, 2006) up to

, 2006) up to MG-132 in vivo 271% in P’an’s most highly-exposed subset (1981). Although the very high values obtained by P’an may be explained by

the rapid increase in saliva lead levels at blood lead levels >50 μg/L (Koh et al., 2003), there is still a great deal of unexplained variation between studies in the literature. This is most likely due to the lack of a standardised sample collection or preparation method for the analysis of lead in saliva; the wide variety of different procedures employed. The method presented in this study, using a new sampling device and a nitric acid digestion step to release protein-bound lead in the matrix, obtained a mean blank-corrected recovery from 10 μg/L spiked saliva of 65.9% (SD: 1.83 μg/L, n = 13). This demonstrates an improvement on the recovery of 30–35% reported by Morton et al. (2014), where a comparable ICP-MS method was used, but with Proteasome structure a different sampling device and no acid-digestion step. This may account for the higher levels of saliva lead observed by

this study than Morton et al. (2014) (median: 17.1 μg/L and 7.3 μg/L, respectively), despite the sample cohort reported in this paper showing lower blood lead levels (median: 8.34 μg/dL and 20 μg/dL, respectively). However, the StatSure device did exhibit a drawback – a significant level of lead contamination was shown to emanate from the device, with a mean result from blank saliva of 2.86 μg/L (SD: 1.13 μg/L, n = 10) using the device, compared to 0.38 μg/L (SD: 0.36 μg/L, n = 10) by direct analysis, i.e. the device contributed 2.48 μg/L of lead to the saliva result. The results from blank saliva aliquots sampled using the device also showed a higher degree of variation than those analysed directly. An investigation of the lead concentration of the device components showed that this contamination originated in the sampling paddle. For this study of occupationally-exposed lead

workers, the median saliva lead was 17.1 μg/L, and therefore the effect of this contamination would be relatively small. However, this would be of concern for the measurement of lower-level environmental exposures. The manufacturers of the device Thymidine kinase have been made aware of the authors’ findings and will endeavour to ensure that this contamination is not present in future batches. Additional analyses will be necessary to confirm this. A weak but significant correlation (r = 0.457) was observed between the log(saliva lead) and log(blood lead) results from the 105 paired samples analysed. This is a stronger correlation than that observed between the same variables by Barbosa et al. (2006) (r = 0.277) or by Nriagu et al. (2006) (r = 0.156), and slightly stronger than the correlation observed by Koh et al. (2003) between log(saliva lead) and blood lead results (r = 0.41). A further study by Thaweboon et al. (2005) reported a poor correlation between saliva lead and blood lead.

A wide range of proposals has been put forward in order to accoun

A wide range of proposals has been put forward in order to account for the cognitive and functional significance of the P3. These include the influential Context Updating account (Donchin, 1981 and Donchin and Coles, 1988; see also Polich, 1985 and Polich, 2007), according to which the P3 reflects memory adaptions following critical events. Another prominent account (Verleger, 1988 and Verleger et al., 2005) assigns a more tactical role to the P3 by proposing that it marks the linkage between critical events and reactions (henceforth: the Linking account of the P3). In a more strongly

biologically-grounded approach, Nieuwenhuis, Aston-Jones and Cohen (2005) associate the P3 with the norepinephrine (NE) neuromodulator system Afatinib ic50 and systemic NE release from the brainstem nucleus Locus Coeruleus (LC), which facilitates general cortical state transitions and thus supports cognitive

reorientation (like response execution or inhibition). All approaches agree that the P3 follows highly salient events such as novel and unexpected events, highly task-relevant expected events, and self-relevant stimuli. In contrast to the Context Updating theory, however, the Linking and LC/NE accounts stress that, if a task requires overt behaviour and elicits a P3, there is a tight temporal coupling between the P3 and the DAPT mouse response. The P3 is therefore often investigated following stimuli to which subjects respond Resveratrol directly. While overt responses are not a necessary precondition for P3 elicitation, if overt responses do occur, they are typically aligned with the P3. Specifically,

a frontal instance of the P3-family peaks slightly before the response, while the P3b typically peaks just at, or rapidly following it (Delorme et al., 2007a, Makeig et al., 1999 and Makeig et al., 2004). However, the P3 is not a motor component. A P3 is found in response inhibition trials (Falkenstein, Hoormann, & Hohnsbein, 1999). Furthermore, direct comparisons between overt and covert tasks have demonstrated that the P3 is also observable in passive (task-free) paradigms (e.g. in response to incorrect sequence endings), with P3 amplitudes typically (but not always) smaller than in the presence of an active task (see Lang & Kotchoubey, 2002, and the references cited therein). In one study, a silent counting task even increased P3 amplitude (Salisbury, Rutherford, Shenton, & McCarley, 2001). One reliable exception to the tight coupling between response timing and P3 latency is found when response selection is rendered complicated, for example by introducing incompatible stimulus–response mappings or complex motor actions (Verleger, 1997). Stressing speed over accuracy (Kutas, McCarthy, & Donchin, 1977) also dissociates RT and P3.

, 2010), this study would have no way of detecting those effects

, 2010), this study would have no way of detecting those effects. The energetic cost of ship noise may be substantial in terms of reduced prey acquisition (through masking or disruption of feeding activities), even if the energetic cost of Selleck DZNeP avoiding ships is relatively low. Similarly, we have not considered any physiological (i.e., hormonal) stress responses to ship noise, which have been shown to be important in other cetaceans (Rolland et al., 2012). It is hoped that this threshold analysis can provide hypotheses to test on other datasets, such as telemetry data from DTAG

deployments on killer whales around the world in the presence and absence of ships. Although the behavioral responses to ships that we documented in this study are subtle and minor, relative to some extreme responses of whales

to some extreme levels of anthropogenic noise (e.g., (Fernandez et al., 2005 and Jepson et al., 2003)), there are several reasons to keep ship noise on the conservation and management agenda for killer whales. In many parts of the industrialized world, ship noise is simply a more important contributor to the ocean soundscape than military sonar or seismic surveys (Croll et al., 2001, Hatch et al., 2008 and McKenna et al., 2012). In critical habitat for southern resident killer whales, a large ship transits the area, on average, every hour of every day of every year, with three transits second per hour observed at the busiest http://www.selleckchem.com/products/fg-4592.html times (Erbe et al., 2012). There is evidence to suggest that northern and southern resident killer whales are already prey-limited, due to natural and anthropogenic stressors

affecting the Chinook salmon that are the whales’ preferred prey (Ford et al., 2010, Ward et al., 2009 and Williams et al., 2011). If ship noise is masking (Bain and Dahlheim, 1994, Clark et al., 2009 and Erbe, 2002) communication signals that killer whales use to find or share prey (Ford and Ellis, 2006), then the ubiquitous nature of global shipping traffic (Halpern et al., 2008) makes it worthwhile to evaluate whether ship noise could cause population-level consequences to whales that are already coping with multiple other natural and anthropogenic stressors. Finally, in practical terms, ship noise lends itself to mitigation much faster than the prey- and contaminant-related threats these killer whales are also facing (Leaper and Renilson, 2012). The authors thank Christopher Clark, Phil Hammond, Patrick Miller, Brandon Southall and Len Thomas for feedback on various technical aspects of this analysis, and Marianne Gilbert, Dom Tollit, Jason Wood and an anonymous reviewer for helpful feedback on an earlier draft of the manuscript. RW collected the theodolite data with support from BC Parks and National Marine Fisheries Service, and he thanks SMRU Canada Ltd, Hemmera and Port Metro Vancouver for support for these analyses.

6b-e correspond to the 65Cu isotope

(μ/(μNI) (65Cu) = 1 5

6b-e correspond to the 65Cu isotope

(μ/(μNI) (65Cu) = 1.5877 compared to μ/(μNI) (63Cu) = 1.484897, [22]). Representative spectra from Cu/EGCG at S-band frequencies are presented in Fig. 7, The spectra from the individual Cu isotopes are seen more clearly at this frequency, and are illustrated in the expanded spectrum in Fig. 7g. The corresponding results for the Cu/GA system are available as supplementary material (Figure learn more S13). Because of incomplete averaging of the spectral anisotropy, only one of the four Cu(II) hyperfine peaks (that at the highest field) is well resolved in the solution spectra of each of the Cu(II) EGCG complexes in fluid solution at X-band frequencies (Fig. 3). The high field peak of Complex I is clearly visible in Fig. 3b, but the spectra of Complexes II and III strongly overlap (Fig. 3c), and their individual components are not resolved from one another. However, the position of the high field peak from Complex III was determined from the spectrum recorded at very high pH where the contribution from Complex II was weak (Fig. 3d). Somewhat better resolution of the component peaks was observed in the fluid solution spectra from the Cu/GA system at X-band frequencies by Ferreira Severino et al. [9] (see also Figure S14 for a full set of data), but even with this smaller ligand the resolution was not good. There are a number of reasons for the lack of resolution

of the component peaks in the spectra. Firstly, the

widths of the four individual hyperfine peaks are unequal because of incomplete averaging of the spectral anisotropy through molecular motion, and in addition the anisotropic data from the CH5424802 order frozen solution spectra show that most samples contain more than one type of complex. Furthermore, the peaks from the individual 63,65Cu isotopes are not resolved from one another. Thus there are considerable Wilson disease protein uncertainties in deriving isotropic parameters from the X-band fluid solution spectra, and these spectra were only able to be analysed by using the parameters obtained from the frozen solution spectra (Table 1) with partial motional averaging. Since the frozen solution spectra provide no information on the relative signs of A// and A⊥, simulations were performed with the A// and A⊥ values having the same and opposite signs. However, only the use of the same signs reproduced the experimental spectra. The copper hyperfine peaks are much better resolved in the fluid solution spectra recorded at S-band frequencies (Fig. 4 and Fig. 5). The magnitudes of the Aiso values derived from these spectra (Table 1) show clearly that A// and A⊥ must have the same signs in Complexes II and III with both the Cu/GA and Cu/EGCG systems, thus providing support for the X-band analyses. Simulated parameters for the spectra of all Cu complexes detected in fluid and frozen solutions are reported in Table 1, the values of Aiso being assumed to be equal to (A// + 2A⊥)/3 for the X-band spectra.


the cognitive processes involved when asses


the cognitive processes involved when assessors evaluate samples using new methodologies can strongly contribute to the development of recommendations for their implementation. This approach has already been addressed by researchers working Galunisertib with different methodologies. Some examples are presented below. Check-all-that-apply (CATA) questions are a type of multiple choice question in which assessors are presented with a list of terms and are asked to select all those are regarded as applicable to describe a focal sample 7 and 8•. Visual attention plays a key role when assessors complete this type of self-administered written questionnaire [9]. In order to select a term they should be aware of its presence on the list of options, that is, they have to fixate their gaze on it when evaluating a focal sample [10]. Recent research has shown that the first time that consumers go through a CATA question they tend to perform a thorough examination of the list of options [11]. However, they usually pay more attention to the terms locate at the top of the list than to those located at the end. Besides, as the task progresses assessors reduce the depth with which they learn more process the list of options. These results suggest that it is necessary to balance the position

of the terms within the list between and within assessors in order to avoid biased results. Another example of how studying cognitive processes can contribute to the development of recommendations for the implementation of new methodologies for sensory characterization is related to the influence of short term memory on the number of samples that can evaluated using holistic methodologies, such as sorting or projective mapping. In these methodologies participants tend to memorize the characteristics of samples when evaluating their similarities

and differences [12]. Considering that short term memory only maintains a limited amount of information for a short period of time [13], results are expected to be strongly affected by the number of samples included in the study. It can be hypothesized that increasing the number of samples reduces assessors’ ability to discriminate among samples. Research on sorting tasks with beers has much shown that the number of samples should be lower than 20, being 12 the optimum 14 and 15. However, it is still necessary to further explore the influence of sample complexity on assessors’ ability to memorize their sensory characteristics and discriminate samples using holistic methodologies. A process of synthesis is necessary for analyzing and processing sensory information in holistic methodologies. Assessors have to determine the relative importance of the different sensory characteristics of the products to reach a judgment on their global degree of similarity/dissimilarity [16].

3 It is observed that this system of faster exchange of zinc aro

3. It is observed that this system of faster exchange of zinc arose and is only found in eukaryotes. The ability of zinc in this exchange between a series of sites allows it to integrate such cellular effects which arise from binding to zinc fingers. One case is the binding of hormones, especially sterol hormones. If this central control role of zinc is confirmed it is then possible to consider zinc as a super-hormone or a second messenger [26] and [30]. Note that the tighter binding to many enzymes is

not related to this exchange directly but their synthesis may still depend on chaperones. Deficiency in the zinc proteins in man leads to slow growth and an inability to go through puberty. Both are cured by giving zinc to the patient. This is an example of what Vallee had hoped to find when he first began to examine zinc biochemistry. The final section of my paper refers to a problem Bert and I discussed in more Selleckchem 3 MA social circumstances. We did no experiments and published no papers together in this area. How did zinc biochemistry evolve AC220 cost [29]? It had to be in two steps. First very early zinc enzymes in a low zinc environment with a high binding constant and no exchange and second a later larger amount of zinc in the environment became associated with message functions either of hormones,

in zinc fingers or free zinc acting itself as a messenger in nerve tissue. The earliest use appears to be at the beginning of life around 3.5 Ga whilst the later uses are related to the beginning of single-cell eukaryotes and multi-cell eukaryotes, Fig. 2, Fig. 4 and Fig. 5. One well-recognised pathway of evolution of zinc proteins and of many other proteins is from an original parent by mutation. It is easy to see that mutational changes, in the evolution of somewhat different organisms and within the increasing diversity of recent organisms, will affect the required efficiency of the enzyme. It does not explain the times when evolution of very different proteins occurred, for example the appearance of zinc fingers in unicellular eukaryotes around 2.5 Ga (billion years ago) and more strongly on the

evolution of multicellular eukaryotes Liothyronine Sodium around the time of the Cambrian Explosion, 0.54 Ga, Fig. 4. Both these faster periods of evolution correspond with the periods of faster chemical change in the sea shown by geochemical evidence of both a rapid rise of oxygen in the atmosphere, of sulphate in the sea and of trace elements including zinc in sediments. Both the second two changes arise from oxidation of sulphides. The sediments are assumed to define the probable nature of the sea which would have a great influence on organisms. Using the comparison of the element content of organisms alive today, from the anaerobes to large plants and animals, it is apparent that the content of zinc (or copper) proteins increases in the series of complexity and that the steps of the most rapid changes, Fig.

” We hypothesized

” We hypothesized Staurosporine that Boston Bowel Preparation Scale (BBPS) scores

could provide a way to standardize the concept of “adequacy”. We performed a retrospective analysis of average-risk screening colonoscopy reports submitted to the Clinical Outcomes Research Initiative (CORI) data repository between 10/2009 and 8/2012. We included only reports documenting a BBPS score and a recommendation for timing of the next colonoscopy and excluded procedures with polyps. We evaluated recommended follow-up intervals stratified by total and segment BBPS scores. We also presented 4 standardized colonoscopy videos with varying degrees of bowel cleanliness to participants of the BBPS Educational Program, a web-based program demonstrating the BBPS, and asked for recommended colonoscopy follow-up intervals. Among 3226 average risk colonoscopies with a BBPS score, 1340 (41.5%) had polyps and 601 (18.6%) lacked follow-up recommendations and were thus excluded. The remaining 1285 procedures, performed by 55 endoscopists, had a median (interquartile range) BBPS score of 8 (7-9). Median recommended follow-up time decreased as BBPS scores decreased, with a sharp drop-off below a BBPS

score of 6 (see Figure). Among reports with total BBPS score of 6 or 7 (n=364), 17 (5%) contained a segment score of 0 or 1 and were associated with shorter median follow-up time compared to reports in which all segment scores were ≥2 (5 vs.10 years, P<.001). Whenever any colonoscopy Sodium butyrate contained a single segment score of 1 (n=55), that segment’s location (right, left, transverse colon) had no impact on recommended follow-up intervals (P=0.955). Video cases were reviewed by 119 endoscopists, including RG7204 chemical structure 39 CORI users, 51 non-CORI US endoscopists and 29 international endoscopists. Recommended follow-up time decreased as BBPS scores decreased (P<.001; see Table). There was no difference in recommended follow-up time by location

of practice, although more US participants (87%) recommended 10 year follow-up compared to international participants (52%) for Case D (P=.0012). BBPS scores correlate with endoscopist behavior regarding follow-up intervals for colonoscopy. Because BBPS scores have previously been shown to have excellent inter-rater agreement, a total BBPS score ≥ 6 and/or all segment scores ≥ 2 provides a standardized definition of “adequate” when describing bowel cleanliness. Recommended follow-up interval for next colonoscopy for video cases among endoscopists who agreed on the Boston Bowel Preparation Scale score for each case “
“Despite advances in bowel preparation methods, the quality of bowel preparation in patients undergoing colonoscopy remains unsatisfactory. The time point chosen for improvement of education may be important for adequate bowel preparation. To evaluate the effect of telephone re-education on the day before colonoscopy (instead of the day of appointment – regular appointment) on the quality of bowel preparation and colonoscopic findings.

3) showed a higher homology of TsNP for both, human and rat CNP

3) showed a higher homology of TsNP for both, human and rat CNP. Based on this result, a higher affinity of TsNP for NPR-B and NPR-C is expected. NPR-B has approximately a 50-fold higher affinity for CNP than ANP. CNP essentially does not bind to NPR-A but, it binds to NPR-B and NPR-C with similar affinities (Schulz, 2005). The effects of TsNP on the isolated perfused rat

kidney were similar to those reported by Evangelista et al. (2008) for a natriuretic peptide from C. durissus cascavella venom. In the same perfusion system, ANP also showed an increase in GFR and urine flow, with no effect in the perfusion pressure. The urinary excretion of sodium, potassium and chloride were increased by the TsNP treatment, as has been shown for ANP, guanylin and urodilatin ( Santos-Neto et al., 2006). The concentration of cGMP in the urine Dasatinib was elevated as would be expected for a guanylate cyclase activating drug. CD-NP, a chimeric natriuretic peptide, has been shown to enhance GFR, selleck producing diuresis and natriuresis ( Lisy et al., 2008). Fonteles

et al. (2009) showed a reduction in NPR-A expression accompanied by an increase in GC-C expression in animals receiving a high-salt diet and treated with uroguanylin (a GC-C agonist). This observation corroborates with our data, which showed that GC-C expression was elevated while NPR-A expression was down regulated following treatment with TsNP. These effects point to a possible activation of GC-C by TsNP. However, Anand-Srivastava (2005) acetylcholine demonstrated another possible relationship, which could explain NPR-A down regulation and link the up regulation of TGFβ-1 and NPR-C, with the down regulation of NPR-A. These results were also observed in our studies. The increased concentration of cGMP in the urine points to the activation of guanylate cyclase, as occurs after NPR-B and GC-C activation. NPR-C could play a role in the increased perfusion pressure caused by TsNP. NPR-C has three distinct signaling pathways, which involve the activation of eNOS, MAPK-1 and phospholipase C (PLC) (Anand-Srivastava,

2005). For this reason, the gene expression of eNOS and MAPK-1 following TsNP treatment were also tested. TsNP treatment resulted in down regulation of eNOS expression, whereas MAPK-1 expression was not changed. This result possibly excludes the involvement of these two NPR-C signaling pathways in the biological actions of TsNP. Thus, activation of PLC downstream might explain the augmentation of the perfusion pressure through direct vascular smooth muscle contraction (Anand-Srivastava, 2005). In conclusion, this work describes the isolation and modeling of the first natriuretic peptide isolated from scorpion venom. In addition, the renal actions and the effects on NPRs mRNA expression in the kidneys are delineated. Our data showed that TsNP might act promiscuously with NPR-B, NPR-C and GC-C. Future binding analyses should elucidate the relative affinities between TsNP and various receptors.

The results of in vitro bacterial genetic mutation assay are summ

The results of in vitro bacterial genetic mutation assay are summarized in Table 1. In all bacterial strains exposed to different doses of EAHE mycelium (5, 2.5, 1.25, 0.625, and 0.3125 mg/plate) with the presence and absence of S9-Mix metabolic activators, the revertant colonies

showed no dose dependency and were similar to those of negative control. These findings suggest that EAHE mycelium displayed no mutagenicity, especially in Salmonella typhimurium strains TA98, TA100, TA102, TA1535, and TA1597. Since the Ames test cannot detect chromosome aberrations induced by chemicals [35], it has been recommended to use in vitro tests as a minimum requirement selleck chemicals llc for mutagenicity testing [36]. According to the guidelines OECD 473 [31], the highest dose used in the in vitro chromosome aberration test should be a concentration above the limit of solubility to avoid false positive results. As the amount of precipitation recorded for EAHE was at 5 mg/ml, dose levels of 2.5, 1.25, and 0.625 mg/ml were selected and exposed to the CHO-K1 cells (BCRC 60006) in the presence and absence of a metabolic activation system derived from rat liver Selleck AZD9291 S9 mix [30]. The cell proliferation of CHO-K1 in the 3 h treatment with the presence and absence of S9 activation was inhibited by <7.7% and <21.4%, respectively. Incubation of these cells under the same condition for 20 h in the absence of S9 activation

resulted in <34.3% decreases of cell growth (data not shown). As the highest concentration showed a significant reduction in the degree of confluency, such doses were then used for chromosome observation. The validity of the tests was observed in the incidence of cells having aberrant chromosomes, which was 0% to 1% and 6.5% to 12.5% in negative groups and positive C-X-C chemokine receptor type 7 (CXCR-7) groups, respectively ( Table 2). Neither 3 h nor 20 h EAHE mycelium treatments induced higher frequency of aberrations that were significantly different from negative controls (p > 0.05, Chi-square test) (Table 2). In summary, these data indicate that exposure to EAHE mycelium does not result in genetic damage in cultured

mammalian cells under the test conditions. The uptake of EAHE mycelium that resulted in chromosomal damage was further investigated by using the in vivo erythrocyte micronucleus test in ICR mice since the in vivo assay takes into account whole animal processes, such as absorption, distribution, metabolism, and excretion. Our earlier study on acute oral toxicity indicated that acute oral LD50 of EAHE mycelium was greater than 5 g/kg (data not shown). Hence, doses of 1.25 g/kg (low dose), 2.5 g/kg (mid dose) and 5 g/kg (high dose) were selected for this study, whereas distilled water and cyclophosphamide were served as negative and positive controls, respectively. During the 72-hr post-treatment, EAHE at all tested doses (1.25, 2.5, and 5 g/kg) did not induce any symptoms of toxicity, morbidity, or mortality in all mice (n = 25).

We evaluated the in vivo myotoxicity of Bothrops snake venoms usi

We evaluated the in vivo myotoxicity of Bothrops snake venoms using two different protocols. First we assessed the activity of creatine kinase (CK) in plasma, which increases in cases of muscle damage. Mice received perimuscular injections of venom, as described above. Two hours after the injection of venom alone or associated with treatments, the animals were lightly anesthetized and ROCK inhibitor blood was collected by orbital puncture. The determination of plasma CK activity was performed as previously described ( Melo and Suarez-Kurtz, 1988b; Melo et al., 1993 and Melo et al., 1994), and

expressed as units per liter of plasma (U/L). We also determined the EDL muscle CK content in the same groups of mice at 24 and 72 h after the injections, which is expected to decrease in cases of muscle damage. Animals were sacrificed under anesthesia, then the EDL muscles were removed, weighed, minced and homogenized in 2 mL PSS + 0.1% albumin and the CK content was determined in the homogenate as described previously ( Melo and Ownby, 1996; Tomaz et al., 2008). The results

were expressed as units per gram see more of muscle tissue (U/g). Inflammation was assessed by multiple parameters. The local edema induced by the venom, and the protection by the treatments, were evaluated using a caliper rule. The diameters of mice legs were measured before and 1 h after the perimuscular venom injections (as described above) and the results are shown as mm of increase in the leg diameter click here (Melo et al., 2010). Blood was collected by orbital puncture under ether anesthesia 24 h after the injections, and treated with Turk’s solution (19:1 v/v) for leukocyte count in a Neubauer chamber. The results are shown as leukocytes per mm3 (×103 cells/mm3). The same animals were

killed under anesthesia and the EDL muscles were removed, weighed, minced and homogenized in 2.0 mL PSS and then centrifuged at 20,000 rpm. We then removed 1.8 mL of the supernatant, resuspended the cell pellet and treated a 20 μL sample with 380 μL of Turk’s solution. The leukocyte count was performed in Neubauer chamber and the results expressed in leukocytes per gram of muscle tissue (×106 cells/g). To determine the myeloperoxidase (MPO) activity in the muscles, EDL muscles were removed, weighed, minced and homogenized in 1.0 mL of 0.5% hexadecyltrimethyl-ammonium bromide (HTAB) in potassium phosphate buffer 50 mM, pH 6.0 (Bradley et al., 1982). After centrifugation at 10,000 rpm for 5 min, 100 μL samples of the supernatant were mixed with potassium phosphate buffer 50 mM containing 1.0 mM O-dianisidine dihydrochloride and 0.001% H2O2. Absorbance was measured at 460 nm taking 4 readings at 60 s intervals. Each MPO unit activity was defined as that degrading 1 μmol of H2O2 per minute at 25 °C and considering that 1 μmol H2O2 gives a change in absorbance of 1.13 × 10−2 nm min−1 (Posadas et al., 2004). The results were expressed as units per gram of muscle tissue (U/g).