Dual inhibition of EZH2 and G9A/GLP histone methyltransferases by HKMTI-1-005 promotes differentiation of acute myeloid leukemia cells.

All-trans-retinoic acid (ATRA)-based differentiation therapy of acute promyelocytic leukemia (APL) represents one of the most clinically effective examples of precision medicine and the first example of targeted oncoprotein degradation. The success of ATRA in APL, however, remains to be translated to non-APL acute myeloid leukemia (AML). We previously showed that aberrant histone modifications, including histone H3 lysine 4 (H3K4) and lysine 27 (H3K27) methylation, were associated with this lack of response and that epigenetic therapy with small molecule inhibitors of the H3K4 demethylase LSD1/KDM1A could reprogram AML cells to respond to ATRA. Serving as the enzymatic component of Polycomb Repressive Complex 2, EZH2/KMT6A methyltransferase plays a critical role in normal hematopoiesis by affecting the balance between self-renewal and differentiation. The canonical function of EZH2 is methylation of H3K27, although important non-canonical roles have recently been described. EZH2 mutation or deregulated expression has been conclusively demonstrated in the pathogenesis of AML and response to treatment, thus making it an attractive therapeutic target. In this study, we therefore investigated whether inhibition of EZH2 might also improve the response of non-APL AML cells to ATRA-based therapy. We focused on GSK-343, a pyridone-containing S-adenosyl-L-methionine cofactor-competitive EZH2 inhibitor that is representative of its class, and HKMTI-1-005, a substrate-competitive dual inhibitor targeting EZH2 and the closely related G9A/GLP H3K9 methyltransferases. We found that treatment with HKMTI-1-005 phenocopied EZH2 knockdown and was more effective in inducing differentiation than GSK-343, despite the efficacy of GSK-343 in terms of abolishing H3K27 trimethylation. Furthermore, transcriptomic analysis revealed that in contrast to treatment with GSK-343, HKMTI-1-005 upregulated the expression of differentiation pathway genes with and without ATRA, while downregulating genes associated with a hematopoietic stem cell phenotype. These results pointed to a non-canonical role for EZH2, which was supported by the finding that EZH2 associates with the master regulator of myeloid differentiation, RARα, in an ATRA-dependent manner that was enhanced by HKMTI-1-005, possibly playing a role in co-regulator complex exchange during transcriptional activation. In summary, our results strongly suggest that addition of HKMTI-1-005 to ATRA is a new therapeutic approach against AML that warrants further investigation.

Memory technology-a primer for material scientists.

From our own experience, we know that there is a gap to bridge between the scientists focused on basic material research and their counterparts in a close-to-application community focused on identifying and solving final technological and engineering challenges. In this review, we try to provide an easy-to-grasp introduction to the field of memory technology for materials scientists. An understanding of the big picture is vital, so we first provide an overview of the development and architecture of memories as part of a computer and call attention to some basic limitations that all memories are subject to. As any new technology has to compete with mature existing solutions on the market, today's mainstream memories are explained, and the need for future solutions is highlighted. The most prominent contenders in the field of emerging memories are introduced and major challenges on their way to commercialization are elucidated. Based on these discussions, we derive some predictions for the memory market to conclude the paper.

Investigating the familiarity effect in texture segmentation by means of event-related brain potentials.

The familiarity effect (FE) refers to the phenomenon that it is easier to find an unfamiliar element on a background of familiar elements than vice versa. In this study, we examined the FE in texture segmentation while recording event-related brain potentials with the aim to find out which processing stages were influenced by familiarity. In two experiments, with different levels of texture homogeneity, the N1, the N2p, and the P3 components were investigated. It was found that the FE in texture segmentation is associated with a modulation of the early N1 and of the intermediate N2p component for homogeneous textures. For inhomogeneous (jittered) textures, the FE was found for the intermediate N2p and for the late P3 components, but not for the N1 component. Our findings suggest that increasing texture inhomogeneity shifts the FE occurrence to later processing stages.

[Hemophagocytic lymphohistiocytosis : A diagnostic challenge on the ICU]. / Hämophagozytische Lymphohistiozytose : Diagnostische Herausforderung auf der ITS.

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH) has well been studied as a genetic disorder in children (primary HLH). Mutations in the regulatory complex of the cellular immune synapse lead to a loss of function of cytotoxic T­cells and natural killer cells with excessive inflammation based on a cytokine storm. During the last decade, an increasing number of adult HLH patients without a family history of HLH (secondary or acquired HLH) have been reported. Various triggers - infections, malignancies or autoimmune diseases - result in an acquired loss of function of these cells and a sepsis-like disease. Missed or late diagnosis is believed to be a major cause of the high mortality. OBJECTIVES: To describe the current knowledge on HLH and to raise awareness. MATERIALS AND METHODS: Analysis of case reports, current studies, and expert recommendations. RESULTS: Increased vigilance in identifying the adult form of HLH resulted in an increasing number of case reports over the past few years. HLH patients typically present with a clinical phenotype resembling severe sepsis or septic shock with fever, cytopenia, and organomegaly, which do not or insufficiently respond to anti-infective treatment. Early recognition of HLH distinction from sepsis, and prompt initiation of treatment - which is fundamentally different from sepsis - are crucial for improved outcome. A promising diagnostic parameter is ferritin, which has gained sufficient specificity, but only in the context of the triad of fever, cytopenia, and organomegaly. Treatment of adult HLH patients requires immunosuppression, with strict therapeutic guidance derived from the triggering disease. CONCLUSIONS: Because of the similar clinical presentation to that of sepsis, HLH is often not recognized, resulting in a fatal outcome. In "sepsis" patients on the ICU with deterioration despite a standard of care, HLH needs to be considered by testing for ferritin when considering differential diagnoses. The complexity of the illness requires interdisciplinary patient care with specific integration of the hematologist in the diagnostic workup and therapeutic management, because of the frequent use of chemotherapy-based immunosuppression.

From sweet to sweat: Hedonic olfactory range is impaired in Parkinson's disease.

INTRODUCTION: Olfactory dysfunction and neuropsychological symptoms like depression and anhedonia are common non-motor symptoms in Parkinson's disease (PD). The assessment of both functional domains includes clinical examination, olfactory testing, and standardized questionnaires. While olfaction is readily assessed by functional tests, the distinction of anhedonia as a separate symptom from other depressive symptoms is challenging. Thus, a test focusing on the assessment of hedonic olfaction may be helpful in the assessment of neuropsychological symptoms in PD. METHODS: We examined anhedonia by evaluating the perception of pleasantness of odors in PD patients (n = 57) and healthy controls (n = 46). Pleasantness of odors was registered on a visual 9-point scale. For the assessment of anhedonia we used the Snaith-Hamilton-Pleasure-Scale (SHAPS). Depression was evaluated with the Zung Self-Rating Depression Scale and the Beck Depression Inventory II. RESULTS: PD patients showed a substantial reduction in hedonic olfaction compared to controls (hedonic score: 1.5 vs. 2.2). Hyposmia, one of the most prevalent non-motor symptoms in PD, was a confounding factor. However, even normosmic PD patients showed a reduced hedonic olfaction compared to controls (hedonic score: 1.6 vs. 2.2). Furthermore, we observed a correlation between hedonic olfaction and the SHAPS-score for PD patients even though positive SHAPS-rating was observed in 9% of PD patients only, while no correlation to depression was present. CONCLUSION: These findings suggest that reduced hedonic olfaction might be an additional neuropsychological feature, probably giving insights into changes in hedonic tone complementary to hyposmia and depression in PD.

Unlocking the potential of retinoic acid in anticancer therapy.

All-trans-retinoic acid (ATRA) is a physiologically active metabolite of vitamin A. Its antitumour activities have been extensively studied in a variety of model systems and clinical trials; however, to date the only malignancy responsive to ATRA treatment is acute promyelocytic leukaemia (APL) where it induces complete remission in the majority of cases when administered in combination with light chemotherapy and/or arsenic trioxide. After decades of studies, the efficacy of ATRA to treat other acute myeloid leukaemia (AML) subtypes and solid tumours remains poor. Recent studies directed to improve ATRA responsiveness in non-APL AML seem to indicate that the lack of effective ATRA response in these tumours may be primarily due to aberrant epigenetics, which negatively affect ATRA-regulated gene expression and its antileukaemic activity. Epigenetic reprogramming could potentially restore therapeutic effects of ATRA in all AML subtypes. This review discusses the current progresses in the understanding how ATRA can be utilised in the therapy of non-APL AML and other cancers.

Limb activation ameliorates body-related deficits in spatial neglect.

Many neglect patients show deficits in the mental representation of their contralesional body side or body parts, termed personal neglect. These deficits include impairments in identifying body parts on schematic drawings of human bodies. Limb activation and alertness cues have been shown to modulate neglect transiently, and are effective treatments for several symptoms of the neglect syndrome. Here, we tested on eight patients with right-hemispheric stroke and left-sided spatial neglect whether these two techniques modulate deficits in the mental representation of hands, assessed with a hand-test in which the subjects had to decide whether a depicted schematic hand belongs to the left or right side of the human body. The results showed that neglect patients made marginally significant (p = 0.065) more errors in left-hand-decisions than right-hand-decisions, indicating a neglect-specific disorder. Moreover, we found that left-sided limb activation but not non-lateralized alertness cueing (a loud noise immediately before patients made their perceptual decision) significantly reduced misidentifications for depicted left hands as compared to baseline. No effect of any intervention was observed on error rates for depicted right hands. We conclude that the amelioration of the performance in the hand task is modulated by the activation of the body schema or other body representations through left-sided limb activation.

Omacetaxine mepesuccinate prevents cytokine-dependent resistance to nilotinib in vitro: potential role of the common ß-subunit c of cytokine receptors.

Overcoming resistance against BCR-ABL-inhibitors in chronic myeloid leukemia (CML) is central to prevent progression to advanced phase disease. Kinase mutations of BCR-ABL and cytokine-mediated modulation of response to tyrosine kinase inhibitors (TKIs) are key mechanisms governing clinical response to imatinib and second generation TKIs. Omacetaxine mepesuccinate is effective in imatinib-resistant CML with reported stem cell activity. We specifically thought to explore omacetaxine in the context of the pan-resistant mutant T315I, and in its potential to modify cytokine-dependent resistance. Omacetaxine was investigated in cell lines and primary CD34+ enriched progenitor cells from patients with CML. Addition of cytokines, shown to revert the efficacy of TKIs in BCR-ABL-positive cells, does not affect omacetaxine mediated antiproliferative activity, neither in cell lines nor in primary CML CD34+ progenitor cells. Looking at potential mechanisms, we found marked downregulation of the common ß-subunit c of the cytokine-receptors (cCRßc) for IL3, IL5 and GM-CSF by omacetaxine in cell lines and primary progenitor cultures. The observed cytokine-independent in-vitro cytotoxicity of omacetaxine may be explained by downregulation of cCRßc. Whether this can be used clinically as a means to optimize the stem cell activity of TKIs merits further evaluation.

Long-term effects of a partially supervised conditioning programme in cystic fibrosis.

Little is known about the long-term persistence of positive effects induced by a physical conditioning programme in cystic fibrosis. Therefore, this study determined the effects of a 6-month conditioning programme on peak oxygen uptake (primary outcome) and other markers of fitness, physical activity, anthropometry, lung function and quality of life (secondary outcomes), 18 and 24 months after the programme was initiated. Patients with cystic fibrosis aged 12-40 yrs were randomly assigned to an intervention (n = 23) and a control (n = 15) group. The intervention group consented to add 3 h of sports per week for > or =6 months to their previous activities. Controls were asked to maintain their level of activity for 12 months. Patients were seen at baseline and after 3, 6, 12, 18 and 24 months. There was no significant difference between groups at baseline. The intervention induced positive effects on peak oxygen uptake (difference in changes from baseline to the 18- and 24-month assessments between groups: 3.72+/-1.23 mL.min(-1).kg(-1); p<0.01), maximal workload (0.37+/-0.11 W.kg(-1); p<0.01), vigorous physical activity (1.63+/-0.82 h.week(-1); p<0.05), forced vital capacity (6.06+/-2.87% predicted; p<0.05) and perceived health (9.89+/-4.72; p<0.05). A home-based partially supervised physical conditioning programme can improve physical fitness, lung function and perceived health long after the intervention has ended.

Harmonization of molecular monitoring of CML therapy in Europe.

The high efficacy of the standard treatment of chronic myeloid leukemia (CML) with imatinib has prompted the need for accurate methods to monitor response at levels below the landmark of complete cytogenetic remission. Quantification of BCR-ABL transcripts has proven to be the most sensitive method available, and has shown prognostic impact with regard to progression-free survival. Until recently, variations in methods used to quantify BCR-ABL made it difficult to compare results between laboratories. An international program is now underway to harmonize the reporting of results according to an international scale (IS). In this review, we consider the background to the IS and the progress that has been made to date, with a particular focus on ongoing harmonization efforts in Europe. We provide recommendations for the propagation of the IS by national or regional laboratory networks.

Artichoke leave extract for chronic hepatitis C - a pilot study.

BACKGROUND: Artichoke leave extracts (ALE) have hepatoprotektive properties and are used by patients with chronic liver disease. Effects in patients with chronic hepatitis C are unclear. METHODS: 17 patients with chronic hepatitis C and persistently elevated aminotransferase levels were treated for 12 weeks with 3200mg standardized ALE per day. Primary outcome parameter was the rate of alanine aminotransferase (ALT) normalisation after 12 weeks. Secondary parameters were the course of ALT, aspartate aminotransferase and gamma glutamyltransferase levels, quantitative HCV RNA, subjective symptoms frequently associated with chronic hepatitis C (fatigue, discomfort upper abdomen, joint problems) and safety. RESULTS: None of the patients had normalized ALT levels after 12 weeks of treatment. There was no significant change of aminotransferase levels or viral load compared to baseline levels. Fatigue and joint problems significantly improved after 4 weeks of treatment. However, after 12 weeks, there was no significant difference to baseline. Tolerability of ALE was rated as good to excellent. Severe side effects did not occur. CONCLUSION: ALE seem not to be effective to improve aminotransferase levels in patients with chronic hepatitis C.

Selective coral mortality associated with outbreaks of Acanthaster planci L. in Bootless Bay, Papua New Guinea.

Population outbreaks of crown-of-thorns sea star (Acanthaster planci L.) remain one of the most significant biological disturbances on tropical coral reefs although the increasing attention given to other threats has greatly limited recent progress in understanding the cause and consequences of this phenomenon. In September 2005 dramatic increases in the abundance of A. planci were observed on reefs within Bootless Bay, Central Province, Papua New Guinea, where few crown-of-thorns have previously been reported. Densities of A. planci peaked at 162 sea stars per hectare and caused extensive coral mortality. This outbreak killed upwards of 55% of live corals, reducing overall coral cover from 42.4% in 2005 down to just 19.1% in March 2006. Declines in coral cover were largely driven by widespread mortality of Acropora spp. which dominated reef assemblages prior to the outbreak. The extensive depletion of Acropora spp. greatly altered the coral composition as well as the physical structure of reef habitats. In the absence of any other major disturbances these coral communities are likely to quickly recover, but this outbreak highlights the ongoing contribution of A. planci to degradation of coral reef environments.

Performance of a polymer-based DNA chip platform in detection and genotyping of human papillomavirus in clinical samples.

Human papillomavirus (HPV) plays a key role in the development of cervical and laryngeal cancers. The aim of our study was to compare the performance of a new hydrogel-based HPV genotyping biochip assay (Biochip) to a commercially available and CE-marked conventional PCR followed by reverse hybridization (GenID-PCR). One hundred twenty-three samples were available for the study. Of these samples, 101/123 were gynecological swabs, 8/123 were swabs or biopsy samples of genital warts, 7/123 were biopsy samples of otorhinolaryngeal lesions, 5/123 were samples of skin warts, and 2/123 were samples of orolabial abnormalities. These molecular methods for HPV genotyping showed comparable sensitivity and specificity. However, 19/123 of the results were discrepant. Specifically, Biochip showed better performance in the detection of multiple infections, especially when more than one high-risk genotype was present. Due to the different probe configurations used in the two assays, GenID-PCR achieves only group-specific detection of many HPV genotypes, whereas Biochip allows for specific identification. Overall, the newly developed HPV chip system (Biochip) proved to be a suitable tool for HPV detection and genotyping; it also proved to be superior for establishing HPV genotyping methods.

Reflexive attention attenuates change blindness (but only briefly).

Humans are remarkably insensitive to large changes in a visual display if the change occurs simultaneously with a secondary perceptual event. A widely held view is that this change blindness occurs because the secondary perceptual event prevents the change from capturing attention. However, whereas some studies have shown that top-down attentional priming can attenuate change blindness, the evidence regarding the effect of bottom-up attentional capture on change blindness is less clear-cut. Here, we compare the effects of attentional capture on change detection with participants' performance on a well-established attentional paradigm (a Posner-style cuing task). Experiment 1 established the time course of attentional capture in our paradigm. Experiment 2 demonstrated that this attentional capture was associated with facilitated change detection at short (150-msec), but not long (480-msec), latencies. These data show that reflexive attentional shifts facilitate change detection and are consistent with the view that shifts of attention are a necessary precondition for visual awareness.

Comparison of nine commercially available assays for quantification of antibody response to hepatitis B virus surface antigen.

Quantitative measurement of anti-HBs is used to evaluate the response to hepatitis B vaccination in health care workers and to optimize postexposure management. The different guidelines for hepatitis B vaccination and booster policy imply that the measurement of anti-HBs levels by different assays is accurate and consistent, yielding comparable quantitative results. We measured anti-HBs levels in 200 serum samples from patients and health care professionals by nine different anti-HBs assays and compared the quantitative results and the performance characteristics of the different test systems. The assay specificity ranged between 96.8 and 100% when sera from individuals without a vaccination history and with negative anti-HBc status were defined as true negatives. Sensitivity ranged between 93.5 and 100%. A high number of sera showed discrepancies between measurements by the different systems. The mean coefficient of variation between the different measurements was 47.1% (range, 15.0 to 201.0%), and the factors of multiplication ranged from 2.8 to 105. Hemolysis or lipemia did not seem to influence the measurement, and there was no difference between anti-HBc-positive and -negative individuals. The classical enzyme immunoassays tend to find lower anti-HBs levels than the automated systems, with higher values by the Abbott AXSYM assay. The serial dilution of the international standard preparation was measured accurately by most of the assays. In conclusion, the quantitative measurement of anti-HBs levels is not reliable, even though an international standard is used for the calibration of the systems. Some systems showed specific problems that should be addressed by the manufacturers.

Harmonization of BCR-ABL mRNA quantification using a uniform multifunctional control plasmid in 37 international laboratories.

Individualized PCR strategies hamper comparability of molecular results between different laboratories in several fields of medicine. To harmonize BCR-ABL mRNA quantification an international multicenter trial involving 37 laboratories in 14 countries was initiated using 10 samples, each containing various dilutions (10, 2, 1 and 0.1%) of b3a2 or b2a2 BCR-ABL positive in normal leukocytes and negative controls. A novel control plasmid (pME-2) was designed for external calibration containing BCR-ABL and glucuronidase-beta (GUS) sequences. Median BCR-ABL/ABL ratios were 9.1, 1.8, 0.85 and 0.11% in b3a2 samples and 9.5, 1.6, 0.84 and 0.11% in b2a2 samples. Median BCR-ABL/GUS ratios were 3.4, 0.77, 0.37 and 0.042% in b3a2 samples and 2.8, 0.48, 0.29 and 0.031% in b2a2 samples. The coefficients of variation were 0.62 for ratios BCR-ABL/ABL and 1.03 for ratios BCR-ABL/GUS. Five of 37 evaluable participating laboratories (13%) detected low BCR-ABL copy numbers in negative control samples; one laboratory failed to detect BCR-ABL in a low-level sample. We conclude that the use of a common control plasmid does indeed improve comparability of BCR-ABL mRNA quantification results. However, further standardizing efforts like introducing a calibrator and regular control rounds are needed.

A new surgical model for hepatectomy in pigs.

BACKGROUND: Anhepatic animal models are suitable for simulating acute liver failure. Hepatectomy in pigs includes en bloc resection of the vena cava, and therefore, a temporary extracorporeal bypass and total clamping of the inferior vena cava are needed. These steps cause severe depression of circulation with impaired survival. METHODS: Previous to en bloc hepatectomy including retrohepatic vena cava in 20 female pigs, a Y-shaped bypass was implanted starting with end-to-side anastomosis between the vena cava and the portal vein, followed by anastomosis to the intrathoracic vena cava. RESULTS: Blood flow was constant during and after hepatectomy because vessels were only partially clamped. No venous stasis of intestinal organs was observed. Hemodynamic parameters like heart rate, mean arterial pressure, central venous pressure, pulse oximetry and intracranial pressure remained extremely stabile during and after hepatectomy. Postoperative survival time was 100% after 12 h. Maximum survival time was 84.9 h and mean survival time 51.2 +/- 18.7 h. All animals died from multiple organ failure. Intracranial pressure remained stable during the surgical procedure and rose continuously until death. The autopsy showed massive brain edema. CONCLUSIONS: This new surgical technique is safe and easy to perform and permits total hepatectomy with minimal blood loss under stable circulation without requiring an extracorporeal bypass.

Enhanced probe discrimination at the location of a colour singleton.

There is ample evidence to suggest that preparing to saccade to a location is sufficient to produce attentional shifts to this location. However, it is not clear whether engagement of the eye-movement system is also a necessary condition for any spatial shifts in attention. Recent neurophysiological data indicates that neurons in the frontal eye field (FEF) can select visual stimuli (a colour singleton) in the absence of a concurrently activated saccade plan in non-human primates (Juan et al. in Proc Natl Acad Sci USA 101:15541-15544, 2004), suggesting that saccade planning and visual selection are dissociable. However, it is still unclear whether the visual selection is accompanied by an attentional enhancement at the target location. To test this, we used a similar paradigm (i.e. an antisaccade task) with humans to the one employed by Juan and colleagues with monkeys. Our paradigm included a probe-discrimination task, which allowed us to test whether attentional facilitation is indeed observed at the location of the selected visual stimuli. Our results confirm that visual selection is accompanied by attentional facilitation.

In situ and real-time probing of quasicrystal solidification dynamics by synchrotron imaging.

Quasicrystal growth remains an unsolved problem in condensed matter. The dynamics of the process is studied by means of synchrotron live imaging all along the solidification of icosahedral AlPdMn quasicrystals. The lateral motion of ledges driving faceted growth at the solid-melt interface is conclusively shown. When the solidification rate is increased, nucleation and free growth of new faceted grains occur in the melt due to the significant interface recoil induced by slow attachment kinetics. The detailed analysis of the evolution of these grains reveals the crucial role of aluminum rejection, both in the poisoning of their growth and driving fluid flow.