Encouraging results with a specificity of 85.7% and a sensitivity of 83.3% did indicate that the Venetoclax in vitro model can effectively discriminate active TB from CRD and HC (Table 3). The demonstration elsewhere suggested that this classification tree model could be a potential diagnostic tool for active TB. Similar research of TB has been performed in the recent years, which set up a diagnostic model containing 20 peaks that can distinguish

TB from other inflammatory diseases and healthy controls [5]. However, the model we established is based on recruitment of several pulmonary diseases with clinical manifestations or laboratory indices that can overlap those of active TB. Apparently, the latter one is more appropriate for clinical utility, but a second dataset, which is prospectively obtained

from patients with respiratory symptoms as Agranoff et al. did [5] should be used to further confirm the model’s specificity and sensitivity for diagnosing Dabrafenib molecular weight active TB. Although we tried our best to rule out patients with latent TB from the non-TB group, some patients or healthy controls with latent TB might still be recruited. As no similar research has been performed between latent TB and active TB, we cannot decide whether latent TB affects the performance of the model or not and this should be further explored. Also, HIV/TB, multidrug TB and ETB restrain the management of TB so strongly that related classification tree models should be set up. Some studies reported that different biomarkers might exist in diverse situations of sputum smear microscopy of patients with

TB [27], while others considered it results from the bias of quality control. To investigate this interesting phenomenon, comparison among peaks of SPP-TB, SNP-TB and non-TB has been performed. There were 54 proteins that can discriminate these three groups (Table 4). Forty of the 54 proteins also showed up in the differential expressed proteins between active TB and non-TB, which suggested that these proteins not only play an important role in the pathogenesis of active TB but also regulate the status of active TB. Surprisingly, both 8561 and 8608 m/z showed up in this Selleck Abiraterone analysis, which further highlighted that the importance of these two peaks and further identification of them are needed. Comparing to the prior study that only recruited 10 patients with pneumonia and three patients with COPD in the non-TB group [28], none of their differential expressed peak was found in our research. Inherent complexity of active TB, technological difference between magnetic beads and protein chips and different composition of non-TB group might result in this inconsistent condition. As we know, identification of meaningful peaks is necessary for understanding the pathogenesis of TB. Furthermore, Agranoff et al. [5] identified two of their differently expressed peaks to be serum amyloid A protein and transthyretin.

The method also combined measurement of changes in Ca2+i using fluo-4 and excitation at 490 nm. Results:  After establishing loading conditions, a linear relationship was demonstrated between Em and fluorescence signal in FRET dye-loaded HEK cells held under voltage clamp. Over the voltage range from −70 to +30 mV, slope (of FRET signal vs. voltage, m) = 0.49 ± 0.07, r2 = 0.96 ± 0.025. Similar data were obtained in cerebral artery SMCs, slope (m) = 0.30 ± 0.02, r2 = 0.98 ± 0.02. Change in FRET emission ratio over the holding potential of −70 to +30 mV was 41.7 ± 4.9% for HEK cells and 30.0 ± 2.3%

for arterial SMCs. The FRET signal was also shown to be modulated by KCl-induced depolarization BAY 73-4506 in vitro in a concentration-dependent manner. Further, in isolated arterial SMCs, KCl-induced depolarization (60 mM) find more measurements occurred with increased fluo-4 fluorescence emission (62 ± 9%) and contraction (−27 ± 4.2%). Conclusions:  The data support the FRET-based approach for measuring changes in Em in arterial SMCs. Further, image-based measurements of Em can be combined with analysis of temporal changes in Ca2+i and contraction. “
“Please cite this paper as: Zhang (2011). Effect

of Suspending Viscosity on Red Blood Cell Dynamics and Blood Flows in Microvessels. Microcirculation 18(7), 562–573. To obtain a better understanding of the beneficial effect of high plasma viscosity observed in hemodilution and resuscitation experiments, we conducted a computational study to investigate

the suspending viscosity effect on red blood cell (RBC) dynamics and blood flow behaviors in microvessels. For single RBCs in simple shear or channel flows, RBCs appear more flexible as indicated by the tank-treading motion in shear flows and the strong transverse migration in channel flows. For the multiple RBC flows in straight channels, our results indicate no significant change with the suspending viscosity in stable flow structure and hemorheologic behaviors, under both constant Calpain flow and forcing conditions. However, due to the increase in apparent cell deformability in a more viscous medium, the cell-free layer (CFL) can be established in a shorter distance along the channel. Considering the multilevel bifurcated structure of the microvascular network, this change in CFL development distance may affect the phase skimming and RBC separation processes at the downstream bifurcation, and therefore the microcirculation performance in the tissue. This may suggest a possible mechanism for the high functional capillary density associated with a high suspending viscosity observed in experiments. “
“Please cite this paper as: Folkesson KT, Samuelsson A, Tesselaar E, Dahlström B, Sjöberg F.

, 2010; Workentine et al., 2010). Phenotypic variants such as mucoid variants (Govan & Deretic, 1996), small colony variants (SCVs) (Häußler et al., 2003; Häußler, 2004) and quorum-sensing (QS) variants (D’Argenio

et al., 2007; Hoffman et al., 2009) are commonly CT99021 clinical trial isolated from chronic infections. It has therefore been suggested that long-term adaptation to the lung of the host results in reduced expression of acute virulence factors to develop a chronic infection type which may facilitate long-term, chronic infection (Proctor et al., 2006; Bragonzi et al., 2009; Hogardt & Heesemann, 2010). We have demonstrated that mucoid clinical strains of P. aeruginosa isolated from the sputa of chronically infected patients with CF exhibit seeding dispersal during in vitro biofilm growth, as do model laboratory strains. Intriguingly,

dispersal of clinical strains resulted in a higher frequency and colony diversity of dispersal variants than the dispersal population of the laboratory strain PAO1 (Kirov et al., 2005, 2007). Colony morphotype is only one indicator of variation, and therefore, this study characterised the biofilm dispersal population of a mucoid, chronic infection CF isolate of P. aeruginosa and the laboratory strain, PAO1, for a variety of functional traits to determine the extent of diversification that occurs during biofilm development. The traits examined included those that are likely to enhance the capacity of P. aeruginosa to establish chronic airway infection, such as the capacity to utilise different carbon sources, as this has been shown to be important FK506 mw for the growth of P. aeruginosa in artificial sputum

and is of functional significance for bacterial survival in the CF lung mucus (Sriramulu et al., 2005; Palmer et al., 2007; Starkey et al., 2009). Additionally, the dispersal population was tested for properties likely to contribute to niche colonisation and persistence in the CF airway, including attachment and biofilm formation, QS signal production and virulence factor production such as general protease and elastase production. The mutation frequencies of the parental strains were quantified under planktonic and biofilm growth to correlate mutation frequency with variant formation. Pseudomonas aeruginosa PAO1 (Holloway, 1955) and the clinical strain 18A, isolated from the sputum of a chronically infected patient Aurora Kinase with CF in Tasmania, Australia, and characterised as previously described (O’May et al., 2006), were used here. The latter strain was selected for further study as it was representative of a number of clinical isolates that showed seeding dispersal and marked heterogeneity in the morphotypes of its dispersal cells (Kirov et al., 2005, 2007). Cultures were stored at −80 °C, and all strains and biofilm-derived isolates were routinely grown on LB10 agar [Luria–Bertani agar: 10 g L−1 of tryptone, 5 g L−1 of yeast extract, 10 g L−1 of NaCl and 15 g L−1 of agar (Research Organics Inc.)] at 37 °C.

Working memory buy IWR-1 processes are closely interrelated to attentional processes as attention permits information to be further stored and processed in working memory. Attentional processes are reflected by the visual N1 event-related potential (ERP)-component. The visual N1 may reflect effects of attention on sensory processing or an integrated process of perception and attention. The visual N1 is an exogenous potential that is modulated by attentional processes modifying the magnitude of neural responses to incoming information. Beste et al.

[136] examined the association of the TNF-α rs1800629 polymorphism with attention and mental rotation performance in an event-related potential (ERP) study in healthy participants. The results show that carriers of rs1800629 A-allele display elevated attentional processes as compared to the GG genotype group. Carriers of the rs1800629 A allele performed Ivacaftor better than the GG genotype group. The finding of enhanced attentional and mental rotation performance in A-allele carriers supports recent findings that the A-allele of this SNP enhances cognitive performance on a general measure of cognitive processing speed. Interferon-alpha increases

the expression of TNF-α. During interferon-alpha therapy in psychiatric symptoms, TNF-α polymorphism played a role in susceptibility to this disorder. Recently role of TNF-α rs1800629 polymorphism in labile anger and depression was investigated by Lotrich et al. [137]. A-allele of rs1800629 was associated with worsened labile anger and fatigue during treatment but not with major depression incidence or increased Beck Depression Inventory Meloxicam II. Labile anger was not predicted by the serotonin transporter polymorphism. During treatment with an exogenous cytokine, vulnerability to worsening labile anger distinct from major depression is associated with genetic variability in TNF-α. Tumour necrosis factor-alpha has been reported to play a role in neuropathic pain. Leung and Cahill [138] described the role of TNF-α in neuropathic pain. Neuropathic pain is pathological pain where nociceptive responses

persist beyond the resolution of damage to the nerve or its surrounding tissue. Animal models of neuropathic pain based on various types of nerve injuries have persistently implicated a pivotal role for TNF-α at both peripheral and central levels of sensitization. Achrol et al. [139] identified SNPs associated with increased risk of new intracranial haemorrhage (ICH) after brain arteriovenous malformation (BAVM). Achrol et al. [125] investigated four promoter SNPs in interleukin-6 and tumour necrosis factor (rs1800629, rs361525). An association has been found between TNF-α rs361525 polymorphism and increased risk of new ICH after diagnosis. The patients with TNF-α rs361525 AG genotype had increased risk of new ICH. No other SNP was found to be associated with new ICH. Genetic factors play role in endometriosis [5, 140].

Together, FCAS, MWS and CINCA syndrome are grouped and called CAPS. These syndromes are characterized by recurrent fevers, leukocytosis, elevated acute phase proteins, myalgias and generalized fatigue. CINCA syndrome is a severe form of CAPS beginning in neonatal life. The term “cryopyrin” was coined by Hoffman during his studies regarding the mutation in FCAS 15. MAPK Inhibitor Library research buy Upon exposure to cold, the affected subjects develop fevers, leukocytosis and generalized flu-like symptoms, hence the use of “cryo” for cold and “pyrin” for fever. Blood monocytes from these patients release more IL-1β upon incubation in the cold as compared with monocytes from persons without the mutation 21. CAPS patients

treated with either anakinra 23, 44, 45, a soluble IL-1 receptor (rilonacept) 17 or a monoclonal

anti-human IL-1β (canakinumab) 29, experience a rapid, sustained and near complete resolution of the disease. Of particular importance is the amelioration of the central nervous system abnormalities in children with CINCA during sustained treatment with anakinra 23 or canakinumab 46. Colchicine is routinely used to prevent attacks of FMF 47. Although the mechanism of action of colchicine in FMF is poorly understood, one effect of colchicine is a reduction in the migration of monocytes into an inflamed area 47. Because oral colchicine is converted in the liver to an active compound by p450 cytochrome C, some patients are resistant to colchicine because they harbor a mutation in p450 cytochrome C. As a result, these patients are treated with anakinra. Other patients are intolerant of the loose stools associated with colchicine Selleck CP 673451 use. Anakinra brings about a rapid cessation of the local and systemic inflammation of an attack. However, periodic anakinra is effective in preventing FMF attacks when administered early during the prodrome and in some patients daily anakinra is used. Colchicine-resistant Etomidate FMF disease severity can present as

bilateral pneumonia; initiation of anakinra therapy in such patients has been shown to result in a rapid improvement in clinical symptoms as well as radiographic resolution within 2 days 48. Since TRAPS was originally believed to be due to a lack of endogenous soluble TNF-α receptor, disease activity was thought to be best controlled by administration of agents that neutralized TNF-α such as etanercept and infliximab. However, TRAPS turns out to be an IL-1β-mediated auto-inflammatory disease and optimally responsive to IL-1β blockade. Blood monocytes from TRAPS patients release IL-1β in greater amounts than cells from healthy subjects 13, a characteristic of auto-inflammatory diseases. In fact, treating patients with TRAPS with infliximab worsened disease severity 13, 49. Another characteristic of patients with auto-inflammatory diseases is the response to reducing IL-1β activity, which is observed in patients who are refractory to corticosteroids, cyclosporine, azathiaprine or colchicine.

Pathophysiological mechanisms by which the risk to develop MS may increase after selleck kinase inhibitor childhood are largely unknown. Much of our current knowledge regarding the assumed auto-immune pathogenesis

of MS derives from EAE, the animal model of MS. Activated, myelin-reactive CD4+ Th1 cells are thought to have a central role in the pathogenesis of both MS and EAE [4]. Initial activation of CD4+ T cells occurs through recognition of Ag presented in the context of MHC class II (MHC II). Processing of Ag and presentation of linearized peptides is provided by MHC II-expressing APCs [5], such as myeloid monocytes and macrophages, DCs as well as B cells. Following Ag recognition, efficient activation of CD4+ T cells requires further ligation with co-stimulatory molecules expressed on the APC surface. Besides the density of MHC II expression [6, 7] and the composition of co-stimulatory molecules CAL101 [8, 9], the fate of the corresponding T cell to either

differentiate into a proinflammatory Th1 or Th17 phenotype or to alternatively develop into an anti-inflammatory Th2 cell or Treg cell is determined by the cytokine milieu present at the site of APC-T-cell interaction [10, 11]. Thus, a variety of signals provided by the APCs is required for efficient development of proinflammatory T cells in vivo. Based on this conception, we tested in the EAE model whether an age-associated alteration of innate immune cell function may determine Ixazomib chemical structure susceptibility to CNS autoimmune

disease. EAE is traditionally induced by active immunization with CNS autoAg in 8- to 20-week-old mice, as EAE susceptibility is maximal at this age [12]. To establish that susceptibility may be lower at an earlier age, EAE was induced in C57BL/6 mice at the age of 2 weeks using an active immunization protocol with MOG p35–55 in CFA and PTx. As indicated in Figure 1A, none of the 2-week-old mice showed any clinical signs of EAE (0/13), whereas 8/8 mice at the age of 8 weeks developed ascending paralysis around day 10 after immunization. Twelve days after immunization, a subgroup of mice was analyzed for development of myelin-reactive T cells. As shown in Figure 1B, splenocytes from 2-week-old mice revealed a strongly reduced proliferation of T cells in response to MOG p35–55. Furthermore, secretion of IFN-γ and IL-17 was decreased suggesting that EAE resistance of 2-week-old mice relates to an inability of younger mice to generate encephalitogenic T cells. In order to elucidate mechanistically why young mice are unable to generate EAE-inducing, proinflammatory T cells, we first confirmed that the frequency of peripheral T cells was unchanged. As indicated in Figure 2A, there was no difference in 2- or 8-week-old mice in the frequency of total CD3+ T cells as well as the ratio of CD4+ to CD8+ T cells.

To exclude unspecific effects of the chitin particles, we included glass beads of comparable size or just PBS as controls.

Chitin did neither induce nor inhibit Th2 polarization under neutral or polarizing conditions, respectively (Fig. 1C). This result led us to conclude that reduced Th2-cell expansion in vivo is not due to a direct inhibitory effect of chitin on T cells. The reduced frequency of TCR-tg cells in lung and LN of OVA/chitin-treated mice (Fig. 1A and B) could be due to inefficient homing, impaired proliferation or reduced survival of transferred T cells. To address the possibility that chitin inhibits T-cell proliferation, we labeled total splenocytes from BALB/c mice with CFSE and stimulated them in vitro with anti-TCR/anti-CD28 in the presence or absence of chitin. Proliferation of CD4+ T cells was inhibited by 30–40% Trichostatin A in the presence of chitin (Fig. 2A and B). Next, we sought to determine whether the inhibitory function of chitin was mediated by binding of chitin to the mannose receptor which has previously been shown to

serve as chitin receptor 11. T cells were cultured in the presence of chitin and soluble mannan in order to block binding of chitin to the mannose receptor as described earlier 11. However, the inhibitory activity of chitin was not reduced in the presence of Lumacaftor mannan and mannan alone did not inhibit T-cell proliferation (Fig. 2A and B). To further determine whether inhibition of T-cell proliferation can be induced by direct interaction of chitin with T cells, we stimulated purified and CFSE-labeled CD4+ T cells on anti-TCR/anti-CD28-coated plates in the presence or absence of chitin or glass. Chitin or glass had no influence on the efficiency of T-cell proliferation, suggesting that the inhibitory effect of chitin is mediated by an accessory cell

type (Fig. 2C). The accessory cell type that mediates chitin-induced inhibition of T-cell proliferation might be a macrophage population. Indeed, cocultures of macrophages and T cells revealed Sorafenib chemical structure that chitin caused a strong inhibitory effect (Fig. 3A and B). We have previously shown that chitin upregulates expression of Arg1, an enzyme which is induced in macrophages by Stat6-mediated signaling from the IL-4 receptors and therefore served as marker to indicate differentiation into AAM 9, 23, 24. However, it has recently been shown that Arg1 can also be induced by TLR-mediated signaling in a Stat6-independent fashion 25. Therefore, other markers like the chitinase-like protein Ym1 or “found in inflammatory zone 1” (Fizz1)/“resistin-like molecule α” appear to be more specific for AAM 26.

For this purpose, human immature DCs were

exposed to fluorescein isothiocyanate (FITC)-labelled AGE-OVA and FITC-labelled regular OVA and uptake was analysed by flow cytometry and fluorescence microscopy. Furthermore, autologous CD4+ T-cell proliferation and cytokine production induced by mature DCs loaded with AGE-OVA were compared with those induced by mature DCs loaded with OVA. Finally, expression of the receptor for advanced glycation endproducts (RAGE) and activation of the transcription factor nuclear factor (NF)-κB by AGE were investigated. Internalization of FITC-AGE-OVA by immature DCs was significantly increased compared with FITC-OVA. Blocking the mannose receptor, macropinocytosis https://www.selleckchem.com/products/Trichostatin-A.html or the scavenger

receptor strongly reduced uptake of both FITC-OVA and FITC-AGE-OVA. In a comparison of CD4+ T cells co-cultured with AGE-OVA-loaded mature DCs versus those co-cultured with OVA-loaded mature DCs, AGE-OVA DCs were found to produce more interleukin (IL)-6 and to induce a stronger T helper type 2 (Th2) and a weaker Th1 cytokine response, while ABT-263 mouse there was no difference in proliferation of CD4+ T cells. The expression of RAGE was higher on immature DCs compared with mature DCs. AGE-OVA-exposed immature DCs showed a stronger expression of RAGE and activation of the transcription factor NF-κB compared with OVA-loaded immature DCs. Our data indicate that AGE-OVA may be more immunogenic/allergenic than regular OVA. In the industrialized nations, the prevalence of food allergy is increasing.1,2 Factors such

as food production, processing, conservation, storage, sterilization and final preparation may play an important role in this increase.3 Although heat treatment of food has many advantages, such as improvements in taste, appearance and smell and the destruction of pathogens, it may produce drastic changes in the allergenicity of proteins.4,5 Most food proteins are denaturated by heat treatment, and this denaturation includes the destruction Phloretin of their three-dimensional structure. Therefore, certain epitopes show a diminished capacity to bind immunoglobulin E (IgE) antibodies and thus reduced allergenic potential. However, there are also examples for the creation of new epitopes by food processing, for example during the Maillard reaction, leading to advanced glycation endproducts (AGEs).6,7 This non-enzymatic reaction of amino acids with non-reducing sugars occurs in the heat treatment during cooking of cakes, biscuits and amylase containing foods or after their long-term storage.8,9 It also takes place in the human body, mainly in aging tissues or in blood vessels of diabetic patients with increased blood sugar levels. Neoantigens induced by the Maillard reaction such as AGEs are more resistant to digestion in comparison to native proteins.

The recombinant antigens, early secretory antigen target 6 (ESAT-6) and culture filtrate protein 10 (CFP-10), are encoded in a region of difference (RD)1, a genomic segment absent from the BCG and most environmental mycobacteria [22]. The advantages of using IGRA for the diagnosis of LTBI and TB disease include the need for only a single patient visit, this website the speed with which results can be obtained and the high sensitivity and near perfect specificity of the diagnosis [23, 24]. The major disadvantages of the IGRA include

the need for a suitably equipped laboratory and trained people and the relatively high cost [25]. However, IGRA has already been shown to be significantly more specific than TST mTOR inhibitor for the diagnosis of LTBI and TB disease, especially in endemic countries such as Brazil [26]. Their performance in the diagnosis of children has, nevertheless, not been extensively investigated [27, 28], and there is thus a lack

of knowledge in this area. In view of this, the aim of this study was to analyse the differences between IFN-γ levels against ESAT-6, CFP-10 and PPD in vitro, in children with LTBI and TB disease and healthy donors from an area where TB is endemic, and to assess the diagnostic potential of these antigens based on these differences. The importance of applying new diagnostic approaches to detect LTBI early in children lies in the fact that it halts progression to TB disease, which causes irreversible damage to the lungs and future respiratory problems in infected children. Selection of patients and control.  BCG-vaccinated Erastin nmr children, aged between 3 and 15 years old, were selected prospectively over the period between the years

2005 and 2007 from the Hospital das Clínicas da Universidade Federal de Pernambuco and the Instituto Materno Infantil Professor Fernando Figueira (IMIP), in Recife, in the Brazilian State of Pernambuco, according to the criteria used by the ATS (American Thoracic Society) [29]. The patients were divided into two groups: (1) children with confirmed tuberculosis (TB disease, n = 21), with an epidemiological history of contact, clinical evidence and/or a chest radiography compatible with tuberculosis and/or TST >10 mm; (2) patients with a high risk of having latent tuberculosis infection (LTBI, n = 17), including children with a history of contact with an individual with tuberculosis or who have TST >10 mm. Both groups were selected prior to treatment. The negative control group (NC, n = 21) was composed of children with a non-reactive TST, with no history of contact with TB and no specific symptoms of tuberculosis. This group was selected from the IMIP cardiology unit. Demographic and clinical data were obtained for each child using a detailed questionnaire. These data included date of birth, TB exposure history, BCG vaccination status (vaccination certificate and/or the presence of the typical scar) and symptoms suggestive of TB.

In the case of splenic macrophages, 10 units/mL IFN-γ was added to the culture medium to prime cells. In addition, 5 μg/mL polymyxin B was also added to avoid cell activation by LPS in the IFN-γ sample. Separately, RAW264.7 cells were incubated with pDNA/LA2000 complex for 2 h, then the cells were washed with RPMI-1640 CCI-779 solubility dmso and incubated with fresh growth medium for an additional 6 h, and the supernatants were collected for ELISA and kept at −80°C until use. PMDC05 cells were incubated with ODNs for 24 h, then the supernatants were collected for ELISA and kept at −80°C until

use. The level of murine TNF-α and murine IL-6 in the media was determined by ELISA using the OptEIA set (BD Biosciences Pharmingen, San Diego, CA, USA). The level of human TNF-α in the media was determined by human TNF-α ELISA set (eBioscience, San Diego, CA, USA). RAW264.7 cells were incubated with Alexa488-labeled ODN1668 with or without ODN1720 or DNase-treated ODN1720 for 4 h and MI-503 mouse washed

three times with PBS. Then, the intensity of cell fluorescence was analyzed by flow cytometry (FACScan; BD Biosciences) using CellQuest software (version 3.1; BD Biosciences). Cellular uptake was estimated by subtracting the mean fluorescence intensity (MFI) at 4°C from that at 37°C (ΔMFI) and was plotted against the incubation time. ODN1668 was mixed with DNase I- or DNase II-treated ODN1720. The mixture containing 0.5 μg ODN1668 was incubated with DNase I (2 units/μg ODN1668) or DNase II (3 units/μg ODN1668) at 37°C. After 0, 5, 15, 30 min of incubation (DNase I treatment) or 0, 20, 40, 80 min of incubation (DNase II treatment), the mixture Progesterone was placed on ice and the reaction was terminated by the addition of 3 μL 0.2 M EDTA solution per 10 μL of samples. The ODN samples were run on a 21% PAGE and stained with ethidium bromide. The image of the gel was recorded using LAS 3000 (Fujifilm Life Science, Tokyo, Japan) and analyzed using Multi Gauge software (Fujifilm Life Science). Differences in the cytokine release were statistically evaluated by Student’s t-test. Differences in the thickness

of mouse footpad were statistically evaluated by one-way analysis of variance (ANOVA) followed by the Tukey–Kramer test for multiple comparisons. A p-value of less than 0.05 was considered to be statistically significant. We wish to thank Dr. Hiroyuki Yoshitomi (Graduate School of Medicine, Kyoto University) for providing technical assistance for subcutaneous injection of ODN into the footpad of mice. This work is partly supported by the 21st Century COE Program “Knowledge Information Infrastructure for Genome Science” and by a grant-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Sciences and Technology, Japan. Conflict of interest: The authors declare no financial or commercial conflict of interest. Detailed facts of importance to specialist readers are published as ”Supporting Information”.