We have recently identified miR-125b upregulation in glioblastoma (GMB). The aim of this study is to determine the correlation between miR-125b expression and malignant grades of glioma and the genes targeted by miR-125b.
Real-time PCR was employed to measure the expression level of miR-125b. Cell viability was evaluated by cell growth and colony formation in soft-agar assays. Cell apoptosis was determined by Hoechst 33342 staining and AnnexinV-FITC assay. The Luciferase assay was used to confirm the actual binding sites of p38MAPK mRNA. Western blot was used to detect the gene expression level.
The expression level of miR-125b is positively correlated with the malignant grade of glioma. Ectopic expression of miR-125b promotes the proliferation of GMB cells. Knockdown of endogenous miR-125b inhibits cell proliferation and promotes cell apoptosis. Further studies reveal that p53 is regulated by miR-125b. However, downregulation of the endogenous miR-125b also results in p53-independent apoptotic pathway leading to apoptosis in p53 mutated U251 cells and p53 knockdown U87 cells. Moreover, p38MAPK is also regulated by miR-125b and downregulation of miR-125b activates the p38MAPK-induced mitochondria apoptotic pathway.
High-level expression of miR-125b is associated with poor outcomes of GMB. MiR-125b may have an oncogenic role in GMB cells by promoting cell proliferation and inhibiting apoptosis.
microRNA; miR-125b; glioblastoma; cell apoptosis; p53; p38MAPK
This phase I study assessed the safety, tolerability, maximum tolerated dose (MTD), pharmacokinetics, and preliminary antitumor effects of sunitinib combined with modified FOLFOX6 (mFOLFOX6).
Patients with advanced solid malignancies received mFOLFOX6 in 2-week cycles with escalating sunitinib doses (25, 37.5, and 50 mg/day) on three schedules: 2 weeks on, 2 weeks off (2/2); 4 weeks on, 2 weeks off (4/2); or continuous daily dosing (CDD). Patients received up to 8 treatment cycles (Schedule 2/2 and CDD schedule) or 6 cycles (Schedule 4/2). An expansion cohort enrolled patients with metastatic colorectal cancer at the Schedule 2/2 MTD.
Overall, 53 patients were enrolled, with 43 evaluable for dose-limiting toxicity (DLT). On Schedule 2/2 (n = 18), DLTs occurred in three patients at 50 mg/day (grade 4 neutropenia [n = 1]; grades 3 and 4 thrombocytopenia [n = 2]) and two patients achieved partial responses (PRs). On Schedule 4/2 (n = 13), 37.5 mg/day exceeded the MTD with two DLTs (febrile neutropenia and grade 4 hypokalemia, respectively). On the CDD schedule (n = 12), the MTD was 25 mg/day; one DLT (grade 3 stomatitis) was reported and two patients achieved PRs. The most common adverse events were neutropenia, fatigue, and thrombocytopenia. No clinically significant drug–drug interactions were apparent between sunitinib, its metabolite SU12662, and mFOLFOX6.
Sunitinib combined with mFOLFOX6 had acceptable tolerability. The MTDs were sunitinib 50 mg/day on Schedule 2/2 and 25 mg/day on the CDD schedule. A MTD for Schedule 4/2 was not established.
Sunitinib; Tyrosine kinase inhibitor; FOLFOX; Solid tumors; Pharmacokinetics; Phase I
Ceramidases are a group of enzymes that catalyze hydrolysis of ceramides to generate fatty acid and sphingosine. In this study, we report the cloning and characterization of the rice small brown planthopper Laodelphax striatellus neutral ceramidase (nCDase), LsnCer. LsnCer was identified by sequencing the transcriptome of Laodelphax striatellus. LsnCer is a protein of 717 amino acids with a predicted molecular weight of 79.3 kDa. Similar to other known nCDases, LsnCer has a pH optimum at 8.0 and a temperature optimum at 37 °C for its in vitro activity. LsnCer activity is inhibited by Zn2+ significantly and Fe2+ slightly. LsnCer has broad substrate specificity with preference for ceramides with a medium acyl-chain or a mono unsaturated long acyl-chain. Infection with the rice strip virus (RSV) or treatment with insecticides significantly increased LsnCer mRNA expression and its enzymatic activity in L. striatellus. These results suggest that LsnCer is a bona fide nCDase that may have a role in adaption of L. striatellus to environmental stresses.
nCDase; LsnCer; activity; mRNA; rive strip virus; insecticide
Macrophages are critical immune effector cells of the tumor microenvironment that promote seeding, extravasation and persistent growth of tumor cells in primary tumors and metastatic sites. Tumor progression and metastasis are affected by dynamic changes in the specific phenotypes of macrophage subpopulations; however, the mechanisms by which tumor cells modulate macrophage polarization remain incompletely understood. Caspase recruitment domain-containing protein 9 (CARD9) is a central adaptor protein of innate immune responses to extracellular pathogens. We report that increased CARD9 expression is primarily localized in infiltrated macrophages and significantly associated with advanced histopathologic stage and the presence of metastasis. Using CARD9-deficient (CARD9−/−) mice, we show that bone marrow-derived CARD9 promotes liver metastasis of colon carcinoma cells. Mechanistic studies reveal that CARD9 contributes to tumor metastasis by promoting metastasis-associated macrophage polarization through activation of the nuclear factor-kappa B signaling pathway. We further demonstrate that tumor cell-secreted vascular endothelial growth factor facilitates spleen tyrosine kinase activation in macrophages, which is necessary for formation of the CARD9–B-cell lymphoma/leukemia 10–mucosa-associated lymphoid tissue lymphoma translocation protein 1 complex. Taken together, our results indicating that CARD9 is a regulator of metastasis-associated macrophages will lead to new insights into evolution of the microenvironments supporting tumor metastasis, thereby providing targets for anticancer therapies.
The physiological mechanisms involved in isoproterenol (ISO)-induced chronic heart
failure (CHF) are not fully understood. In this study, we investigated local changes
in cardiac aldosterone and its synthase in rats with ISO-induced CHF, and evaluated
the effects of treatment with recombinant human brain natriuretic peptide (rhBNP).
Sprague-Dawley rats were divided into 4 different groups. Fifty rats received
subcutaneous ISO injections to induce CHF and the control group (n=10) received equal
volumes of saline. After establishing the rat model, 9 CHF rats received no further
treatment, rats in the low-dose group (n=8) received 22.5 μg/kg rhBNP and those in
the high-dose group (n=8) received 45 μg/kg rhBNP daily for 1 month. Cardiac function
was assessed by echocardiographic and hemodynamic analysis. Collagen volume fraction
(CVF) was determined. Plasma and myocardial aldosterone concentrations were
determined using radioimmunoassay. Myocardial aldosterone synthase (CYP11B2) was
detected by quantitative real-time PCR. Cardiac function was significantly lower in
the CHF group than in the control group (P<0.01), whereas CVF, plasma and
myocardial aldosterone, and CYP11B2 transcription were significantly higher than in
the control group (P<0.05). Low and high doses of rhBNP significantly improved
hemodynamics (P<0.01) and cardiac function (P<0.05) and reduced CVF, plasma and
myocardial aldosterone, and CYP11B2 transcription (P<0.05). There were no
significant differences between the rhBNP dose groups (P>0.05). Elevated cardiac
aldosterone and upregulation of aldosterone synthase expression were detected in rats
with ISO-induced CHF. Administration of rhBNP improved hemodynamics and ventricular
remodeling and reduced myocardial fibrosis, possibly by downregulating CYP11B2
transcription and reducing myocardial aldosterone synthesis.
Chronic heart failure; Aldosterone; Aldosterone synthase; Recombinant human brain natriuretic peptide
Tissue engineering encapsulated cells such as chondrocytes in the carrier matrix have
been widely used to repair cartilage defects. However, chondrocyte phenotype is
easily lost when chondrocytes are expanded in vitro by a process
defined as “dedifferentiation”. To ensure successful therapy, an effective
pro-chondrogenic agent is necessary to overcome the obstacle of limited cell numbers
in the restoration process, and dedifferentiation is a prerequisite. Gallic acid (GA)
has been used in the treatment of arthritis, but its biocompatibility is inferior to
that of other compounds. In this study, we modified GA by incorporating
sulfamonomethoxine sodium and synthesized a sulfonamido-based gallate, JJYMD-C, and
evaluated its effect on chondrocyte metabolism. Our results showed that JJYMD-C could
effectively increase the levels of the collagen II, Sox9, and aggrecan genes, promote
chondrocyte growth, and enhance secretion and synthesis of cartilage extracellular
matrix. On the other hand, expression of the collagen I gene was effectively
down-regulated, demonstrating inhibition of chondrocyte dedifferentiation by JJYMD-C.
Hypertrophy, as a characteristic of chondrocyte ossification, was undetectable in the
JJYMD-C groups. We used JJYMD-C at doses of 0.125, 0.25, and 0.5 µg/mL, and the
strongest response was observed with 0.25 µg/mL. This study provides a basis for
further studies on a novel agent in the treatment of articular cartilage defects.
Sulfamonomethoxine sodium; Gallic acid; Pro-chondrogenic agent; Chondrocyte; Rabbit articular cartilage; Dedifferentiation
Prognostic factors for progression-free survival (PFS), overall survival (OS), and long-term OS (⩾30 months) were investigated in sunitinib-treated patients with metastatic renal cell carcinoma (RCC).
Data were pooled from 1059 patients in six trials. Baseline variables, including ethnicity, were analysed for prognostic significance by Cox proportional-hazards model.
Median PFS and OS were 9.7 and 23.4 months, respectively. Multivariate analysis of PFS and OS identified independent predictors, including ethnic origin, Eastern Cooperative Oncology Group performance status, time from diagnosis to treatment, prior cytokine use, haemoglobin, lactate dehydrogenase, corrected calcium, neutrophils, platelets, and bone metastases (OS only). Characteristics of long-term survivors (n=215, 20%) differed from those of non-long-term survivors; independent predictors of long-term OS included ethnic origin, bone metastases, and corrected calcium. There were no differences in PFS (10.5 vs 7.2 months; P=0.1006) or OS (23.8 vs 21.4 months; P=0.2135) in white vs Asian patients; however, there were significant differences in PFS (10.5 vs 5.7 months; P<0.001) and OS (23.8 vs 17.4 months; P=0.0319) in white vs non-white, non-Asian patients.
These analyses identified risk factors to survival with sunitinib, including potential ethnic-based differences, and validated risk factors previously reported in advanced RCC.
sunitinib; prognostic factors; renal cell carcinoma; metastatic; survival
To evaluate the feasibility and safety of a revised technique of botulinum toxin type A (BTA) injections for the treatment of infantile esotropia.
Forty-seven patients with infantile esotropia were randomly divided into two groups. In group A, 23 cases were treated with a bilateral injection of 2.5–3.75 U BTA combined with sodium hyaluronate (SH) to the medial rectus muscle. In group B, 24 cases were treated with a bilateral injection of 2.5–3.75 U BTA solution alone to the medial rectus muscle. Electromyography was not used in the study. All patients received one injection and were evaluated 2 weeks, 3 months, and 6 months following injection.
The measured changes between groups A and B included the frequencies of good alignment 6 months after injections (30.4% vs 37.5%), complicated ptosis (2.2% vs 20.8%), and vertical deviation (2.2% vs 2.1%).
BTA injections combined with or without SH in the absence of electromyography demonstrated effectiveness and feasibility in the treatment of infantile esotropia. A relative decrease in the frequency of complicated ptosis resulted from injections of BTA+SH.
botulinum toxin; infantile esotropia; sodium hyaluronate; complication
Soil degradation has been a very serious problem for sustainable production, especially by a re-cropping of greenhouse-cultivated cucumber (Cucumis statirus L.). The aim of this research was to expound the actuality for soil degradation, at the same time, put forward some suggestion for preventing from soil degradation and maintain sustainable production in greenhouse basic on the two experiments conducted in a solar greenhouse during 2001-2008 suburb area of Yan'an, Shaanxi province in North China. The result shown that cucumber fruit productivity increased as the increasing of re-cropping years, but decreased after 5years continuously cropping. As increasing of re-cropping years, the population of fungus and bacteria increased, which was assumingly main factor of soil degradation. There was significant difference in cropping models on soil bio-characteristics and system productivity. The productivity were the highest in cropping model between cucumber and greengrocery, cucumber and cowpea (Vigna sinensis L), the second higher were in cropping model between cucumber and maize (Zea mays) for green manure, cucumber and kidney bean (Phaseolus vulgaris). That was the best way to reduce soil bacteria and epiphyte amount to follow lasting three or four months during summer season after cucumber harvest, the better method was planting cowpea or other leguminous crops. Basic on the experiment, the optimums approaches to preventive soil degradation were put forward.
Soil degradation; greenhouse production; cropping model; soil bio-characteristics; preventing from soil degradation
Few data are available about intakes and food sources of trans-fatty acids (TFAs) or their associations with cardiometabolic outcomes in Asian people who consume a prudent diet but are experiencing rapid nutritional transitions. We aimed to investigate the relationships between TFA biomarkers and type 2 diabetes and cardiovascular risk factors in Chinese individuals.
Erythrocyte fatty acids were measured by gas chromatography among 3,107 men and women (50–70 years) recruited from urban and rural areas in Beijing and Shanghai, China.
Total trans-18:1 and two trans-18:2 isomers were detected and accounted for 0.37% of the total fatty acids in the erythrocytes. Concentrations of TFAs were higher in women than men, and in urban than rural residents. Of the TFAs, trans-18:1, but not trans-18:2, showed a modest association with dairy consumption (β=0.27), but not with other foods. After adjustment for BMI, social-demographic, lifestyle and dietary factors and other TFAs, erythrocyte trans-18:1 was shown to be associated with a lower risk of type 2 diabetes (OR comparing extreme quartiles=0.68, 95% CI=0.48, 0.97, ptrend=0.02), as well as 20–50% lower odds of central obesity, dyslipidaemia, hyperglycemia, insulin resistance and chronic inflammation. In contrast, trans-18:2 fatty acids were positively associated with high triacylglycerol (ptrend<0.001) and LDL-cholesterol (ptrend=0.03) levels, but not with diabetes and other cardiometabolic risk factors.
Among middle-aged and older Chinese individuals with overall low erythrocyte TFAs levels, trans-18:1 might serve as a marker of dairy intake. Higher trans-18:1 levels were associated with a lower risk of type 2 diabetes, whereas higher trans-18:2 levels were associated with dyslipidaemia.
Biomarkers; Diet; Dyslipidaemia; trans-Fatty acids; Type 2 diabetes
The aim of this article was to prospectively evaluate the accuracy and radiation dose of 320-detector row dynamic volume CT (DVCT) for the detection of coronary artery disease (CAD) in a high-risk population.
60 patients with a high risk of CAD underwent DVCT without preceding heart rate control and also underwent invasive coronary angiography (ICA), which served as the standard reference.
On a per segment analysis, overall sensitivity was 95.3%, specificity was 97.6%, positive predictive value was 90.6%, negative predictive value was 98.8% and Youden index was 0.93. In both heart rate subgroups, diagnostic accuracy for the assessment of coronary artery stenosis was similar. The accuracy of the subgroup with an Agatston score ≥100 was lower than that for patients with an Agatston score <100. However, the difference between DVCT and ICA results was not significant (p=0.08). The mean estimated effective dose of CT was 12.5±9.4 mSv. In those patients with heart rates less than 70 beats per minute (bpm), the mean radiation exposure of DVCT was 5.2±0.9 mSv. The effective radiation dose was significantly lower than that of ICA (14.1±5.9 mSv) (p<0.001). When the heart rate was >70 bpm, a significantly higher dose was delivered to patients with DVCT (22.6±5.2 mSv, p<0.001) than with ICA (15.0±5.3 mSv, p<0.001).
DVCT reliably provides high diagnostic accuracy without heart rate/rhythm control. However, from a dosimetric point of view, it is recommended that heart rate should be controlled to <70 bpm to decrease radiation dose.
Periodic blood exchange transfusion is a treatment modality commonly used to manage pediatric sickle cell anemia (SCA) at the University of California Davis Medical Center. The goal of exchange transfusion therapy is to ameliorate vasoocclusion and improve tissue perfusion by removing sickled red blood cells (RBCs) and introducing normal RBCs. Using computer-assisted intravital microscopy, pre- and post-transfusion microvascular characteristics were analyzed. In this study, the bulbar conjunctiva exhibited a “blanched” avascular appearance in all six pediatric SCA patients prior to transfusion, indicative of tissue hypoperfusion and ischemia. Immediately following transfusion, substantial improvement in vascularization and tissue perfusion resulted, reflected by the enhanced appearance of capillaries and arterioles. In addition, a decrease in red cell velocity was observed. These observations provide evidence that exchange transfusion therapy is beneficial in ameliorating vasoocclusion and improving tissue perfusion. However, with the paradoxical post-transfusion decrease in red cell velocity presumably due to induced hyperviscosity from the large transfusion volume, blood flow is still impaired. This decreased velocity may thwart efforts to improve oxygen delivery via transfusion and may, to some extent, promote vasoocclusion instead. This paradoxical result warrants further investigation on the effects of transfusion volume and viscosity in the exchange transfusion process.
Sickle cell anemia; microcirculation; exchange transfusion; vasoocclusion; tissue perfusion; intravital microscopy
The abundances, compositions, and activities of microbial communities were investigated at bog and fen sites in the Glacial Lake Agassiz Peatland of northwestern Minnesota. These sites contrast in the reactivity of dissolved organic matter (DOM) and the presence or absence of groundwater inputs. Microbial community composition was characterized using pyrosequencing and clone library construction of phylogenetic marker genes. Microbial distribution patterns were linked to pH, concentrations of dissolved organic carbon and nitrogen, C/N ratios, optical properties of DOM, and activities of laccase and peroxidase enzymes. Both bacterial and archaeal richness and rRNA gene abundance were >2 times higher on average in the fen than in the bog, in agreement with a higher pH, labile DOM content, and enhanced enzyme activities in the fen. Fungi were equivalent to an average of 1.4% of total prokaryotes in gene abundance assayed by quantitative PCR. Results revealed statistically distinct spatial patterns between bacterial and fungal communities. Fungal distribution did not covary with pH and DOM optical properties and was vertically stratified, with a prevalence of Ascomycota and Basidiomycota near the surface and much higher representation of Zygomycota in the subsurface. In contrast, bacterial community composition largely varied between environments, with the bog dominated by Acidobacteria (61% of total sequences), while the Firmicutes (52%) dominated in the fen. Acetoclastic Methanosarcinales showed a much higher relative abundance in the bog, in contrast to the dominance of diverse hydrogenotrophic methanogens in the fen. This is the first quantitative and compositional analysis of three microbial domains in peatlands and demonstrates that the microbial abundance, diversity, and activity parallel with the pronounced differences in environmental variables between bog and fen sites.
Most genetic variants identified for type 2 diabetes have been discovered in European populations. We performed genome-wide association studies (GWAS) in a Chinese population with the aim of identifying novel variants for type 2 diabetes in Asians.
We performed a meta-analysis of three GWAS comprising 684 patients with type 2 diabetes and 955 controls of Southern Han Chinese descent. We followed up the top signals in two independent Southern Han Chinese cohorts (totalling 10,383 cases and 6,974 controls), and performed in silico replication in multiple populations.
We identified CDKN2A/B and four novel type 2 diabetes association signals with p < 1 × 10−5 from the meta-analysis. Thirteen variants within these four loci were followed up in two independent Chinese cohorts, and rs10229583 at 7q32 was found to be associated with type 2 diabetes in a combined analysis of 11,067 cases and 7,929 controls (pmeta = 2.6 × 10−8; OR [95% CI] 1.18 [1.11, 1.25]). In silico replication revealed consistent associations across multiethnic groups, including five East Asian populations (pmeta = 2.3 × 10−10) and a population of European descent (p = 8.6 × 10−3). The rs10229583 risk variant was associated with elevated fasting plasma glucose, impaired beta cell function in controls, and an earlier age at diagnosis for the cases. The novel variant lies within an islet-selective cluster of open regulatory elements. There was significant heterogeneity of effect between Han Chinese and individuals of European descent, Malaysians and Indians.
Our study identifies rs10229583 near PAX4 as a novel locus for type 2 diabetes in Chinese and other populations and provides new insights into the pathogenesis of type 2 diabetes.
Electronic supplementary material
The online version of this article (doi:10.1007/s00125-013-2874-4) contains peer-reviewed but unedited supplementary material, which is available to authorised users.
Chinese; Diabetes; East Asians; Genetics; Genome-wide association study
The objective of this study was to examine the effect of the off axis retinoscopy on objective central refractive measurement in adult clinical practice.
In all, 40 subjects underwent undilated retinoscopy in a randomly selected eye both on and off the visual axis by a single masked examiner. Off axis retinoscopy was defined as retinoscopy performed with the testing eye of the examiner aligned with the contralateral (non-test) eye of the subject resulting in an off axis deviation in the nasal horizontal visual field. Retinoscopy was performed in negative cylinder only and spherocylindrical measurements were converted to power vectors for analysis. Paired t-test was used to assess differences in M, J0 and J45 power vectors including differences between mean aided and unaided LogMar acuities.
In all, 14 subjects were myopic (SE≤−0.5 D), 13 subjects were emmetropic (SE between −0.49 and 1.0 D) and 13 subjects were hyperopic (SE >1.0 D). Mean angle of deviation was 5.58° in the nasal horizontal visual field. J0 showed a significant negative shift in those with myopia (P<0.001) and emmetropia (P=0.049) following off axis retinoscopy. No significant differences in M, J0 and J45 were found in the hyperopes. Mean aided LogMar acuities after on and off axis retinoscopy were both significantly better than mean unaided LogMar VA (P<0.001 in both cases).
Small degrees of off axis retinoscopy encountered in everyday clinical practice can induce errors in objective central refractive measurement.
off axis; retinoscopy; refractive; error; power; vectors
Ischaemic heart failure remains a significant health and economic problem worldwide. This paper presents a user-friendly software system that will form a part of the virtual pathological heart of the Virtual Physiological Human (VPH2) project, currently being developed under the European Commission Virtual Physiological Human (VPH) programme. VPH2 is an integrated medicine project, which will create a suite of modelling, simulation and visualization tools for patient-specific prediction and planning in cases of post-ischaemic left ventricular dysfunction. The work presented here describes a three-dimensional interactive visualization for simulating left ventricle restoration surgery, comprising the operations of cutting, stitching and patching, and for simulating the elastic deformation of the ventricle to its post-operative shape. This will supply the quantitative measurements required for the post-operative prediction tools being developed in parallel in the same project.
heart; heart failure; surgical planning; virtual surgery; visualization; computer graphics
To determine the supporting role of a novel foldable capsular vitreous body (FCVB) with magnetic resonance imaging (MRI) in the treatment of severe retinal detachment in human eyes.
The study examined nine eyes of nine patients. Among the nine eyes, five had suffered penetrating injuries while four had suffered contusions of the eyeball involving large defects of the retina or choroids. A standard three-port pars plana vitrectomy was performed, FCVB was triple-folded and sent into the vitreous cavity; balanced salt solution (BSS) was injected into the capsule to support the retina. Three cardinal axes of nine eyes were examined using MRI at baseline and at the 3-month follow up.
MRI revealed that the signal intensity of the FCVB was similar to the normal vitreous body, with low-signal intensity on T1-weighted image and high-signal intensity on T2-weighted image. In three pre-operative silicone oil- or heavy silicone oil-filled eyes, FCVBs were not fully inflated, and eyeball deformation was observed in one eye. Shifts of three cardinal axes of three eyes (horizontal, anteroposterior, and vertical) according to MRI, were −4.33, −4.67, and −2.67 mm. In the remaining six eyes, FCVBs were well distributed in the vitreous cavity and evenly supported the retina; the cardinal axes of the eyes were similar to pre-operation. Shifts of three cardinal axes of six eyes were −0.34, −0.34, and −0.34 mm. In a total of nine eyes, shifts of three cardinal axes were −1.67, −1.77, and −1.11 mm. Statistically significant difference showed only between the horizontal axis of nine eyes pre-operatively and post-operatively (P1=0.041, P2=0.058, P3=0.123).
This study demonstrated the effectiveness of MRI to monitor the supporting role of an FCVB in the treatment of severe retinal detachment in human eyes.
FCVB; human eyes; magnetic resonance imaging
The Wnt signaling pathway is capable of self-regulation through positive and negative feedback mechanisms. For example, the oncoprotein c-Myc, which is upregulated by Wnt signaling activity, participates in a positive feedback loop of canonical Wnt signaling through repression of Wnt antagonists DKK1 and SFRP1. In this study, we investigated the mechanism of Wnt inhibitory factor-1 (WIF-1) silencing. Mapping of CpG island methylation of the WIF-1 promoter reveals regional methylation (–295 to –95 bp from the transcription start site) that correlates with transcriptional silencing. We identified Miz-1 as a direct activator of WIF-1 transcriptional activity, which is found at WIF-1 promoter. In addition, we show that c-Myc contributes to WIF-1 transcriptional repression in a Miz-1-dependent manner. Although the transient repression mediated by Miz-1/c-Myc is independent of de novo methylation, the stable repression by this complex is associated with CpG island methylation of the critical –295 to –95-bp region of the WIF-1 promoter. Importantly, Miz-1 and c-Myc are found at WIF-1 promoter in WIF-1 non-expressing cell lines DLD-1 and 209myc. Transient knockdown or somatic knockout of c-Myc in DLD-1 failed to restore WIF-1 expression suggesting that c-Myc is involved in initiating rather than maintaining WIF-1 epigenetic silencing. In a genome-wide screen, DNAJA4, TGFβ-induced and TRIM59 were repressed by c-Myc overexpression and DNA promoter hypermethylation. Our data reveal novel insights into c-Myc-mediated DNA methylation-dependent transcriptional silencing, a mechanism that might contribute to the dysregulation of Wnt signaling in cancer.
WIF-1; Wnt signaling; c-Myc; Miz-1; DNA methylation
FTO harbours the strongest known obesity-susceptibility locus in Europeans. While there is growing evidence for a role for FTO in obesity risk in Asians, its association with type 2 diabetes, independently of BMI, remains inconsistent. To test whether there is an association of the FTO locus with obesity and type 2 diabetes, we conducted a meta-analysis of 32 populations including 96,551 East and South Asians.
All studies published on the association between FTO-rs9939609 (or proxy [r2 > 0.98]) and BMI, obesity or type 2 diabetes in East or South Asians were invited. Each study group analysed their data according to a standardised analysis plan. Association with type 2 diabetes was also adjusted for BMI. Random-effects meta-analyses were performed to pool all effect sizes.
The FTO-rs9939609 minor allele increased risk of obesity by 1.25-fold/allele (p = 9.0 × 10−19), overweight by 1.13-fold/allele (p = 1.0 × 10−11) and type 2 diabetes by 1.15-fold/allele (p = 5.5 × 10−8). The association with type 2 diabetes was attenuated after adjustment for BMI (OR 1.10-fold/allele, p = 6.6 × 10−5). The FTO-rs9939609 minor allele increased BMI by 0.26 kg/m2 per allele (p = 2.8 × 10−17), WHR by 0.003/allele (p = 1.2 × 10−6), and body fat percentage by 0.31%/allele (p = 0.0005). Associations were similar using dominant models. While the minor allele is less common in East Asians (12–20%) than South Asians (30–33%), the effect of FTO variation on obesity-related traits and type 2 diabetes was similar in the two populations.
FTO is associated with increased risk of obesity and type 2 diabetes, with effect sizes similar in East and South Asians and similar to those observed in Europeans. Furthermore, FTO is also associated with type 2 diabetes independently of BMI.
Electronic supplementary material
The online version of this article (doi:10.1007/s00125-011-2370-7) contains peer-reviewed but unedited supplementary material, which is available to authorised users.
Asians; FTO; Meta-analysis; Obesity; Type 2 diabetes
Polycythemia vera (PV), essential thrombocythemia and primary myelofibrosis, are myeloproliferative neoplasms (MPNs) with distinct clinical features and are associated with the JAK2V617F mutation. To identify genomic anomalies involved in the pathogenesis of these disorders, we profiled 87 MPN patients using Affymetrix 250K single-nucleotide polymorphism (SNP) arrays. Aberrations affecting chr9 were the most frequently observed and included 9pLOH (n=16), trisomy 9 (n=6) and amplifications of 9p13.3–23.3 (n=1), 9q33.1–34.13 (n=1) and 9q34.13 (n=6). Patients with trisomy 9 were associated with elevated JAK2V617F mutant allele burden, suggesting that gain of chr9 represents an alternative mechanism for increasing JAK2V617F dosage. Gene expression profiling of patients with and without chr9 abnormalities (+9, 9pLOH), identified genes potentially involved in disease pathogenesis including JAK2, STAT5B and MAPK14. We also observed recurrent gains of 1p36.31–36.33 (n=6), 17q21.2–q21.31 (n=5) and 17q25.1–25.3 (n=5) and deletions affecting 18p11.31–11.32 (n=8). Combined SNP and gene expression analysis identified aberrations affecting components of a non-canonical PRC2 complex (EZH1, SUZ12 and JARID2) and genes comprising a ‘HSC signature' (MLLT3, SMARCA2 and PBX1). We show that NFIB, which is amplified in 7/87 MPN patients and upregulated in PV CD34+ cells, protects cells from apoptosis induced by cytokine withdrawal.
myeloproliferative neoplasms; JAK2V617F; NFIB; SNP
Photovoltaic functions in organic materials are intimately connected to interfacial morphologies of molecular packing in films on the nanometer scale and molecular levels. This review will focus on current studies on correlations of nanoscale morphologies in organic photovoltaic (OPV) materials with fundamental processes relevant to photovoltaic functions, such as light harvesting, exciton splitting, exciton diffusion, and charge separation (CS) and diffusion. Small molecule photovoltaic materials will be discussed here. The donor and acceptor materials in small molecule OPV devices can be fabricated in vacuum-deposited, multilayer, crystalline thin films, or spin-coated together to form blended bulk heterojunction (BHJ) films. These two methods result in very different morphologies of the solar cell active layers. There is still a formidable debate regarding which morphology is favored for OPV optimization. The morphology of the conducting films has been systematically altered; using variations of the techniques above, the whole spectrum of film qualities can be fabricated. It is possible to form a highly crystalline material, one which is completely amorphous, or an intermediate morphology. In this review, we will summarize the past key findings that have driven organic solar cell research and the current state-of-the-art of small molecule and conducting oligomer materials. We will also discuss the merits and drawbacks of these devices. Finally, we will highlight some works that directly compare the spectra and morphology of systematically elongated oligothiophene derivatives and compare these oligomers to their polymer counterparts. We hope this review will shed some new light on the morphology differences of these two systems.
organic photovoltaics; perylene; thiophene; charge transport; transient absorption; grazing incidence x-ray scattering
We investigated receptor activator of nuclear factor-κB ligand (RANKL) expression by B lymphocytes during early and late aspects of the immune response to Aggregatibacter actinomycetemcomitans, a gram-negative, anaerobic bacterium associated with aggressive periodontal disease.
Expression of messenger RNA transcripts (tumor necrosis factor-α, Toll-like receptors 4 and 9, interleukins 4 and 10, and RANKL) involved in early (1-day) and late (10-day) responses in cultured rat splenocytes was examined by reverse transcription–polymerase chain reaction (RT-PCR). The immune cell distribution (T, B, and natural killer cells and macrophages) in cultured rat splenocytes and RANKL expression in B cells were determined by flow cytometric analyses. B-cell capacity for induction of osteoclast differentiation was evaluated by coculture with RAW 264.7 cells followed by a tartrate-resistant acid phosphatase (TRAP) activity assay.
The expression levels of interleukins 4 and 10 in cultured cells were not changed in the presence of A. actinomycetemcomitans until cultured for 3 days, and peaked after 7 days. After culture for 10 days, the percentages of B and T cells, the overall RANKL messenger RNA transcripts, and the percentage of RANKL-expressing immunoglobulin G-positive cells were significantly increased in the presence of A. actinomycetemcomitans. These increases were considerably greater in cells isolated from A. actinomycetemcomitans-immunized animals than from non-immunized animals. RAW 264.7 cells demonstrated significantly increased TRAP activity when cocultured with B cells from A. actinomycetemcomitans-immunized animals. The addition of human osteoprotegerin-Fc to the culture significantly diminished such increases.
This study suggests that B-lymphocyte involvement in the immune response to A. actinomycetemcomitans through upregulation of RANKL expression potentially contribute to bone resorption in periodontal disease.
Aggregatibacter actinomycetemcomitans; B lymphocytes; immune response; periodontal disease; receptor activator of nuclear factor-κB ligand
The s allele variant of the serotonin transporter gene (5-HTT) has recently been observed to moderate the relationship of stress to depression and anxiety. To date no study has considered interactive effects of 5-HTT genotype, stress and HPA function on cognition in healthy, older adults, which may reflect developmental, functional, or neurodegenerative effects of the serotonin transporter polymorphism.
We investigated whether 5-HTT genotype interacts with cumulative life stress and HPA-axis measures of waking and diurnal cortisol slope to impact cognition in 154 non-depressed, older adults. Structural images of hippocampal volume were acquired on a subsample of 56 participants.
The 5-HTT s allele was associated with both significantly lower delayed recall and higher waking cortisol levels. Presence of the s allele interacted with higher waking cortisol to negatively impact memory. We also observed a significant interaction of higher waking cortisol and the s allele on lower hippocampal volume. Smaller hippocampi and higher cortisol were associated with lower delayed recall only in s allele carriers. No impact or interactions of cumulative life stress with 5-HTT or cortisol were observed.
This is the first investigation to identify an association of the 5-HTT s allele with poorer memory function in older adults. The interactive effects of the s allele and waking cortisol levels on reduced hippocampal volume and lower memory suggest that the negative effect of the serotonin polymorphism on memory is mediated by the HPA axis. Further, given the significant association of the s allele with higher waking cortisol in our investigation, future studies may want to evaluate the impact of the serotonin transporter polymorphism on any neuropsychiatric or behavioral outcome which is influenced by HPA axis function in older adults.
Serotonin transporter; 5-HTT; Stress; HPA; Memory; Hippocampus; Aging
Epstein-Barr virus nuclear antigen EBNA1, the one viral protein uniformly expressed in nasopharyngeal carcinoma (NPC), represents a prime target for T-cell-based immunotherapy. However, little is known about the EBNA1 epitopes, particularly CD4 epitopes, presented by HLA alleles in Chinese people, the group at highest risk for NPC. We analyzed the CD4+ T-cell responses to EBNA1 in 78 healthy Chinese donors and found marked focusing on a small number of epitopes in the EBNA1 C-terminal region, including a DP5-restricted epitope that was recognized by almost half of the donors tested and elicited responses able to recognize EBNA1-expressing, DP5-positive target cells.