on many factors including single dose intensity, duration of infusion, cumulative dose, prior or concurrent treatment with cisplatin, and co-existing conditions such as diabetes and alcohol abuse. Oxaliplatin, a third-generation platinum-based chemotherapy drug, is a key drug in the treatment of colorectal cancer.

The fraction of the mineralised phosphorus is adsorbed to sediment particles but the rest is instantly released

to the water column. In this study, this pathway was simplified by excluding the desorption process. The model equations and parameter values are described in detail in  and . Calibration of the new N flux model and a simplified version of P flux model presented by Müller-Karulis & Aigars (2011) against median PO43−, NOx− and NH4+ flux measurements was performed using a simulated annealing routine (SANN) in statistical analysis software R v.3.0.2. ABT-199 The average fluxes of PO43− (42–115 μmol m−2 d−1) were always directed out of the sediments. Although PO43− fluxes tended to decrease with increasing O2 concentration in the near-bottom water,

they exhibited no significant differences (ANOVA; p < 0.01) among treatments, most likely due to the substantial variability of fluxes within the treatments selleck chemicals ( Figure 3). The simulated values of PO43− flux (Figure 3) are in good agreement with the median values of the experimental data set and show nearly constant maximum values (105–106 μmol PO43− m−2 d−1) at an O2 concentration range of 1–2 mg l−1 and a smooth decline with increasing O2 concentrations, reaching the lowest fluxes (57 μmol PO43− m−2 d−1) at oxygen concentrations in the range between 5 and 10 mg l−1. Sediment-water fluxes of NH4+ are always positive and exhibit large variability within and among O2 treatments, ranging on average from Dehydratase 1800 μmol m−2 d−1 at an O2 concentration of 2 mg l−1 to 140 μmol m−2 d−1 at an O2 concentration of 10 mg l−1 ( Figure 4). At this latter O2 concentration the observed fluxes vary between –734 and 528 μmol NH4+ m−2 d−1 (the highest observation

is treated as an outlier) with 90 μmol NH4+ m−2 d−1 as the median value. Although there is no significant difference in NH4+ fluxes between treatments 1 and 3, the significant differences between treatments 2 and 3 (ANOVA; p < 0.01) and 3 and 4 (ANOVA; p < 0.01) clearly demonstrate increasing NH4+ fluxes when O2 concentrations are < 4 mg l−1. Larger oxygen concentrations do not result in a further decrease of NH4+ fluxes, however. The modelled NH4+ fluxes (Figure 4) show a smooth decline with increasing concentration, reaching the lowest value (2.3 μmol NH4+ m−2 d−1) at an O2 concentration of 10 mg l−1. The model fits the data well at low (1 mg l−1) and intermediate to high (≥ 4 mg l−1) O2 concentrations, but does not correspond with the high fluxes observed at an O2 concentration of 2 mg l−1, which vary between 1051 and 2467 μmol NH4 m−2 d−1. In contrast to NH4+, NOx− fluxes are mostly directed into the sediments, although, like NH4+, these fluxes exhibit a considerable variability within and among treatments, ranging on average from –390 μmol m−2 d−1 at an O2 concentration of 1 mg l−1 to 85 μmol m−2 d−1 at an O2 concentration of 10 mg l−1 ( Figure 5).

(2012). MVPA, especially Searchlight methods (Kriegeskorte and Bandettini, 2007a, Kriegeskorte and Bandettini, 2007b and Kriegeskorte et al., 2006), should be useful for elucidating neural representation of

language switching in the functional mapping of bilingual brains. A Searchlight analysis primarily aims at identifying brain regions that carry information for the given experimental conditions, without assuming local homogeneity in activations. It enables us to decode fMRI data by focussing the analysis around a single voxel at a time, while combining the signals within a certain radius from the centred voxel to compute a multivariate effect statistic at every location (Haynes and Rees, 2006, Alink et al., 2012 and Corradi-Dell’Acqua Navitoclax mw et al., 2011; Bode et al., 2011, Gilbert, 2011, Kahnt et al., 2011, Kotz et al., 2012 and Momennejad and Haynes, 2012). Based on the methodological research regarding univariate Searchlight (Jimura & Poldrack, 2012), MVPA is more sensitive to distributed coding of information than GLM, which seems better at identifying global engagement in ongoing tasks. Therefore, MVPA might also be useful for detecting some aspects of the cortico-cortical and cortico-subcortical networks that subserve the functions in bilingual language switching, while still Epigenetics Compound Library clinical trial being sensitive to the contiguous areas of homogenous activation that

might be detected by the GLM. Hence, in the current study, we focused on highly proficient Korean–Chinese early bilinguals (Bai et al., 2011) by using language-switching tasks with written stimuli to explore the neural basis of their bilingual behaviour. We also considered

the Age of Acquisition and the language proficiency of the bilinguals. The tasks were subdivided into two-day sessions with different levels Phenylethanolamine N-methyltransferase of difficulty: situational non-translation language switching condition (abbreviated as ‘SnT’) and focused simultaneous translation language switching condition (abbreviated as ‘FST’). The SnT refers to the conventional language switching task used in previous studies in which subsequent trials switch from L1 to L2 and vice versa, without interlingual translation being required within a trial. In the FST condition, switching is required within the trial, and the direction of translation is randomly varied from trial to trial. We applied the univariate Searchlight and GLM in a complementary manner as methods to identify the informative regions of fMRI activity for different types of language switching. Our findings from Korean–Chinese early bilinguals, especially under the focused simultaneous translation language (FST) condition, supported the new ‘hodological’ view of language switching by detecting several regions of interest that play important roles in the network for executive control and in the cortico-subcortical sub-networks (Abutalebi and Green, 2008 and Moritz-Gassera and Duffau, 2009). Fig.

The BIOPEP database developed at University of Warmia and Mazury in Poland is unique in that it focuses primarily on peptides of food origin [17]. It offers the user the ability to generate profiles of potential biological activity of the protein of interest as

well as the frequency of occurrence of bioactive fragments in the protein. For example, in silico analysis was applied to assess the potential of different food commodities to serve as sources of peptides with inhibitory activity against the enzyme DPP-IV, which acts on incretin hormones that play a role in blood glucose regulation buy PCI-32765 [19]. One limitation is that the DPP-IV inhibitors reported in the literature at the time

of that study consisted primarily of di-and tri-peptides, in contrast to the much longer physiological substrates of the DPP-IV enzyme, GLP-1 and GIP. Higher frequency of occurrence of bioactive sequences in a protein molecule does not necessarily correlate with the potential of that protein to serve as a good source of bioactive peptides unless the potency of each bioactive fragment and any overlaps of bioactive Vemurafenib in vivo sequences are taken into account. To address these limitations, Nongonierma and FitzGerald [20] developed an in silico approach incorporating protein coverage and potency indices, and applied a peptide alignment strategy to investigate the relationship between sequence and activity. Potency is represented in the BIOPEP database by EC50 values, that is, the concentration of the bioactive fragment corresponding to its half-maximal activity. Unfortunately, EC50 values are not always reported in the literature and moreover, may vary for identical sequences if assayed under different conditions. For example

the concentration of a peptide required to inhibit an enzyme to its half-maximal activity (referred to as the IC50 value), can be influenced by assay conditions including enzyme and substrate concentrations. Thus unless the inhibitory activity is reported as the inhibitor affinity constant (Ki), potency of different peptides reported by different researchers may not always be comparable. Molecular docking simulations N-acetylglucosamine-1-phosphate transferase have also been applied to elucidate which peptide sequences, either experimentally identified or predicted from bioinformatics investigation, may actually be able to interact with the proteins that are the target of the biological activity [21]. Acharya et al. [22] noted that the dynamic conformational changes induced in both the bioactive peptide and the receptor target protein upon binding impose limitations on computational docking studies, and advocated for a 4D structural database documenting these changes. Nongonierma et al.

In most but not all studies, elevated levels of MVs of endothelial origin are reported in plasma from ACS patients compared to non-ACS patients.[95] and [96] To which extent these endothelial MVs contribute to the hypercoagulable status of these patients, however, is unknown. MVs isolated from blood of patients with essential

thrombocythemia, a chronic myeloproliferative disease that is characterized by an increased risk of both arterial and venous thrombosis, are mostly derived from platelets and ECs. The MVs in these patients are thought to contribute to the hypercoagulable state that is observed in vivo.97 Plasma from patients with certain types of cancer contains higher numbers of vesicles than plasma from healthy subjects.[13], [14] and [98] Furthermore, MVs exposing coagulant TF in blood of cancer patients have been associated not only Stem Cell Compound Library order with thrombosis but also with disease progression.[13] and [15] Interestingly, in some cancer patients Alectinib cost with a detectable level of coagulant TF

present within the blood, a minor fraction of MVs exposes the epithelial marker, MUC-1.13 To which extent these MUC-1-expressing vesicles, i.e. vesicles likely to originate from the tumor, are exposing coagulant TF and to which extent such vesicles are associated with development of VTE, however, remain to be determined.99 Furthermore, tumor cells may elicit a host response that leads to expression of TF by monocytes and possibly ECs, and

to the shedding of MVs bearing TF. Recently, in a study comprising over 200 cancer patients, we found a subpopulation of vesicles in one patient exposing TF, VE-cadherin (CD144) and E-selectin (CD62e), both specific markers of endothelial origin. How much TF exposed by this subpopulation is coagulant or how TF contributes to coagulation activation in vivo has not been investigated yet (A. Kleinjan, MD, personal communication). One has to bear in mind that TF can also induce angiogenesis and transmembrane signaling, each processes important for cancer growth and development. To which extent vesicle-exposed TF contributes to such functions in cancer patients is unknown. It is still unknown whether exosomes are coagulant. This is a relevant question because most vesicles selleck chemicals llc present in body fluids are within the size range of exosomes rather than of MVs, and thus may have a relatively large contribution to coagulation because formation of tenase and prothrombinase complexes requires a membrane surface which both MVs and exosomes could provide. The membrane surface has to expose negatively charged lipids such as PS to enable the formation of the coagulation factor complexes and the PS can be detected by binding of annexin V. Heijnen et al.20 showed that only a relatively low number of exosomes, supposed to originate from platelets, bound annexin V. Furthermore, MVs but not exosomes bound factor X and prothrombin in this study.

Protease activity has been detected in various species of scorpion venoms (Morgenstern et al., 2011; Seyedian et al., 2010). However, little information about their primary structure has been available. In our study, we were unable to find gelatinase activity in the venoms analysed. In an early study from Almeida et al. (2002), a gelatinase activity associated with serine proteases was observed in venoms from T.

serrulatus and T. bahiensis. In addition, a gelatinase activity attributed to the presence of a metalloproteinase was recently observed in the venom of Hemiscorpius lepturus, a scorpion found in Iran ( Trametinib ic50 Seyedian et al., 2010). These discrepancies might be due to the sensitivity of the methods of measurement or to intraspecific/interspecific variations in venom composition. A FRET substrate, a dynorphin analogue peptide, was used in our proteolytic studies. Using this fluorometric method, it was possible to demonstrate that the Tityus spp. venoms studied were able to hydrolyse the substrate (Abz-FLRRV-EDDnp), with optimal hydrolysis efficiency learn more at pH 8.5 and 10. Under these conditions,

venom from T. bahiensis demonstrated more than two times greater proteolytic activity compared to venom from T. serrulatus and T. stigmurus. Furthermore, the proteolytic activity was completely inhibited by the metalloproteinase inhibitor 1,10-phenanthroline Y-27632 in vitro but not by PMSF, a serine protease inhibitor. The first metalloproteinase from the venom of T. serrulatus was recently identified and characterised ( Fletcher et al., 2010). This enzyme, named antarease, exhibits action on the protein vesicle-associated membrane proteins 2 and 8 (VAMP2 and VAMP8), also known as synaptobrevins. Antarease has a molecular mass of 25.5 kDa. The cleavage

sites in VAMP2 were identified as L//KRK//Y and those in VAMP8 as A//RK//F. The antarease VAMP2 cleavage site is similar to that of the metalloproteinase cleavage site of dynorphin 1-13 (L//RR) from T. serrulatus, T. bahiensis and T. stigmurus venoms found in this study. This result suggests that dynorphin-cleaving metalloproteinases detected in T. serrulatus, T. bahiensis and T. stigmurus venoms might be antarease-like molecules. Further studies will be performed to purify and characterise the dynorphin-cleaving metalloproteinases from Tityus spp. venoms. The dynorphin-degrading capacity of Tityus spp. venoms, resulting in the generation of the biologically active peptide leu-enkephalin, might be implicated in the hypotension and bradycardia symptoms ( Feldman et al., 1996), as observed in patients stung by Tityus scorpions.

, 2010). The drought and wetness hydrological behavior reproduced by the linear combination of the first components of PCA applied to SPI field at scale of 18 months is considered satisfactory since, in almost the totality of the NEA, the proportion selleck products of the total variance explained at each grid point was higher than 60%. The implementation of SSA allowed us to find a common oscillatory cycle for SPI and drought/wetness spatial coverage series with a dominant period of about 6.5 years, both for dry and wet events, so we could infer that EPE, in spatial extent and in intensity have the same leading periodicities. Consistently with

these results, Krepper and Garcia (2004) reported a cycle of T ≈ 6 years in monthly precipitation series for the whole LPB and for gauging stations on the Uruguay River. Another important hydrological cycle in SPI series at 18 month time scale was the oscillatory mode with dominant period of T = 8.7 years. This result is consistent with a quasi-decadal cycle found in the annual streamflow of the Paraná and Paraguay rivers reported by García and Mechoso (2005) and Robertson and Mechoso (1998), who associated this cycle with SST anomalies

situated over the tropical North Atlantic. In addition, Venencio and García (2012) detected a similar cycle (close to 8 years) in annual precipitation in the South of the province of Santa Fe. As could RO4929097 purchase be expected, extremely dry and wet periods that affect the largest areas of the NEA, considering the percentage of grid points that exceed the thresholds of extremely wet or dry months, were the same as those showing EPE of higher intensity and duration according to the temporal behavior given by SPI field at different time scales. The analysis of historical events with large portions of the entire region under water excess/deficit at critical months enabled us to determine the most vulnerable zones to

extreme drought/wetness. The implications of these results depend on which time scales are used. The shorter scales (n = 6 months) provide valuable information for decision-making in livestock and crop production, while the longer period time scales (n = 12 or 18 months) describe the hydrological behavior of the region. The exploration at all time scales buy Atezolizumab indicates that the Central-West portion of NEA seems to be the most vulnerable area to extraordinary extreme drought/wetness. We have presented the spatiotemporal behavior of EPE observed throughout the 20th century and up to the year 2010 in the NEA. Dry and wet events were characterized by means of the SPI applied to monthly precipitation series at different time scales (6, 12 and 18 months). Given that the stations in NEA are not homogeneously distributed in space we used gridded precipitation datasets on high-resolution. The dataset CRU TS 3.

In a previous experiment it was noticed a significant increase of the hemagglutination activity upon leaf injury (data will be published elsewhere). After this, the leaves were powdered in the presence of liquid nitrogen and stored at −80 °C until required. DEAE-cellulose column was obtained from Whatman International

Ltd., Maidstone, England; Phenyl-Sepharose 6-Fast Flow column was obtained from GE Healthcare, Uppsala, Sweden. Morphine was purchased from Sigma Aldrich Chemical (Saint Louis, MO, USA). www.selleckchem.com/screening/protease-inhibitor-library.html The other chemicals were all of analytical grade and obtained from local suppliers. The soluble proteins were extracted from the leaf powder with three volumes of 25 mM Tris–HCl, pH 7.5, supplemented with 3% (w/v) polyvinylpolypyrrolidone (PVPP) and 5 mM ascorbic acid, for 2 h at 4 °C, under gentle shaking. After filtration through nylon cloth, the filtrate was centrifuged at 10,000 × g for 30 min, at 4 °C, and the supernatant (crude extract) recovered. The crude extract was precipitated with ammonium sulfate at 30% saturation (176 g/L) and the suspension maintained at 4 °C for 12 h. The precipitate obtained (Fraction 0–30%, shortly F030) after centrifugation (10,000 × g, Selumetinib 40 min, 4 °C) was dialyzed exhaustively against Milli-Q grade water, lyophilized, and suspended in 25 mM Tris–HCl, pH 7.5. After centrifugation (10,000 × g, 20 min, 4 °C), the fraction

F030 was submitted to ion-exchange chromatography on a DEAE-cellulose column equilibrated with 25 mM Tris–HCl, pH 7.5. The through fraction was eluted from the column with the equilibrating buffer. The retained material was eluted with 25 mM Tris–HCl, pH 7.5, containing 200 mM NaCl, at a flow rate of 1 mL/min, dialyzed exhaustively against water and lyophilized. Next, it was suspended in 25 mM why Tris–HCl, pH 7.5, containing 420 mM of ammonium sulfate, centrifuged (10,000 × g, 20 min, 4 °C), and the supernatant obtained chromatographed on a Phenyl-Sepharose 6-Fast Flow column, equilibrated with the above buffer.

The protein fraction obtained after elution with 25 mM Tris-HCl, pH 7.5, containing 100 mM of ammonium sulfate, at a flow rate of 1 mL/min, was dialyzed against Milli-Q grade water and lyophilized. This material represented the lectin-enriched fraction (LEF) that was characterized and used to assess toxicity. It was determined as previously described (Bradford, 1976). Absorbance at 280 nm was also used to monitor protein elution profiles during chromatographies. Protein fractions were analyzed by polyacrylamide gel electrophoresis (15% running gel, 3.5% stacking gel) (Laemmli, 1970). The samples were solubilized in 125 mM Tris–HCl buffer, pH 6.8, containing 2.6% (w/v) SDS, 0.5 mM EGTA, 0.5 mM EDTA, 12.6% (w/v) glycerol. Gels were stained with silver (Blum et al., 1986).