Core-binding factor leukemia hijacks the T-cell-prone PU.1 antisense promoter.

The blood system serves as a key model for cell differentiation and cancer. It is orchestrated by precise spatiotemporal expression of crucial transcription factors. One of the key master regulators in the hematopoietic systems is PU.1. Reduced levels of PU.1 are characteristic for human acute myeloid leukemia (AML) and are known to induce AML in mouse models. Here, we show that transcriptional downregulation of PU.1 is an active process involving an alternative promoter in intron 3 that is induced by RUNX transcription factors driving noncoding antisense transcription. Core-binding factor (CBF) fusions RUNX1-ETO and CBFß-MYH11 in t(8;21) and inv(16) AML, respectively, activate the PU.1 antisense promoter that results in a shift from sense toward antisense transcription and myeloid differentiation blockade. In patients with CBF-AML, we found that an elevated antisense/sense transcript and promoter accessibility ratio represents a hallmark compared with normal karyotype AML or healthy CD34+ cells. Competitive interaction of an enhancer with the proximal or the antisense promoter forms a binary on/off switch for either myeloid or T-cell development. Leukemic CBF fusions thus use a physiological mechanism used by T cells to decrease sense transcription. Our study is the first example of a sense/antisense promoter competition as a crucial functional switch for gene expression perturbation by oncogenes. Hence, this disease mechanism reveals a previously unknown Achilles heel for future precise therapeutic targeting of oncogene-induced chromatin remodeling.

Functional and radiological outcome of distal radius fractures stabilized by volar-locking plate with a minimum follow-up of 1 year.

INTRODUCTION: Distal radius fractures (DRF) are the most common fractures of the upper extremities and incidence is expected to continue rising as life expectancy increases. Palmar locking plate stabilizing has since become the standard treatment for dorsally displaced DRF. Main aim of this study was to investigate correlation between radiological and clinical outcome in patients stabilized by palmar locking plate with a minimum follow-up of one year. METHODS: A total of 524 patients with DRF, stabilized using palmar angular stable locking plate fixation were included in the study. Of these, 117 patients had to be excluded and another 177 were not accessible. The study group thus compromised 230 patients who returned for the follow-up investigation and were followed-up clinically and radiologically with a mean follow-up interval of 20 months. Outcome was evaluated using pain, range of motion (ROM) and grip strength parameters. In addition, self-assessment by patients was registered on the QuickDASH, PRWE and Mayo Score. The immediate postoperative and final checkup radiographs were scrutinized for alignment and intra-articular step-off. RESULTS: Bivariant correlation analysis showed a significant correlation between ulnar variance and QuickDASH (r = 0.18, p = 0.01), grip strength (r = - 0.18, p = 0.04) and Mayo Score (r = - 0.23, p = 0.001). No significant differences could be found between an unacceptable (> 2 mm) and acceptable (< 2 mm) ulnar variance in respect of pain, ROM, grip strength and patient-reported outcome measurements. Age, gender, additional fracture to the ulnar styloid, or type of postoperative immobilization showed no significant or clinical important impact on the final patient-reported outcome. No significant differences in incidence of complications, ROM or loss of reduction could be found in any patients over or under 65 years of age. CONCLUSIONS: Stabilization of DRF by palmar angular stable locking plate is a safe form of treatment and results in a good clinical and radiological outcome with low complication rate. Ulnar variance showed a significant correlation to grip strength, QuickDASH and Mayo Score, but an unacceptable ulnar variance (> 2 mm) was not associated with a worse clinical important outcome. Age (< 65/> 65 years), gender and type of immobilization had no impact on the complication rate or in the final functional or radiological outcome.

Ventilation time and prognosis after stroke thrombectomy: the shorter, the better!

BACKGROUND AND PURPOSE: The aim was to investigate the clinical impact of the duration of artificial ventilation in stroke patients receiving mechanical thrombectomy (MT) under general anaesthesia. METHODS: All consecutive ischaemic stroke patients who had been treated at our centre with MT for anterior circulation large vessel occlusion under general anaesthesia were identified over an 8-year period. Ventilation time was analysed as a continuous variable and patients were grouped into extubation within 6 h ('early'), 6-24 h ('delayed') and >24 h ('late'). Favourable outcome was defined as modified Rankin Scale scores of 0-2 at 3 months post-stroke. Pneumonia rate and reasons for prolonged ventilation were also assessed. RESULTS: Amongst 447 MT patients (mean age 69.1 ± 13.3 years, 50.1% female), the median ventilation time was 3 h. 188 (42.6%) patients had a favourable 3-month outcome, which correlated with shorter ventilation time (Spearman's rho 0.39, P < 0.001). In patients extubated within 24 h, early compared to delayed extubation was associated with improved outcome (odds ratio 2.40, 95% confidence interval 1.53-3.76, P < 0.001). This was confirmed in multivariable analysis (P = 0.01). A longer ventilation time was associated with a higher rate of pneumonia during neurointensive care unit/stroke unit stay (early/delayed/late extubation: 9.6%/20.6%/27.7%, P < 0.01). Whilst stroke-associated complications represented the most common reasons for late extubation (>24 h), delayed extubation (6-24 h) was associated with admission outside of core working hours (P < 0.001). CONCLUSIONS: Prolonged ventilation time after stroke thrombectomy independently predicts unfavourable outcome at 3 months and is associated with increased pneumonia rates. Therefore, extubation should be performed as early as safely possible.

Early complications and radiological outcome after distal radius fractures stabilized by volar angular stable locking plate.

INTRODUCTION: Distal radius fractures (DRF) are the most common fractures of the upper extremities. The incidence is expected to continue rising in the next years due to the increased life expectancy. Palmar locking plate stabilizing has since become the standard treatment for dorsally displaced DRF with a complication rate of 8-39% reported in the literature. Main aim of this study was to investigate the incidence of complications after DRF stabilization using palmar angular stable locking plate. METHODS: A retrospective medical records review conducted from January 2013 to December 2016 included a total of 392 patients with DRF, that were stabilized using palmar angular stable locking plate and showed a minimum follow-up of 3 months. The group comprised 259 female and 133 male patients with a mean follow-up interval of 11 months (range 3-52 months). All recorded complications were documented. Range of motion (ROM) in extension, flexion, supination, pronation, radial- and ulnar deviation of the last follow-up was noted. Age was divided into younger than 65 years (< 65 years) and older than 65 years (≥ 65 years). The primary, immediate postoperative and final checkup radiographs were scrutinized for alignment and intra-articular step-off. RESULTS: A total of 51 (13%) early and 17 late (4%) complications were recorded in 392 patients. The most common complications included carpal tunnel syndrome (3%), complex regional pain syndrome (3%) and loss of reduction (2%). Of the 68 complications, only 25 (6%) were directly related to the plate. 73% of all complications occurred in AO type C fractures. Patients without complications showed a significantly better ROM in extension, flexion, pronation and supination than patients with complications. No significant differences in incidence of complications, ROM or loss of reduction could be found between patients over and under 65 years of age. Gender and type of immobilization showed no significant influence on the complication rate. CONCLUSIONS: Stabilization of DRF by palmar angular stable locking plate is a safe form of treatment. In the majority of the cases a good clinical and radiological outcome with no complications was documented. Gender and type of immobilization had no impact on the complication rate and an age over 65 years is not associated with an increased risk for complications or restricted ROM.

Experimental search for the violation of Pauli exclusion principle: VIP-2 Collaboration.

The VIolation of Pauli exclusion principle -2 experiment, or VIP-2 experiment, at the Laboratori Nazionali del Gran Sasso searches for X-rays from copper atomic transitions that are prohibited by the Pauli exclusion principle. Candidate direct violation events come from the transition of a 2p electron to the ground state that is already occupied by two electrons. From the first data taking campaign in 2016 of VIP-2 experiment, we determined a best upper limit of [Formula: see text] for the probability that such a violation exists. Significant improvement in the control of the experimental systematics was also achieved, although not explicitly reflected in the improved upper limit. By introducing a simultaneous spectral fit of the signal and background data in the analysis, we succeeded in taking into account systematic errors that could not be evaluated previously in this type of measurements.

Serum netrin-1 in relation to gadolinium-enhanced magnetic resonance imaging in early multiple sclerosis.

BACKGROUND: Netrin-1, a secreted laminin-related protein, is known to regulate not only axonal guidance and neuronal cell migration, but also blood-brain barrier integrity and inflammation. Two preliminary studies reported altered serum netrin-1 levels in multiple sclerosis; however, associations with longitudinal clinical and magnetic resonance imaging activity have not been investigated. OBJECTIVES: We aimed to assess serum netrin-1 in multiple sclerosis and controls with respect to disease activity and its temporal dynamics. METHODS: Serum netrin-1 was assessed by enzyme-linked immunosorbent assay in 79 patients with clinically isolated syndrome or multiple sclerosis, and 30 non-inflammatory neurological disease controls. In patients, serum samples were collected immediately prior to gadolinium-enhanced 3 T magnetic resonance imaging at two time points (initial contrast-enhancing gadolinium+ n = 47, non-enhancing gadolinium- n = 32; reference gadolinium- n = 70; median time-lag 1.4, interquartile range 1.0-2.3 years). RESULTS: Serum netrin-1 levels were similar in clinically isolated syndrome, multiple sclerosis and controls, and gadolinium+ and gadolinium- patients. Among gadolinium+ patients, serum netrin-1 was decreased in clinically active (n = 8) vs non-active patients (n = 39; p = 0.041). Serum netrin-1 showed no temporal dynamics in multiple sclerosis and was unrelated to clinical data. CONCLUSIONS: Serum netrin-1 levels show no multiple sclerosis specific changes and are not sensitive for detection of subclinical disease activity. Netrin-1 changes during relapses may deserve further examination.

Cerebrospinal fluid lipocalin 2 in patients with clinically isolated syndromes and early multiple sclerosis.

BACKGROUND: Lipocalin 2 (LCN2) may be involved in the immunopathogenesis of multiple sclerosis (MS) and might further impact on iron homoeostasis. Brain iron accumulates in MS; however, the association to iron-related proteins is still unsolved. OBJECTIVE: To investigate cerebrospinal fluid (CSF) and serum LCN2, transferrin (Trf) and ferritin in early MS in relation to disease evolution and longitudinal brain iron accumulation. METHODS: We analysed CSF and serum LCN2 by enzyme-linked immunosorbent assay (ELISA) and Trf and ferritin by nephelometry in 55 patients (45 clinically isolated syndrome (CIS), 10 MS, median clinical follow-up 4.8 years) and 63 controls. In patients, we assessed sub-cortical grey matter iron by 3T magnetic resonance imaging (MRI) R2* relaxometry (median imaging follow-up 2.2 years). RESULTS: Compared to controls serum (p < 0.01), CSF (p < 0.001) LCN2 and CSF Trf (p < 0.001) levels were reduced in the patients. CSF LCN2 correlated with CSF Trf (r = 0.5, p < 0.001). In clinically stable patients, CSF LCN2 levels correlated with basal ganglia iron accumulation (r = 0.5, p < 0.05). In CIS, higher CSF LCN2 levels were associated with conversion to clinically definite MS (p < 0.05). CONCLUSION: We demonstrate altered LCN2 regulation in early MS and provide first evidence for this to be possibly linked to both clinical MS activity and iron accumulation in the basal ganglia.

Combined analysis of global and compartmental brain volume changes in early multiple sclerosis in clinical practice.

BACKGROUND: The extent and clinical significance of brain volume changes in different phases of multiple sclerosis (MS) is still under discussion. OBJECTIVE: To determine the rate of global and compartmental brain volume changes in patients with a clinically-isolated syndrome (CIS) compared to patients with definite MS, by long-term follow-up and as a predictor of conversion to MS in a routine clinical setting. METHODS: We investigated 120 patients (63 CIS and 57 MS) at baseline and after a mean follow-up period of 43 months, including detailed clinical examination and 3-Tesla magnetic resonance imaging (MRI). Our imaging analyses comprised the normalized brain volume (NBV), cortical grey matter (cGMV) and white matter (WMV) volumes using SIENA/X, the percentage of brain volume change (PBVC) using SIENA and the change in the volume of the thalami (TV) and basal ganglia (BGV). We also determined the amount and change of T2-lesion load (T2-LL). RESULTS: At baseline, all the brain volume metrics, except cGMV, were significantly lower; and the T2-LL was significantly higher, in patients with MS rather than CIS. During the follow-up, only the PBVC was higher in MS (p = 0.008) and this difference was driven by converters from CIS to MS. Quartiles of PBVC did not allow us to predict conversion to MS, but were associated with the degree of disability. CONCLUSIONS: PBVC is the most sensitive marker of progressing atrophy and a higher PBVC was generally associated with more active disease; however, it did not serve to predict the course of MS on an individual basis, in this study.

Differences and similarities in the evolution of morphologic brain abnormalities between paediatric and adult-onset multiple sclerosis.

BACKGROUND: Paediatric-onset multiple sclerosis (pMS) is multiple sclerosis (MS) occurring before the age of 18 years and may present and develop differently from adult-onset MS (aMS). Whether there are also differences regarding the accrual of brain changes is largely unknown. METHODS: We compared the evolution of the T2- and T1-lesion load (LL), the black hole ratio (BHR), and annualised brain volume change (aBVC) between 21 pMS patients (age at onset: 14.4±2.3 years) and 21 aMS patients (age at onset: 29.4±6.5 years) matched for disease duration (pMS: 1.0±1.8 years; aMS: 1.6±1.7 years, p=0.27). Follow-up was for 4.2±3.7 years in pMS and 3.1±0.6 years in aMS. Clinical comparisons included the course of disability assessed with the Expanded Disability Status Scale (EDSS) score and annualised relapse rate (ARR). RESULTS: At baseline, pMS and aMS had similar EDSS, T1-LL, BHR, whereas T2-LL was higher in aMS (aMS: 9.2±11.6 ccm; pMS: 4.1±6.2 ccm, p=0.02). The change of T2-LL and T1-LL during the observation period was similar in both groups. At follow-up, disability was lower in pMS (EDSS score in pMS: 0.9±0.9; aMS: 1.7±1.3, p=0.04), despite a significantly higher accrual of destructive brain lesions (BHR in pMS: 23.7±23.7%; aMS: 5.9±4.0%, p=0.02) and a similar rate of brain volume loss. CONCLUSION: Our observation of a morphologically more aggressive disease evolution paralleled by less disability in pMS than in aMS (defined using EDSS) suggests a higher compensatory capacity in pMS. This fact may obscure the need for treatment of pMS patients with disease modifying treatments (DMTs) based solely on clinical observation.

Predicting the severity of relapsing-remitting MS: the contribution of cross-sectional and short-term follow-up MRI data.

BACKGROUND AND OBJECTIVE: Predicting the long-term clinical course of multiple sclerosis (MS) is difficult on clinical grounds. Recent studies have suggested magnetic resonance imaging (MRI) metrics to be helpful. We wanted to confirm this. METHODS: Contactable individuals (N=84) from an initial 99 patients with relapsing-remitting MS (RRMS) who had undergone careful baseline and 2-year follow-up examinations including MRI were reassessed after a mean of 10.8±2.7 years. We investigated using multivariate linear regression analyses if clinical and MRI data obtained at the prior time-points and the rates of change in morphologic variables over a mean observational period of 2.5 years could have served to predict a patient's MS severity score (MSSS) 11 years later. Conversion to secondary progressive MS (SPMS) was a further outcome variable. RESULTS: In univariate analyses, the 'black hole ratio' (BHR) at baseline (p=0.017, beta=0.148) and at first follow-up (p=0.007, beta= -0.154) was the only MRI parameter showing a significant correlation with the MSSS. In a multiple regression model, the independent predictive value of imaging variables became statistically non-significant and the latest MSSS was predicted primarily by the baseline EDSS (r (2)=0.28; p<0.001). The BHR at baseline explained 9.4% of variance of conversion to SPMS (p=0.033). Over the observational period the MSSS remained stable in patients remaining RRMS, but increased in converters to SPMS from 4.0 to 6.4. CONCLUSIONS: We failed to confirm a clear independent contribution of cross-sectional and short-term follow-up MRI data for the prediction of the long-term clinical course of MS. The MSSS is not a stable indicator of disease severity but may increase in converters to SPMS.

A foodborne norovirus outbreak due to manually prepared salad, Austria 2006.

BACKGROUND: Norovirus is increasingly being recognized as a leading cause of foodborne disease. Nevertheless, well documented foodborne outbreaks due to norovirus are rarely found in the literature. MATERIAL AND METHODS: A retrospective cohort study was conducted for identifying the source of a gastroenteritis outbreak. A total of 325 persons were identified as the at risk group. RESULTS: The overall attack rate was 56% (182/325). Of the four working days of possible foodborne exposure to norovirus (Monday till Thursday), Wednesday (risk ratio [RR]: 18.82; 95%CI 11.82-29.96) and Thursday (RR 2.14; 95%CI 1.65-2.79) turned out to be the most likely days on which infections with norovirus occurred. The day-by-day food specific cohort analyses yielded consumption of salad on Wednesday (adjusted RR 2.82; 95%CI 1.0-7.94) to be associated with highest risk of illness. The most likely source of food contamination is a kitchen assistant having prepared salad manually. She fell ill with symptoms of gastroenteritis on Wednesday during the early working hours and continued working. Human stool samples obtained from five out of six outbreak cases, including the sick kitchen assistant, were RT-PCR positive for norovirus genotype GGII.7 (Leeds-like). CONCLUSION: This foodborne norovirus outbreak underlines the drastic consequences of neglecting the rules of basic kitchen hygiene. Food handlers working despite manifest diarrhea or vomiting--often in fear of job loss--are a common cause of foodborne norovirus outbreaks.

Salmonella Enteritidis phage type 21 outbreak in Austria, 2005.

We report an outbreak of gastroenteritis due to Salmonella Enteritidis PT 21 associated with attending an annual traditional fair in a small Austrian village on 4 May 2005. The outbreak lasted from 4 to 8 May. Descriptive and analytical epidemiological investigations were conducted in order to determine the extent of the outbreak and to identify outbreak risk factors. Of the 115 persons who visited the fair, 85 persons fulfilled the criteria of an outbreak case (attack rate = 73.9%). Stool specimens from 52 patients, including two kitchen staff, were tested for salmonella, and 20 specimens were positive for Salmonella Enteritidis PT 21. The cohort study revealed mixed salad (which included potatoes) as the likely cause of the outbreak (RR: 10.4, 95%CI 2.8 - 39.1; P = < 0.001). The causative agent of the outbreak was cultured from the stock of eggs used at the fair and from all three drag swabs and one barn dust sample collected from the responsible egg laying flock. Molecular subtyping by pulsed-field gel electrophoresis of genomic DNA after XbaI digestion showed that isolates from eggs, from the flock and from humans were indistinguishable. We hypothesise that cross contamination from eggs to boiled potatoes occurred in the kitchen area, where raw eggs were handled by village residents preparing a traditional Viennese egg dressing. Unrefrigerated storage of peeled potatoes may have favoured bacterial growth. Eggs from small rural flocks of laying hens kept in a traditional 'natural' way should not be assumed to be salmonella-free.

Salmonella Enteritidis phage type 21 outbreak in Austria, 2005.

We report an outbreak of gastroenteritis due to Salmonella Enteritidis PT 21 associated with attending an annual traditional fair in a small Austrian village on 4 May 2005. The outbreak lasted from 4 to 8 May. Descriptive and analytical epidemiological investigations were conducted in order to determine the extent of the outbreak and to identify outbreak risk factors. Of the 115 persons who visited the fair, 85 persons fulfilled the criteria of an outbreak case (attack rate=73.9%). Stool specimens from 52 patients, including two kitchen staff, were tested for salmonella, and 20 specimens were positive for Salmonella Enteritidis PT 21. The cohort study revealed mixed salad (which included potatoes) as the likely cause of the outbreak (RR: 10.4, 95%CI 2.8 - 39.1; P=<0.001). The causative agent of the outbreak was cultured from the stock of eggs used at the fair and from all three drag swabs and one barn dust sample collected from the responsible egg laying flock. Molecular subtyping by pulsed-field gel electrophoresis of genomic DNA after XbaI digestion showed that isolates from eggs, from the flock and from humans were indistinguishable. We hypothesise that cross contamination from eggs to boiled potatoes occurred in the kitchen area, where raw eggs were handled by village residents preparing a traditional Viennese egg dressing. Unrefrigerated storage of peeled potatoes may have favoured bacterial growth. Eggs from small rural flocks of laying hens kept in a traditional 'natural' way should not be assumed to be salmonella-free.

PIASy, a nuclear matrix-associated SUMO E3 ligase, represses LEF1 activity by sequestration into nuclear bodies.

The Wnt-responsive transcription factor LEF1 can activate transcription in association with beta-catenin and repress transcription in association with Groucho. In search of additional regulatory mechanisms of LEF1 function, we identified the protein inhibitor of activated STAT, PIASy, as a novel interaction partner of LEF1. Coexpression of PIASy with LEF1 results in potent repression of LEF1 activity and in covalent modification of LEF1 with SUMO. PIASy markedly stimulates the sumoylation of LEF1 and multiple other proteins in vivo and functions as a SUMO E3 ligase for LEF1 in a reconstituted system in vitro. Moreover, PIASy binds to nuclear matrix-associated DNA sequences and targets LEF1 to nuclear bodies, suggesting that PIASy-mediated subnuclear sequestration accounts for the repression of LEF1 activity.

Timing of peritoneal dialysis catheter removal after pediatric renal transplantation.

BACKGROUND: A peritoneal dialysis (PD) catheter is in place at the time of kidney transplantation in children receiving PD. Removal of the catheter eliminates the risk of catheter-related infections. However, the patient benefits from leaving the catheter in place if dialysis is necessary posttransplantation. There is currently no consensus on the proper timing of PD catheter removal after kidney transplantation in children. OBJECTIVE: To identify the risks and benefits of an in-dwelling PD catheter after renal transplantation in children. DESIGN: Retrospective single-center study of infectious complications and posttransplantation PD catheter use in 31 renal transplantations in 26 children. RESULTS: Peritoneal dialysis catheters were used postoperatively in 13 of the 31 transplantations. In 12 instances the catheter was needed during the first month after transplantation, and 2 of the patients involved did not have a catheter in place when needed. Six catheter-related infections occurred in 5 patients posttransplantation, with only 1 infection taking place within 1 month after transplantation. CONCLUSION: Our data suggest that the need for catheter use occurs predominantly during the first month, while infectious complications usually happen later. This strongly suggests that PD catheters should not be removed until approximately 1 month after kidney transplantation.

Helix morphology changes in B-DNA induced by spontaneous B(I)<==>B(II) substrate interconversion.

Investigations of spontaneous, i.e. not forced, B-DNA's B(I)<==>B(II) substate transitions are carried out on the d(CGCGAATTCGCG)2 EcoRI dodecamer sequence using Molecular Dynamics Simulations. Analysis of the resulting transition processes with respect to the backbone angles reveals concerted changes not only for backbone angles epsilon, zeta, and beta, but also for the 5'-delta and 5'-chi angles. For alpha and delta inside the interconverting base step, a change is seen in short lived B(II) conformers. With respect to base morphology distinct changes are observed for buckle, propeller twist, shift, roll and twist, as well as x-displacement and tip. The base mainly involved in the changes is identified as the base preceding the interconverting phosphate. Altogether single B(I)<==>B(II) interconversions result only in local distortions represented by the larger spread of most parameters. Comparison of the atomic positional fluctuations derived from the simulation with those obtained from the static X-ray structure results in striking similarities.

Unexpected BII conformer substate population in unoriented hydrated films of the d(CGCGAATTCGCG)2 dodecamer and of native B-DNA from salmon testes.

Conformational substates of B-DNA had been observed so far in synthetic oligonucleotides but not in naturally occurring highly polymeric B-DNA. Our low-temperature experiments show that native B-DNA from salmon testes and the d(CGCGAATTCGCG)2 dodecamer have the same BI and BII substates. Nonequilibrium distribution of conformer population was generated by quenching hydrated unoriented films to 200 K, and isothermal structural relaxation toward equilibrium by interconversion of substates was followed by Fourier transform infrared spectroscopy. BI interconverts into BII on isothermal relaxation at 200 K, whereas on slow cooling from ambient temperature, BII interconverts into BI. Our estimation of the dodecamer's BI-to-BII conformer substate population by curve resolution of the symmetrical stretching vibration of the ionic phosphate is 2.4 +/- 0.5 to 1 at 200 K, and it is 1.3 +/- 0.5 to 1 between 270 and 290 K. Pronounced spectral changes upon BI-to-BII interconversion are consistent with base destacking coupled with migration of water from ionic phosphate toward the phosphodiester and sugar moieties. Nonspecific interaction of proteins with the DNA backbone could become specific by induced-fit-type interactions with either BI or BII backbone conformations. This suggests that the BI-to-BII substate interconversion could be a major contributor to the protein recognition process.