In the second study, GSTP1 overexpression was observed in the synaptosomal fraction of PD cases and was suggested to protect cells against rotenone-induced neurotoxicity via oxidative and ER stress attenuation in a PD cell model [152]. Three other studies by Choi et al. proved useful for elucidating some of the PTMs associated

with PD. Using 2-DE, they demonstrated oxidation in multiple proteins previously linked to PD, including the chaperone DJ-1, superoxide dismutase Cu/Zn, as well as the de-ubiquitinating ERK inhibitor in vitro protein UCH-L1 in the frontal cortex of PD patients compared to controls [238], [239] and [240]. Recently, van Dijk et al. performed a proteomic analysis of the locus ceruleus, one of the earliest affected brain regions in PD [241]. By comparing PD patients (n = 6) versus controls (n = 6) with a label free approach, they identified 2′ 495 proteins of which 87 were differentially expressed between groups. In particular, a pathogenic role for aminoacyl-tRNA-biosynthesis was highlighted. Overall, these proteomics studies were successful in confirming existing theories about PD pathogenesis (Fig. 3). The majority of the differential proteins were indeed implicated in buy Pexidartinib mitochondrial dysfunction, energy metabolism

impairment, oxidative stress, protein aggregation, cytoskeleton impairment, or inflammation. Whereas some of the observed protein alterations were previously associated to PD pathogenesis (i.e., ferritin), others were novel candidates such as CNDP2, mortalin, regucalcin, or seipin. Curiously, α-SYN overexpression did generally not show up significantly in these studies [196], [232] and [241]. The most probable explanation comes from the fact that in a tissue-based approach, the overexpression of synaptic α-SYN in surviving DA neuronal PD cells may be compensated by the higher number of healthy neuronal cells in control patients. These studies also suggested some less conventional

pathways such as defects in protein translation, ER stress, blood brain barrier or extracellular matrix abnormalities (Fig. 3). Of note, it was tuclazepam sometimes unclear whether the observed protein changes were a cause or a consequence of the neurodegenerative process. In tissue-based approach, the decrease in neuronal protein levels may simply reflect PD associated neuronal loss. Further biological evaluation of the pathogenic mechanisms underlying these protein alterations may provide new therapeutic targets for PD. During the past 10 years, only a small number of human tissue based proteomics studies have been published due to limitations in their availability, number, quality and complexity. In the context of a worldwide decline in autopsy rate, some of these issues can be partially overcome through a facilitated access to existing brain banks which ensure the collection of well characterized and preserved brain tissues.

, 1997). Tagged termini might influence binding and postbinding processes of TDH and could be an explanation for the weaker hemolytic activity of tagged proteins. One disulfide bridge is formed in the subunits (Tsunasawa et al., 1987 and Nishibuchi et al., 1989), however, the disulfide bond is possibly not important for hemolytic activity of TDH (Baba et al., 1992 and Yanagihara et al., 2010). It has been shown that forming of disulfide bonds is generally possible in a simple batch reaction of the E. coli based cell-free protein synthesis system ( Kim and Swartz, 2004). The addition selleck screening library of tags offers a great advantage for the

purification of the toxin and its further application, therefore we decided to analyze the toxin variant synthesized with an additional

C-terminal His-tag more intensively. His-tagged toxins were purified using His-tag Dynabeads®. Selleck Dabrafenib Aliquots of CRMs and purified toxin derivatives were loaded on SDS-PAGE and transferred to nitrocellulose membrane after electrophoresis (Fig. 7). The Western Blot revealed again that only one protein was synthesized when the DNA construct encoding preTDH-His (protein band I, Fig. 7B lane 1) was used as a template, whereas from the PCR product encoding mTDH-His two His-tagged proteins were produced (bands II and III in lane 2). This result was also visible after purification with His-tag Dynabeads® (lanes 3 and 4). The purified His-tagged mTDH displayed hemolytic activity as did the unpurified TDH from the supernatant fraction (data not shown). To confirm the identities of the cell free expressed TDH proteins, three protein bands were excised from SDS-PAGE gels and subjected to tryptic in-gel digestion followed by tandem MALDI-TOF mass spectrometry (MS/MS) protein identification. Protein database searches for band I retrieved thermostable direct hemolysin A (TDH2) of O3:K6 reference strain V. parahaemolyticus RIMD 2210633. MALDI-TOF MS/MS spectrometry analysis revealed two diagnostic peaks corresponding to peptides DTTFNTNAPVNVEVSDFWTNR (m/z 2427.1) and SDQVQLQHSYDSVANFVGEDEDSIPSK

(m/z 2994.4) (see Suppl. Fig. S2). The same peaks were found in band II. Interestingly, Carnitine palmitoyltransferase II the peak at m/z 2994.4 was missing in the MS analysis of band III, but instead a peak at m/z 2877.6 was observed, which is diagnostic for thermostable direct hemolysin S (TDH1) of V. parahaemolyticus from RIMD 2210633 and corresponds to peptide SGQVQLQHSYNSVANFVGEDEGSIPSK. This peptide contains three amino acid exchanges compared to the corresponding sequence in TDH2, which could be confirmed by MALDI-TOF MS/MS analyses. Thus we concluded that protein I corresponded to the preprotein of TDH2 containing the signal peptide, while proteins II and III were derived from the chromosomal genes tdh1 and tdh2 lacking the sequence encoding the signal peptide. As shown above, two proteins were synthesized when primers containing gene specific sequences for the mature toxins were used.

4 × 106 seedlings per hectare. The sandy loam soil [Typic Fluvaquent, Entisols (US taxonomy)] contains 12.58 g kg− 1

of organic material and 75.19, 45.52 and 99.3 mg kg− 1 of available N, phosphorus and potassium, respectively. Plot dimensions were 4 × 5 m and plots Entinostat supplier were separated by an alley 1 m wide with plastic film inserted into the soil. Each of the treatment had three plots as repetitions in a complete randomized block. The treatment plots received 240 kg ha− 1 at the booting stage. The control plots received no N at the booting stage. All other field conditions and cultivation managements were kept uniform. During the period of wheat anthesis, the anthesis dates were recorded by dotting the glumes and hanging time tags on the wheat plants. Caryopses that bloomed on the same day but developed on different days for the two treatments were chosen for experimentation. Samples were harvested at 15 and 45 DAA. First, 2 mm cubic blocks were

cut by cross-sectioning from wheat caryopses harvested at 15 DAA. The specimens were then fixed with 2.5% glutaraldehyde and 1% paraformaldehyde in a 0.05 mol L− 1 cacodylate buffer solution (pH 7.2) and post-fixation treatment in 1% osmic acid in a 0.15 mol L− 1 sodium cacodylate buffer solution (pH 7.2) for 3 h was applied. The blocks were washed, dehydrated through an ethanol series of 30%–100%, and embedded click here in Spurr’s low-viscosity embedding medium. Sections of 1 μm thick were cut with a glass knife on a Leica Ultracut R (Leica Microsystems, Inc., Wetzlar, Germany), and stained with 0.5% toluidine blue O for 5 min. The sections were visualized and photographed with a Leica Dmls microscope (Leica Microsystems, Inc.). To reflect the nature of caryopsis structure, the findings were compared and confirmed in numerous sections made from

developing grains. Five representative regions of transverse sections of the endosperm were observed for every specimen: subaleurone in dorsal endosperm (SDE), center in dorsal endosperm (CDE), modified aleurone (MA), subaleurone in ventral endosperm (SVE), and center in ventral endosperm (CVE), using three replications Phosphoprotein phosphatase and 20 micrographs representing ten blocks from different regions. Mature grains were harvested at 45 DAA and fractured by applying slight pressure on the middle of the caryopsis with a razor blade. The sample thickness was ~ 3 mm. Caryopses were mounted with the fractured surface facing upwards on a specimen stub and sputter-coated with gold before viewing with a scanning electron microscope (XL30 ESEM, Philips, The Netherlands) at 20 kV to observe the distribution of SGs. The samples at 15 DAA were used to determine the numbers and percentages of SGs. SGs observed in the image were first marked with a specified color using Photoshop CS4 software (Adobe, U.S.A.) and the image was then analyzed to determine the numbers and percentages of SGs using software Image-Pro Plus 6.0 (Media Cybernetics, U.S.A.).

Fletcher and Frid (1996) systematically manipulated the amount of walking on different communities (often referred to as “trampling” in the literature) and found

that the abundance of some species increased whilst others declined as a consequence. There is a vast amount of literature examining recreational ecology, the study of the ecological relationships in recreational Selleckchem Bleomycin contexts between human and nature; however many of the empirical studies focus on one particular activity (e.g. trampling; Beauchamp and Gowing, 1982 and Brosnan and Crumrine, 1994; or four-wheel driving; Priskin, 2003a) and/or on one particular species (e.g. mussels; Smith et al., 2008). Consequently, apart from descriptive review articles (e.g. Branch et al., 2008 and UK CEED, 2000), there appears to be little research simultaneously examining the impacts caused by a range of activities on this particular environment (rocky shores), or focussing on the benefits such activities may have on the visitor. Priskin’s paper (2003b) is one exception that examined the detrimental effects of different activities. Using a survey completed by visitors as they left the shore, Priskin examined tourists’ perceptions of twelve activities according to their impact on sandy shores and compared this with her personal knowledge guided by the literature. Some activities were seen as more damaging

than others, for instance fishing was seen as very harmful whilst swimming Nintedanib research buy was rated as slightly harmful. Visitors were generally aware of some of the impacts activities had on the environment but rated these consistently as less harmful than the expert did. Priskin’s contribution is important as it compared visitor and expert perceptions, which helps work towards consensual solutions, and

it compared a range of activities, which improves our understanding of the relative harm of individual activities. However, several questions remain. First, Priskin found preliminary differences between Ribose-5-phosphate isomerase the public and her own ratings, but conclusions would be more powerful if perceptions from the general public were compared with a larger sample of experts within the coastal field. Second, the ratings in Priskin’s study assumed that all activities were similar in frequency; hence it would be useful to see if conclusions differ when commonness is taken into account. Third, it is unknown whether these findings would be similar in other habitats, such as rocky shores. Finally, and perhaps most importantly, Priskin examined the negative impacts associated with a visit to the coast, but what are the benefits associated with the different activities, for instance on the visitor’s wellbeing? Only considering both together will allow us to properly understand the impacts, which could then potentially help inform management techniques.

2 as the first schizophrenia-associated CNV [21 and 22], analyses of rare CNVs involving >20,000 cases have revealed associations at more than 15 loci [20, 23 and 24] (Figure 2). The majority of these CNVs substantially increase the risk of developing schizophrenia, with odds ratios (OR) between two and 60 [24]. As their frequency among patients is often less than one in 500,

their individual contribution to the total population variation in schizophrenia genetic liability is small [25], although collectively they are found in around 2.5% of patients [24]. Most schizophrenia-associated CNVs are large and recurrent, meaning multiple mutation events have occurred at the exact same, or near identical, genomic location. The breakpoints HDAC inhibition of recurrent CNVs are usually flanked by repetitive genomic elements such as low copy repeats (LCRs), which mediate mutation through non-allelic CDK activation homologous recombination [26]. 10 recurrent CNVs have been associated with schizophrenia at a level of statistical support that survives correction for the multiple testing of 120 potential recurrent CNV loci in the human genome (Figure 2). Drawing biological insights from recurrent CNVs remains a challenge, largely because multiple genes and regulatory elements are often disrupted. However, single-gene disrupting non-recurrent CNVs have also been associated with schizophrenia at NRXN1, VIPR2 and PAK7. These mutations have the potential

to offer clearer insights into disease pathogenesis, although only the NRXN1 Rapamycin order association survives correction for the multiple testing of all human genes (∼20,000). NRXN1 encodes a synaptic cell adhesion molecule neurexin 1 that links presynaptic and postsynaptic neurons [ 27]. Gene-set analyses have

shown rare CNVs in schizophrenia to be enriched among biological pathways previously implicated in schizophrenia, such as the NMDAR and metabotropic glutamate receptor 5 (mGluR5) components of the post synaptic density (PSD), calcium channel signalling (see single nucleotide polymorphisms below) and FMRP targets [20]. Additional gene-sets recently implicated in rare CNV studies include signalling components within the immune system, chromatin remodelling complexes and targets of microRNA miR-10a [20]. Schizophrenia-associated CNVs have been shown to increase risk for additional neuropsychiatric disorders [28• and 29]. For example, schizophrenia-associated duplications of the Williams-Beuren and Prader-Willi/Angelman syndrome regions are also implicated in ASD [9 and 30], deletions of 15q11.2 and 15q13.3 in epilepsy [31 and 32] and duplications of 16p13.11 in attention-deficit hyperactivity disorder (ADHD) [33]. Up to 72 pathogenic CNVs, which include the majority of those presented in Figure 2, are enriched in large cohorts of patients with early onset neurodevelopmental phenotypes, such as ID, ASD and congenital malformations (CM) [34 and 35].

2007); often, there are also diverse changes in water levels, habitat structures and water residence times (Jones & Elliot 2007). The trend of increasing water temperatures and longer ice-free periods in recent decades, confirmed in Lake Onega, was also found to apply to various small lakes in north-western Russia, Finland, Sweden, Norway (Weyhenmeyer et al. 1999, Adrian et al.2009, Finland’s Fifth National Communication under the United Nations Framework

Conventions on climate change, 2010 and Efremova et al., 2010) and other regions (Austin & Colman 2008). For example, it was found in Lake Superior, the largest GSK1120212 solubility dmso and coldest of the North American Great Lakes, that the summer water temperature had increased Selleckchem 17-AAG by 3.5°C over the previous 100 years (Austin & Colman 2008): this is the greatest warming of any lacustrine ecosystem in the last three decades. Significant correlations between physical parameters (ice-free period, water temperature, precipitation) and different characteristics of biota (Chl a, zoobenthos), revealed by the present study of the Petrozavodsk Bay ecosystem, were also found for other shallow and relatively unpolluted small lakes in northern Russia ( Maksimov et al. 2012). The expected impacts

on biota, however, can differ strongly between ecosystems depending on the climatic region. One of the first studies of the impact of climate on biota was done by Adrian et al., 1995 and Adrian et al., 1999 and showed that the composition, timing and maximum abundance of the phytoplankton and zooplankton communities that start to develop in the spring

were strongly dependent on the duration of the winter ice-cover. In different lakes climate warming leads to greater primary productivity with intense algal blooms Baf-A1 nmr (Blenckner et al., 2007 and Jeppesen et al., 2009). As far as Lake Onega is concerned, we also found a close correlation between the abundance of phytoplankton and, in particular, between the abundance of Cyanobacteria and climatic variables (especially NAO). The positive correlations between NAO and summer Cyanobacteria abundance found for the study area may be mediated by the precipitation rate. This rate increases significantly in years with a high positive NAO, resulting in an increase of nutrient loading from the catchment area. The Cyanobacteria bloom, a common summer phenomenon, has been observed in Petrozavodsk Bay since the 1980s (Sharov 2008). Results from Swedish lakes (Weyhenmeyer 2004) and Lake Pääjärvi, Finland (Järvinen et al. 2006) suggest, moreover, that temperature-sensitive phytoplankton groups such as Cyanobacteria and Chlorophyta would benefit from the earlier warming-up of the lake water and the earlier onset of temperature stratification. Water temperature was distinguished as the most important factor reflecting climatic variability in different studies (Adrian et al. 2009).

, 2007). The introduction of the meta-nitrophenylamine group in C-3 of nor-beta, leading to QPhNO2, could increase its growth inhibitory effects toward HL-60 cells, regardless of the incubation period. It is interesting to note that while QPhNO2 presents a higher Sirolimus in vitro IC50 value (0.91 μM)

after 72 h incubation, the other quinones, nor-beta and doxorubicin, presented a clear time-dependency, increasing their activity after 72 h. Quinones are redox active molecules that form semiquinones and hydroquinones that can redox cycle in the presence of oxygen, leading to the formation of reactive oxygen species (ROS) (Asche, 2005, de Abreu et al., 2002a, Hillard et al., 2008 and Ferreira et al., 2009). In fact, it is postulated that ROS generation selleckchem and the alkylation of cellular nucleophiles, including DNA and enzymes with a –SH group, account for the mechanism of cytotoxic action of drugs containing a quinone moiety (Asche, 2005, de Abreu et al., 2002a, Hillard et al., 2008 and Ferreira et al., 2009). In view of this fact, we measured the cytotoxic effect of QPhNO2 in the presence of NAC, an antioxidant that acts as a ROS scavenger (Zafarullaha et al., 2003). The IC50 value for QPhNO2 increased from 0.32 to 1.03 μM and that for nor-beta increased from 2.01 to 2.72 μM (Table 1, column 3). While these data suggest the participation of ROS in QPhNO2 cytotoxic effects,

they do not exclude other direct targets. Doxorubicin effects, in contrast, were not affected by NAC treatment (Table 1). ROS production was also evaluated in HL-60 cells by flow cytometry using the oxidation-sensitive fluorescent dye H2-DCF-DA after 1 h of incubation. QPhNO2 and nor-beta stimulated ROS generation, while doxorubicin was inactive (Fig. 2). ROS generation was higher in the first hour than after 3 h of incubation (data not shown). The pre-incubation with

NAC protected the cells from oxidative stress, reducing intracellular ROS generation. Cell death can be classified according to its morphological appearance, which may be apoptotic, necrotic, Lck autophagic or associated with mitosis catastrophe (Melino, 2001 and Okada and Mak, 2004). Cells undergoing apoptosis show typical well-defined morphological changes, including plasma membrane blebbing, chromatin condensation with margination of chromatin to the nuclear membrane, karyorrhexis (nuclear fragmentation), and formation of apoptotic bodies (Kerr et al., 1972), as already described. Cells treated with QPhNO2 displayed those features, suggesting that this compound induces apoptosis in HL-60 leukemia cells (data not shown). Considering the observed morphological features, we conducted a flow cytometry analysis of several cellular and biochemical events to assess the mechanisms involved in cell death induced by the tested compounds.

Because of this, it is suggested here that DPSIR should perhaps more accurately become DAPSI(W)R. In order to control those State changes and Impacts (or Impacts on human Welfare), we therefore require Responses. Those Responses may include bringing in technological advances (such as better

fishing gear, habitat re-creation or water treatment plants), economic instruments (such as quotas or penalties) or laws administered by statutory bodies. Hence we need a management framework to accommodate and describe all the linked processes in this framework. Such a framework must then be aimed at what we may term the ‘big idea’ – ‘that marine management is designed to protect and enhance the natural INK 128 manufacturer structure and functioning of the seas while at the same time ensuring the marine processes which deliver ecosystem services from which we then obtain societal goods and benefits’ ( Elliott,

2011). Hence many of the Impacts in Table 1 relate to a loss of ecosystem services buy Ku-0059436 and societal benefits. Given the adage that ‘if you don’t know where you are going then any road will take you there’, then in order to set down the ultimate aim as a readily communicable message, this should be encapsulated in a vision for the seas, for example to achieve ‘clean, healthy, safe, productive and biologically diverse oceans and seas’ as adopted by the UK government and others ( Defra 2010). Furthermore, it is argued that sustainable and successful marine management can then only be obtained by including all facets and players in the system, the so-called 10-tenets ( Elliott, 2013) in which the major players and responses are included. The latter suggest that our actions should be: Ecologically sustainable (identified as ecol. in the figures below), Technologically feasible (Tech.), Economically viable (Econ.), Socially desirable/tolerable (Soc.), Legally

permissible (Leg.), Administratively achievable (Admin.), Politically expedient (Pol.), Ethically defensible (morally correct) (Ethic.), Culturally inclusive (Cult.) and Effectively communicable (Comm.). This discussion and its diagrams will therefore try to indicate the major steps in an integrated marine management framework while cross-referring to the elements D, P, S, I(W) and R and the Doxacurium chloride 10-tenets. The Pressures on the marine environment (e.g. Kennish and Elliott, 2011) can be regarded as coming from three sources – activities which remove materials and space from the system, activities which place materials into the system, and thirdly, external and wider pressures, such as global climate change, which emanate from outside the system (Fig. 1). The materials extracted include fish, shellfish, water, and seabed sands and gravels, and space is also removed, for example by occupying the seabed with harbours, windfarms, etc.

While macroH2A produces a signal by ChIP after analysis of formaldehyde crosslinked chromatin surrounding the

DSB, it does not produce a signal by ChIP after analysis of uncrosslinked chromatin, suggesting only a transient interaction as part of the DDR pathway. Thus, histone variants represent a crucial player in the proper repair of double strand breaks and maintenance of the genome. While histone variants Trametinib datasheet generally aid the DNA repair process, there are examples where histones can serve as an obstacle. In vitro experiments demonstrate that when an oxidized abasic site, one of the most common lesions resulting from oxidative damage, is present in the nucleosome, the lesion is not merely removed from the DNA, but can be transferred to the closest histone tail, usually the lysine rich tails

of H3 or H4, creating a DNA/protein crosslink [ 40]. By monitoring the length of 32P-labeled substrates before and after incorporation into a nucleosome, the formation of single strand breaks (SSBs) was determined to increase between 130 and 550 fold, depending on the location of the lesion within the nucleosome, with lesions positioned near the entry/exit site of the DNA displaying the highest rates of SSB formation. While these experiments were conducted using recombinant, canonical histones, the effect of histone variants on the rate of SSB and DNA/protein crosslink formation is completely ERK inhibitor purchase unknown. Histones play an important role in cellular aging; histone levels decrease as part of the natural aging process in yeast [41]. Upon inactivation of the Hir histone chaperone complex or overexpression of histone proteins in S. cerevisiae, lifespan can be artificially increased, indicating that regeneration of cellular chromatin is vital for extending lifespan [ 42]. Histone variants

are also implicated in cancer. A recent study has shown that specific splice variants of macroH2A are correlated with the known invasiveness of cancer cell lines [ 43]. While the total macroH2A content is consistent between the cell lines studied, when a cell has a greater amount of macroH2A1.1 as compared to macroH2A1.2, the cell is less invasive, as measured by migration through a porous membrane. Conversely, when the cell has a greater amount of macroH2A1.2, the cell tends to be more invasive. Mechanistically, it is not known if this Avelestat (AZD9668) correlation reflects an increase in fragile chromatin structure imparted by macroH2A1.2 versus macroH2A1.1, or whether the increase in macroH2A1.2 is an indirect downstream effect of other factors. Indeed, the potential for interaction of upregulated macroH2A1.2 with other histone variants remains a completely unexplored arena in the study of cancer invasiveness. Interestingly, alterations in histone genes are not just associated with diseases of age. In pediatric glioblastomas, mutations such as K27M and G34R/V are found clustered on the tail of histone variant H3.

While it may become necessary to give up on the idea of a specific EEG index of structural or combinatorial processing, a reliable measure for these fundamental mechanisms

and how they contribute to language processing may be won instead. Parts of the research reported here were supported by the German Research Foundation (BO 2471/3-2), the LOEWE programme of the German state of Hesse and by the University of Mainz as part of the research initiative “Pro Geisteswissenschaften”. We would like to thank Karin Hollerbach, Miriam Burk, Alexander Dröge and Phillip Alday for help with the stimulus materials and Brita Rietdorf for the data acquisition. We are also grateful to Herman Kolk and several anonymous reviewers for valuable comments on previous versions of the manuscript. “
“In everyday communication, we typically link our utterances to the discourse environment of the interlocutor check details in order to efficiently achieve our communicative objectives. Besides other factors, the speaker considers background information SGLT inhibitor and feedback of the listener. Linguistic (e.g., information structure, stress) as well as extra-linguistic features (e.g., gestures, eye-gaze) are dynamically used to clarify what the utterance is about and ultimately guide the cooperative listener to the communicative intention of the speaker. It has been proposed that the listener structurally represents all relevant aspects of information

(e.g., participants, events) delivered via language and MRIP perception within a mental model in which further incoming discourse information is integrated (e.g., Cowles, 2003 and Johnson-Laird, 1980). Information structure (cf. information packaging) is concerned with how information is packaged within a discourse to optimize information transfer ( Chafe, 1976). In this regard the idea of efficient communication was defined by Clark and Haviland (1977) as: “The speaker tries, to the best of his ability, to make the structure of his utterances

congruent with his knowledge of the listener‘s mental world” (p. 4). Ordering of information at the sentence-level is thought to be influenced by information structural concepts, such as topic-comment, given-new, or focus-background (e.g., Büring, 2007, Halliday, 1967, Krifka, 2008 and Lenerz, 1977). However, these information structural concepts lack a uniform definition and depend on the field of research and respective theoretical framework. For the purposes of our study, we use the following definitions: The TOPIC of a sentence is typically understood as the information that the speaker intends to increase the listener’s knowledge ( Gundel, 1985). Hence, topic is defined as what the sentence is about; COMMENT is what is said about the topic ( Gundel, 1988 and Reinhart, 1981; see Section 1.4 for a more detailed definition of topic). GIVEN INFORMATION constitutes information the speaker expects to be already known by the listener (e.g.