Likewise a portion of catechol gene 1 27 kb (C23O) was pulled out

Likewise a portion of catechol gene 1.27 kb (C23O) was pulled out using F: 5′- ATG AGC AAC AAA

TAC GAA TT- 3′ and R: 5′- TCA AAC GGT CAA TCT GAT AT- 3′ primers, with 1.5 U of Taq DNA polymerase in a 25 μL reaction mixture, consisting of 100 ng of genomic DNA, 20 pmol of each primer, 200 μM dNTPs and 1X Taq GSK2118436 datasheet buffer with 1.5 mM MgCl2. PCR was conducted using the following temperature profile: initial denaturation at 93 °C for 2 min, then 30 cycles of 1 min at 93 °C, 35 s at 45 °C, and 1.5 min at 72 °C; and finally an extension reaction of 5 min at 72 °C. PCR products were analyzed by electrophoresis on 1% agarose TAE gels. The expected DNA bands of 0.26/1.27 kb were excised from Gefitinib gel and purified using the Gel Extraction Kit (Sigma–Aldrich, USA) as per the manufacturer’s protocol. Sequencing reactions were carried out with a Big Dye Terminator cycle sequencing kit by using ABI Prism 3100 genetic analyzer (Applied Biosystems, Foster City, CA, USA). Fig. 1(a–c) illustrates the morphology, SEM image and phylogenic profile of the isolate MTCC 5514 employed in the present study. The bacterial colony has irregular margin, rough

surface with pink pigmentation. The staining studies revealed the Gram +ve nature of the isolate and the SEM analysis suggested the short rod nature of the isolate. The phylogenic profile infers the isolate MTCC 5514 belongs to Bacillus licheniformis. The distance matrix showed the genetic distance value between MTCC 5514 with B. licheniformis ATCC 14580 was 0.004. Anthracene biodegradation study carried out at 37 °C under shaking conditions using MTCC 5514 displayed an interesting observation. The physical

observations made during the growth suggested that from day 1 to till day 7 most of the anthracene molecules (irrespective of the concentrations studied) were settled at the bottom of the flask, despite, much turbidity in the external medium due to the growth of the organisms. However, after day 15, deposition of only fewer anthracene molecules at lower concentration than higher concentrations was observed. Further, after 22 days, no P-type ATPase deposits were found at lower concentration, however, a fewer deposits were at higher concentration. Samples withdrawn at scheduled time intervals (10, 16 and 22 days) were subjected to various analyses after extracting with ethyl acetate. However, before extraction, analysis such as pH, biomass and surface activity were made for all the concentrations. The percentage of degradation of anthracene was calculated based on the absorption displayed in UV–visible spectral analysis at 254 nm and using standard graph. Fig. 2a displays the growth profile of the isolate MTCC 5514 in the presence of anthracene at 100–1000 ppm concentration. The chosen isolate MTCC 5514 showed a bi-phasic growth profile in the presence of anthracene at 100 and 300 ppm concentration.

Additionally, catch levels may have experienced

a certain

Additionally, catch levels may have experienced

a certain amount of resiliency if fishers started using other, lower-value species or smaller individuals that were previously discarded. The species composition of the fin trade has not been assessed for more than a decade [9], hence this should become a research priority. Further, the apparent failure of anti-finning laws to curb global mortality may indicate that these laws have yet to be adequately enforced [24]. On the other hand, anti-finning laws primarily address animal welfare selleck screening library and food security issues (i.e. to reduce waste). Although an important first step, these policies are not explicitly designed to reduce catch or ensure sustainability. The premise that anti-finning Belinostat cell line legislation would contribute to sustainable fisheries rests on the assumption that most fishermen target sharks for their fins only, and would refrain from targeting sharks if they had to retain the carcass. This assumption is weak. Many

countries consume shark meat [25] and fishermen opt to land whole sharks, even if the meat is not as valuable as the fins. Several at-risk shark species are generally kept rather than being finned in certain pelagic fisheries where freezer space is limited [24]. It is not surprising that anti-finning measures have been introduced widely given the intense public pressure that arose, especially since anti-finning laws are more palatable

to industry than stringent catch reductions when local markets for the meat exist. In contrast, the monitoring, assessment and enforcement capacity required to sustainably manage shark fisheries is often perceived by regulatory agencies as being prohibitively costly relative to the simple adoption of anti-finning legislation. Regardless, some nations have recently invested in sustainable shark fisheries management, introducing catch limits, effort Non-specific serine/threonine protein kinase control, time-area closures, and other protective measures for the most vulnerable species. In some cases, such local measures appear to have been successful in halting declines [8]. The findings reported here highlight the fact that shark conservation policies generally need to focus on sustainability, as there is no evidence that a legislative focus on anti-finning has reduced global landings and shark mortality rates. From a legislative perspective, an important question to consider is what proportion of shark species may be at risk from extinction? According to the International Union for the Conservation of Nature (IUCN) Shark Specialist Group, 28% of assessed and non-data deficient shark species are globally at risk of extinction, i.e. classed as vulnerable, endangered or critically endangered (Table 6). A small number of these species are now receiving protection through national and international agreements.

There were no mesh-related

There were no mesh-related selleck compound complications and no operative mortality. Objective follow-up was available in 69 patients at a median of 5 months

postoperatively, and in 15 patients at 1 or more years. The follow-up was by videoesophagram in 79%, upper endoscopy in 52%, and both in 48% of patients. Two patients underwent conversion from a Nissen to a Toupet for protracted dysphagia. A small recurrent hernia was found in 3 patients (4%) by upper endoscopy, but no patient has required reoperation. All recurrences developed after primary laparoscopic repair of a PEH (n = 2) or sliding hiatal hernia (n = 1). One recurrence was in a patient who had a Collis gastroplasty and a right relaxing incision; no adjunct procedures were performed in the other 2 patients. A recurrent hiatal hernia is the most common form of anatomic failure after laparoscopic hiatal hernia repair and fundoplication.1

Hernia recurrence is particularly common after laparoscopic PEH repair; the rate exceeds 50% at 5 years when objective studies such as barium swallow or upper endoscopy are done to evaluate the repair.1 and 2 These recurrence rates www.selleckchem.com/products/PF-2341066.html are higher than those in historic reports with open repairs.1 and 8 The explanation for the higher recurrence rate with laparoscopic repair is unclear, but theories include the lack of deep bites during crural closure with the use of laparoscopic suturing devices and reduced adhesions associated with a laparoscopic compared with an open procedure. However, an alternative explanation is that during laparoscopic repairs there may be an underappreciation of tension on the repair. This tension can come from 2 directions: axial tension related to esophageal shortening and lateral tension related to widely splayed crura that must be reapproximated as part of the repair. The consequences of tension on hernia recurrence are well documented at other SB-3CT sites including inguinal and ventral hernias.9 In an effort to reduce tension and improve outcomes with laparoscopic hiatal hernia repair, we adopted adjunct techniques to reduce tension when encountered. These techniques included a diaphragm relaxing incision or a wedge-fundectomy Collis

gastroplasty. In this series, a crural relaxing incision was performed in 12% and a Collis gastroplasty in 28% of patients. These numbers increased to 21% and 45%, respectively, in those undergoing PEH repair. In part, these high numbers are related to the addition of patients undergoing reoperations when tension was likely a contributing factor to the initial failure, but also to the complexity of patients who are sent to a tertiary referral center. When a relaxing incision was deemed necessary, it was most commonly performed on the right side. This is the easiest of the diaphragmatic relaxing incisions. If the right side relaxing incision was inadequate, or if the right crus was too thin to allow a relaxing incision, then a left-sided diaphragmatic relaxing incision was used.

This is particularly clear for the Simon task In this task, part

This is particularly clear for the Simon task. In this task, participants are asked to respond with a left or right hand response to a certain stimulus feature, typically the color of a circle [11]. The stimuli are presented left or right of a central fixation cross. This

spatial outline creates a condition in which the location of the stimulus (e.g. left of the fixation cross) is congruent with the required response (e.g. the color blue should yield a left button press), and a condition in which the location of the stimulus and the required response are incongruent (e.g. a blue circle to the right of the fixation cross, requiring a left button press). The Simon task is demanding because on incongruent trials, participants are confronted with interfering information in the form of the spatial location, but have to respond to the task-relevant feature only. The response time distributions of the Simon task deviate from other http://www.selleckchem.com/products/azd9291.html conflict tasks in that the time cost of resolving the interfering information is mostly associated with relatively fast http://www.selleckchem.com/products/nu7441.html responses. That is, most conflict tasks show a greater interference effect for slower responses, but the Simon task shows the greater interference effect for faster responses

[13]. This pattern of interference is atypical for most conflict tasks. It suggests that in terms of the cause of interference, more processes are involved than in for example a flanker task [14]. While it is clear that an accumulator model of spatial interference control such as in the Simon task would greatly increase our understanding of cognitive control, as yet no such Niclosamide model has been published. The aim of the current article is to pave the way to use accumulator models to understand latent processes in spatial interference control. That is, based on previous model-based approaches towards the neural basis of perceptual decision

making, and previous functional Magnetic Resonance Imaging (fMRI) studies in spatial interference control, we aim to outline the important processes in an accumulator model of spatial interference. In this section we review basic perceptual decision making experiments that have sought to relate diffusion model properties to Blood Oxygen Level Dependence (BOLD) responses. In fact, the two most important properties of the diffusion model can be identified in the brain [3••]. The rate of evidence accumulation has been shown to be positively related to BOLD in dorsolateral prefrontal cortex (DLPFC, e.g. 15, 16 and 17) and anterior Insula (aI, e.g. 15, 16, 17 and 18). Thus, a high accumulation rate elicits a stronger BOLD response than a low accumulation rate, suggesting that easier perceptual decisions yield a stronger BOLD response in DLPFC than hard perceptual decisions. In addition, some studies report a correlation between the rate of evidence accumulation and the BOLD signal in right inferior frontal gyrus (rIFG, 19 and 20•).

Bob told me that his choice only involved substituting immunology

Bob told me that his choice only involved substituting immunology for endocrinology. Of course, two other reasons for thinking of Bob as the founder of psychoneuroimmunology were that he established the journal Brain, Behavior and Immunity 7 and assumed a leadership role in GSK2118436 purchase forming, and then guiding, the Psychoneuroimmunology Research Society (PNIRS) during its early years as its President. Bob was highly, but fairly, critical of scientific submissions to BBI but never brutally so, even when he received a manuscript that was unchanged

from one that he had previously rejected for another journal. Neither was Bob overly concerned when some disgruntled colleagues whose manuscripts were repeatedly rejected claimed that the journal was being run by the “Rochester mafia”. Nick Hall: You

also demanded that PNI remain on the high road by establishing an exceptionally high standard for the study of the brain, behavior and immune system. It would have been so easy to accept the large number of poorly conceived papers that were submitted in the early days of BBI. Instead, you insisted on rejecting more papers than were accepted even though the continuation of the journal was in jeopardy when deadlines for various issues were missed due to lacking enough articles. Thank you, Bob, for nurturing Gefitinib research buy PNI into an endeavor we can all be proud of. Steve Cole: …the role you played as founder and editor of the field’s defining journal really consolidated PNI as an endeavor – creating a new scientific “community on the ground” to help realize the implications of the new “facts on the ground” that you and the others began to recognize in the late 1970’s”. Bob knew the vital role he played in establishing a new field. Yet he never flaunted this role even when it might have served him personally. He

did not have to – his scientific contributions were known worldwide, as were his honesty and integrity. Formal recognition included: his appointment P-type ATPase as the George L. Engel Professor in Psychiatry and as the Distinguished University Professor in Psychiatry at the URMC; receipt of an honorary medical degree from the University of Trondheim in Norway (1992) and an honorary D.Sc. degree from Tulane University (2002); and the establishment of the Robert Ader New Investigator Award by the PNIRS. Bob wrote with a simple elegance–clarity was all-important. Data-rich publications, including Bob’s, are formulaic and therefore, rather dull from a literary perspective. But given the opportunity to break away from the format of a scientific paper, Bob’s writing became, at least to me, an engrossing narrative. For those of you interested in this facet of his writing, I suggest you read two papers. The first is his presidential address to the American Psychosomatic Society (Ader, 1980).

Persistent

inappropriate tachycardia has been demonstrate

Persistent

inappropriate tachycardia has been demonstrated to induce an impairment of left ventricular function both in animal models and in humans [39]. Clozapine induces a rise in plasma catecholamines that correlates with the degree of myocardial inflammation [40]. Moreover, the histopathology of clozapine-treated mice showed a significant dose-related increase in Selleckchem R428 myocardial inflammation that correlated with plasma catecholamine levels. Propranolol, a beta-adrenergic blocking agent, significantly attenuated these effects [12]. The clozapine-induced increase in serum levels of catecholamines increases the myocardial oxygen demand,

both directly and indirectly via direct myocardial stimulation and increasing cardiac loads [41] in addition it decreases myocardial oxygen perfusion [42]. Moreover, increased serum level of catecholamines stimulates renin-angiotensin-aldosterone system leading to further increase in cardiac loads, the fact that explains the protective role of angiotensin converting enzyme inhibitors as captopril against clozapine cardiotoxicity [43]. Increased cardiac loads with decreased perfusion myocardial ischemia and increased generation of free reactive oxygen species, leading to increase in myocardial lipid peroxidation, inflammation and cell injury. These effects were reflected in our results in form of increased myocardial lipid peroxidation Oligomycin A cost product MDA and 8-OHdG, the marker of oxidative

DNA damage. Our results showed that clozapine significantly increased the cardiac level of nitrites, a stable product and indirect marker of NO. In addition, Montelukast Sodium the immunohistochemical study showed increased immunoreactivity to 3-nitrotyrosine, the marker of peroxynitrite, in cardiac tissues of clozapine-treated animals. The myocardial cytotoxicity of peroxynitrite involves direct oxidative injury to cardiac cells and damage to proteins, lipids and DNA [44] and the nitration of tyrosine residues of pro-apoptotic proteins in cardiomyocytes [45]. Previous studies showed increases in cardiac NO levels following exposure to clozapine, an effect that can be related to the drug itself or to its metabolite N-desmethylclozapine via its agonistic activity toward M1 receptors on cardiac vagal preganglionic fibres [46]. NO is an immune regulator and an effector molecule that mediates tissue injury. Increased formation of NO may induce negative inotropic effects and become deleterious to the heart. Where, excess amounts of NO produced by inducible nitric oxide synthase (iNOS) appeared to contribute to the progression of myocardial damage in myocarditis [47].

Such loss of control may range from the comparatively minor incid

Such loss of control may range from the comparatively minor incidents Carfilzomib in vivo within a workplace to major events where people well beyond the workplace may be exposed. Biological monitoring can serve several purposes in the aftermath of a chemical incident. For instance, it can confirm the presence or absence of internal exposure in subjects

potentially exposed; it can help relate clinical symptoms to an exposure or can support medical care (Scheepers et al., 2011). The important need to consider the use of biological monitoring in the response phase of an incident was recognised by the World Health Organisation (WHO, 1997 and WHO, 2009). Depending on the type and scale of the incident it may be necessary to assess the exposure of the workers, first responders

or the public. Critical to the utility of biological monitoring is the availability of quality assured analytical methods from accredited laboratories and guidance values to put the results in perspective. There are no biological monitoring guidance values specifically for chemical incident scenarios. The two major sources of biological monitoring guidance values relate to either occupational or environmental exposure. Examples of workplace guidance values are those produced by the American Conference of Governmental Industrial Hygienists (ACGIH, 2013) the German Science foundation (DFG, 2012), the UK Health & Safety Executive (HSE), the French Agency for Food, Environmental and Occupational Health & Safety (ANSES, 2013) and the European ITF2357 solubility dmso Scientific Committee on Occupational Exposure limits (SCOEL, 2014). Biological monitoring guidance values for occupational exposure are usually derived from peer-reviewed ethically-approved volunteer and workplace studies that enable a relationship to be derived between

a biomarker and an absence of ill-health, airborne occupational exposure limit and/or acceptable level of exposure. Guidance on environmental exposures comes from studies of biomarkers in the general population like the US national Health and Nutrition Survey (CDC, 2013) and the German Human Biomonitoring Commission (Umwelt Bundesamt, 2014). These are often expressed as 95th percentile reference ranges or, increasingly, based on the “Biomonitoring Equivalents” concept (Hays et Ketotifen al., 2007) where “acceptable exposures” are identified from, for example, tolerable daily intake doses. Biological monitoring guidance values for both environmental and occupational exposures are derived for specific purposes and have limitations when applied outside these. One of the major limitations is the relatively small number of them in comparison to the number of substances to which people may be exposed. This is in part due to the costs of population studies and the availability of studies in the peer-reviewed literature linking biomarkers to health or exposure.

Participants received a monetary reward Procedures were approved

Participants received a monetary reward. Procedures were approved by the local Psychology ethics committee. Laboratory

apparatus comprised an Apple Mac Mini, with Labtec speakers positioned either side of a 17″” Sony HMD-A420 cathode ray tube (CRT) display, viewed in darkness from 70 cm. Mobile apparatus for older participants and PH comprised a Sony Vaio SZ1XP PC with built-in speakers Bcl-2 inhibitor and 13.3″” liquid crystal display (LCD) display, viewed from approximately 57 cm. In both cases video mode was 1200 × 800 with a 60 Hz refresh rate. Subjects responded using the cursor keys on the standard keyboard. McGurk stimuli were based on Soto-Faraco and Alsius (2007), which were kindly provided by the authors (see Fig. 2 for dimensions, and Video 1 and Video 2). Auditory /ba/ and /da/ phonemes (with white noise at 15% of maximum

amplitude) were combined with visual lip-movements for [ba], [da] and [ga]. The two incongruent pairings for eliciting the McGurk effect were /ba/ + [ga] = ‘da’ and /da/ + [ba] = ‘ba’ or ‘bda’. The other two ‘congruent’ pairings /ba/ + [ba] and /da/ + [da] tend to be heard correctly. Background was set to the average red green blue (RGB) value across all pixels and frames. For the Selleckchem ZVADFMK Stream–Bounce experiment, visual stimuli were two yellow circular at maximum contrast on a black background. Each moved from positions left and right above fixation, via the central fixation point, to opposite positions below fixation (see Fig. 2 for dimensions, and Video 1 and Video 2). Animations were accompanied by a 400 Hz tone of 100 msec duration, with the same manipulation Exoribonuclease of asynchrony

as for the McGurk stimuli. Movies were followed by 9 pt white text prompting responses, displayed centrally. The following are the supplementary videos related to this article: To view the video inline, enable JavaScript on your browser. However, you can download and view the video by clicking on the icon below Video 1.   McGurk stimulus demo: Four combinations, played consecutively: 1. Auditory /ba/ with visual [ba]: congruent. 2. Auditory /ba/ with visual [ga] (incongruent: McGurk effect sounds like “da”). 3. Auditory /da/ with visual [ba] (incongruent: McGurk effect sounds like “ba”). 4. Auditory /da/ with visual [da]: congruent. We also tested PH with various biological and/or non-speech stimuli. Finger-click movies, of 3000 msec duration, showed a hand with the middle finger clicking against the thumb. Sequences began with either the hand open (to provide predictive information) or closed. For scrambled-speech stimuli, the soundtrack from the original McGurk stimuli was passed through a three-channel noise vocoder using Praat software (version 5.1.21, http://www.praat.org), rendering the speech unintelligible but without affecting the spectral composition of the sound or the temporal sequence of amplitude modulations. The video sequence remained the same. Non-biological stimuli comprised a white square (1.

Cell numbers were determined by the trypan blue (Gibco) dye exclu

Cell numbers were determined by the trypan blue (Gibco) dye exclusion method and they were reported by considering the number Ibrutinib solubility dmso of expanded cells cultivated in the differentiation stage. (i) In order to assess the degree of megakaryocytic differentiation, CD41 (Mk lineage cells) expression was analyzed by flow cytometry (FACSCalibur, BD) using an anti-CD41 antibody (Biolegend). CD34 and CD33 expression was also determined using appropriate antibodies and isotype controls. (ii) Mk ploidy was determined by double-staining technique with flow cytometry (FACSCalibur, BD) and using CellQuest Pro software (BD) by choosing CD41+ events as a respected gate

[13]. Briefly, the cell cultures incubated 15 min with anti-CD41 antigen (Biolegend) and then fixed by 70% cold ethanol (4 °C). Cells were re-suspended in a staining solution

containing propidium iodide (50 μg/mL; Sigma), sodium citrate (4 mM; Sigma), RNase A (0.1 mg/mL; Sigma), Triton X-100 (0.1%; Sigma) in pH 7.8 1 h before performing the flow cytometry. (i) For scanning electron microscopy imaging, cell population were first fixed in a solution of glutaraldehyde (Sigma) 1.5% (v/v) in PBS (Gibco), then post-fixed in a solution of osmium tetroxide (0.05%; Sigma) www.selleckchem.com/products/dinaciclib-sch727965.html in PBS (Gibco); both for 30 min at room temperature. Cells were then dehydrated by gradually increase of ethanol (Sigma) concentration (50%, 75% and 100% in distilled water). Finally, cell populations were coated with gold and observed using scan electron microscope (Hitachi S2400). (ii) In order to observe internal structure of Mks by transmission electron microscopy (TEM), culture-derived cells were fixed in a solution containing 2% paraformaldehyde (Sigma) and 2.5% glutaraldehyde

(Sigma) in 0.1 M sodium cacodylate Phospholipase D1 buffer (Sigma) (pH 7.4) for 1 h at room temperature (22 °C). After rinsing with cacodylate buffer (Sigma), cells were post-fixed with a 1% osmium tetroxide (Sigma) in 0.1 M cacodylate buffer (Sigma) for 1 h at room temperature. Cells were then fixed with uranyl acetate (Sigma) (0.5%) in citrate–acetate acid buffer (pH: 5–6) and dehydrated by graduate increasing ethanol (Sigma) concentration (50%, 75% and 100% in distilled water). Finally, cell populations were embedded in Epon (Sigma), cut and Mks ultrastructure observed with TEM apparatus (Hitachi 8100). Results are presented as a mean ± standard error of mean (SEM). Results were statistically analyzed using two-sided non-paired Student’s t-test by Microsoft Excel and considered significant when p < 0.05. CD34+-enriched cells from UCB were expanded using a previously optimized protocol [12] and differentiated toward Mk lineage using a simple protocol containing only two cytokines (TPO and IL-3). Expanded cells were also differentiated, as a control, using the same protocol but without any supplemented cytokines.

The chromatographies were done using a Shimadzu 20A

HPLC

The chromatographies were done using a Shimadzu 20A

HPLC modular equipment with an SPD-20A detector, buy Nutlin-3a and the data were generated and stored using the Shimadzu LC-Solutions Software, acquiring the absorbance data at a rate of 1 Hz, resulting in a total of 9005 points which were processed for each chromatography. The fractal dimension (D) analysis on the chromatographic profiles of the venoms was calculated for the initial 60 min of venom fractioning. This analyses is an alternative to study inter and intraspecific venom variability taking advantage of the multiple waveforms of these and comparing them point to point in all or in partial intervals of elution time. By this study, it is also possible to calculate the probability (P) of the difference Etoposide between two values of D (ΔD) that is used to indicate the contortedness of waveforms, inter venoms. The data were transformed to ASCII format (American

Standard Code for Information Interchange) in data pairs, and analyzed under VeFractDim software according to D’Suze and Sevcik (2010). For the phase plot of D analysis a sliding window sequences (SWS) of the 500 continuous points starting from time 0 of the chromatography was used and with a recursive displacing by 1 s for 60 min of elution stored as [(ti,Di) sets]. For this data sets the contortedness represented by Q = D − 1 was calculated, and a phase plot with their (ti,Qi) sets was constructed. The determination coefficient (ds) was calculated squaring the Spearman rank correlation coefficient as suggested previously ( D’Suze and Sevcik, 2010). The data were plotted using the GraphPad Prism 5 software.

this website All fractions obtained from Ts-DF and Ts-MG venom chromatography separation were analyzed by mass spectrometry performed on a MALDI-TOF AutoFlex III (Bruker Daltonics®, Germany) in linear and reflector modes and the spectra were processed with MassLynx™3.5 (Manchester, UK) and FlexAnalysis 3.3 (Bruker Daltonics®, Germany). Briefly, solubilized fractions (0.5 μL of sample, variable concentrations) were spotted onto the target followed by 0.5 μL of CHCA (α-cyano-4-hidroxycinnamic acid) matrix solution (60% acetonitrile/0.3% TFA), and allowed to dry at room temperature (dried-droplet method). Peptide Calibration Standard II (700 − 4000 Da) and Protein Calibration Standard I (3000–25,000 Da) (Bruker Daltonics®, Germany) were used as external calibrates. Mass spectra from the average of 256 laser pulses from m/z 600 to 39,400 were obtained. The experimental procedure was approved by the Ethical Committee for Animal Experimentation of Brasilia University (CEUA/UnB) under protocol number 133424/2009.