Chl.a turned out to be a suitable indicator across the gradient from land to sea. In several coastal waters winter dissolved inorganic nutrient concentrations have only a low value as quality indicator. The used model revealed several weaknesses that require attention and improvement. However, it became obvious that the higher the spatial and temporal resolution,

the more important becomes quality as well as spatial and temporal resolution of input data, namely discharge and nutrient concentrations. Further the bio-availability of compounds and the N/P ratio in loads requires attention. It seems that in some coastal waters similar chl.a targets can be reached with alternative management approaches either focussing on N or on P load reductions. Selleckchem CAL-101 selleck chemicals llc Additionally, the role of extreme events on the state of ecosystems requires more attention. The MAI for Germany and the updated nutrient reduction targets of the Baltic Sea Action Plan HELCOM [25] are, according

to our results, not sufficient to reach a good ecological status in German Baltic coastal waters. The BSAP has a focus on the open sea. The suggested low N load reductions into the western Baltic Sea in general, and the focus on a reduction of atmospheric deposition, allows much too high N loads into German coastal waters to meet the WFD targets. Future updates of the Baltic Sea Action Plan should take coastal waters and their specific demands and conditions into account. At present, transport pattern and spatial distribution as well as amount and bio-availability of atmospheric N and P deposition to the Baltic Sea are not well known, generate uncertainty in the results and require further attention and additional research. The work has been funded by the German Federal Ministry for Education

and Research within Project SECOS (03F0666A) and partly supported by Projects RADOST (01LR0807B) and MOSSCO (03V01246B). We thank all members of the national BLANO UAG ‘Nutrient reduction targets and eutrophication in the German Baltic Sea’ working-group members for feedback and fruitful discussions. Supercomputing power was provided by HLRN (North-German Supercomputing Alliance). “
“Breakthroughs in technology that facilitate efforts by scientists to monitor the movements of marine migratory species L-NAME HCl and collect and transmit environmental data gives rise to new questions in the law of the sea [1]. The law of the sea recognizes the special importance of highly migratory species as critical shared resources, although this list is no longer comprehensive. (Appendix A1). Rules for deployment of research vessels and the conduct of traditional MSR are set forth in the United Nations Convention on the Law of the Sea (UNCLOS).1 Coastal states have the right to regulate and authorize MSR in offshore areas under their sovereignty and jurisdiction, including a 12-nautical mile (nm) territorial sea and 200-nm EEZ.

0 ( Delcher et al., 2007). The tRNAs and rRNAs were identified using tRNAscan-SE version 1.21 (http://lowelab.ucsc.edu/tRNAscan-SE/), RNAmmer (http://www.cbs.dtu.dk/services/RNAmmer/) and Rfam database (http://www.sanger.ac.uk/resources/databases/rfam.html). KAAS server was used to assign translated amino acid sequences (with genetic code in table 11) into KEGG Orthology using the SBH (single-directional best hit) method ( Kanehisa et al., 2008). Translated Akt inhibitors in clinical trials genes were aligned

with COG database using NCBI blastp (hits should have scores no less than 60, e value is no more than 1e− 6) ( Tatusov et al., 2001). SignalP 4.1 (http://www.cbs.dtu.dk/services/SignalP/) was used to identify genes with signal peptides with default parameters except “-t gram +”. Genes with transmembrane helices were identified using TMHMM 2.0 (http://www.cbs.dtu.dk/services/TMHMM/). The draft genome sequence of B. flexus strain T6186-2 revealed a genome size of 4,254,248 bp and a G + C content of 37.51%. These contigs contain 4700 coding sequences (CDSs), 36 tRNAs and 3 rRNAs. Moreover, 2923 genes

were categorized into COG functional groups ( Table 1). Analysis of ORFs indicated that T6186-2 possesses at least 46 putative ARGs (Table S1), which is consistent with the phenotype that this isolate showed regarding resistance to erythromycin, gentamicin, vanomycin, fosfomycin, fosmidomycin, tetracycline and teicoplanin. Interestingly, 10 putative MarR family transcriptional regulators were found in the selleck inhibitor genome (Table 2), which is a widely conserved multiple antibiotic resistance regulator in response to diverse antibiotics, toxic chemicals

and many other important biological processes (Hao et al., 2014). In light of the fact that T6186-2 was isolated from a deep-subsurface oil reservoir, thus it has the relatively low probability of being exposed to anthropogenic antibiotics. So it is possible that T6186-2 possesses some novel antibiotic resistance genes and/or Metalloexopeptidase resistance mechanisms. Further studies are required to elucidate the resistance mechanisms, and information on these mechanisms could potentially aid in antibiotic development. The genome project is deposited in the Genome Online Database and the draft genome sequence is deposited in GenBank under the accession JANV00000000. This study was sponsored by the National Natural Science Foundation of China (Grant No. 81301461, 50974022 and 51074029), the 863 Program (Grant No. 2008AA06Z204 and 2013AA064402) of the Ministry of Science and Technology, the Zhejiang Provincial Natural Science Foundation of China (Grant No. LQ13H190002) and the Scientific Research Foundation of Zhejiang Provincial Health Bureau (Grant No. 2012KYB083). “
“Methane is considered as a clean and environmentally compatible fuel. Methane hydrate is an ice like structure comprised of methane trapped in a lattice of water molecules.

However, because of increased urbanization and land use changes, the nutrient loading in wetlands learn more far exceed their capacity to retain pollutants and remove them through nitrification, sedimentation, adsorption, and uptake by aquatic plants. This adversely affects the wetland

water quality and its biodiversity. Such wetlands show drastic changes in nutrient cycling rates and species lose (Verhoeven et al., 2006). Various scholars in India have mainly focused on the usefulness and potential of constructed wetlands in pollution abatement on experimental scale (Billore et al., 1999, Juwarkar et al., 1995 and Kaur et al., 2012). Also, role of wetland plants in ameliorating heavy metal pollution both in a microcosm and natural condition is well

established (Dhir et al., 2009). Typha, Phragmites, Eichhornia, Azolla, and Lemna are some of identified potent wetland plants for heavy metal removal ( Rai, 2008). Constructed wetlands are considered to be a viable option for treatment of municipal wastewater. A well designed constructed wetland should be able to maintain the wetland hydraulics, namely the hydraulic loading rates (HLR) and the hydraulic retention time (HRT), as it affects the treatment performance of a wetland (Kadlec and Wallace, 2009). However, one of the major constraints to field-scale constructed wetland systems selleck inhibitor in India is the requirement of a relatively large land area that is not readily available. Thus, for Indian conditions, batch-fed vertical sub-surface flow wetlands that require just about 1/100th of land area and 1/3rd HRT than the surface flow systems have been suggested (Kaur et al., 2012). Wetlands play an important role in flood control. Wetlands help to lessen the impacts of flooding by absorbing water and reducing the speed at which flood water flows. Further,

during periods of flooding, they trap suspended solids and nutrient load. Thus, streams flowing into rivers through wetlands will transport fewer suspended solids and nutrients to the rivers than if they flow directly into the rivers. In view of their effectiveness associated with flood damage Chorioepithelioma avoidance, wetlands are considered to be a natural capital substitute for conventional flood control investments such as dykes, dams, and embankments (Boyd and Banzhaf, 2007). Based on the study in Rat River Watershed (Canada), it is estimated that with 10% increase in wetland area, there was a reduction of 11.1–18.6% in the total flood volume (Juliano and Simonovic, 1999). The flood protection value of human-made wetlands along the Nar and Ancholme rivers in the UK was estimated to be around 8201 USD/ha/year and 8331 USD/ha/year (Ghermandi et al., 2010). In India too, researchers have worked on estimating the value of flood protection function of the wetlands.

Kaplan-Meier plots were generated PLX3397 order and patients were

divided into groups on the basis of gene expression. Statistical power analysis for determining the sample size effective for the result was performed by using Time to an Event, a sample-size calculator (http://hedwig.mgh.harvard.edu/ sample_size/time_to_event/para_time.html). All statistical tests were two-sided and conducted at the .05 significance level. Median survival was defined as the time after which 50% of patients with ACC were living. The median survival ratio (> 1) was calculated by dividing one group’s smallest median survival time by the other group’s smallest median survival time. Table summarizes the clinical attributes of the patients in whom the 27 ACC tumor Ku-0059436 in vivo samples were obtained. All tumors had arisen sporadically; 16 occurred in women; and the median age at presentation was 58 years (range, 33 to 91 years). Tumors arose at the following sites: maxillary sinus (9 tumors), submandibular gland (6 tumors) or (6), parotid gland (5 tumors) or (5), sublingual gland (2 tumors) or (2), and one each in the nasal cavity, mandibular mucosa, nasopharynx, base of tongue, and tongue. Tumors were classified by morphologic subtype: tubular (4 tumors) or (4), cribriform (3 tumors) or (3), solid (1 tumor) or (1), combined

cribriform and tubular (10 tumors) or (10), combined solid and tubular (8 tumors) or (8), and combined cribriform and ZD1839 solid (1 tumor) or (1). We performed hematoxylin and eosin (H&E) staining (Figure 1A) and antibody-based IHC for c-Kit on tumor sample sections ( Figure 1B [case 17] and Supplemental Figures 1B [case 2] and 1F [case 7]). Mast cell staining was a positive internal control with the antibody (data not shown). c-Kit staining was estimated as described in Methods, and Table 1 shows our

results. c-Kit expression occurred in the inner luminal (duct-type epithelial) cells of all the tumors (see Figure 1B and Supplemental Figure 1B and F), as reported previously [3]. In searching for genomic alterations, we examined exons 8, 9, 11, 13, and 17, which encode domains for dimerization (exons 8 and 9), the juxtamembrane region (exon 11), and protein kinase activity (exons 13 and 17). We chose them for this study because gain-of-function mutations are recurrently found in these regions in other neoplasms [6]. We performed direct sequencing of each exon’s PCR product. Each sample was confirmed by at least three different sets of mutation analyses. No missense, frameshift, nonsense, synonymous missense, or splice mutations were detected. In light of the results of our mutational analysis, we hypothesized that c-Kit was activated by receptor dimerization upon stimulation by SCF and used IHC to determine levels of SCF protein in the salivary glands in tumor sample sections (Figure 1, C and D [case 17] and Supplemental Figure 1C [case 2] and 1G [case 7].

More information is needed to determine if repeated exposure is toxic, if animals become selleck inhibitor habituated, or if consumption is only related to the availability of alternative forage. This work was funded by the National Institute of Science and Technology for Control of Plants Poisoning, CNPq grant number 573534/2008-0. The author acknowledges Prof. Odaci de Oliveira, Federal University of the Semiarid (UFERSA), for identifying the Jatropha species.

“
“The 17th World Congress of the International Society on Toxinology (IST) and Venom Week 2012 (4th International Scientific Symposium on All Things Venomous) are being combined into a multi-disciplinary scientific meeting on animal, plant and microbial toxins. The meeting will be held July 8 – 13, 2012, in Honolulu, Hawaii at the Hilton Hawaiian Village, a world-class hotel, right on Waikiki beach, and with special conference rates. The meeting will contain state-of-the-art toxinological research and practice, with platform

and poster sessions on animal, plant and microbial toxinology, proteomics, genomics, pharmacology, pathophysiology, venoms, antivenoms, clinical toxinology, veterinary RG7420 cell line toxinology, venomous animal collections issues, and more! The meeting website can be found at: http://www.istworldcongress17-venomweek2012.org/ “
“The 17th World Congress of the International Society on Toxinology (IST) and Venom Week 2012 (4th International Scientific Symposium on All Things Venomous) are being combined into a multi-disciplinary scientific meeting on animal, plant and microbial toxins. The meeting will be held July 8 – 13, 2012, in Honolulu, Hawaii at the Hilton Hawaiian Village, a world-class hotel, right on Waikiki ifenprodil beach, and with special conference rates. The meeting will contain state-of-the-art toxinological research and practice, with platform and

poster sessions on animal, plant and microbial toxinology, proteomics, genomics, pharmacology, pathophysiology, venoms, antivenoms, clinical toxinology, veterinary toxinology, venomous animal collections issues, and more! The meeting website can be found at: http://www.istworldcongress17-venomweek2012.org/ “
“Bothrops alcatraz is a pitviper found only in the Alcatrazes Archipelago, off the northern coast of São Paulo state in southeastern Brazil. This species feeds primarily on invertebrates (centipedes) and vertebrates (amphibians) since these islands are devoid of small rodents, the main prey of mainland Bothrops spp. B. alcatraz differs from Bothrops jararaca primarily by its darker coloration, lower number of ventral, subcaudal, and infralabial scales, number and shape of anterior cephalic scales, shape of hemipenal spines and body size ( Marques et al., 2002). As a geographically and ecologically isolated species, B. alcatraz has a high potential for morphological variation and divergence in its venom composition compared to other Bothrops species.

It should be acknowledged, however, that the earlier-described hypothesis is not supported

by the results of several studies that have shown the benefits of increasing the infliximab dose in some patients with IBD or rheumatoid arthritis who lose response.27 and 28 Nonetheless, these findings suggest that clinicians should carefully weigh the need for a dose adjustment when applicable, considering that not all patients may benefit from such a dose increase. The current results had some limitations. First, although the relationship between serum Ku-0059436 molecular weight infliximab concentrations and efficacy outcomes appears to be both consistent and robust, these results are retrospective and may have been confounded by other determinants of both drug concentrations and outcomes. These data, however, were generated from large randomized trials in which known confounders were identified prospectively and in which randomization would be expected to result in an even distribution of confounding variables between the experimental groups. Nevertheless, prospective studies designed to assess the value Bak protein of infliximab concentration–guided dose escalation in terms of efficacy and potential impact on safety could provide valuable information pertaining to a more optimized approach toward infliximab therapy of UC. One such study suggests that therapeutic drug monitoring may predict the likelihood of

achieving mucosal healing after infliximab dose escalation in IBD.29 The positive findings of a small randomized trial of therapeutic drug monitoring in patients with Crohn’s disease who experienced a loss of response to infliximab also are encouraging.30 It also generally is held that a therapeutic drug monitoring approach using treatment algorithms in the management of secondary loss of response may be more rational than empiric dose escalation.31 The results of these analyses also may be limited by the fact that Ribonucleotide reductase the serum infliximab concentrations were measured using

a proprietary assay not presently available commercially. As a result, research is needed to cross-validate the assay with those available to physicians to help translate these identified infliximab concentrations into practice settings. In addition, the low NPV and low positive likelihood ratio associated with the identified threshold imply that false-negative results may be common and additional clinical judgment will be needed to manage patients who show concentrations less than the target threshold. Conversely, the magnitude of the PPV at the identified thresholds may reassure a clinician that a given patient is not undertreated if serum drug concentrations are greater than these thresholds. In summary, we have established that there is a strong association between serum infliximab concentration and efficacy outcomes in patients with UC.

In fact, reaction time often fails to detect differences between monolinguals’ and bilinguals’ responses to competition, even when other behavioral measures (such as eye-tracking or mouse-tracking) indicate group differences (e.g., Bartolotti and Marian, 2012 and Blumenfeld and Marian, 2011). Instead, more sensitive measures, such as eye-tracking or functional neuroimaging are needed to highlight meaningful differences in how monolinguals and bilinguals manage phonological competition. Here, we demonstrate that, even in the absence of behavioral differences between groups, monolinguals

and bilinguals differ in the cortical resources recruited to manage phonological competition. In contrast to the increased recruitment Navitoclax of language and executive

control regions observed by Righi et al. (2010) in competitor trials, participants in our current study showed limited activation in response to direct manipulations of competition. This is likely due to differences between the populations tested in the two studies. Although Righi et al.’s sample was not explicitly controlled for language experience, all participants were native English speakers. In contrast, our current study includes both native English speakers (monolinguals) and native Spanish speakers (bilinguals). When we consider only monolingual subjects, the group likely most analogous www.selleckchem.com/products/gsk1120212-jtp-74057.html to the participants used by Righi et al., competitor effects emerge in executive control regions such as the anterior cingulate (Milham

et al., 2001) and superior frontal gyrus (du Boisgueheneuc et al., 2006), though activation in linguistic areas remained unaffected by competition. The most striking finding from the current study is that bilinguals displayed substantially less cortical activation compared to monolinguals throughout the duration of the task. A main effect of group illustrated that monolinguals (but not bilinguals) recruited a network of executive control areas (e.g., left superior frontal gyrus: du Boisgueheneuc et al., 2006; anterior cingulate: Milham et Evodiamine al., 2001; left inferior frontal gyrus: e.g., Swick, Ashley, & Turken, 2008; left middle frontal gyrus: e.g., Milham et al., 2002) and primary visual cortex while completing the task. This broad activation in monolinguals is also supported by a significant group by condition interaction and planned comparisons showing that, specifically in response to competition, monolinguals recruited anterior cingulate and left superior frontal gyrus. Such extensive reliance on executive control regions, particularly when confronted with linguistic competition, suggests that monolinguals’ management of phonological competition is not automatic, but rather requires the allocation of domain-general cognitive resources.

44 ppm; Ribeiro et al., 2011). Under this condition, HQ exposure did not alter the number of circulating mononuclear cells but it did reduce the migration of mononuclear cells into the BALF after LPS inhalation, with a consequent reduction in the

number of macrophages. Leukocyte migration to the inflammatory site depends on the highly controlled, sequential expression of adhesion molecules and inflammatory mediators (Borregaard, Mitomycin C concentration 2010 and Ley et al., 2007). It was reported that in vivo HQ exposure increased the physiological expression of β2 and β3-integrins and PECAM-1 and reactive oxygen species (ROS) production by circulating neutrophils. These effects appeared to be connected to impairments to leukocyte migration to the LPS-inflamed lung due to the lack of a neutrophilic response under a challenge ( Ribeiro et al., 2011). However, see more in the current study, adhesion molecules expression on the mononuclear cell membranes was not altered, suggesting that other mechanisms may be involved. It has been clearly demonstrated that mononuclear cell traffics is effectively influenced by MCP-1/CCR2 interactions, mainly under inflammatory conditions (Huffnagle et al., 1995, Melgarejo et al., 2009, Yadav et al., 2010 and Young and Arndt, 2009). Interestingly, reduced levels of

MCP-1 were found in the BALF of HQ-exposed animals after LPS inflammation. The effect depended on functional alterations in AMs and tracheal tissue as reduced MCP-1 levels were found in the supernatant of these cultures. Since a limited number of AMs is found

in the BALF of mice, rendering total RNA extraction unfeasible, an RT-PCR assay was only performed on the tracheal tissue, which showed that the reduction in MCP-1 was defined by impaired mRNA synthesis. Inappropriate MCP-1 secretion was also detected Interleukin-3 receptor when naive mononuclear cells and tracheal tissue were incubated in vitro with HQ, indicating a direct action of the phenolic compound in these cells/tissues. In support of our data, it was recently shown that in vitro HQ exposure impairs MCP-1 secretion by human epithelial cells via the inhibition of mRNA synthesis and by human neutrophils via unknown mechanisms ( Pons and Marin-Castaño, 2011 and Yang et al., 2011). Monocyte chemoattractant protein-1 is a fundamental chemotactic molecule that is mainly released following cell stimulation. It is transcriptionally induced after NF-κB, AP-1 and/or STAT activation in a highly controlled process, which is tissue and stimulus specific (Ding et al., 2010, Tanimoto et al., 2008 and Yadav et al., 2010). Unlike other cytokines synthesized via NF-κB activation ( Ribeiro et al., 2011), only MCP-1 levels were reduced in the respiratory system by HQ exposure. We believe that this effect could be related to the following: (1) the partial activation of transcription factors; (2) reduced interaction between transcription factors and their specific gene promoter region or (3) diminished mRNA stability ( Ding et al.

The supernatants were collected and used to determine the MCP-1 levels. A subgroup of animals was exposed to inhaled LPS (E. coli 026:B6; 0.1 mg/ml; 10 min) or sterile saline (control) for 1 h following the

last in vivo HQ or vehicle exposure using an ultrasonic nebulizer; 8 h later, blood and BALF were obtained in order to quantify total and differential cell numbers. Leukocytes collected from the abdominal aorta blood of vehicle and HQ, exposed or not to LPS were used to quantify the expression levels of l-selectin, β2-integrin, β3-integrin and PECAM-1. Briefly, erythrocytes were lysed by adding ammonium chloride solution (0.13 M) to the samples and leukocytes were recovered after washing with Hank’s balanced salt solution (HBSS). In order to quantify the expression of adhesion molecules,

Selumetinib chemical structure leukocytes (1 × 105) were incubated for 20 min in the dark at 4 °C with monoclonal antibody (β2 or β3-integrin conjugated with FITC or l-selectin or PECAM-1 conjugated with PE). Following this, the cells were analysed in a FACSCalibur Flow Cytometer (Becton & Dickinson, San Jose, CA, USA). Data from 10,000 events were obtained and only the morphologically viable mononuclear Ruxolitinib purchase cells were considered for analysis. Flow cytometry standard (FCS) files were analysed using FlowJo software 8.7.1 (Treestar, Ashland, OR, USA). The results were presented as arbitrary units of fluorescence. The concentrations of MCP-1 were measured in the BALF and the supernatant of tracheal tissue or AM cultures using enzyme-linked

immunosorbent assay (ELISA) kits according to the manufacturer’s specifications. The results were expressed as pg/ml. Total RNA was extracted from in vitro N-acetylglucosamine-1-phosphate transferase LPS-stimulated trachea using Trizol reagent and following the manufacturer’s instructions. The RNA extraction was carried out in an RNAse-free environment and quantified by reading the absorbance at 260 nm. The cDNA was synthesized from total RNA (2 μg) using an oligo(dT)15 primer (20 μg/ml) after incubation (70 °C, 5 min) in the presence of a deoxynucleotide triphosphate mixture (dNTP, 2 mM), a ribonuclease inhibitor (20 U) and Moloney murine leukaemia virus reverse transcriptase (200 U) in reverse transcriptase buffer (25 μl final volume). The reverse transcription occurred during incubation at 42 °C (60 min). For PCR, the cDNA obtained was incubated with Taq DNA polymerase (2.5 U), 3′- and 5′-specific primers (0.4 μM) and dNTP mix (200 μM) in buffer-thermophilic DNA polymerase containing MgCl2 (1.5 mM).

When the GenBank Hyal amino acid sequence for Pp-Hyal (ADLO9135) from this study was aligned with the same allergen of V.

vulgaris (PDB 2ATM), P. annularis (HUGA_POLAN), and A. mellifera (PDB 1FCQ_A), high levels of similarity were revealed (75%, 90%, and 54%, respectively). In Fig. 2, shaded blue areas indicate several regions of similarity mainly among the three first molecules. In addition, the amino acids DFE (highlighted by a red rectangle), which are present in the active site, are also highly conserved. The two proteins – Ves v 2 (PDB ID: 2ATM) and Api m 2 (PDB ID: 1FCQ) – used for building the model of the 3D-structure of the Pp-Hyal had their 3D-structures already determined by X-ray crystallography at a resolution of 2.0 Å ( Skov et al., 2006) and 2.7 Å ( Markovic-Housley et al., 2000), respectively. Despite the greater similarity among sequences have been found between Fluorouracil nmr the proteins of P. paulista and V. vulgaris, only the 3D-structure of the Api m 2 was solved with HA as its substrate, reason why the latter was used in this study to identify the Pp-Hyal active site and points of contact with the substrate. Based on its model (Fig. 3A,B), Pp-Hyal displays a structure comprised of a central barrel (β/α)7 containing seven α-helix and seven beta-sheets, in agreement with the

expected structure for all hyaluronidases CP-868596 belonging to family 56 of glycoside hydrolases ( Henrissat and Bairoch, 1996; Markovic-Housley et al., 2000; Skov et al., 2006). This model also reveals two important characteristics of the Pp-Hyal structure: the presence of two disulfide bonds between Cys 19–308 and Cys 185–197 ( Fig. 3A) and putative glycosylation sites on residues Asn79, Asn187, and Asn325 ( Fig. 3B). The sites Asn79 (5′ NITI 3′) and Asn325 (5′ NITI 3′) are also found in Hyal of V. vulgaris venom ( Skov et al., 2006), indicating that they exert a direct influence on the immunogenicity of the molecule. Glycosylation is the most common post-translational modification of many eukaryotic intracellular proteins, contributing to biological

activity, immunogenicity, solubility, stability, and protease resistance. Thiamet G Carbohydrate residues may be enzymatically attached to proteins through the N-glycoside bond via the amide nitrogen of asparagine, or through the O-glycoside bond via the hydroxyl group of serines, threonines, hydroxylysines or hydroxyproline, or by a glycosylphosphatidylinositol anchor, which is subsequently removed ( Steinberg et al., 2001). Fig. 4 shows the topology of the Pp-Hyal molecule ( Fig. 4A), making evident its active site position when compared to that of Hyal from A. mellifera and the predicted amino acid residues in the model that establish interaction with the substrate Ser299, Asp107 and Glu109 ( Fig. 4B).