Studies on CEBPA mutations in acute myeloid leukaemia

Acute myeloid leukaemia (AML) is a highly heterogeneous disease with regard to clinical outcome, and molecular markers with prognostic impact can be used to stratify patients for risk-adapted therapy. CEBPA mutations have been associated with a favourable prognosis, however several questions remained, in particular whether one (CEBPA-single) or two (CEBPA-double) mutations were necessary for this benefit, and their interaction with other molecular markers. A method of detecting CEBPA mutations in patient samples using denaturing HPLC was developed and the CEBPA status of 1427 young adult AML patients (median age 43 years, range 15-68 years) determined. Overall, 107 (7%) were CEBPA-mutant: 48 (45%) CEBPA-single and 59 (55%) CEBPA-double. The majority of CEBPA-double patients (83%) had an out-of-frame insertion/deletion in the N-terminus and a mutation in the C-terminal DNA-binding/leucine zipper domains (DBD/LZD) that were on different alleles as determined by cloning. By contrast, mutations in CEBPA-single cases were distributed across the gene. CEBPA-double patients were less likely to have a FLT3/ITD (P=.04) and highly unlikely to have an NPM1 mutation (P<.0001) compared to CEBPA-WT/CEBPA-single cases. Eight year overall survival (OS) was higher in CEBPA-double patients compared to CEBPA-WT and CEBPA-single cases (54%, 34%, 31%, respectively, P=.004). In multivariate analyses, CEBPA-double, but not CEBPA-single, was an independent favourable factor for OS (P=.004) and relapse (P=.02). However, this benefit was completely lost in the presence of a FLT3/ITD. The mutant level of 101 mutations was determined by fragment analysis and the majority were of a level consistent with a heterozygous mutation present in most cells. The impact of ten atypical CEBPA mutations on C/EBPα transactivation activity was explored by a luciferase reporter assay. Only mutations affecting the DBD or LZD functional domains had an impact on transactivation activity. This work provides insight into the biology of CEBPA mutations and their use as clinical markers

Impact of handgrip exercise intensity on brachial artery flow-mediated dilation.

PURPOSE: Previous studies that have examined the impact of exercise intensity on conduit artery endothelial function have involved large muscle group exercise which induces local and systemic effects. The aim of this study was to examine flow-mediated dilation (FMD) before and after incremental intensities of handgrip exercise (HE), to assess the role of local factors such as blood flow and shear rate on post-exercise brachial artery function. METHODS: Eleven healthy men attended the laboratory on three occasions. Subjects undertook 30 min of handgrip exercise at three intensities (5, 10 or 15 % MVC). Brachial artery FMD, shear and blood flow patterns were examined before, immediately after and 60 min post exercise. RESULTS: Handgrip exercise increased mean and antegrade shear rate (SR) and blood flow (BF) and reduced retrograde SR and BF (all P < 0.01). Exercise intensity was associated with a dose-dependent increase in both mean and antegrade BF and SR (interaction, P < 0.01). Post-hoc tests revealed that, whilst handgrip exercise did not immediately induce post-exercise changes, FMD was significantly higher 60 min post-exercise following the highest exercise intensity (5.9 ± 2.8-10.4 ± 5.8 %, P = 0.01). CONCLUSIONS: Handgrip exercise leads to intensity-and time-dependent changes in conduit artery function, possibly mediated by local increases in shear, with improvement in function evident at 1 h post-exercise when performed at a higher intensity

Imaging the cold molecular gas in SDSS J1148 + 5251 at z = 6.4

We present Karl G. Jansky Very Large Array (VLA) observations of the CO ($J = 2 \rightarrow 1$) line emission towards the $z = 6.419$ quasar SDSS J$114816.64+525150.3$ (J$1148+5251$). The molecular gas is found to be marginally resolved with a major axis of $0.9"$ (consistent with previous size measurements of the CO ($J = 7 \rightarrow 6$) emission). We observe tentative evidence for extended line emission towards the south west on a scale of ~$1.4"$, but this is only detected at $3.3\sigma$ significance and should be confirmed. The position of the molecular emission region is in excellent agreement with previous detections of low frequency radio continuum emission as well as [C ii] line and thermal dust continuum emission. These CO ($J = 2 \rightarrow 1$) observations provide an anchor for the low excitation part of the molecular line SED. We find no evidence for extended low excitation component, neither in the spectral line energy distribution nor the image. We fit a single kinetic gas temperature model of 50 K. We revisit the gas and dynamical masses in light of this new detection of a low order transition of CO, and confirm previous findings that there is no extended reservoir of cold molecular gas in J$1148+5251$, and that the source departs substantially from the low $z$ relationship between black hole mass and bulge mass. Hence, the characteristics of J$1148+5251$ at $z = 6.419$ are very similar to $z$~$2$ quasars, in the lack of a diffuse cold gas reservoir and kpc-size compactness of the star forming region.IIS thanks the Science & Technology Facilities Council for a studentship.This article has been accepted for publication in Monthly Notices of the Royal Astronomical Society (c): 2015 The Authors. Published by Oxford University Press on behalf of the Royal Astronomical Society. All rights reserved

Evidence for Shear Stress-Mediated Dilation of the Internal Carotid Artery in Humans.

Increases in arterial carbon dioxide tension (hypercapnia) elicit potent vasodilation of cerebral arterioles. Recent studies have also reported vasodilation of the internal carotid artery during hypercapnia, but the mechanism(s) mediating this extracranial vasoreactivity are unknown. Hypercapnia increases carotid shear stress, a known stimulus to vasodilation in other conduit arteries. To explore the hypothesis that shear stress contributes to hypercapnic internal carotid dilation in humans, temporal changes in internal and common carotid shear rate and diameter, along with changes in middle cerebral artery velocity, were simultaneously assessed in 18 subjects at rest and during hypercapnia (6% carbon dioxide). Middle cerebral artery velocity increased significantly (69±10-103±17 cm/s; P<0.01) along with shear in both the internal (316±52-518±105 1/s; P<0.01) and common (188±40-275±61 1/s; P<0.01) carotids. Diameter also increased (P<0.01) in both carotid arteries (internal: +6.3±2.9%; common: +5.8±3.0%). Following hypercapnia onset, there was a significant delay between the onset of internal carotid shear (22±12 seconds) and diameter change (85±51 seconds). This time course is associated with shear-mediated dilation of larger conduit arteries in humans. There was a strong association between change in shear and diameter of the internal carotid (r=0.68; P<0.01). These data indicate, for the first time in humans, that shear stress is an important stimulus for hypercapnic vasodilation of the internal carotid artery. The combination of a hypercapnic stimulus and continuous noninvasive, high-resolution assessment of internal carotid shear and dilation may provide novel insights into the function and health of the clinically important extracranial arteries in humans

Novel gallium(III) complexes transported by MDR1 P-glycoprotein: potential PET imaging agents for probing P-glycoprotein-mediated transport activity in vivo

AbstractBackground: Multidrug resistance (MDR) mediated by expression of MDR1 P-glycoprotein (Pgp) represents one of the best characterized barriers to chemotherapy in cancer patients. Positron emission tomography (PET) agents for analysis of Pgp-mediated drug transport activity in vivo would enable noninvasive assessment of chemotherapeutic regimens and MDR gene therapy.Results: Candidate Schiff-base phenolic gallium(III) complexes were synthesized from their heptadentate precursors and gallium(III)acetylacetonate. Crystal structures demonstrated a hexacoordinated central gallium with overall trans-pseudo-octahedral geometry. Radiolabeled 67Ga-complexes were obtained in high purity and screened in drug-sensitive (Pgp−) and MDR (Pgp+) tumor cells. Compared with control, lead compound 6 demonstrated antagonist-reversible 55-fold lower accumulation in Pgp-expressing MDR cells. Furthermore, compared with wild-type control, quantitative pharmacokinetic analysis showed markedly increased penetration and retention of 6 in brain and liver tissues of mdr1a/b(−/−) gene disrupted mice, correctly mapping Pgp-mediated transport activity at the capillary blood–brain barrier and hepatocellular biliary cannalicular surface in vivo.Conclusions: These results indicate that gallium(III) complex 6 is recognized by MDR1 Pgp as an avid transport substrate, thereby providing a useful scaffold to generate 68Ga radiopharmaceuticals for molecular imaging of Pgp transport activity in tumors and tissues in vivo using PET

Reproducibility of four frequently used local heating protocols to assess cutaneous microvascular function.

BACKGROUND: Skin microvascular responses to local heating are frequently used to assess microvascular function. Several local heating protocols have been developed, all varying slightly in execution. The aim of this study was to determine the inter-day reproducibility of the four most commonly used local heating protocols in healthy young subjects. METHODS: Fifteen, healthy males (28±5yrs, BMI 25±2kg/m(2)) attended two experimental trials 2-7days apart. During each trial, baseline and maximal thermally stimulated forearm skin responses were examined simultaneously at four sites on the dominant forearm using laser Doppler flowmetry (LDF). The following heating protocols were adopted: 1. Rapid 39°C (0.5°C/5s), 2. Rapid 42°C (0.5°C/5s) 3. Gradual 42°C (0.5°C/2min 30s) and 4. Slow 42°C (0.5°C/5min). The coefficient of variation (CV) was calculated for absolute flux, cutaneous vascular conductance (CVC; flux/mean arterial pressure, MAP) and CVC expressed as a percentage of maximal CVC at 44°C (%CVCmax) at three different time points; baseline (33°C), plateau (39/42°C) and maximal (44°C). RESULTS: Reproducibility of baseline flux, CVC and %CVCmax was 17-29% across all protocols. During the plateau, Rapid, Gradual and Slow 42°C demonstrated a reproducibility of 13-18% for flux and CVC and 5-11% for %CVCmax. However, Rapid 39°C demonstrated a lower reproducibility for flux, CVC and %CVCmax (all 21%). Reproducibility at 44°C was 12-15% for flux and CVC across all protocols. CONCLUSION: This is the first study examining inter-day reproducibility across four local heating protocols. The good-to-moderate reproducibility of the Rapid, Gradual and Slow 42°C protocols support their (simultaneous) use to assess microvascular function. Using Rapid 39°C may require a greater number of subjects to detect differences within subjects

The CPTPP trade deal is a major threat to public health and warrants a health impact assessment.

The UK’s decision to join one of the world’s largest free trade agreements has implications for health that need urgent assessment, argue Courtney McNamara and colleague

Decision for reconstructive interventions of the upper limb in individuals with tetraplegia: the effect of treatment characteristics

Objective: To determine the effect of treatment characteristics on the\ud decision for reconstructive interventions for the upper extremities (UE) in\ud subjects with tetraplegia. - \ud Setting: Seven specialized spinal cord injury centres in the Netherlands. - \ud Method: Treatment characteristics for UE reconstructive interventions were\ud determined. Conjoint analysis (CA) was used to determine the contribution\ud and the relative importance of the treatment characteristics on the decision\ud for therapy. Therefore, a number of different treatment scenarios using these\ud characteristics were established. Different pairs of scenarios were presented\ud to subjects who were asked to choose the preferred scenario of each set. - \ud Results: forty nine subjects with tetraplegia with a stable C5, C6 or C7\ud lesion were selected. All treatment characteristics significantly influenced\ud the choice for treatment. Relative importance of treatment characteristics\ud were: intervention type (surgery or surgery with FES implant) 13%, number\ud of operations 15%, in patient rehabilitation period 22%, ambulant\ud rehabilitation period 9%, complication rate 15%, improvement of elbow\ud function 10%, improvement of hand function 15%. In deciding for therapy\ud 40% of the subjects focused on one characteristic. - \ud Conclusion: CA is applicable in Spinal Cord Injury medicine to study the\ud effect of health outcomes and non-health outcomes on the decision for\ud treatment. Non-health outcomes which relate to the intensity of treatment\ud are equally important or even more important than functional outcome in the\ud decision for reconstructive UE surgery in subjects with tetraplegia

Boundary entropy of supersymmetric Janus solutions

In this paper we compute the holographic boundary entropy for half-BPS Janus deformations of the $AdS_3\times S^3\times T^4$ vacuum of type IIB supergravity. Previous work \cite{Chiodaroli:2009yw} has shown that there are two independent deformations of this sort. In one case, the six-dimensional dilaton jumps across the interface, while the other case displays a jump of axion and four-form potential. In case of a jump of the six-dimensional dilaton, it is possible to compare the holographic result with the weak-coupling result for a two-dimensional interface CFT where the radii of the compactified bosons jump across the interface. We find exact agreement between holographic and CFT results. This is to be contrasted with the holographic calculation for the non-supersymmetric Janus solution, which agrees with the CFT result only at the leading order in the jump parameter. We also examine the implications of the holographic calculation in case of a solution with a jump in the axion, which can be associated with a deformation of the CFT by the $Z_2$-orbifold twist operator.Comment: 35 pages, pdf-LaTeX, 5 figures, v2: minor changes, typos corrected, reference adde

Antibiotic resistance profiles and relatedness of enteric bacterial pathogens isolated from HIV/AIDS patients with and without diarrhoea and their household drinking water in rural communities in Limpopo Province South Africa

Antibiotic resistance profiles and the correlation of enteric bacterial pathogens from HIV positive individuals with and without diarrhoea and their household drinking water were determined using the KirbyBauer disk diffusion and polymerase chain reaction methods respectively. The sef gene of Salmonella enteritidis was amplified with the primer pair sefA-1 and sefA-2. The fliC gene of Salmonella typhimurium was amplified with the primer pair flicA-1 and flicA-2. Heat-labile toxin (LT) primers (Lta and LTb) were used to amplify Escherichia coli isolates and VirA1 and VirA2 for the Vir A gene of Shigella dysenteriae. Results of antibiotic resistance profiles of enteric bacterial pathogens isolated from stool samples of HIV positive and negative individuals with and without diarrhea and their household drinking water showed very similar drug resistance patterns. Over 90% of all the organisms isolated from the various study cohorts showed resistance to penicillin, cloxacillin and amoxicillin. Conversely, almost all the organisms were sensitive to ciprofloxacin, gentamycin, meropenem and imipenem. About 50% of E. coli isolated from the various study cohorts showed multiple antibiotic resistance to penicillin, amoxicillin, ampicillin, erythromycin, tetracycline, doxycycline and cotri-moxazole ( PR, AR, APR, ER, TR, DXTR, and TSR ) whereas less than 10% resistance was consistently reported for ofloxacin, gentamycin, meropenem cefotaxime, cefuroxime and imipenem ( OFXS, GMS, MEMS, CTXS, CXMS and IMIS ). The majority of Salmonella and Shigella isolates from all the groups were sensitive to ciprofloxacin, gentamicin, amikacin, meropenem, imipenem, nalidixic acid, kanamycin, piperacillin-tazo bactam, cefuroxime, doxycyclin, cefepime and ceftazidime (CIPS, GMS, AKS, MEMS, IMIS, NAS, KNS, DXTS, CXMS, CPMS, CAZS and PTZS). For Campylobacter, over 30% of the isolates were resistant to erythromycin, ampicillin, tetracycline,cotrimoxazole and ceftazidime (ER, APR TSR and CAZR) whereas over 85% were susceptible to ciprofloxacin, ofloxacin, gentamycin, amikacin, mero-penem, and nalidixic acid (CIPS, OFXS, GMS, AKS,MEMS and NAS). In addition to penicillin, amoxicillin, ampicillin and erythromycin, Aeromonas and Plesiomonas spp were more resistant to chloramphenicol, but were susceptible to ciprofloxacin, gentamycin, amikacin, meropenem, imipenem and nalidixic acid (CIPS, GMS, AKS, MEMS, IMIS and NAS). Polymerase Chain Reaction (PCR) experiments using targeted species genes of S. enteritidis, S. typhimurium, E. coli, Sh. dysenteriae showed that isolates from stool samples of HIV positive and HIV negative individuals with and without diarrhoea were also present in the household drinking water of the same study cohorts, suggesting that drinking water may have been the sources of the organisms in stool sample. Furthermore, by showing that the primers were able to amplify the genes in both clinical and environmental isolates, the link between the virulence of the pathogens was established