Bacteria conjugated to pHrodo™ show a very low fluorescent signal

Bacteria conjugated to pHrodo™ show a very low fluorescent signal at the neutral pH present on the cell find more 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, VX-809 price 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

Phosphatidylinositol diacylglycerol-lyase 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.

Following single-cell isolation, the epigenome and the transcript

Following single-cell isolation, the epigenome and the transcriptome may also be studied [21]. While epigenomics of single cells remains challenging [52, 53, 54 and 55], methods for single-cell transcriptomics have flourished (Figure 4) and delivered baffling new insight into the (functional) heterogeneity of cell populations [56, 57 and 58]. A single cell contains less than 1 pg of mRNA. To characterize it via array [59 and 60] or sequencing [56, 57 and 58] approaches, whole-transcriptome amplification (WTA) is required. Methods for WTA are grounded on PCR-based [61, 62, 63, 64, 65•, 66, 67, 68 and 69], MDA-based

[67] or in vitro transcription (IVT)-based [ 70] amplification LBH589 purchase of reverse transcribed single-cell mRNA, whereby IVT likely results in a more linear amplification. However, WTA and subsequent analysis methods struggle with reliable amplification and detection of transcripts expressed at less than 10 copies per cell. In addition, the majority of the methods only selectively amplify the polyadenylated RNAs of a cell’s transcript repertoire [ 61, 62, 63, 64, 65•, 66 and 70], and may be biased to the 3′-end [ 70] or the 5′-end [ 61 and 62]

of a transcript ( Figure 4). Full-length mRNA-characterization from a IBET762 single cell can only be achieved by a few WTA-methods [ 63, 64, 65•, 66, 67 and 68] ( Figure 4). To the best of our knowledge, no large-scale single cancer cell transcriptome sequencing studies have been reported, although

this is on the horizon [71, 72 and 73]. In a recent elegant proof-of-principle experiment, Ramsköld et al. found differences between melanoma CTCs and primary melanocytes, giving insight into the disease [ 65•]. Additionally, the technology allowed defining potent plasma membrane CTC biomarkers and discovering expressed coding mutations. This and other studies [ 60, 73, 74 and 75] show that single-cell transcriptomics will illuminate further insights into oncogenesis, tumour subclonal architecture and cell lineage diversity. Single-cell sequencing studies currently only process either WGA-products or WTA-products of a cell, although protocols for combined approaches are under development [76• and 77]. The ability to profile both the genome and the transcriptome of the same cell has enormous potential to elucidate Astemizole heterogeneity at the genome, epigenome and transcriptome level. In addition, such techniques would allow mutations of the genome in a single cell to be confirmed in that cell’s transcriptome, opening avenues to detect mutations at high confidence, even if they are observed only in one single cell. The emerging field of single cell genomics opens new avenues that may have far-reaching implications for cancer research and clinical practice. It allows characterization of intra-tumour genetic heterogeneity genome-wide to single-cell resolution, and thereby offers a unique viewpoint into tumour evolution.

We would like to thank Vincent Récamier, Raphaël Voituriez, Leoni

We would like to thank Vincent Récamier, Raphaël Voituriez, Leonid Mirny, Yitzhak Rabin, Lana Bosanac and Benjamin Guglielmi for stimulating discussions. We also acknowledge financial support from the following grants: ANR-12-BSV8-0015 and ANR-10-LABX-54. “
“Modification of cysteine residues by reactive oxygen species (ROS), reactive nitrogen species (RNS) and electrophiles has emerged as a significant means of altering the structure and function of many proteins [1, 2, 3, 4, 5 and 6]. Reversible oxidation

of certain protein thiol groups plays key signaling INCB024360 supplier roles in a range of physiological processes, for example in the regulation of tyrosine phosphatase activity [7], the redox regulation of transcription factors [8] and in T cell activation during the immune response [9]. The reactivity of protein thiols with ROS, RNS and electrophiles additionally underlies Omipalisib their important role in defense against oxidative damage and xenobiotics [1, 2, 3, 4, 5 and 10].

In all of these processes there are a broad range of reactions that can occur to the cysteine thiol (Figure 1). Whether a modification occurs depends on a number of factors including the local environment of the cysteine residue, its proximity to the relevant reactive species, its pKa, solvent exposure and subcellular location [ 1, 6, 11 and 12••]. Additionally, some of these cysteine modifications are reversible by the action of reductive processes through the thioredoxin

and glutathione systems [ 13 and 14]. Reversible thiol modifications include glutathionylation [ 15], mixed disulfide formation with low molecular weight thiols, sulfenic acid formation [ 3], S-nitrosation (S-nitrosylation) [ 16], S-acylation [ 17], sulfenylamide formation [ 18], and the generation of intraprotein and interprotein disulfides [ 19 and 20]. In addition to reversible modifications, there are a number of cysteine adducts that can form irreversibly due Amine dehydrogenase to reactions with electrophiles, which generally produce thioether products [ 10]. Similarly, the prolonged exposure of cysteine residues to ROS and RNS can also lead to the formation of irreversibly modified forms, such as sulfinic or sulfonic acids [ 21 and 22]. These protein modifications may contribute to oxidative damage, to the defense against oxidative stress and xenobiotics, or be part of redox signaling pathways. Consequently, it is of interest to be able to identify both the proteins and the cysteine residues affected, to determine the nature of the modification to the cysteine residue and to quantify the extent of the modification occurring during pathology or redox signaling.

msc org/track-a-fishery/fisheries-in-the-program/certified/pacifi; accessed 25th July 2013). If this sets

a precedent for certification of purse seine fisheries this may mark a move away from FAD fishing with renewed focus on pursuing free schools. The increase in the use of FADs over the past two decades has given rise to concern over their associated ecological impacts, yet management of FAD fishing is complicated by the compromise between achieving a reduction in these impacts and allowing the sustainable exploitation of healthy tuna stocks, namely skipjack tuna. This is complicated further by the current reliance of the purse seine fishery on this highly efficient fishing practice, which is likely to only increase further under a business-as-usual scenario as fishing operations

become more expensive buy PFT�� and shrinking profit margins require an ever greater use of FADs. CDK inhibitor However, continued growth in FAD fishing might be expected to result in diminishing returns as the relative benefit of each FAD in the fishery is diluted. Explicit management of the use of FADs is undoubtedly a necessity to ensure future sustainability of the fishery. Whilst there are several options available to manage the use of FADs, each option is expected to produce a different response from the purse seine fleet. Time-area closures have already been implemented but with mixed success in reducing juvenile mortality due to the flexibility of the fleet in reallocating effort. Whilst larger (and longer) closures may achieve greater reductions in juvenile catch this would be at the expense of significant reductions in skipjack catch. This has major implications on the fishing and processing industries based in the Indian Ocean, with a realistic danger that many purse seiners would choose

to leave the Indian Ocean altogether. On the other hand, input controls such as limiting Lenvatinib manufacturer the number of actively monitored FADs or the number of sets made on floating objects directly address concerns about FAD fishing, if designed and implemented appropriately, but are likely to be challenging to negotiate within the IOTC and difficult to enforce. We are grateful to the Economic and Social Research Council and the Natural Environment Research Council for funding this research. This paper is a contribution from Imperial College’s Grand Challenges in Ecosystems and the Environment initiative. Generous thanks are also given to J. Pearce, L. Dagorn and A. Fonteneau for informative discussion on the current and future management of FAD fishing, and to J.J. Areso, several members of staff at the Seychelles Fishing Authority and a number of anonymous skippers who gave up their time to offer invaluable insight into the practical aspects of purse seine fishing. “
“Coastal communities throughout the developing world are recognised as being particularly vulnerable to environmental change [1], [2] and [3].

The Harvard Educational Review published an entire issue consisti

The Harvard Educational Review published an entire issue consisting of critiques of Art’s work and Art himself faced many personal attacks and not a few physical attacks. When I was Art’s graduate student in 1980 the Selleckchem PARP inhibitor campus police still opened all of his mail to ensure that none contained a bomb. These attacks notwithstanding, Art unflinchingly responded to his critics with sound research evidence to support him. Though there are still some who consider his work to be “race science”, in the worst sense, the rigor of Art’s research eventually convinced many others that he was correct. In recognition of this, Art was elected a fellow of the American Association for the Advancement of Science, he

was awarded the prestigious Kistler prize, and both the International Society for the Study of Individual Differences and the International Society for Intelligence Research gave him their lifetime achievement awards. In addition to his academic life, Art had several other passions and interesting hobbies. As a teenager he caught snakes which he gave to the San Diego Zoo. Also as a teen he wrote a book-length manuscript about Gandhi: a figure he had the utmost admiration for.

Art was also an accomplished clarinetist and at one point considered pursuing a career as a musician. Although he didn’t do this, music was undoubtedly a major passion of his: he had season’s tickets to the San Francisco Opera and could talk for hours about his favorite conductor: Arturo Toscanini. He was also a skilled chess player. At his house on Clear Lake, Art enjoyed sailing and he would swim for selleckchem up to an hour at a time every day until quite late in his life. He also trained a flock of ducks to swoop onto his lawn at precisely 4 pm each afternoon for a reward of bread crumbs and bird seed! Last but by no means least Art was a wonderful cook who specialized in East Indian cuisine. Even when his Parkinson’s made it very difficult for him to talk and move about, Art continued working and writing right up to his death. In one of the chapters in The Scientific

Study of General Intelligence: A Tribute to Arthur R. Jensen, edited by Helmuth Nyborg and presented Metformin manufacturer to Art at the ISSID meeting in Graz when he was given his lifetime achievement award, another of his former graduate students wrote that he was “Inspiring…scientifically rigorous…a wonderful mentor…deeply committed to his students…a formative influence on my values as a researcher, and a model of courage in pursuing the truth regardless of the opposition encountered”. I couldn’t sum up his legacy any better. Art is survived by his daughter, Roberta. “
“The authors regret that in the 3.2. Structural model section, the standardized path coefficient from social support to life satisfaction was reported (b = .01, p < .05). In fact, the standardized path coefficient should be from EI to life satisfaction and be non-significant (b = .01, p < .05). The authors would like to apologize for any inconvenience caused.

In order to describe the thalamic data more accurately we next cl

In order to describe the thalamic data more accurately we next classified neurons by location into those in Vim versus Vop. The Vim mean rates were significantly greater in postural Dabrafenib chemical structure ET than in cerebellar tremor (P<0.01, shown in Fig. 2A), but not different from intention ET or from controls with pain (not shown). The mean Vim firing rate for intention ET was not significantly different from cerebellar tremor (not shown). The Vop mean rates of subjects with postural ET were higher than in cerebellar tremor (P=0.002, not shown). Therefore, firing rates in

postural ET were consistently higher than those in cerebellar tremor. The activity of all neurons included was studied for a response to joint movements during mapping of the thalamus, as described in the Methods. These cells were located in the region anterior ( Fig. 1C: P2) and dorsal to the region in which cells respond to cutaneous stimuli

( Fig. 1C: P1, and Lenz et al. (1988)). The proportions of cells responding to deep sensory stimuli and those not responding to such stimuli are shown by tremor type in Table 2. The proportion of neurons in Vim responding was greater for postural ET than cerebellar tremor (P=0.00012, Chi square with Bonferroni correction) and controls with pain (P=0.048). The number of sensory cells in Vop was different only between intention ET and cerebellar tremor (P=0.02, Fisher with Bonferroni). Since sensory inputs may be an important factor in the relationship of cerebellar tremor and cortical MS-275 ic50 activity (Flament et al., 1984, Hore and Flament, 1986 and Vilis and Hore, 1977), we next examined the mean spontaneous rates for sensory cells across the four groups (Fig. 2B). There was a clear and significant change in the firing rate of sensory cells according to patient groups (1-way ANOVA, F=3.47, P<0.05). Post-hoc testing showed that the firing rate of sensory cells in the postural ET group was significantly higher than that of cerebellar tremor and controls with pain. The

rate for intention ET was not different from postural ET. We next examined how the thalamic signal qualitatively differed between groups of patients. The frequency at which peak spike activity these occurred was found for each neuron within the tremor range (1.9–7 Hz) (Lenz et al., 2002). The mean “frequency of peak spike power” occurred at a different frequency for each group of tremor patients (1-way ANOVA, F(3,259)=8.75, P<0.0005). The mean frequency of this peak is significantly higher in postural ET patients (4.8 Hz+0.25, mean+SEM) as compared to cerebellar tremor (3.4 Hz+0.2, post-hoc Newman–Keuls test, P=0.0057) and intention ET (3.7 Hz+0.4, P=0.032). The frequency was not significantly different between intention ET and cerebellar tremor (Newman–Keuls test P=0.34).

0001) Fig 3D represents the enzyme activity levels measured for

0001). Fig. 3D represents the enzyme activity levels measured for CYP1A2. The results showed that the levels of activity detected in BEAS-2B cells were equivalent to those observed when the CYP1A2 inhibitor fluvoxamine was present in the cultures. These results concurred with the results obtained for CYP1A2 gene expression that indicate that without induction there is no expression of the CYP1A2 gene in

BEAS-2B cells based on a Ct > 36. HepaRG cells (positive control cell line) showed a reduction in enzyme activity (1.6-fold) in the presence of inhibitor, however, this reduction was not statistically significant (p = 0.127). The lung-derived cell line BEAS-2B has been identified as a cell line of interest in the in vitro toxicological testing of inhaled toxicants ( Veljkovic et al., 2011 and Ansteinsson et al., 2011). However, to date the metabolic capabilities of this cell line have not been thoroughly investigated. In this study, we employed Selleck FG 4592 high throughput technology to provide a rapid screening for gene expression and inducibility for a panel of 41 metabolism-related genes, producing a profile of gene expression and gene inducibility. Then, four key CYP enzymes involved in the bioactivation of some smoke pro-toxicants were selected for functional enzyme activity assay. The data obtained from both analysis would confirm if enzyme activity was consistent with gene expression. The scientific approach used

in this study is a working example of our proposed strategy for the metabolic characterization of cell systems used in the context of in vitro toxicology testing. Our gene expression results show that non-induced BEAS-2B cells have high and moderate mRNA selleck screening library expression for, GSTM1 and SULT1A1 respectively, both related to conjugative reactions which mainly act as detoxification mechanisms (Castell et al., 2005). As expected, when cultures were pre-induced with TCDD, CYP1A1, CYP1B1

and CYP1A2 genes showed an up-regulation of 25-fold, 6-fold and 4-fold respectively compared with non-induced cultures. The up-regulation of CYP1A1 and CYP1B1 genes after pre-incubation with TCDD has also been reported in normal G protein-coupled receptor kinase human primary bronchial epithelium (NHBE) cells (Newland et al., 2011). Surprisingly, in a recent publication Courcot and colleagues report high levels of CYP1B1 gene expression in non-induced cultures of BEAS-2B cells and high levels of CYP1A1/1B1 gene expression in non-induced cultures of human primary bronchial epithelium cells (HBEC), among other lung cell systems (Courcot et al., 2012). This contrasts with our gene expression results and other published results (Newland et al., 2011 and Castell et al., 2005). It is possible that the high levels of CYP1A1 and CYP1B1 reported in HBEC by Courcot and colleagues could be as a result of the smoking habit of the donor; cigarette smoke is known to activate these enzymes in the lung (Nishikawa et al., 2004 and Anttila et al., 2011). However, this information is not disclosed in their methodology.

During the post-transplant follow-up phase, patients were followe

During the post-transplant follow-up phase, patients were followed up for 48 weeks for evidence of recurrent HCV infection. All participating sites planned to use a standard post-transplantation immunosuppressive regimen of solumedrol/prednisone, tacrolimus,

and/or mycophenolate mofetil (up to 2 g/day) for the first 12 weeks after transplantation. selleckchem Antibody induction was prohibited during the study. The primary efficacy end point was post-transplantation virologic response (pTVR), defined as HCV-RNA level less than the lower limit of quantification (LLOQ, 25 IU/mL) at 12 weeks post-transplant in patients who had HCV-RNA levels less than the LLOQ at their last assessment before transplantation. According to the original study analysis plan, only patients who received at least 12 weeks of treatment before transplantation were to be included in the efficacy analysis. However, this restriction was not used in the analysis, therefore the efficacy population includes patients who received any duration of treatment (Table 2 shows the overall results for both populations). Other secondary efficacy end points included an evaluation of safety and tolerability. Plasma HCV-RNA levels were measured with the COBAS TaqMan HCV

Test, version 2.0, for use with the High Pure System (Roche Molecular Systems). Population sequencing PI3K Inhibitor Library chemical structure of the HCV NS5B-encoding region of the viral polymerase was performed using standard sequencing technology on all baseline (pretreatment) viral samples. Deep sequencing with an assay cut-off filipin value of 1% was performed for all patients who qualified for resistance testing as a result of an incomplete virologic response on treatment, post-treatment relapse, post-transplant recurrence, or early termination with HCV-RNA levels greater than 1000 IU/mL. Nucleoside inhibitor-associated variants were defined as N142T, L159F, L230F, and V321A, and any substitutions at position S282 of NS5B. Drug susceptibility testing was performed using a replicon system with either patient population samples or site-directed mutants. Assuming an observed week 12 pTVR rate of 50%, we calculated that a sample size

of 31 would be sufficient to show that the 1-sided 95% upper bound of the confidence interval (using a normal approximation of the binomial) for the recurrence rate would be 65%. See the Supplementary Appendix for a detailed description of the statistical methods. The study was approved by the institutional review board or independent ethics committees at participating sites and was conducted in compliance with the Declaration of Helsinki, Good Clinical Practice guidelines, and local regulatory requirements. The study was designed and conducted according to protocol by the sponsor (Gilead) in collaboration with the principal investigators. The sponsor collected the data, monitored study conduct, and performed the statistical analyses.

Several studies have investigated interactivity in self-managemen

Several studies have investigated interactivity in self-management educational programs [10] and [11]. In addition, in artificial intelligence, interactivity is a current topic of interest in automated systems for health communication [12]. However, less research on interactivity on consumer health websites is available [8]. Overall,

the dimensions of interactivity remain unclear [8], [13] and [14]. In light of this, the objective of this paper is to present the conceptual design of PARAFORUM (, a consumer-oriented website in the field of SCI developed by Swiss Paraplegic Research on behalf of Swiss Paraplegic Foundation. PARAFORUM will be launched in November 2013 and will implement several dimensions of interactivity. This paper examines these ZD1839 datasheet dimensions and discusses the potential benefits and challenges of websites like PARAFORUM from the point of view of consumers and organizations. This paper presents the design of the website PARAFORUM in the

context of its conceptual roots. These roots have been identified by analyzing, confronting, and merging different streams of research in online communication [15] and [16] with a specific focus on health [13] and [14], web-based communities [17], [18] and [19] with a specific focus on the production of health knowledge [20], [21], [22] and [23], open innovation communities [24], [25] and [26], and through formative research with peer counselors, health professionals, and this website board members who work for the Swiss Paraplegic Group ( MYO10 PARAFORUM is a website that enables interaction with and among main stakeholders in the field of SCI, namely, individuals with SCI, their families, health professionals, and researchers active in the field of SCI. PARAFORUM supports information and interaction

in four languages (German, Italian, French, and English). In designing PARAFORUM, the main emphasis was on operationalizing the roles of the users and the directions of their interactions. More specifically, PARAFORUM aims to capture and share among the members of its community three different types of expertise: • the expertise of individuals with SCI and their families who have years of experience in managing SCI; PARAFORUM provides interactions in five main directions: • from health professionals to individuals with SCI and their families (the professional-to-consumer interaction, when the lay community requests information from professionals); These last two dimensions aim to promote the growth of a collaborative online community where the expertise of individuals with SCI and their families is identified and translated to enhance new ideas for research and organizations’ practice. Through interactions regarding specific problems and challenges, users can co-design solutions. By exploiting communication in the Web 2.

MaβFS1 transgenic lines showed a unique peak compared with the co

MaβFS1 transgenic lines showed a unique peak compared with the control ( Fig. 6-A, B). The peak was identified as EβF with a retention time and mass spectrum identical to that of authentic EβF ( Fig. 6-C, D). EβF emission levels of the Ma1, Ma4, and Ma10 transgenic lines were 2.81, 4.85 and 2.62 ng d− 1 g− 1 in fresh tissues, respectively. To test the efficacy of the transgenic lines in control of aphids, two independent evaluations were conducted in a setup as indicated in Fig. 7-A.

In the repellence test, the numbers of aphids on transgenic and control tobacco plants were counted 12 h after the aphids were released. Compared with the control, aphids on transgenic lines Ma1, Ma4, and Ma10, were reduced by approximately 8.8%, 10.4% Etoposide in vivo and 7.7%, respectively, whereas about 25.5% of the aphids stayed on the net cover or died (Fig. 7-B; Table 2). When 400 alate aphids and 10 lacewing larvae were simultaneously introduced into the setup, the numbers of aphids after 12 h were reduced by 19.2% in Ma1, 29.5% in Ma4 (P < 0.05) and 16.7% in Ma10, compared with the control. Most of the surviving aphids were preyed upon by the lacewing larvae or stayed on the net cover ( Fig. 7-C; Table 3). Therefore, MaβFS1 transgenic tobacco selleck chemicals llc plants showed a pleiotropic

effect on aphid behavior, including repellence to aphids and attraction to aphid predators. Notably, in the presence of lacewing larvae, transgenic line Ma4 could recruit lacewing larvae that significantly affected aphid infestation. In this study, we isolated MaβFS1 and MaβFS2 genes from Asian peppermint and showed that MaβFS1 was functional in tobacco. Although MaβFS1 was identical to the published EβF synthase gene from black peppermint (AF024615), it shared only 34.1% and 28.2% similarities at the amino acid sequence level with EβF synthases from Yuzu and Douglas fir, respectively, and only 34.0% similarity with that of the gene we isolated from sweet wormwood and characterized in vivo ( Fig. 1) [39]. So far, the EβF synthase genes from Douglas fir, Yuzu, sweet wormwood and black

peppermint have been isolated Cepharanthine and characterized in vitro [37], and only the genes from black peppermint and sweet wormwood were successfully introduced into plants and proved to be functional in vivo [38] and [39]. Furthermore, remarkable sequence differences were observed among the EβF synthase genes from various plant species/varieties. However, some regions, including the Asp-rich motif known as DDXXD (at positions 301 to 305 in MaβFS1) and the NSE/DTE motif known as (N/D)DXX(S/T)XXX(E/D) (at positions 444 to 452 in MaβFS1), are highly conserved among the so far isolated plant-derived EβF synthase genes ( Fig. 1). These two conserved domains are supposed to be responsible for divalent metal ion–substrate binding during catalysis [47].