Unless infection occurs, a slight inflammation at the puncture site can arise. Spiders of the family Theraphosidae have ALK inhibitor urticating hairs covering their bodies, which are brushed off by the spider as a mechanism of defense to deter predators. These hairs were found to induce local dermatitis in vertebrates, including humans ( Shrum et al., 1999). The puncture wounds from the spider’s fangs require local wound care, follow-up

for signs of infection, short-term analgesia and a tetanus booster ( Kelley and Wasserman, 1998; Shrum et al., 1999). The spider venom is a diverse mixture of low molecular mass compounds (16% of all compounds), acylpolyamines (11%), linear peptides (6%), cysteine-knotted mini-proteins (60%), neurotoxic www.selleckchem.com/products/fg-4592.html proteins (1%) and enzymes (6%) (Jackson and Parks, 1989; Kuhn-Nentwig et al., 2011). It is mainly used to paralyze prey and for defense, and contains toxins that affect the central or peripheral nervous systems. These neurotoxins have been identified mostly as acylpolyamines and peptides or proteins that act on membrane receptors or ion channels (see review Estrada et al., 2007). The acylpolyamine toxins

are low molecular mass compounds (<1 kDa) that appear to have evolved to specifically provoke rapid paralysis. Their complex structures are composed by a polyamine chain with a primary amino or a guanidine group at one end and an aromatic ring at the other. These compounds interact with multiple targets in the central and peripheral nervous systems of insects, and also in the CNS of mammals, whereas the main targets are ionotropic

glutamate and nicotinic acetylcholine receptors (Kawai et al., 1982; Herold and Yaksh, 1992; Bixel et al., 2001). Eight hundred curated sequences of protein toxins have been described for spider venom to date, among them approximately 20% corresponds to Theraphosidae spiders (available at ArachnoServer 2.0; Herzig et al., 2011). Most of these 200 peptides has 30–40 amino Gefitinib acid residues, three disulfide bridges and basic character (Escoubas and Rash, 2004), and are modulators of ion-channels, such as calcium, sodium, and potassium. In this communication we report the results of proteomic and pharmacological characterizations of the venom extracted from the Brazilian spider Acanthoscurria paulensis. A. paulensis (Theraphosidae, Mygalomorphae) is a dark brown colored spider widely distributed in three Brazilian regions: South, Southeast and Midwest ( Mello-Leitão, 1923; Lucas et al., 2010).

3). Total proteins were extracted from the first expanded leaves of salt-treated seedlings of T349 and Jimai 19. The profiles of wheat leaf proteins were established at a pI range of 3.5 to 10.0 and with a molecular

learn more mass range of 13 to 110 kDa ( Fig. 4). Compared with Jimai 19, 17 protein spots (S1-1 to S1-17) were up-regulated in T349 ( Fig. 5), and all of these proteins were identified by mass spectrometry ( Table 3). The significant differences between Jimai 19 and T349 leaves corresponded to their different protein responses to salt stress. The functional classification analysis according to gene ontology (GO) annotations and PubMed references revealed that the proteins were clustered into several categories. Those 17 differential proteins were involved in osmotic stress, oxidative stress, photosynthesis, and lipid metabolism. Osmotic stress-related proteins include methionine synthase (S1-11) and glyceraldehyde-3-phosphate dehydrogenase (GPD) (S1-6). Oxidative stress-related proteins include NADP-dependent malic enzyme (S1-12), glutathione transferase (S1-3) and 2-cys peroxiredoxin (S1-10). Photosynthesis-related

proteins include Rubisco large subunit (RLS), Rubisco activase (S1-16) and chlorophyll a–b binding proteins (S1-9). Spots S1-7, S1-8, S1-13, S1-14, and S1-15 were all identified as Rubisco large subunits with different molecular masses and isoelectric points corresponding to their spot positions on the gel. Lipases (S1-17) (-)-p-Bromotetramisole Oxalate directly

catalyze the hydrolysis or synthesis of lipids. Spots S1-1, S1-2, S1-4, and S1-5 were identified as predicted proteins of barley. According to NCBI BLAST results, spot S1-1 (gi|326503994) Alpelisib mouse contains the region PLN00128, which is annotated as a succinate dehydrogenase (ubiquinone) flavoprotein subunit, and has 94% identity with the Triticum urartu protein succinate dehydrogenase (ubiquinone) flavoprotein subunit (sequence ID: gb|EMS46614.1|). Spot S1-2 (gi|326511988) contains the region MopB_Res-Cmplx1_Nad11, which is annotated as the second domain of the Nad11/75-kDa subunit of the NADH-quinone oxidoreductase, and has 98% identity with the T. urartu protein NADH-ubiquinone oxidoreductase 75 kDa subunit (sequence ID: gb|EMS48685.1|). Spot S1-4 (gi|326493416) contains the region PLN02300, which is annotated as lactoylglutathione lyase, and has 98% identity with the Aegilops tauschii protein lactoylglutathione lyase (sequence ID: gb|EMT08036.1|). Spot S1-5 (gi|326491885) contains the region WD40, a domain found in many eukaryotic proteins that cover a wide variety of functions, including adaptor/regulatory modules in signal transduction, pre-mRNA processing and cytoskeleton assembly. The coleoptile length, radicle length, and radicle number of the GmDREB1 transgenic wheat lines were significantly higher than those of the wild type, suggesting that the overexpression of the GmDREB1 gene improves the growth of wheat seedlings under saline conditions.

042). Cortical perimeter increased significantly from baseline in both the ELD group (2.63 ± 7.52%, p = 0.008) and the ALF group (3.86 ± 6.28%, p < 0.001). Thus, although there was no significant difference between the effects of the two drugs on the increased cortical perimeter, ELD prevented the decrease in cortical thickness. Cortical vBMD of the femoral neck increased

significantly in both the ELD group (1.82 ± 4.78%, p = 0.004) and the ALF group (2.21 ± 4.98%, p < 0.001), with no difference between the two groups ( Fig. 1). Trabecular vBMD of the femoral neck significantly decreased in both the ALF group (− 7.49 ± 8.82%, p < 0.001) and the ELD group (− 3.99 ± 7.83%, p < 0.001), and there was a significant difference between the two groups (p = 0.020). Total vBMD EPZ015666 of the femoral neck decreased from baseline in the ALF group (− 2.25 ± 5.32%, p < 0.001), whereas it was maintained in the ELD group. Accordingly, the percentage changes in total vBMD differed significantly between the ELD and ALF groups (p = 0.009). Regarding cortical CSA, the ELD group showed a non-significant trend for an increase (1.73 ± 7.62%,

p = 0.082) and the ALF group showed a non-significant trend for a decrease (− 0.96 ± 6.14%, p = 0.212) ( Fig. 1). Thus, the percentage changes from the baseline in cortical CSA showed a significant difference between the ELD and ALF groups (p = 0.031). Trabecular CSA of the femoral Megestrol Acetate neck increased significantly in the ALF group (2.92 ± 7.74, p = 0.003), but not in the ELD group (1.92 ± 7.61%, p = 0.054). Total CSA increased from the baseline in both the Caspase pathway ELD group (1.69 ± 6.78%, p = 0.056) and the ALF group (1.51 ± 5.77%, p = 0.039), with no difference between the two groups. Cortical bone mass of the femoral neck increased significantly from baseline in both the ELD group (3.68 ± 7.51%, p < 0.001) and the ALF group (2.45 ± 9.64%, p = 0.045) ( Fig. 1). Total bone mass of the

femoral neck increased significantly only in the ELD group (1.93 ± 5.89, p = 0.013). Trabecular bone mass significantly decreased in the ALF group (− 3.96 ± 9.39, p < 0.001), whereas it did not change from baseline in the ELD group, and there was no significant difference between the two groups (p = 0.268). Thus, in the ELD group, both total and cortical bone mass increased from baseline, and trabecular bone mass was maintained. Biomechanical properties (CSMI, SM, and BR) of the femoral neck were compared between the ELD group and the ALF group (Fig. 2). CSMI and SM improved significantly in the ELD group (5.30 ± 11.56%, p < 0.001 for CSMI; 4.33 ± 11.92%, p = 0.006 for SM), whereas these parameters did not change in the ALF group. Thus, there were significant differences between the ELD and ALF groups in the percentage changes of CSMI and SM from baseline (p = 0.037 and p = 0.023, respectively).

Bacteria conjugated to pHrodo™ show a very low fluorescent signal at the neutral pH present on the cell Cyclopamine manufacturer surface, but emit a bright red fluorescence in the acidic environment of phago-lysosomes. This level of discrimination eliminates washing and quenching

steps that are necessary with other non pH-dependent indicators of bacterial uptake. Moreover the fOPA here described takes advantage of the introduction of specific markers of HL-60 differentiation to neutrophils, which allow keeping under control the variability of effector cells. The method was evaluated for sensitivity and specificity, by testing a panel of sera from mice immunized with different GBS glycoconjugate vaccines against polysaccharide Ia. kOPA titers were compared with fOPA titers, and a confocal microscopy analysis was conducted to study bacterial localization inside neutrophils, XL184 cell line in the presence or in the absence of specific antibodies and

complement. GBS strains 515 (serotype Ia) (Baker et al., 1982) and COH1 (serotype III) (Wessels et al., 1992) were used in this work. Bacteria were grown in Todd–Hewitt Broth (THB) to an optical density at 600 nm (OD600 nm) of 0.45. Ten percent glycerol was added to the culture before dispensing 1 ml aliquots in cryo-vials for flash freezing in a 95% ethanol-dry ice bath. Frozen cultures were kept at − 70 °C until use. OPAs were performed with rabbit and mouse sera. Rabbit sera were raised by immunizing one animal with three doses of monovalent CRM197-conjugated polysaccharide Ia, Ib and III in presence of aluminum hydroxide (Alum). Mouse sera were pooled from animals immunized with a GBS vaccine composed by polysaccharide Ia, Ib and III conjugated to CRM197, formulated with Alum or MF59 (Podda, 2001). Animal treatments were performed in compliance with the Italian laws and approved by the institutional review board (Animal Ethical Committee) of Novartis Vaccines and Diagnostics, Siena, Italy. Bacteria were grown in THB

VAV2 to OD600 nm = 0.6, washed twice with Phosphate Buffered Saline (PBS, pH 7.2–7.4,Gibco) and suspended in half volume of PBS-0.08% paraformaldehyde (PFA, Sigma). Cells were incubated at 37 °C for 30 min and kept at 2–8 °C in PBS-0.08%PFA. Immediately before labeling, cells were washed with PBS, suspended at 20 mg (wet weight)/ml using a freshly prepared 100 mM Sodium Hydrogen Carbonate solution pH 8.5 (Merck) and split into aliquots of 750 μl. A 10 mM stock solution of PHrodo™ Succinimidyl Ester (Invitrogen) in dimethyl sulfoxide (Sigma) was diluted in the bacterial suspension at a final concentration of 0.1 mM. Each sample was incubated for 45 min at room temperature in the dark and then added with 750 μl of Hank’s Balanced Salt Solution with Ca2 + and Mg2 + (HBSS, Gibco), then spin down with a bench top centrifuge for 60 s at 14,000 ×g. The supernatant was aspirated and the pellet suspended in HBSS and stored in the dark at 4 °C for two months.

In the rare case of a patient with severe pain an analgesic review with their GP or consultant may be required in order to allow participation in rehabilitation. Many people believe that activities that cause pain must be harmful. Clinicians need to gain a clear understanding of the patient’s pain experience and beliefs about pain (Eccleston and Eccleston, 2004) and counter those which are mal-adaptive. Clinicians should reinforce messages

which reduce fear or anxiety about pain, e.g. that the presence of pain should mTOR inhibitor not prevent most patients from safely participating in therapeutic exercise (Waddell et al., 2004) and may lead to reduction in symptoms (Guzman et al., 2002), improved function and return to work (van

Tulder et al., 2000). Those who participate in regular exercise are also less likely to experience progressive problems (McLean et al., 2007). Patients should be encouraged to start exercise gently and advised to progress to moderate or even high intensity levels of this website exercise over a period of time (Pernold et al., 2005). This evidence could counter the fears held by many pain sufferers that movement could be damaging or lead to re-injury. Low levels of physical activity at baseline (Minor and Brown, 1993, Rejeski et al., 1997, Stenstrom et al., 1997 and Schoo et al., 2005) or in previous weeks (Rejeski et al., 1997 and Oliver

and Cronan, 2002) and low in-treatment adherence with exercise (Alewijnse et al., 2003, Schoo et al., 2005 and Dobkin et al., 2006) were barriers to treatment adherence. Physiotherapists need to recognise and be ready to mitigate the many barriers to initiating and adhering to exercise programmes; these include poor programme aminophylline organisation and leadership, poor education, poor history of exercise, perceived physical frailty, perceived poor health and readiness to change (Duncan and McAuley, 1993, Courneya and McAuley, 1995, Boyette et al., 1997, Hellman, 1997 and Rhodes et al., 1999). Several strategies may be employed to improve patient adherence. Firstly providing explicit verbal instruction, checking the patient’s recall and supporting this with additional written instructions may be effective at improving exercise adherence (Schneiders et al., 1998). Secondly, employing motivational techniques such as counselling sessions, positive feedback, reward, written treatment contracts and exercise diaries may also be helpful (Friedrich et al., 1998). Setting goals and drawing up action plans and coping plans which have been agreed collaboratively between the clinician and patient may be effective with patients who intend to participate in exercise (Bassett and Petrie, 1999, Evans and Hardy, 2002 and Ziegelmann et al., 2006).

The use of MTL or MTI in the United States has not received as much attention as in other regions. It is possible that this is because the United States is not party to the Convention on Biological Diversity. As such, the country is not obligated to the conventional laws therein see more and the subsequent recommendations and calls for action. Although the U.S. has not been obliged to explore the use of MTL or MTI in management decisions, the National Marine Fisheries Service (NMFS) has been an active supporter of a shift toward an ecosystem based approach to management. In a 1998 report to Congress developed by a NMFS Ecosystem Principles Advisory

Panel (EPAP), the department outlined the importance of developing an ecosystem based management (EBM) plan as well as guidelines in the development of this strategy. The report highlights that species within an ecosystem are linked trophically and accepts the trend of decreasing MTL, citing “Fishing down food webs… disrupts natural predator-prey relationships and may lead first to increasing catches, but then to stagnating or declining catches” [22]. Among the recommendations issued in the report is the determination of total removals and their relationship to trophic structure. The authors cite Pauly et al., claiming that the

relationship between landings and trophic structure has “potential negative effects on sustainability” [22]. Additionally, the report recommends the development of ecosystem health indices and incorporation of these indices find protocol into regional Fishery Ecosystem Plans (FEP). The Advisory Panel highlighted the use of mean trophic level as such an index, noting that a specific FEP goal could be the maintenance

of a predetermined MTL [22]. More recently, in a 2009 Report to Congress the NMFS reaffirmed their recommendation of an EBM approach to fisheries and the need for “fundamental knowledge of basic ecosystem principles…as outlined by the EPAP” [23]. Ultimately, the use of marine Digestive enzyme trophic indices in policy development and ecosystem management has received sporadic acceptance and adoption. Large intergovernmental and transnational bodies have readily accepted the measure as a suitable indicator of ecosystem health and stability. Several national and regional governing bodies, however, have concluded that the index is only reliable at a larger scale, and not applicable at smaller-scale national levels [17] and [18]. Adoption of MTI as an indicator of sustainable fisheries, however, has been accepted by the CBD, EU, and CLME Project, and many suspect that it will be adopted as a tool for policy development in the European Marine Strategy and Common Fisheries Policy [17]. In 1998, Daniel Pauly and colleagues published a revolutionary study examining change in MTL over time. An examination of global catch data between 1950 and 1994 revealed a startling trend of decreasing MTL over time.

, 2001; Boehm, 2003; Liu et al., 2006 and Thupaki et al., 2010). Recreational beach use, especially in California (where surfing is common), is not limited to the shoreline. This

makes it Natural Product Library purchase important to evaluate FIB contamination and the processes controlling it over wider recreational domains where physical processes are different, and FIB survivorship may also change (Davies-Colley et al., 1994 and Kim et al., 2004). Here we present results from an along and cross-shore resolved field program with joint physical and bacterial observations designed to identify the dominant mechanisms controlling FIB variability within (and seaward) of the surfzone. By directly measuring currents out to 300 m cross-shore, we both enable the evaluation FIB flow fields PTC124 cost over appropriate recreational domains, and avoid estimating current velocity from wave direction or alongshore drift, which has increased uncertainty in other models (Boehm, 2003; Kim et al., 2004). In the present paper we focus on quantifying the contribution of physical processes (advection and diffusion) to observed FIB patterns, and developing a best-fit physical model from this analysis. The contribution of biological processes to nearshore FIB variability is addressed in Rippy et al. (2012). Southern California’s Huntington State Beach is ∼3.2 km long, with chronically poor surfzone water

quality (Grant et al., 2001 and Kim

et al., 2004). At its southern end, the beach receives brackish flows from the Talbert Marsh (TM) and the Santa Ana River (SAR), both of which have been implicated as sources of surfzone FIB (Kim et al., 2004). In fall 2006, a multi-institutional field campaign (“HB06”) focused on observing nearshore waves, currents, temperature, phytoplankton, and FIB at this beach. The present study concerns the bacterial component of HB06, a 5-h FIB survey with high spatial and temporal resolution conducted on October 16th along transects extending 1 km north of the TM/SAR outlets, and 300 m offshore. FIB concentrations were measured at 8 stations: 4 in knee-deep water along a 1000 m alongshore transect north of SAR (SAR, TM, FHM, F1; Fig. 1), and 4 along a 300 m cross-shore transect starting at F1 (knee-deep Cetuximab mw water), and terminating at an offshore Orange County Sanitation District mooring (OM) in ∼8 m mean water depth (F1, F3, F5, F7, OM; Fig. 1). Every 20 min, from 0650 h to 1150 h PDT, 100 ml water samples were taken at all stations. Samples were stored on ice and transported to the Orange County Sanitation District (OCSD) within 6 h of collection. All samples were analyzed for Escherichia coli (IDEXX Colilert) and Enterococcus (EPA method 1600) concentrations by OCSD personnel. Temporal rates of FIB loss were estimated for each station from regressions of log (FIB) versus time.

During the study period, no clinical signs, ophthalmological abnormalities, or deaths were observed in either the control or the treatment groups (data not shown). The animals showed no significant

differences in body weight and food consumption between the control and treatment groups. Body weight increased gradually throughout the study period in males and females of all groups (Fig. 1). At the end of the study, all animals were euthanized selleck chemicals and subjected to a necropsy. Data in Table 4 indicate that there were no significant differences in hematological parameters between the control and Vigiis 101-treated groups. Data in Table 5 show some statistically significant differences (p < 0.05) in clinical chemistry check details parameters between the control and Vigiis 101-treated groups. In male rats, blood chemistry parameters, including potassium (K), aspartate aminotransferase (AST), and triglyceride (TG) were significantly different but were within the physiologically acceptable range (K: 3.82∼5.55 mg/dl; AST: 74∼143 U/l; TG: 20∼114 mg/dl) in the treatment groups. In Vigiis 101-treated female rats, statistically significant changes in AST also resulted in the values that was within

the acceptable range (AST: 65∼203 U/l). The data in Table 6 and Table 7 indicate that no significant organ weight and relative organ weight changes were noted in either the male or female rats. Fig. 2 and Fig. 3 demonstrate the results of histopathological examination

of the rats. The results showed that no significant lesions were present in Methane monooxygenase the liver, kidneys, heart, spleen, adrenal glands, epididymis, testes, uterus, and ovaries of the control or high-dose Vigiis 101 groups. Probiotic products fermented by L. paracasei subsp. paracasei NTU 101 have been shown to have various beneficial effects on humans and animals, such as hypolipidemic, immunomodulatory, osteoprotective, and antiobesity effects. In the literature, there are no available classical toxicology data on Vigiis 101. To our knowledge, there are no published studies on traditional genotoxicity or mutagenicity of L. paracasei subsp. paracasei NTU 101 or L. paracasei subsp. paracasei strains in general. Hence, we decided to evaluate the safety of Vigiis 101 powder made from L. paracasei subsp. paracasei NTU 101 provided by SunWay Biotech Co., Ltd. (fermentation by means of typical industrial equipment). We used two in vitro genotoxicity tests of Vigiis 101, one in vivo genotoxicity test, and a 28-day oral toxicity assay in Wistar rats. The Ames test in Salmonella strains TA98, TA100, TA102, TA1535, and TA1537 showed that Vigiis 101 does not induce a greater than two-fold increase in the number of reverse mutations at the doses 0.3–5.0 mg/plate. Nor does metabolically activated Vigiis 101 (with an S9 mix) exhibit mutagenicity.

The three fields emerged from an unusual concentration

in space and time of a handful of seminal experimental observations. In just a few years, we learned that heterotopic transplantation of transitional epithelium into skeletal muscle induces heterotopic bone formation [1]; that heterotopic transplants of bone marrow also do so [[2] and [3]], but that the two phenomena are radically distinct from one another: the former is dependent on the release of a soluble factor, while the latter is not. Identification of BMPs [[4], [5], [6] and [7]] and perisinusoidal reticular cells as the specific factor and cell type generating bone in heterotopic transplants of transitional http://www.selleckchem.com/products/GDC-0980-RG7422.html epithelium and bone marrow, respectively, PLX3397 in vivo represents the ending point of two long and diverging journeys that originated from those seminal experiments. Likewise, the definition of the bone marrow microenvironment as the host of signals provided by stromal cells and required for hematopoiesis, and the pursuit of a “niche” for hematopoietic stem cells proper represent the developments over time of a third seminal observation; that is, that grafting of bone

marrow in closed systems (diffusion chambers) would generate bone but bar the development of hematopoiesis, whereas transplantation in open systems would allow for both bone formation and development of marrow [2]. That all of these fundamental observations, which not only withstood the test of time, but also represented the seed for the subsequent flourishing of major fields of investigation, arose from the practice of heterotopic transplantation cannot escape notice. Considering the tremendous impact of establishing quail–chick chimeras (a kind of heterotopic transplantation in embryos) [8] and [9]in developmental

Adenosine triphosphate biology and how much it contributed to further developments in lineage tracing, one is tempted by foolishly wondering what magic is inherent in putting tissues and cells where they do not belong (ectopic transplantation), and why is this practice so instructive. Perhaps all this simply highlights the fundamental link between space (and time) and development (lineage, commitment, differentiation), a notion we owe, ultimately, to Alan Turing (the father, among many other things, of the diffusion–reaction model which established the chemical basis of morphogenesis [10]), and before him, to D’Arcy Thompson (a classicist and a morphologist renowned for his attention to the physical and mathematical laws underpinning morphogenesis) [11].

However, if RSC represents

each permanent item in a given view, then it could play a key role in detecting and mapping individual landmarks as we encounter them in our surroundings. This operation could be crucial for successful navigation, as the very building blocks of any representation of an environment Ponatinib ic50 are the most stable items within it. To test the nature of RSC processing, we had good and poor navigators view quartets of outdoor items (Fig. 1). The stimuli differed in terms of how many of their four items were permanent, i.e., with a fixed location in the environment – they contained either no, 1, 2, 3, or 4 permanent items. We used multi-voxel pattern analysis (MVPA; Chadwick et al., 2012, Haynes and Rees, 2006 and Norman et al., 2006) to assess whether information about the number of permanent items in view could be decoded from activity in RSC and, if so, whether this differed between good and poor navigators. The quartets were carefully designed such that variations in landmark size and visual salience could be assessed by the same method, allowing us to determine

whether any patterns of response observed in RSC were specific to item Cyclopamine solubility dmso permanence. Thirty-two, right-handed, healthy participants (16 females, mean age 23.5 years, SD 2.5) took part in the experiment. All had normal or corrected to normal vision, were highly proficient in English and gave written informed consent in accordance with the local research ethics committee. None of the participants had

taken part in any of our previous studies of item permanence. clonidine Each stimulus comprised four different everyday outdoor items, with each item enclosed by a grey outline on a white background, and laid out in a grid (Fig. 1). The stimuli differed in terms of how many of their four items were permanent – they contained either no, 1, 2, 3, or 4 permanent items (giving 5 category types). Permanent items were defined as those consistently rated as ‘never moving’ by an independent set of participants from previous behavioural experiments (Auger et al., 2012). There were 20 stimuli for each of the 5 category types, giving 100 stimuli in total. We ensured that across the trials of each condition, the non-permanent elements were sampled from the full range of permanence ratings (excluding those that ‘never moved’). The stimuli not only varied according to the number of permanent items they contained; their items also varied in terms of real-world size and visual salience. The size and visual salience of items was also determined by an independent set of participants from the previous behavioural experiments (Auger et al., 2012). In designing the stimuli we ensured a full range of values of these two other landmark features, from the very smallest to largest, and from least to most salient items. This allowed us to also group the 100 stimuli into 5 categories for size and 5 for visual salience.