Table 2 gives specific data of the study group and shows the relationship between clinical data and the presence or

absence of cerebral embolism. Table 2 shows that cerebral embolism in this patient cohort was associated with a high-grade internal carotid artery stenosis. Retinal events and aphasia were more frequently seen in patients who experienced cerebral embolism. Table 3 shows the epidemiology of cerebral embolism. It showed a wide range of frequencies of emboli during the 30 min monitoring. Most emboli were short lasting, low intensity events that occurred in the diastolic phase of the cardiac Selleck Gefitinib cycle. The emboli had a very prominent musical sound expressed by the low zero-crossing index. The most prominent source of the embolus was an internal carotid artery stenosis. In most patients the internal carotid artery stenosis was located at the origin of the vessel. In two out of eleven patients the stenosis was located at the level of the carotid syphon. The embolic activity decreased after therapeutical interventions such as carotid surgery, angioplasty and a drug switch from aspirin to clopidogrel. Table 4 shows the outcome of the study protocol in relation to positive and negative embolism. Table 5 shows the outcome of both the control and study group. Table 4 shows that the diagnosis and treatment of

patients with positive cerebral embolism was performed much faster than the diagnosis and treatment of patients without cerebral embolism. Stroke and TIA

recurrence rate in both groups were very low (respectively 0.0% and 3.2%). In the study AZD9291 cell line group, one patient experienced a stroke recurrence in the ipsilateral posterior cerebral artery resulting in a permanent hemi-anopsia. In the control group four recurrent strokes were observed. All these events occurred in the ipsilateral middle cerebral artery territory; two of these events occurred in the post-operative phase of carotid surgery. One of these Thiamine-diphosphate kinase events was classified as a possible cerebral hyperperfusion syndrome. Spencer was the first investigator who showed that detection of cerebral embolism was possible with TCD [8]. His initial study describes the ongoing cerebral embolism in patients scheduled for carotid surgery. Soon after his publication the first reports appeared about MES signals in TIA and stroke patients. In the last ten years a number of studies showed unequivocal that ongoing cerebral embolism in carotid artery disease is a strong independent predictor of stroke [1] and [2]. The current clinical study tried to explore the potential of embolus detection to enhance the outcome of patients with symptomatic carotid artery disease. Briefly summarized this study revealed a non-significant reduction in recurrent events in the study group. Probably sample size in this pilot study was insufficient to detect a significant decline.

For example, the median serum infliximab concentration at week 8 in clinical responders was 35.0 μg/mL compared with 25.8 μg/mL in clinical nonresponders for the 5-mg/kg group at week 8. Similar results were observed selleck for clinical response and mucosal healing during maintenance at week 30 and week 54 (Table 1). For example, in patients who received the 5-mg/kg regimen, the median trough serum infliximab concentration

in clinical responders was several-fold that of clinical nonresponders (eg, 3.9 vs 1.2 μg/mL at week 30 and 5.0 vs 0.7 μg/mL at week 54, respectively). With respect to clinical remission among patients in the 5-mg/kg group, the median serum infliximab concentration at week 8 was not significantly higher in week-8 remitters than in nonremitters (35.1 vs 30.8 μg/mL; P = .097). By comparison, the difference in serum infliximab concentrations between remitters and nonremitters at week 8 was statistically significant for the 10-mg/kg dose group (P = .0002) ( Table 1). The median

serum infliximab concentration was significantly higher in remitters than Dasatinib nonremitters at week 30 (P < .0001) and week 54 (P < .005), regardless of infliximab dose ( Table 1). Although median serum infliximab concentrations were consistently higher in patients with positive efficacy outcomes than those who failed to achieve these outcomes, there was some overlap of the distribution of serum infliximab concentrations between these groups. The overlap, however, was greater during induction at week 8, but less prominent during maintenance at week 30 or week 54. It also appears that there was more variability of serum infliximab concentrations in patients P-type ATPase who failed to respond during maintenance

(Figure 3). When assessed by infliximab concentration quartiles, the proportions of patients with treatment success as defined by multiple outcome measures (ie, clinical response, mucosal healing, and/or clinical remission) generally increased with increasing infliximab concentration for the 5-mg/kg dose regimen. In each case, a significantly positive association was observed for the relationship between serum infliximab concentration quartiles and clinical outcomes (Supplementary Figure 3). Patients with serum infliximab concentrations in the lowest quartile consistently were less likely to show clinical response, clinical remission, or mucosal healing and had rates of success approaching those observed in patients assigned to placebo.2 Notably, this finding was still evident when the quartiles were examined for the 10-mg/kg dose regimen, as illustrated for the end point of clinical response in Supplementary Figure 4.

, 2005 and Olli and Trunov, 2010). This may be due to the fact

that the depositional behaviour of dinoflagellate cysts is like that of fine particles, and that their abundance increases in sediments with higher mud contents (Dale 1983). The present study also showed that most dinoflagellate cysts identified in Saudi sediments germinated successfully, with germination rates varying significantly among cyst types at different temperatures. This finding thus concurs with the conclusions drawn from previous studies that temperature is the major factor regulating the germination of marine phytoflagellate cysts (Dale, 1983, Pfiester and Anderson, 1987, Ishikawa and Taniguchi, 1996 and Ishikawa and Taniguchi, 1997), and that cyst germination is stimulated in different organisms by different water temperatures (Meksumpun et al. 2005). find more Our results showed that an increase in temperature from 15 to 25°C lowered the germination rates of dinoflagellate (Alexandrium) cysts from Saudi sediments. These results are in agreement with those of Meksumpun et al. (2005), who reported that some dinoflagellate cysts (but not Alexandrium cysts) can germinate well at temperatures between 10 and 28°C. Also, Ishikawa & Taniguchi (1996) found that Scrippsiella cysts can germinate

at temperatures between 5 and 25°C. Therefore, the increase in temperature may act to prevent MS-275 ic50 seeding or the maintenance of blooms in the water column during summer periods ( Genovesi et al. 2007). Unlike other cyst types, the germination of Alexandrium cysts was not affected by the difference in temperatures, with maximum germination rates reaching as high as 95.6%. Perez et al. (1998) reported that temperature had no significant effect on the germination of Alexandrium cysts collected from the St. Lawrence Estuary, Canada. The germination rate of Alexandrium cysts from Saudi

sediments Janus kinase (JAK) is higher than that obtained (48–52%) by Bravo et al. (2006), but is comparable with that reported by Garcés et al. (2004) (up to 91%). Such a remarkable difference in the germination rates of Alexandrium cysts between the two studies may be explained by the presence of some distinctive internal features, such as globular content, or other, genetic or external, factors ( Bravo et al. 2006). Germination success can also be affected by excystment medium conditions, where higher rates of germination were found for A. catenella cysts isolated in seawater than in L1 medium ( Figueroa et al. 2005). Overall, such information on the germination of dinoflagellate cysts may be helpful for understanding the mechanism of the outbreak of dinoflagellate red tides along Saudi coasts, as cyst bank germinations contribute to the initial seeding of blooms ( Genovesi et al. 2007). Our study also highlighted the presence of harmful marine dinoflagellate cysts in Saudi marine sediments.

It is now generally agreed that NO has a highly context-dependent dose–response stimulation-inhibition relationship with cytotoxicity at high doses and mitogenicity at low doses [22]. Thus, NO has the ability to both

promote and suppress cancer. However, these binary either/or descriptions are an oversimplification. At low constitutive levels induced by hypoxia in tumors, NO levels are optimal for the mediation of aberrant, proliferative signaling. In contrast, levels either above or below this optimal range can have the opposite effect and activate signal transduction pathways that contribute to/result in growth inhibition or cell death. NO is a radical with a free electron capable click here of interacting with reactive oxygen species (ROS) such as the superoxide anion to form a variety of highly reactive

nitrogen oxides (NOx). The term nitrosative stress refers to the formation of NOx compounds such as peroxynitrite (ONOO−), nitrogen dioxide (NO2), and dinitrogen trioxide (N2O3) that are responsible for cytotoxic nitration and oxidation reactions  [23] leading to apoptosis and cell death. In particular, the formation of peroxynitrite is a first-order reaction  [23] dependent on the concentrations of NO and the superoxide anion and, therefore, on oxygen tension, because in the presence of hypoxia, both NO and ROS such as the superoxide anion will be less prevalent. Xie et al [24] demonstrated that transfection of murine K-1735 melanoma cells with inducible NOS leading to the generation of high levels of NO resulted in suppression

of tumorigenicity and metastasis. The cytotoxicity of CYC202 cell line higher concentrations of NO is consistent with the assumption that the toxic effect becomes apparent above a threshold dose of NOx. This balance between mitogenic and toxic effects of NO in tumor cells is potentially attributable to an increased susceptibility to free radical damage due to severe impairment of the antioxidant defense system [25] compared with healthy cells. In cancer cells, reactive oxygen/nitrogen species “reprogram” the cellular metabolism toward a dependence on glucose use, termed the Warburg effect, a signature of virtually all tumors and the basis of fluorodeoxyglucose positron emission tomography imaging, to support anabolic proliferation. The fact that this core feature of tumors, metabolic reprogramming, Rho is dependent on redox signaling implies that ROS/reactive nitrogen species (RNS) levels are higher in tumors than in healthy tissue, resulting in a differential sensitivity to oxidant stress [26]. Indeed, the presence of high levels of ROS in tumors has been linked with cell cycle arrest and apoptosis [27]. However, NOx cytotoxicity may not require superelevated doses but rather approximate “normalization” to physiological levels [27], because shifts in a particular direction can have important consequences. For example, Frederiksen et al.

The effect of these SNPs on HbF levels

have been investigated mainly in patients with predominantly African or European ancestry. This study aimed to validate SNPs commonly studied (HBG2, rs748214; BCL11A, rs4671393; and HBS1L-MYB, rs28384513, rs489544 and rs9399137) and to analyze the effect of genetic admixture on the distribution of these SNPs in a sample of SCA patients from Belém, capital of Pará State, Brazilian Amazon. The sickle cell mutation GDC-0199 cell line is absent among Native American populations and was introduced into the American continent by gene flow from Africa during the Atlantic slave trade from the 16th to the 19th century. Africans mixed with Native Americans and Europeans to various extents across the continent, so that SCA patients exhibit different levels of admixture mainly European and Native American origin, as observed in the general population. In Brazil, although the populations of all geographic regions are the result of interbreeding between Europeans, Africans and Native Americans, there are slight differences in admixture proportions. European ancestry is the most prevalent GPCR & G Protein inhibitor in all urban populations, but is higher in the southeast and south, while in the Northeast, Midwest and Southeast, the African ancestry in general is the second most

prevalent. The Native American ancestry is higher in the North and in general more prevalent than the African contribution [5]. Thus, if genetic modifiers are associated with genetic ancestry then the level of mixing in SCA patients has obvious implications on

the distribution of SNPs, and therefore on the levels of HbF and clinical manifestations. Blood samples from Rucaparib SCA patients attended at the Center for Hemotherapy and Hematology of Pará Foundation — HEMOPA, in Belém, capital of Pará state, Northern Brazil, were selected for this study. HEMOPA is the reference center for diagnosis and treatment of hemoglobinopathies in the region. The frequency of the HBB*S gene in this population is estimated at 0.016 and the expected number of SCA patients in this population (384) is in accordance with the number of patients registered at HEMOPA, approximately 400 patients, at the time the samples were selected. Of the 240 patients initially selected those younger than 5 years and those under treatment with hydroxyurea™ were excluded resulting in a sample of 167 patients (47% of registered patients). Of the 167 study subjects, 89 (53.2%) were female. The mean (SD) age was 18.0 (10.6) years and the median age was 15.0 (IQR 10.0–24.0) years. HbF was measured by high performance liquid chromatography (HPLC) using equipment D10-Hemoglobin A1C Testing System (Bio Rad ®, France). The mean HbF level of the participants was 7.6% (SD 5.2) and the median was 6.6% (IQR 3.6–9.8%).

In conclusion our data indicate that – during medium-term follow-up (3 years) and using self-reported clinical risk factors – more complex tools as FRAX® did not perform better in the fracture risk prediction compared with simpler tools such as OST, ORAI, OSIRIS and SCORE or even age alone in a screening scenario where BMD was not measured. These findings suggest that simpler tools based on fewer risk factors, which would be easier to use in clinical practice

by the GP or the patient herself, could just as well as FRAX® be used to identify women with increased risk of fracture and therefore should be referred to a DXA scan. This study is a part of the Risk-stratified Osteoporosis Strategy Evaluation study (ROSE study) which is supported by INTERREG 4A (JNR 08/4177), the Region of Southern Denmark (JNR 08/8133 and JNR 11/5761) and Odense University Hospital. The funding agencies had no direct role in the conduct of the study; collection, management, analysis Protein Tyrosine Kinase inhibitor and interpretation of the data; and preparation, review or final approval of the manuscript. Conflicts of interest None. “
“Parathyroid hormone (PTH) is the major regulator of calcium homeostasis through its actions on bone and kidney. PTH is critical for bone remodeling, exerting both anabolic and catabolic effects on bone in vivo by activating the PTH1 receptor, BYL719 a G-protein coupled receptor, on osteoblast (OB) lineage cells [1] and [2]. Intermittent PTH was the first anabolic agent approved

for osteoporosis therapy Avelestat (AZD9668) in the USA [1] and [3]. For reasons still not completely understood, daily injections of PTH increase bone formation more than resorption, thereby increasing bone mass, while continuous infusion increases bone resorption more than formation, resulting in bone loss [4], [5] and [6]. Despite the anabolic effects of PTH in vivo and the demonstration that PTH can stimulate OB precursors or mesenchymal stem cells (MSCs) to differentiate into OBs [2] and [7], it has been difficult to demonstrate

osteogenic effects of PTH in vitro. A number of in vitro studies have reported that PTH present continuously in culture inhibits OB differentiation [8], [9], [10] and [11]. These observations suggest that the bone loss associated with continuous PTH is not simply the result of increased resorption but may also involve suppressed differentiation of bone-forming cells. PTH is also a potent inducer of cyclooxygenase-2 (COX-2) and prostaglandin (PG) production, especially PGE2, in OB lineage cells [12] and [13]. PGs are locally produced lipids that have receptors on both OB and osteoclast (OC) lineage cells [14] and [15]. PGE2 is abundantly expressed in bone and can have important roles in skeletal metabolism. Although originally identified as a resorption agonist, PGE2 also increases bone formation in vivo [16] and OB differentiation in vitro [14] and [15]. Multiple regulators of bone metabolism induce COX-2, the major enzyme responsible for PG production.

3 and Fig. 4. A similar plot for the equimolar 24-hour BChE is presented in Fig. 6. The fact that ChE activity in the blood did not correlate well with lethality can be seen by contrasting Fig. 5 and Fig. 6 with Fig. 3. There was a clear division of oxime efficacy in animals challenged with tabun (GA), cyclosarin (GF), and phorate oxon (PHO). Against GA and GF, MMB4-DMS, HLö-7 DMS, and HI-6 DMS showed statistically significant protection Selleckchem RG7422 against lethality (lethality ≤ 38%) relative to control animals, while obidoxime Cl2 was statistically significant against lethality (25% lethality) only for GA. MMB4-DMS, HLö-7 DMS, 2-PAM Cl, and obidoxime Cl2 showed statistically significant protection

against lethality (lethality ≤ 38%) against both phorate oxon and CPO-challenged animals. All other oximes (TMB-4, RS194B, and MINA), when used as therapy against GA, GF, CPO, or phorate oxon, demonstrated low efficacy with generally ≥ 50% lethality. GB and VX had much higher ChE reactivation rates and no more than 50% lethality in all oxime-treated groups. BKM120 cost For oxime therapy against GB, MMB4-DMS, HLö-7 DMS, HI-6 DMS, 2-PAM Cl, and RS194B all exhibited statistically significant improvement in lethality relative to the controls. In pesticide oxon-challenged animals, MMB4-DMS, HLö-7 DMS, and obidoxime Cl2 were

each significantly efficacious relative to lethality in control animals. Obidoxime Cl2 demonstrated the best overall protection for pesticide oxons with significant ChE reactivation, improvement in QOL scores, and survival through the 24 hour period. Obidoxime Cl2 has been shown to be one of the most efficacious reactivators against OP pesticides by other laboratories (Worek Edoxaban et al., 2007), and, in the present study, the oxime performed well against the nerve agents GA, GB, and VX as well. MMB4 DMS and

HLö-7 DMS were the two most consistently efficacious oximes across all challenge OPs. MMB4 DMS treatment resulted in an average 80% survivability while HLö-7 DMS treatment resulted in an average of 77%. The 24 hour average QOL score was ≤ 3.0 for MMB4 DMS and ≤ 5.4 for HLö-7 DMS on a 0 to 12 scale, excluding GD data. Additionally, reactivation of both AChE and BChE was more than 50% with MMB4 DMS for all OP challenges with the exception of chlorpyrifos oxon and GD. Peripheral blood ChE inhibition by GF and VX was significantly mitigated by MMB4 DMS, with total cholinesterase reactivation at 88 to 100%. Reactivation of AChE and BChE among survivors with HLö-7 DMS was not as significant when compared to that of MMB4 DMS; however, both enzymes were reactivated above 30% for all OPs with the exception of chlorpyrifos oxon and GD. Oxime efficacy is an amalgamation of somewhat unrelated physicochemical and pharmacologic factors. A favored, effective oxime has a relatively high therapeutic/safety index, quickly biodistributes to organs targeted by OPs (Voicu et al.

In a mountainous region like the Hornsund area, mountains additionally limit the horizontal path of photons, especially when the cloud base is below the mountain

peaks. This attenuates the irradiance transmittance, both the increase over the fjord waters and the decrease over the land, which is shown in Figure 7 for the cases of h = 200 m and h = 1800 m (τ = 12, spring albedo pattern, ϑ = 53° and λ = 469 nm). For h = 200 m, the irradiance transmittance over the fjord nearly reaches its ‘oceanic’ value within 2 km from a straight TSA HDAC concentration shore, while for h = 1800 m the ocean value is never reached over the ca 10-km-wide fjord. The transmittance enhancement over the near-shore plots ( Figure 8a) is 1.5–3 times lower for h = 200 m than it is for h = 1800 m. ΔTE drops 7 times with diminishing cloud layer height in plot 11 (the fjord mouth), and 3 times over the whole fjord. The radiative conditions are more local for lower clouds, and dark water diminishes irradiance

transmittance at the coast. Hence, irradiance transmittance at the station drops with increasing cloud base height. The transmittance enhancement over the fjord due to 3D effects (photon transport) weakens in the infrared. It is practically negligible for λ = 1640 nm (Figure 8b), the absolute value of ΔTE is lower than 0.005 for all the plots. In this spectral channel the surface albedo is almost uniform and very low (< 0.11). Because the 3D effects depend strongly on wavelength, they must modify the irradiance spectrum on the fjord surface. The behaviour of the ratio TE (λ = 469 nm)/TE (λ = 858 nm) with increasing τ CP-868596 in vitro is presented in Figure 9. The differences in the ratio between the fjord and the ocean are the highest for inner fjords (plots 5 and 8) and they range from 0.08 for a cloudless sky to 0.66 for clouds of τ = 30 (h = 1 km, spring albedo pattern, ϑ = 53° and Palmatine λ = 469 nm). The respective ratio differences for the whole fjord are 0.05 and

0.29. The variability of TE (λ = 469 nm)/TE (λ = 858 nm) over the fjord are caused mainly by a decrease in snow albedo with the wavelength between λ = 468 nm and 858 nm. All the runs/simulations discussed so far represent radiative transfer through water clouds. So as to simulate 3D effects under ice clouds, the asymmetry factor g was changed from 0.865 used for water cloud simulations with λ = 469 nm to 0.75 (e.g. Zhang et al., 2002, Baran et al., 2005 and Fu, 2007). An ‘ice cloud’ run was performed for the spring albedo pattern, τ = 12, ϑ = 53°, h = 1 km and λ = 469 nm (not shown in the figure). It was found that for ice clouds the 3D effect is stronger than for water clouds of the same height and optical thickness. Lowering factor g increases cloud albedo and decreases its transmittance. Thus it reduces TE but increases ΔTrelE from 19% for g = 0.865 to 25% for g = 0.75 for the whole fjord, and from 40% to 55% for the inner fjords (plots 5 and 8).

2), as well as a significant increase in RANKL immunolabelling (Fig. 1). OVX/E2 group started to show a decreasing in OPG immunolabelling for osteoblasts and osteocytes.

OVX/O group presented expressive labelling against RANKL. Raloxifene administration caused a reduction in RANKL immunolabelling at 28 days and absence immunolabelling at 42 days. TRAP immunolabelling was kept intense to moderate for OVX/O and OVX/E2 groups respectively and reduced for the other groups, primarily for the OVX/RLX group (Fig. 3) (Table 1). Oestrogen deficiency systemically affects bone remodelling through OPG/RANKL signalisation during the Raf inhibitor events that modulates osteoclasts cellular differentiation and lymphocytes development. In the experiments realized in our laboratory, the osteoprotective effect of oestrogen in inhibits bone resorption is confirmed after Veliparib ic50 treating OVX rats with 17β-estradiol. Which increased bone mass in the middle third of the alveolar bone, however the action of raloxifene was not as pronounced as E2.11 and 12 The intense immunolabelling for RANKL

and TRAP observed in OVX animals showed the signalling action of the members of the tumour necrosis factor (RANKL) on osteoclastic responses (TRAP). The oestrogen deficiency following ovariectomy leds to a high bone turnover during the alveolar healing process after tooth extraction whilst, oestrogen and raloxifene treatments led to bone formation. However TRAP expression at 28 and 42 days post-extraction in OVX animals treated with raloxifene was very low, whilst this expression was more expressive in OVX animals treated with oestrogen. Our results suggest that raloxifene treatment may compensate the changes induced by ovariectomy reducing the number of pre-osteoclasts and mature osteoclasts. Studies have shown that oestrogen deficiency leads to an increase of osteocytes apoptosis in human beings13 check and in female rats14 and the osteocytes apoptosis can be reverted through oestrogen replacement therapy14 and 15 or through raloxifene therapy.16 Studies have suggested

an autocrine mechanism, through a Fas ligand (FasL), in which oestrogen-induced osteoclast apoptosis17 and a paracrine mechanism in which oestrogen affects osteoclast survival through FasL upregulation in osteoblast cells leading to pre-osteoclasts apoptosis,18 this may explain the osteoprotective function of oestrogen as well as of SERMs. However, Kawamoto et al.19 evaluated the effects of oestrogen deficiency state in osteoclastogenesis of the periodontal tissue at 7 postoperative days and did not find any difference in the number of osteoclasts between oestrogen replacement therapy and sham groups. The authors also observed a significant increase of TRAP expression at 14 postoperative days on OVX group compared to the others, these finding are in agreement to our findings.

The genotypic and phenotypic data were collected from 60 tobacco leaf samples of three cultivars, from four development stages and three positions in the plants and grown in two cultivation environments. The potential application of the QTX results in breeding practice was also discussed. In 2010, three tobacco cultivars (K326, Hongda and Zunyan 6) were grown in farm fields at Guiding (26.58° N, 107.23°

E) and Xingyi (25.08° N, 104.90° E) under normal condition for crop production in Guizhou province, China. The plants of three cultivars were planted in 10 rows with 30 plants per row and in three blocks. The plant-to-plant spaces between and within rows were 100 cm and 50 cm, respectively. For each of PTC124 ic50 the three cultivars, leaves of 25 plants from 5 points were pooled for 1) the combination of upper or middle leaves from two locations within the plant at four developmental time points with sampling

every 12 days, and 2) the lower leaves from two locations within the plant at two developmental time points, thus resulting in a total of 60 samples. The pooled leaves were immediately frozen in liquid nitrogen and stored at − 80 °C for further use. A methylation DArT chip for tobacco was developed by Diversity Array Technology Ltd. (Canberra, Australia) as PARP inhibition described at http://www.diversityarrays.com/dnamethylation.html. Total DNA was extracted and hybridization followed the DArT methylation profiling protocol as described by Lu et al. [23]. The program

Montelukast Sodium DArT Soft was used to determine whether the fragments in the arrays tested for each sample were methylated or not. A custom-designed microarray platform was used for the analysis of total RNA extracted from the tobacco leaf samples. The microarray was comprised of three 60-mer probes for each of 44,873 unigenes derived from public Expressed Sequence Tags (ESTs) of tobacco and was made following a protocol provided by Roche Co. (http://www.nimblegen.com/). The 60-mer probes were chosen from a group of six to seven non-overlapping probes designed against different parts of each gene model. The probes with E-values most similar to the average of the six to seven non-overlapping experimental probes were assumed to be the most reliable for transcript level estimation. Total RNA was extracted with the RNeasy Mini Kit (Qiagen Corp, Valencia, CA, United States) and DNase treated in-column with the RNase-Free DNase set (also from Qiagen). Double-stranded cDNA was synthesized using the SuperScript Double Strand cDNA Synthesis Kit (Invitrogen Inc., Carlsbad, CA, United States) with oligo (dT) primers following the manufacturer’s protocol. Cy-3 and Cy-5 labeling and hybridization steps were performed by NimbleGen using standard procedures (http://www.nimblegen.com/). Expression values were generated by Roche NimbleGen proprietary software using quantile normalization [24] and the Robust Multichip Average algorithm [25].