0 software package for Windows. Lodging resistance was used as the dependent variable, while lignin, cellulose, AOVB, NOVB, AOT and WOMT were used as independent variables. Potential microsatellite markers linked to stem solidness genes were identified by screening the F2 population using bulked segregant analysis. DNA was extracted from young leaf tissues using the CTAB method. The solid and hollow stem DNA pools were composed of 5 solid and 5 hollow stemmed F2 plants, respectively. Along with the parental DNA, the bulked DNA samples were used to screen 607 SSR markers (210 GWM [19] and 397 BARC [20]). The PCR mixture

(20 μL) consisted of 2.0 μL of 10 × buffer, 1.6 μL of Mg2 + (25 mmol L− 1), 2.0 μL of dNTP (2 mmol L− 1), 2.0 μL of DNA (10–20 ng μL− 1), 2.0 μL of primer (2 μmol L− 1), 0.2 μL of Taq DNA polymerase (5 www.selleckchem.com/products/ve-821.html U μL− 1), and 10.2 μL of ddH2O and was subjected to a thermocycler program of 94 °C for 5 min; IPI-145 research buy followed by 30 cycles at 94 °C for 1 min, 60, 55, or 50 °C for 1 min (depending on each primer set), and 72 °C for 1 min; with a final extension at 72 °C for 5 min. The PCR products were electrophoresed in 4% polyacrylamide gels and detected by silver staining [21]. Marker-trait associations were identified by single factor ANOVA and the proportion of phenotypic variation explained by single marker loci was determined as

the ratio of sum of squares for marker class divided

by sum of squares of entries [12]. The characteristics Thymidine kinase of stem pith varied significantly among the four genotypes examined (Fig. 1). Solid stemmed XNSX showed the greatest amount of pith material (Fig. 1B), whereas CS and Line 3159 had hollow stems (Fig. 1A and C), and the characteristics of F1 plants were similar to the solid-stemmed parent except for the third and fourth internodes (Fig. 1D). Significant differences were also detected in the anatomical characteristics of the four genotypes, especially the transverse sections of solid stemmed wheat XNSX, which had more mechanical and parenchyma tissues (Fig. 2C and D) than the other three genotypes (Fig. 2A, B, E and F); F1 plants were almost intermediate between their parents in the corresponding values (Fig. 2G and H). The morphological data for the four wheat genotypes are shown in Table 1. AOT in the solid stemmed and F1 plants were significantly larger than that of CS. In contrast, there were only minor differences in AOVB among the four genotypes (Table 1). The widths of stem walls in XNSX and F1 were 2.7- and 2.6-fold that in CS, and WOMT values were 2.1- and 1.7-fold that in CS. Only slight differences were observed in TNVB among the four genotypes, but the WOL of XNSX and F1 plants were significantly higher than those of Line 3159 and CS (Table 1). The contents of cellulose and lignin showed slight differences among the four genotypes.

Alicyclobacillus acidocaldarius DSM 446T was used as an outgroup. The scale bar, 0.02 substitutions per nucleotide position. This study was sponsored by the National Natural Science Foundation of China (Grant No. 81301461, 50974022, and 51074029), and the

863 Program of the Ministry of Science and Technology (Grant No. 2008AA06Z204 and 2013AA064402). The authors wish to thank the technical personnel in the oil field under study for kindly collecting the samples. “
“Extremely halophilic bacteria produce Selleck NVP-BGJ398 enzymes that have potential biotechnological applications; for instance, hydrolytic enzymes that tolerate high temperatures and salt concentrations and are stable in the Icotinib presence of organic osmotic solutes (Ventosa et al., 1998). There have been relatively few studies on halophilic enzymes; however, haloarchaea are known to produce enzymes such as DNase, amylase, esterase/lipase, inulinase, pullulanase,

protease, chitinase, cellulase, and xylanase (Litchfield, 2011). Recently, β-agarase was purified from the extremely halophilic archaeon Halococcus sp. 197A and was characterized as a thermophilic and halophilic enzyme, representing the first agarase identified in haloarchaeon ( Minegishi et al., 2013). We previously isolated Halolamina rubra CBA1107T (= CECT 8421T, JCM 19436T) from non-purified solar salt ( Cha et al., 2014). While investigating extremozymes from haloarchaea, H. rubra CBA1107T was found to have agarose-degrading activity. Agarase

(EC 3.2.1.81) has important laboratory and industrial applications for liberating DNA and other embedded molecules from agarose and producing bioactive neoagarosaccharides ( Fu and Kim, 2010). This is the first report of the genome sequence of H. rubra CBA1107T, which is expected to provide general sequence information for halophilic carbohydrate-active enzymes (CAZymes). The draft genomic sequence for H. rubra CBA1107T was obtained from 1,695,985 reads spanning 153 Mb (257-fold coverage of the genome) using the 400-bp library Ion Torrent PGM sequencer ( Rothberg et al., 2011) with a 318D sequencing chip, according to the manufacturer’s instructions. Sequences were assembled into 71 contigs > 1 kb in size with an N50 contig size of approximately 77 kb using GABA Receptor the CLC Genomics Workbench 6.5 for de novo assembly (CLC Bio, Aarhus, Denmark). Gene prediction and contig annotation were carried out using RNAmmer 1.2 ( Lagesen et al., 2007), tRNA scan-SE 1.21 ( Lowe and Eddy, 1997), and the Rapid Annotation using Subsystem Technology (RAST) pipeline ( Aziz et al., 2008). The genome features of H. rubra CBA1107T are summarized in Table 1. The genome is 2,955,064 bp in length, with a G + C content of 69.0%. Single 16S and 23S rRNA genes and 47 tRNA genes were identified. The genome contains 3046 coding sequences and 257 subsystems based on RAST results.

Stepping through the diverse interactions between coronary artery disease, congestive heart failure, cerebrovascular disease, peripheral vascular disease, and cardiac arrhythmias, the review analyzes thoughtfully the epidemiologic and pathophysiologic interrelationship among these diseases independent of their often common and shared risk factors. Discussion of multiple population studies showing the link between carotid arterial intimal medial thickness and numerous pulmonary function parameters—including FEV1, diffusing capacity of carbon monoxide see more (DLCO), residual

volume (RV), and peak expiratory flow rate—help the reader understand the relationship between the pathogenesis of atherosclerosis and COPD. Genetic studies illustrate links between matrix metalloproteinases and glutathione-S-transferase and the development of emphysema and plaque rupture. In addition, discussion of the epidemiologic outcomes of patients with both COPD and cardiovascular disease illustrate the poor prognostic implication of these overlapping clinical entities and provide opportunities for future research and public health interventions.

Overall, this in-depth review of COPD steps through emerging research of risk factors for disease, attempts and challenges of better describing and categorizing this disease, as well as the comorbid conditions associated with COPD as we continue to learn about the complexities of this trans-isomer ic50 systemic syndrome. This series not only celebrates how far we have come since our early descriptions and definitions of disease, but also it highlights how much is still unknown, and reveals many potential areas for future research. It is likely that in the decades to come our current understanding of COPD will continue to change and evolve just as it has during the past decade. Many different areas of research as well as varied study designs will be required to understand more completely this disease that varies throughout a population and among individuals during their life span. The current set of reviews provides a framework for areas that are ripe for future investigation as well as points out

the challenges with which clinical and translational research communities are faced as we further our understanding of this complex, common clinical entity. “
“The affiliation for Brian T. Layden in the article entitled “Short chain HSP90 fatty acids and their receptors: new metabolic targets” was incomplete. Dr. Layden is also affiliated with the Jesse Brown Veterans Affairs Medical Center, Chicago, IL. His complete affiliation is as follows: From the Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL, and Jesse Brown Veterans Affairs Medical Center, Chicago, IL. “
“Thomas C. Kwee and Habib Zaidi Jannie P. Wijnen and Dennis W.J. Klomp Magnetic resonance spectroscopy (MRS) is a noninvasive technique that provides in vivo information about tissue metabolism.

S1). The PCR products for each variable region were pooled according to the natural distribution as described on V-Base. The light chain variable regions were cloned first using restriction digest with SfiI and AvrII for Vλ and SfiI and BsiWI for Vκ and transformed into electrocompetent TG1 cells (48 μg DNA in 48 200 μL Apoptosis Compound Library cost transformations for Vκ and 65 μg DNA in 65 200 μL transformations for Vλ). Transformations were spread on 2xYT medium with 2% glucose and 100 μg/mL carbenicillin, which were incubated overnight at 30 °C. The following morning the bacteria were scraped from the plates, combined and plasmid DNA purified

with the GenElute™ HP Maxiprep Kit (Sigma-Aldrich). The resulting DNA was prepared for cloning VH with NcoI-HF and NheI-HF. The ligated DNA was cleaned with the Wizard® SV Gel and PCR Clean-up system (Promega) and transformed into electrocompetent TG1 cells (66 μg DNA in 66 200 μL transformations for Vκ and 100 μg DNA in 100 200 μL transformations for Vλ). Transformations were spread on 2xYT medium PD98059 manufacturer with 2% glucose and 100 μg/mL carbenicillin, which were incubated overnight at 30 °C. The following morning the bacteria were scraped from the plates, combined, and stored in 15% glycerol 2xYT at − 80 °C. The scFv library was constructed similarly to the above described Fab library with the following changes. Primer sequences are listed in Table S3 and Table S4. cDNA from 20 PBMC samples, 8 bone marrow

samples, 1 lymph node sample, and 1 spleen sample ifenprodil were used. The reverse secondary PCR primers for VH and forward secondary primers for Vκ and Vλ had complementary extensions for an AST(G4S)3 linker and the forward secondary PCR

primers for VH and reverse secondary primers for Vκ and Vλ had sequences to add flanking SfiI restriction sites. A tertiary PCR step was then done to assemble the full length scFv fragment, which was next cloned into pXHMV-scFv ( Fig. S1) using the SfiI sites. The ligated DNA was transformed into electrocompetent TG1 cells (147 μg DNA in 120 200 μL transformations for Vκ and 44 μg DNA in 40 200 μL transformations for Vλ). Transformations were spread on 2xYT medium with 2% glucose and 100 μg/mL carbenicillin, which were incubated overnight at 30 °C. The following morning the bacteria were scraped from the plates, combined, and stored in 15% glycerol 2xYT at − 80 °C. Both XFab1 and XscFv2 phage libraries were rescued using a modification of the standard protocol (Marks et al., 1991). XFab1 was rescued in four batches (two for XFab1λ and two for XFab1κ) each starting with 5-fold more bacteria than the sub-library size. XscFv2 was rescued in five batches (two for XscFv2λ and three for XscFv2κ) with XscFv2λ starting 5-fold more bacteria than the sub-library size and XscFv2κ starting with 3.33-fold more bacteria than the sub-library size. For all rescue batches, cultures were seeded at a starting density of 0.

SW480 colon carcinoma cells were treated with complexes 1–4 for 48 h with concentrations between 5 and 40 μM, and cells were then collected for annexin V–FITC and propidium iodide staining. Exemplarily, dot plots of cell populations treated selleck chemical with 5 μM of each compound from one representative experiment are shown in Fig. 6. Complex 1 shows the strongest impact on cell viability, only 15% cells remain viable, whereas cells in early and late apoptosis amount to 72% in total.

Complex 2 shows a much more moderate impact on cell viability, indicated by 63% viable cells and only 31% apoptotic cells. The same applies for complex 3, yielding a slightly lower amount CB-839 of viable

cells (56%) and a slightly higher amount of apoptotic cells (35%). Complex 4 is the least potent compound and has hardly any impact on the cells at a concentration of 5 μM. Percentages of necrotic cells remain generally low (with a maximum of 14% in the case of 1). The concentration dependence of apoptosis/necrosis induction is illustrated in Fig. 7, and the corresponding values are listed in Table 2. They provide further evidence for the differences in cytotoxic potencies of the compounds, correlating with those observed in the MTT assay. Whereas 5 μM of compound 1 is sufficient for near-maximum effect, even 40 μM of compound 4 is insufficient for comparable effects. Compounds 2 and 3 require concentrations MycoClean Mycoplasma Removal Kit of 20 μM to induce 57% and 61% apoptosis, respectively, taking intermediate positions. Furthermore, compounds 2–4 induce higher proportions

of necrotic cells relative to those undergoing apoptosis, making compound 1 the one with the most favorable properties. Binding paullone ligands to ruthenium(II) and osmium(II) arene moieties led to a considerable improvement of solubility compared to the uncomplexed compounds, enabling biological studies. A comparison with previous results for Sadler’s ruthenium complex with ethylenediamine (instead of the paullone ligand), [(η6-p-cymene)RuII(en)Cl](PF6), (IC50 values of 7.1, 3.5 and 4.4 μM in A549, SW480 and CH1 cells, respectively) under the same experimental conditions  [17] reveals that the presence of the paullone ligand causes a 2.3- to 6.6-fold (complex 1) and a 1.2- to 2.9-fold (complex 3) increase in cytotoxicity, depending on the cell line. In general, complexes with L1 show stronger cytotoxic effects than those with L2 in all human carcinoma cell lines tested. In the most sensitive cell lines SW480 and CH1, IC50 values of complex 1 are in the nanomolar range, whereas in the least sensitive cell lines A549 and LNCaP IC50 values are in the low micromolar range.

In total, 427 subjects were referred to the interdisciplinary assessment. Of those, 160 were not eligible: 79 (48%) did not have WADs; 46 (28%) had a WC of 100%; 17 (10%) had insufficient German language skills or were unable to complete the questionnaires; 6 (4%) had other medical reasons; 5 (3%) had acute comorbidity that limited testing (fracture or severe psychiatric disorder); 3 (2%) were younger than 18 years

or older than 65 years; 2 (1%) had WADs grade III or IV; and 2 (1%) were pregnant. All participants agreed PD0325901 to participate in this study. The Medical Ethics Committee of Canton Aargau granted ethical approval for this study (EK AG 2010/055). A rehabilitation physician performed a review of the medical history and a physical examination (approximately 60min), followed by FCE tests administered by a physiotherapist. After determination of eligibility, patients completed questionnaires

and carried out FCE tests (60min). This was followed by a brief educational intervention and a trial therapy that included a combination of strength exercises, education (ergonomic), and home exercises. The interdisciplinary rehabilitation assessment ended with a face-to-face discussion with the patient about strategies to facilitate recovery. Fitness-for-work certificates or WC settlements were explicitly not part of this interdisciplinary assessment. A sample of 21 physiotherapists (11 women) from the rehabilitation clinic served IPI-145 molecular weight as FCE assessors. Nineteen had attended a 2-day FCE training course of the Swiss Association of Rehabilitation.17 Before the study, all had performed at least ten 1-day FCEs in the previous year (median, 30; interquartile range [IQR], 20–33), had a minimum of 1-year experience in work rehabilitation (median, 3; IQR, 2–3), and had a minimum professional practice experience of 1 year (median, 5y; IQR, 3–12.5). In this study, inter- and intratester

reliability of the FCE assessors was good for the 2-point scale used to determine submaximal effort.18 WC was used as a measure of ability to work. WC was assessed at Florfenicol baseline and at the 1-, 3-, 6- and 12-month follow-ups. WC was determined by the treating physician, usually a general practitioner, and represents the proportion ability to work regarding the preinjury work. Estimation of WC may be determined by suggested measures of WC and based on current national guidelines.19 and 20 WC is expressed in a percentage (0–100%) and is translated into days or hours of modified work. For example, if a worker is deemed to have a WC of 50%, he/she will work for 2.

Further investigation into the importance of this cytokine family in disease models is necessary to determine whether they play a central role in progressive joint degeneration in OA. Chemokines are small molecules that play an important role PD 332991 in mediating recruitment and trafficking of inflammatory cells and mesenchymal progenitors. Many chemokines are produced in the joint tissues of patients with OA [28] and [41].

Thus, they represent potential therapeutic targets to either enhance repair mechanisms or decrease inflammation in patients with OA. We recently demonstrated that synovial inflammatory infiltrates were associated with expression of a distinct mRNA chemokine signature in patients with meniscal injury [87]. The signature included IL-8, CCL5, CCL19 and its receptor CCR7. Expression of CCL19 and CCR7 was also associated with greater pre-operative symptoms, and based on these observations, their expression may have utility as

biomarkers of early synovial inflammation. CCR7 is expressed by synovial fibroblasts, and mediates upregulation of VEGF in response to its ligand, CCL19 [14], suggesting a role in synovial angiogenesis. Other chemokines, PD0332991 specifically MCP-1 and MIP-1β, have been associated with knee pain in patients undergoing arthroscopic procedures [24]. An important role for MIP-1γ produced by T helper cells in the synovium was demonstrated in a murine model of OA induced by ligament transection [94]. Similar to the cytokines discussed above, chemokines can induce matrix metalloproteinase (MMP)-3 and proteoglycan loss from articular cartilage [11]. Furthermore, many chemokines may directly affect osteoclast-mediated remodeling of peri-articular bone. In summary, chemokines represent a class of soluble inflammatory mediators that have pleiotropic effects on multiple joint tissues, and may contribute to inflammation and clinical symptoms in patients with OA. Further studies are needed to investigate the utility of targeting specific chemokines for symptom control or disease modification in OA. There is increasing evidence that synovial inflammation

plays Chlormezanone a critical role in the symptoms and structural progression of OA. Importantly, synovitis has been shown to correlate with symptom severity, rate of cartilage degeneration and osteophytosis (Fig. 2). The synovial response in OA is complex and variable with regard to histologic pattern (Fig. 1), and in part this complexity can be attributed to changing patterns as disease evolves and progresses. Although structural joint damage in OA is a constant feature, the clinical syndrome of OA is quite variable, with differences in affected joint patterns, risk factors, rates of progression, and severity of symptoms. Further efforts to understand the cellular and molecular variability of OA-associated synovitis may provide insights into the clinical heterogeneity of the disease.

The protein www.selleckchem.com/products/Sunitinib-Malate-(Sutent).html structural modeling together with the CEP Server (Kulkarni-Kale et al., 2005) are trustworthy bioinformatics tools which allow to achieve this knowledge with great accuracy. Using these procedures, in this study we identified in the Pp-Hyal 3D-structural model the location of five conformational and seven linear predicted epitopes, thus corroborating with the results observed by Western blotting and contributing for a better understanding of the immunogenic potential of this Pp-Hyal venom allergen. The structural superposition of the three molecules (data not

shown) revealed that the folding of rVes v 2 (Skov et al., 2006), Api m 2 (Markovic-Housley et al., 2000), and Pp-Hyal-3D structures were similar as well as the active site location, but as described by Skov et al. (2006), the Hyal proteins from bee and wasps have significant structural differences in its surfaces related to topology and also in charge distribution, what may explain the unlikely occurrence of cross-reactivity between them. These data could be confirmed in our study since cross-reaction was only observed between wasp venoms of the same genus, Polybia, and no reaction with the venoms of A. mellifera, S. invicta, A. pallipes pallipes, or P. lanio lanio. Meanwhile, these results differ from some reports of wasps in temperate climates, in which cross-reactivity

has been observed between the venoms of wasps and bees, as an example the recent study of Eberlein et al. (2012) that estimated that approximately 59% of patients

allergic to Hymenoptera trans-isomer chemical structure venom show positive results for both bee and wasp. This is mainly due to the IgE-specificity of hyaluronidase, being that this allergen is the most conserved venom component. The absence of cross-reactivity is important, as it allows identification of the insect responsible for sensitization of the victim (or at least the phylogenetically closest insect), which is crucial to develop immunotherapy for allergic patients. The production and use of allergen-specific antibodies (native and/or recombinant), such HSP90 as the Pp-Hyal-specific antibody produced here, has been an ongoing strategy to overcome difficulties in the diagnosis and treatment of allergies. In this context, experiments for the production of the major allergens from the P. paulista venom (Hyal, Ag5 and PLA1) in the recombinant forms and the obtaining of its specifics antibodies are being conducted. This work was supported by FAPESP (Proc. N° 2009/51539-1) through a Doctoral fellowship to Débora Laís Justo Jacomini. The authors also thank the support by PROAP-CAPES from the Post-Graduation Program of Biological Sciences (Cellular and Molecular Biology) at the Univ Estadual Paulista, UNESP, SP, Brazil. We thank Nora Hersoug Nedberg for the typewriting revision of the manuscript.

The width of the stenotic canal can often be

measured in higher degrees of stenosis as well with B-mode imaging. The diameter can then be related to the distal one for measuring the degree of stenosis following the NASCET method, but this is only possible with excellent conditions for insonation. Color Doppler is helpful in delineating plaques of low echogenicity or proving Cobimetinib order absence of flow in the occluded ICA. But it does not allow precise diameter measurements due to its low frame rate and a huge influence of the gain. Grading of stenoses above 50% is the basis of clinical decisions. Combining morphologic and several hemodynamic features allows a reliable description of at least four classes of stenosis. Such a multiparametric approach avoids severe misclassification as is done with a simplified PSV criterion or its derivates alone (end diastolic velocities in the stenosis, ratio of velocities ICA/CCA). Secondary criteria may be helpful in supporting the diagnosis as the extend of flow disturbances being most pronounced in a 70–80% stenosis and diminishing LY294002 research buy together with a reduced flow volume in very a high degree stenosis In a high degree stenosis the hemodynamic effect is shown by the appearance of collateral flow, which is driven by the poststenotic pressure drop. Another effect is a poststenotic decrease of velocity and pulsatility of flow. All these effects can be measured reliably by extra-

and intracranial Doppler duplex sonography. The question is whether the trial result that surgery is highly beneficial in case of a symptomatic ≥70% NASCET stenosis as measured by angiography can be translated into: beneficial in case of a “hemodynamically relevant stenosis” because 70% stenosis is the threshold from which a pressure drop and decreased poststenotic flow can be observed. This seems reasonable but is so far not accepted as level

one evidence. [8]. A meta-analysis of studies correlating PSV and percent of stenosis as measured by angiography showing a considerable disagreement was the background of not accepting ultrasonography. The old concept of a multiparametric diagnosis was not considered. However it has been used and taught over decades. New technical elements have been continuously introduced. But there Y-27632 clinical trial is a lack of well designed and large studies for this concept, including all these new techniques. In older publications e.g. the definition for measuring the degree of stenosis (NASCET or ECST) is missing. This is one of the reasons why, they do not add very much to the evidence. Even with such new studies some disagreement between methods will persist as explained above. Clinically most useful would be to repeat randomized carotid surgery trials with ultrasonography as criterion for decision in symptomatic patients. However it is ethically not justified to randomize for this question again.

g., Chafe, 1976 and Schwarzschild, 1999). In contrast, NEW INFORMATION describes information the speaker expects to introduce to the listener in the sense of “newly activating” it in the

listener‘s consciousness ( Chafe, 1976). FOCUS refers to the new/informative or contrastive part of an utterance. Whereas, BACKGROUND denotes less relevant information (e.g., Vallduvi & Engdahl, 1996). Experimentally, focus is often induced as contrastive focus, where the newness of the information is emphasized by its contrast to previously focused information (e.g., Jacobs, 1988). A special type of contrastive IPI-145 cost focus is corrective focus, where an assumption is explicitly corrected. These information structural concepts are thought to be realized by distinct prosodic (i.e., accenting) and/or syntactic (e.g., sentence position) phenomena (see e.g., Chafe, 1976, Féry and Krifka, 2008, Skopeteas and Fanselow, this website 2010 and Steedman, 2000). In the present study, we aim to investigate how a previously presented context, in particular a context introducing all characters of a fictitious scene with emphasis on one of them as the aboutness topic, affects the comprehension of a subsequent canonical (subject-before-object) or non-canonical (object-before-subject) declarative sentence in German.

Before we present the two experiments (Experiment 1: offline comprehensibility judgments, Experiment 2: Event-related potentials (ERPs) during online sentence processing) we first give a brief overview of German word order, the underlying neurocognitive mechanisms of sentence and discourse

processing, as well as previous findings concerning information structural concepts and sentence processing relevant to understanding the motivation and predictions of the present study design. Word order in German is relatively flexible. Reordering of constituents within a sentence can be used to highlight the communicatively Fossariinae relevant part of the utterance. German has a strong subject-first preference (e.g., Gorrell, 2000), but reordering of constituents within a sentence is possible, because syntactic roles can still be assigned correctly due to morphological case marking at the respective determiner or determiner and noun. Case marking of the subject by nominative (NOM) and object by accusative (ACC) case is ambiguous for feminine, neuter, and plural noun phrases, but unambiguous for masculine singular noun phrases. The example sentences (1a, b) illustrate case marking for masculine subjects and objects in German with the finite, transitive verb in the second sentence position. (1a) depicts a canonical declarative sentence with typical subject-before-object (SO) word order. (1b) depicts a non-canonical sentence with object-before-subject (OS) word order. (1a) Der Uhu malt den Igel. [the[NOM] owl[NOM]]subject [paints]verb [the[ACC] hedgehog[ACC]]object.