Detailed knowledge of the molecular mechanisms underlying ubiquitin processing provided an inroad for designing molecular

probes targeting conjugating and deconjugating enzymes. This approach has regained interest also because it allows small molecule inhibitor development within the UPS [42]. Whereas ubiquitin processing enzymes (USPs, UCHs, OTUs) can be readily profiled using ubiquitin based chemical probes targeting proteolytic catalysis, the application of this activity-based approach towards the ubiquitin conjugating NU7441 cell line cascade has proven to be challenging [43 and 44], although chemical crosslinking was successfully used for HECT domain E3 ligases [45]. In the case of deubiquitination, further progress has been made to create molecular probes mimicking different isopeptide linkages between ubiquitin and protein substrates including ubiquitin itself by integrating peptides at the P′ side of the scissile bond with an electrophilic moiety in the center, which appear to selectively target subsets of DUBs in crude cell extracts [46]. This approach will complement predominantly in vitro studies on how DUBs can distinguish between different poly-ubiquitin chains for processing, currently achieved using model substrates [47] or fluorescence resonance techniques [48]. Also, more recent

probes on the basis of the ubiquitin scaffold are now available with Cobimetinib price C-terminal PTK6 fluorescent moieties via ‘Click Chemistry’ [49], or N-terminal fluorescent or photoreactive moieties through total synthesis [50 and 51•]. Such tools will undoubtedly be used to profile ubiquitin processing enzymes such as DUBs, but also related enzymes specifically recognizing ubiquitin-like proteins, and thereby contribute to our understanding the role of these enzymes within the ubiquitin network in normal physiology as well as disease pathogenesis (Figure 4). The author declares no conflict of interest

in relation to the work described in this manuscript. Papers of particular interest, published within the period of review, have been highlighted as: • of special interest I am grateful for all the helpful discussions with colleagues and would like to apologise for all references that were not cited because of space constrains. “
“Current Opinion in Chemical Biology 2013, 17:73–82 This review comes from a themed issue on Omics Edited by Matthew Bogyo and Pauline M Rudd For a complete overview see the Issue and the Editorial Available online 6th January 2013 1367-5931/$ – see front matter, © 2013 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.cbpa.2012.11.025 Proteomics has made astonishing advances in all areas including peptide enrichment, fractionation, mass spectrometry and data analysis — many of which are reviewed in this issue.

rs2043055 was significantly associated with peak and area under the curve (AUC) triglycerides after an OFTT in the subjects classified as ‘cases’ (P = 0.001 for both). Homozygotes of the less frequent C allele had 17.63 mg/dl lower peak triglycerides and 0.7 less AUC compared to common T allele homozygotes. Selleckchem GSI-IX The same trend was observed in the ‘controls’ but did not reach statistical significance ( Table 4). There was no interaction of case: control status with rs2043055 and no effect on post-prandial insulin and glucose. There was no association of rs2043055 with post-prandial measures after an OGTT (Data not shown). No associations were observed at the haplotypic level (

Table 3). rs2043055 C allele homozygotes exhibited higher insulin levels (P = 0.054), a higher HOMA-IR estimate (P = 0.035) and a lower QUICKI estimate (P = 0.048) in comparison to carriers for the common T allele ( Appendices Table 3). However, these associations did not reach statistical significance. Highly significant associations were observed after haploytpe analysis with plasma insulin levels, HOMA-IR, HOMA-β-cell, and QUICKI estimates (Global P < 0.0001 for all associations) ( Table 3). After further

analysis and Bonferroni correction, it was observed that the phenotypic mean for insulin, HOMA-IR and HOMA-β-cell was significantly higher for Hap6 in comparison to the common haplotype, Hap1 (Bonferroni P < 0.001 for all associations). Insulin levels were 5.56 μIU/ml higher; HOMA-IR and HOMA-β-cell GSK126 estimates

were 1.34 and 20.75 higher, respectively. Furthermore, QUICKI was significantly lower in Hap6 in comparison to Hap1 (Bonferroni P < 0.001). The FDR for these associations was between 0.001 and 0.003% (q-value 0.001–0.003). The major findings from this study are the effects of variation within IL-18 using combined haplotypes analysis, on insulin levels and estimates of insulin resistance. Furthermore, this is the first report over of the influence of a variation within IL18 on post-prandial triglyceride levels, supporting the idea of IL-18 playing a role in metabolic processes. Examining the SNPs individually, by univariate analysis in all three studies, associations were seen only with rs2043055. In EARSII there was a significant association of rs2043055 with peak and AUC triglycerides after an OFTT in the subjects classified as cases in EARSII. Homozygotes of the less frequent C allele had significantly lower peak triglycerides and smaller AUC compared to T allele homozygotes. These results suggest carriers of the C allele clear or absorb triglycerides faster than TT individuals and support the idea of IL-18 playing a role in metabolic processes. Post-prandial measures were not available in the GrOW study and therefore the associations observed in EARSII could not be replicated.

The antibody allowed the isolation of an almost > 95% pure population of osteocytes from calvariae of 18-day-old chicken fetuses

using immunomagnetic separation [2], and the study of characteristics and properties of these osteocytes [4]. Using this antibody it was shown for the first time that isolated Cyclopamine price osteocytes are much more responsive to mechanical load in the form of pulsating fluid flow than osteoblasts or periosteal fibroblasts [5]. Osteocytes are the pivotal cells orchestrating the biomechanical regulation of bone mass and structure for efficient load bearing [5], [6], [7], [8] and [9]. The mechanosensitive osteocytes comprise 90-95% of the whole bone cell population in the adult animal [10]. Within the hard mineralized matrix, osteocyte cell bodies reside

in the spaces called the lacunae. From each osteocyte cell body, approximately 50–60 cell processes originate and radiate through the mineralized matrix via spaces called the canaliculi. Together these structures are called the lacuno-canalicular system (LCS). These cell processes radiate in different directions and form an intricate intercellular network of osteocytes (Fig. 2), which is directly connected to the cells lining the bone surface and cells within the bone marrow [11], [12] and [13]. How the osteocytes sense the mechanical loads on bone and coordinate adaptive alterations in bone mass and architecture is not yet completely understood. However, it is widely accepted that mechanical http://www.selleckchem.com/products/17-AAG(Geldanamycin).html loads placed on bones as an organ drive a flow of interstitial fluid through the unmineralized 3-oxoacyl-(acyl-carrier-protein) reductase pericellular matrix surrounding osteocytes and their dendritic processes [9] and [14]. This

flow is then thought to somehow activate the osteocytes, which produce signaling molecules that can regulate the activity of the effector cells [15] and [16], the osteoclasts and the osteoblasts, leading to adequate bone mass and architecture [17] (Fig. 3). Over the past two decades theoretical and experimental studies have contributed in delineating the role of osteocytes in mechanosensation and their subsequent biological response. New insights have emerged from an enhanced understanding of the anatomical details of the primary osteocytes [4], [11], [13] and [18], osteocyte isolation [2] and [19], mechanosensation [20], and signal transduction [21], [22], [23] and [24], to name just a few of these advances. Computational models have demonstrated the importance of mechanical loading as a potent and stable regulator of complex biochemical processes involved in maintenance of bone architecture [17]. If osteocytes, acting as the bone mechanosensors, indeed orchestrate the adaptation of bone to mechanical loading, the question arises how this biological action is performed.

The percentage of wet deposition was highest over the northern subbasins, around 65% over B1 and B2 in winter and autumn. Nitrogen deposition to the Baltic Sea is very episodic. The number of high deposition events in 1993–1998 (Hongisto & Joffre 2005, Figure 13) shows clear differences in the annual variation of the oxidized and reduced nitrogen depositions. The annual and seasonal numbers of wet episodes

(defined here as the 6 h deposition over a sub-basin exceeding 10-fold the 10-year average 6 h deposition of the month for that sub-basin) in 2000–2009 are presented in Figures 5 and 6. The frequency of NOy deposition episodes had distinct minima in the periods 1995–1997 and 2001–2005, and there was another decrease GSK2118436 price in 2009. The correlation coefficient R of the number of episodes with the total annual NOy deposition was R > 0.55 over B1-B3, the index of determination R2 was 31–33% but the P-value was higher than 0.05, indicating only a statistically suggestive correlation.

The winter episodes depend on the ice conditions: in 2008, when the Gulf of Bothnia and the Gulf of Finland were ice-free most of the time, the episode frequency increased, whereas in the more southerly sea areas seasonal differences in the number of episodes were not so much in evidence. The average MBL conditions have interannual, seasonal, diurnal and very this website short term variations, different in different BS sub-basins. Over all the sub-basins, precipitation was most intensive in the winters of 2007–2008 and 2001–2002, as well as in summer 2007 and autumn 2000–2001; during these seasons, the Baf-A1 concentration pressure was lower than the periodic average. The wind velocity was lowest over the narrower gulf areas. One can notice a rather high interannual variation in the seasonal averages. The MBL height has a north-south gradient, and there is generally a rather high annual variation in seasonal average MBL heights. The correlations R of the 6 h values of wet and dry deposition of NOy over B3 and B1 with wind speed, precipitation, surface pressure, mixing height, friction velocity and temperature

in 2000–2009 are presented as seasonal averages in Figure 7, while the corresponding explanation factors (R2) are shown in Figure 8. The annual correlations are small because opposing stability conditions prevail over BS in spring and autumn: there are > 14 000 time periods, and dispersion of all parameters was high, especially during the peak deposition events. The correlation coefficients indicate only if a linear regression between the variables exists. However, from the scatterplots one can conclude that deposition is nonlinearly dependent on most of the meteorological parameters, and this seems to be the case even for the dependence of wet deposition on precipitation. If we study 6 h correlation averages over shorter periods, e.g.

On receiving a trigger signal a final pH was reported. A software option was available to gate whether the injection proceeded if the pH was within a predefined range, e.g. pH 7 ± 1. The reported pH was estimated to be accurate to ±0.1. Other devices requiring external device control could be connected to the available Y-27632 research buy external digital output pins of the Arduino board. User-programed software functions controlled the activity of the pump, permitting multistep flow rates, volumes (absolute or calculated volume as a function of animal weight for a given dose) and flow direction. Volume calculations were made

by the software using a calibration volume based on a single revolution by the stepper motor. Using a software function, the calibration volume could be determined based on the mass of water delivered after ten revolutions of the stepper motor or by adaptive volume calibration, in which the calibration volume was internally adjusted based on the measured volumes and compared against the requested volume by the software. Additional features included a cleaning routine using flow control to flush the pump and cannula whilst still connected to the animal. The pump was first calibrated

by three measurements of the mass of distilled water delivered through 1100 mm of 0.96 mm O.D., 0.58 mm I.D. tube into a glass vial after ten revolutions of the pump at 80 rpm. The selleck chemicals llc average water mass divided by ten was then entered into the software as a volume per revolution. All subsequent volumes

were calculated by the software based on this calibration. The accuracy and scalability of the injection system delivered volumes were measured against programed volumes in the range 0.100–10.000 ml for an arbitrarily chosen constant flow rate of 7.0 ml/min. The delivered volume for a given demanded volume was similarly measured at least three times (range 3–5) by mass of distilled water. The delivery of hyperpolarized substrate was tested firstly in vitro and subsequently in vivo. In both types of experiments 13C1 pyruvic acid (PA) (Sigma Aldrich, Gillingham Ltd., UK) was Reverse transcriptase mixed with 15 mM OX63 trityl radical (Oxford Instruments, Abingdon, UK) and 1.5 mM DOTAREM (Guerbet, Roissy, France). 45 mg (12.7 mg for in vitro tests) of PA was hyperpolarized using a HyperSense DNP polarizer, operating between 1.2 and 1.4 K, using a microwave frequency 94.150 GHz and 30 mW for approximately 1 h. The hyperpolarized frozen sample was transferred to the receive vessel using 3.4 ml superheated buffer solution containing 40 mM HEPES buffer solution, 0.269 mM disodium EDTA and 50 mM NaCl (all obtained from Sigma Aldrich). The receive vessel contained a predetermined aliquot of 2.0 M sodium hydroxide solution (Sigma Aldrich) required to neutralize the PA and 2.0 ml HEPES/EDTA buffer solution to ensure that the receive vessel outlet pipe was submerged. Final concentration of PA was ∼100 mM.

Perhaps a method of Spiral Array block generation would be of even better use for heterogeneity determination [38]. Nevertheless, it was our study that indicated clearly the heterogeneity

of which proteins might be of use in EC. Another problem was the lack of a unified system that would serve accessing the heterogeneity within the studied markers. However, the analyzed proteins have different functions this website within cells, which means that they differ in terms of localization and quantity. Ergo, different scoring criteria had to be assumed and unified evaluation and cutoff determination were simply not feasible. The studies concerning intratumor heterogeneity were primarily performed at the genomic or transcriptomic level [2], [39], [40] and [41] and the contribution of tumor diversity to disease progression has so far received rather

scarce attention. Nevertheless, effective cancer treatment requires a complex idea about tumor structure and intratumor heterogeneity needs to be taken into account [23]. To the best of our knowledge, we are the first to present tumor heterogeneity distribution measured by IHC in such a wide context. We show that heterogeneity degree in EC might serve as a clinically valid molecular marker and IHC could be a fast and simple method of its determination. The following are the supplementary data related to this Erastin article. Help with ZIP files Options Download file (2510 K) Help with ZIP files Options Download file (2686 K) Help with selleck ZIP files Options Download file (2451 K) Help with ZIP files Options Download file (2836 K) Figure W1.   Consecutive cores of Patient No. 276 illustrating the tumor heterogeneity in the context of estrogen receptor

staining. The research has been financed by the Ministry of Science and Higher Education under grant N407571538. The research has been co-financed by the European Commission in the framework of the European Social Fund, by the European Social Fund, by the State Budget, and by the Pomorskie Voivodeship Budget according to the Operational Programme Human Capital, Priority VIII, Action 8.2, Under-action 8.2.2: ‘Regional Innovative Strategy’ within the system project of the Pomorskie Voivodeship “InnoDoktorant – Scholarships for PhD students, Vth edition”. “
“Gastrointestinal stromal tumors (GISTs) primarily arise from mesenchymal tissue in the gastrointestinal (GI) tract and abdomen. Although GISTs are rare, representing only an estimated 0.1% to 3% of all GI tract tumors [1], they account for the most common mesenchymal malignancy of the GI tract [2]. GISTs appear to be related to the interstitial cells of Cajal [3] and express the cell surface transmembrane receptor KIT, which has tyrosine kinase activity.

All procedures were carried out at 4 °C. The efficiency of this renal fractionation procedure was verified by measurement of lactate dehydrogenase in MF and SF, as previously described by Yamasaki et al. (2008). Total protein was measured, photometrically (Bio-Tek Power Wave XR spectrophotometer), at 630 nm, in triplicates, in samples of SF (diluted 50-fold), MF (diluted 20-fold), plasma (individual and pool of animals of the same experimental group) (diluted 500-fold) and urine (diluted 75-fold), by the method of Bradford (1976), using a Bio-Rad protein assay reagent (Hercules, USA). Protein content was

extrapolated by comparison with standard curves of bovine serum albumin in the same diluent. AP activities were fluorometrically BMS-354825 supplier measured in Epacadostat ic50 pools of plasma and urine, and in individual samples of SF and MF of renal medulla and cortex, as previously described by Yamasaki et al. (2008). AP activities were expressed as picomoles of hydrolyzed substrate/min/mg of protein.

Osmolality was determined in 10 μL individual plasma and pooled urine samples in a cryoscopic osmometer (Osmette II Fisher). Creatinine was photometrically quantified at 500 nm, in 20 μL of individual plasma and pooled urine samples, by the method of Biggs and Cooper (1961) modified by Marinho et al. (2006), with Creatinine Fast kit (Laborlab, Brazil). Uric acid was photometrically quantified at 505 nm, in 5.4 μL of individual plasma and urine samples, by the method of Prencipe et al. (1978) modified by Marinho et al. (2006), with Uric Acid UOD-PAD kit (Laborlab, Brazil). Urea was photometrically quantified, at 340 nm, in 3 μL of individual plasma and pooled urine samples, by the method of Talke and Schubert (1965) modified by Yamasaki et al. (2008), with Urea UOD-PAD kit (Laborlab, Brazil). Oxidative stress was evaluated

on renal cortex and medulla from dissected kidneys stored at −80 °C, by measurements of oxidized (GSSG) and reduced (GSH) glutathione as described by Yamasaki et al. (2008). For this purpose, cortex and medulla were homogenized in 0.1 M phosphate buffer, with 0.005 M EDTA, pH 8.0 (PBEDTA) plus 5.26% HPO3 (0.1 g tissue/1.5 mL PBEDTA plus 0.4 mL check details 25% HPO3), at 800 rpm for 3 min. These homogenates were ultracentrifuged at 100,000×g for 30 min. The pellet was discarded and the supernatant was immediately used as described below. All steps were carried out at 4 °C. GSH was measured in 100 μL of supernatant diluted in PBEDTA (1:10), mixed with 170 μL PBEDTA and 30 μL o-phthalaldehyde (OPT) (100 μg OPT/100 μL ethanol 2%), and incubated for 15 min at 25 °C. Blank values for GSH were obtained by reading 100 μL deionized water with 170 μL PBEDTA plus 30 μL OPT, 15 min after incubation at 25 °C. GSSG was evaluated in 56.7 μL of supernatant incubated with 22.7 μL 0.1 M ethylmaleimide (Sigma) for 30 min at 25 °C. After this incubation, 487 μL 0.

tygodnia życia zarodka, jest procesem wieloetapowym i skomplikowanym. Jego poznanie BMS-354825 molecular weight pozwala nie tylko na zrozumienie mechanizmów powstawania wad wrodzonych, ale również umożliwia interpretację

obrazu poszczególnych malformacji pod kątem ich współistnienia u pacjenta. Komórki biorące udział w rozwoju serca pochodzą z mezodermy trzewnej oraz puli sercowej komórek grzebieni nerwowych [1, 2]. Te pierwsze stanowią podstawę dla uformowania dwóch odrębnych skupisk tworzących pierwsze i drugie pole sercowe. Do niedawna niedoceniana rola komórek grzebieni nerwowych w formowaniu niektórych części serca została potwierdzona w związku ze współistnieniem szczególnych postaci wad wrodzonych serca z zaburzeniami w obrębie innych tkanek i narządów 3., 4., 5. and 6.. Ponieważ stanowią one jednocześnie główną pulę komórek, z których wywodzą się m. in. grasica, przytarczyce oraz niektóre kości trzewioczaszki, zaburzenia ich migracji skutkują powstawaniem charakterystycznych grup wad wrodzonych, zwanych zespołami twarzowo-podniebienno-sercowymi (velocardiofacial syndromes), do których należy m. in. zespół DiGeorge’a (mikrodelecja 22q11) [7, 8]. Jego cechami charakterystycznymi

są m.in.: wada serca (zaburzenia rozwoju odpowiednich dla komórek grzebieni nerwowych struktur – przede wszystkim drogi odpływu i łuku aorty), dysmorfia twarzy, której może towarzyszyć rozszczep wargi i/lub podniebienia, niedorozwój bądź agenezja grasicy i związane z tym pierwotne zaburzenia odporności oraz hipokalcemia będąca efektem niedoczynności

przytarczyc [8]. Jak zatem można spostrzec, istnieją liczne zależności między rozwojem serca i struktur this website nie tylko sąsiednich, ale również znajdujących się w odległych okolicach ciała. Rola poszczególnych populacji komórek w rozwoju serca została przedstawiona na rycinie 1. Pierwsze pole sercowe bierze udział głównie w tworzeniu przedsionków, kanału przedsionkowo-komorowego i lewej komory. Populacja komórek drugiego pola sercowego dzieli się zaś na trzy odrębne grupy – pole sercowe przednie, tylne i wtórne. O ile prawa komora wywodzi się z przedniego i wtórnego pola sercowego, o tyle RAS p21 protein activator 1 tylne pole sercowe stanowi źródło komórek tylnej ściany przedsionków (tej, która nie wywodzi się z pola pierwszego), pierwotnej zatoki żylnej, żył płucnych, układu przewodzącego, a także żył serca, w tym zatoki wieńcowej [1, 9]. Należy zaznaczyć rolę tylnego pola sercowego, a dokładniej wywodzącego się zeń narządu przednasierdziowego, w powstawaniu tętnic wieńcowych [10]. Wspomniane wcześniej komórki grzebieni nerwowych, które migrują w kierunku pierwotnej cewy sercowej, biorą udział w tworzeniu tętnic łuków gardłowych, drogi odpływu prawej komory, zwojów serca, a wraz z narządem przednasierdziowym również układu przewodzącego [6, 11]. Początkowo prosta cewa sercowa ulega w kolejnych etapach zapętlaniu (Ryc. 2). Spowodowane jest to szybszym wzrastaniem jej strony brzusznej w stosunku do grzbietowej.

Inulin Anti-infection Compound Library purchase is a storage carbohydrate found mainly in chicory root (Cichorium intybus) and Jerusalem artichoke (Helianthus tuberosus) and, structurally, is composed of β-d-fructofuranose polymers joined by β(2 → 1) links, with a degree of polymerization that can reach 70 ( Roberfroid & Delzenne, 1998). Oligofructose is obtained through partial hydrolysis of chicory inulin and subsequent purification, and its degree of polymerization

ranges from 2 to 8 ( Biedrzycka & Bielecka, 2004; Roberfroid, 2005). Prebiotics can be applied to a variety of foods. Inulin and oligofructose present, respectively, 10 and 35% of the sweetness power of sucrose (Franck, 2002), allowing them to partially replace sucrose in some formulations (De Castro, Cunha, Barreto, Amboni, & Prudencio, 2009; Villegas, Tárrega, Carbonell, & Costell, 2010; Wang, 2009). Because of gelling characteristics, inulin allows the development of low-fat foods through the replacement of significant amounts of fat and the stabilisation of the emulsion, without compromising texture (Franck, 2002; González-Tomás, Coll-Marqués, & Costell, 2008; O’Brien, Mueller, Scannell, & Arendt, 2003; Paseephol, Small, & Sherkat, 2008). Prebiotics can also increase product flavours, such as citrus HDAC assay aroma

and flavour perception of probiotic fermented milks (Sendra et al., 2008), lemon flavour of dairy desserts (Arcia, Costell, & Tárrega, 2011) and vanilla flavour intensity of custards (Tárrega, Rocafull, & Costell, 2010). However, prebiotics can also impair some sensory characteristics of food, such as a thickening in dairy desserts (Arcia et al., 2011), hardness and cohesiveness in cakes (Moscatto, Borsato, Bona, Oliveira, & Hauly, 2006) and higher firmness and lower FER acceptability of sponge cakes (Ronda, Gómez, Blanco, & Caballero, 2005). Gonzalez, Adhikari, and Sancho-Madriz (2011) found that peach-flavoured yogurts with fructooligosaccharide show similar sensory profile and acceptability, but fructooligosaccharide with added Lactobacillus acidophilus (synbiotic ingredient) present a negative

impact on sensory acceptability. Incorporation of prebiotics into baked goods allows the replacement of sugar, enriches fibre and improves moisture retention properties (Franck, 2002; Wang, 2009). Some studies have been conducted on adding fructans to cakes (Devereux, Jones, McCormack, & Hunter, 2003; Moscatto et al., 2006; Ronda et al., 2005), in which the cakes were evaluated regarding physical properties (texture, colour and volume) and sensory acceptability, but no studies evaluated the effects of prebiotic addition on the sensory profile of cakes. Certain health benefits can be claimed for products containing inulin and oligofructose as prebiotics, but the official rules about the use and exact wording of these claims vary from country to country.

78 Mb region was identified by MLM. However, the mapping resolution was not improved beyond α < 0.05 by MLM with an increased threshold ( Fig. 3). All of the markers identified with a strong association with cob

and pericarp color phenotypes in this study were located within a region of 0.78 Mb, ~ 0.73 Mb upstream and ~ 0.05 Mb downstream of the P1 gene. Among the identified markers, a significant positive correlation between associations (− log10P) with these traits and genetic effects (R2) on these traits was found. The strongest association and the Caspase pathway highest genetic effect were found at marker PZE-101064790, which is located upstream of the P1 gene. The distance from P1 and the surrounding sequence showed that PZE-101064790 is located within the P1 enhancer, which plays a key role in regulating P1 gene expression and conferring its tissue-specific pattern [15] and [16]. The identified locus and associated

markers might be the best targets for potential regulation of cob glume color and also good targets for developing marker-assisted selection tools. Regional LD and the LD decay pattern were analyzed. For the temperate GWAS panel, a clear LD block with a set of markers surrounding the P1 locus was found. It is located Thiazovivin concentration at the P1 gene and includes 22 markers upstream (box shown in Fig. 4) and four markers downstream of the gene ( Fig. 4). LD decay of the P1 locus and its adjacent region was very rapid. The R2 value decreased from 0.83 to 0.30 within this 14 kb region. Outside the target region of

the LD block, the P-values increased rapidly to a pattern similar to that of the genomic background. To compare LD at the target region between temperate maize and tropical maize and to analyze regional LD at better resolution, 10-fold deep sequencing at the target region was performed on 87 lines, which included 40 temperate lines that overlap with the 283 lines in the temperate GWAS panel and 47 tropical lines with white cob glume color (Table 2). Marker density increased from ~ 45 kb/marker for the GWAS panel genotyped via maize SNP50 to ~ 207–271 bp/marker by deep sequencing. A number of markers within the significant LD region were found upstream of P1 in the temperate maize lines ( Fig. 5). Florfenicol Among those markers, two clear and novel structured LD blocks were found in the temperate maize lines, but not in the tropical maize lines. In addition, a new LD block was found downstream of the P1 locus only in the tropical maize lines. These results suggest that the accuracy of LD analysis can be improved, and when marker density increased, more specific information around the locus useful for positional cloning and functional identification of genes was revealed. To study the effects of genetic diversity and artificial selection involved in the development of inbred lines, the markers spanning chromosome 1 were analyzed for genetic diversity among the temperate GWAS lines.