Noncanonical effector functions of the T-memory-like T-PLL cell are shaped by cooperative TCL1A and TCR signaling.

T-cell prolymphocytic leukemia (T-PLL) is a poor-prognostic neoplasm. Differentiation stage and immune-effector functions of the underlying tumor cell are insufficiently characterized. Constitutive activation of the T-cell leukemia 1A (TCL1A) oncogene distinguishes the (pre)leukemic cell from regular postthymic T cells. We assessed activation-response patterns of the T-PLL lymphocyte and interrogated the modulatory impact by TCL1A. Immunophenotypic and gene expression profiles revealed a unique spectrum of memory-type differentiation of T-PLL with predominant central-memory stages and frequent noncanonical patterns. Virtually all T-PLL expressed a T-cell receptor (TCR) and/or CD28-coreceptor without overrepresentation of specific TCR clonotypes. The highly activated leukemic cells also revealed losses of negative-regulatory TCR coreceptors (eg, CTLA4). TCR stimulation of T-PLL cells evoked higher-than-normal cell-cycle transition and profiles of cytokine release that resembled those of normal memory T cells. More activated phenotypes and higher TCL1A correlated with inferior clinical outcomes. TCL1A was linked to the marked resistance of T-PLL to activation- and FAS-induced cell death. Enforced TCL1A enhanced phospho-activation of TCR kinases, second-messenger generation, and JAK/STAT or NFAT transcriptional responses. This reduced the input thresholds for IL-2 secretion in a sensitizer-like fashion. Mice of TCL1A-initiated protracted T-PLL development resembled such features. When equipped with epitope-defined TCRs or chimeric antigen receptors, these Lckpr-hTCL1Atg T cells gained a leukemogenic growth advantage in scenarios of receptor stimulation. Overall, we propose a model of T-PLL pathogenesis in which TCL1A enhances TCR signals and drives the accumulation of death-resistant memory-type cells that use amplified low-level stimulatory input, and whose loss of negative coregulators additionally maintains their activated state. Treatment rationales are provided by combined interception in TCR and survival signaling.

Prognostic significance of present atrial fibrillation on a single office electrocardiogram in patients with atrial fibrillation.

BACKGROUND: Evidence for the association of atrial fibrillation (AF) present on the ECG and cardiovascular outcomes in AF patients is limited. OBJECTIVE: To investigate the prognostic significance of AF on a single surface ECG for cardiovascular outcomes in AF patients. METHODS: A total of 3642 AF patients were prospectively enrolled. Main exclusion criteria were rhythms other than sinus rhythm (SR) or AF. The primary end-point was a composite of all-cause death and hospitalizations for congestive heart failure (CHF). Secondary end-points were all-cause death, CHF hospitalizations, cardiovascular death, myocardial infarction, any stroke and stroke subtypes. Associations were assessed with multivariable Cox proportional hazards models. RESULTS: Mean age was 71 years, 28% were female, and mean follow-up was 3.4 years. Patients with SR on the ECG at study enrolment (56%) were younger (69 vs. 74 years, P < 0.0001), had more often paroxysmal AF (73 vs. 18%, P < 0.0001) and fewer comorbidities. The incidence of the primary end-point was 1.8 and 3.1 per 100 person-years in patients with SR and AF, respectively. The multivariable-adjusted hazard ratio was 1.4 (95% confidence intervals 1.1; 1.7; P = 0.001) for patients with AF on the ECG compared to patients with SR. The hazard ratios (95% confidence intervals) were 1.4 (1.1; 1.8; P = 0.006) for all-cause death, 1.5 (1.2; 1.9; P = 0.001) for CHF and 1.6 (1.1; 2.2; P = 0.006) for cardiovascular death. None of the other associations were statistically significant. CONCLUSIONS: The presence of AF in a single office ECG had significant prognostic implications with regard to mortality and CHF hospitalizations in patients with AF. These patients present a high-risk group and might benefit from intensified treatment.

Evaluation of the inflammatory markers CCL8, CXCL5, and LIF in patients with anastomotic leakage after colorectal cancer surgery.

PURPOSE: Anastomotic leakage constitutes a dreaded complication after colorectal surgery, leading to increased morbidity and mortality as well as prolonged hospitalization. Most leakages become clinically apparent about 8 days after surgery; however, early detection is quintessential to reduce complications and to improve patients' outcome. We therefore investigated the significance of specific protein expression profiles as putative biomarkers, indicating anastomotic leakage. METHODS: In this single-center prospective cohort study serum and peritoneal fluid samples-from routinely intraoperatively inserted drainages-of colorectal cancer patients were collected 3 days after colorectal resection. Twenty patients without anastomotic leakage and 18 patients with an anastomotic leakage and without other complications were included. Protein expression of seven inflammatory markers in serum and peritoneal fluid was assessed by multiplex ELISA and correlated with patients' clinical data. RESULTS: Monocyte chemoattractant protein 2 (CCL8/MCP-2), leukemia-inhibiting factor (LIF), and epithelial-derived neutrophil-activating protein (CXCL5/ENA-78) were significantly elevated in peritoneal fluid but not in serum samples from patients subsequently developing anastomotic leakage after colorectal surgery. No expressional differences could be found between grade B and grade C anastomotic leakages. CONCLUSION: Measurement 3 days after surgery revealed altered protein expression patterns of the inflammatory markers CCL8/MCP2, LIF, and CXCL5/ENA-78 in peritoneal fluid from patients developing anastomotic leakage after colorectal surgery. Further studies with a larger patient cohort with inclusion of different variables are needed to evaluate their potential as predictive biomarkers for anastomotic leakage.

[Diagnostic strategy and differential therapeutic approach for cystic lesions of the pancreas]. / Diagnostische Strategie und differenzialtherapeutisches Vorgehen bei zystischen Läsionen des Pankreas.

CLINICAL/METHODICAL ISSUE: Cystic pancreatic lesions (CPL) are diagnosed with increasing frequency. Because up to 60% of CPL are classified as malignant or premalignant, every CPL should be fully investigated and clarified. Serous CPL with low risk of malignancy must be differentiated from mucinous CPL with relevant potential malignancy (intraductal papillary mucinous neoplasm IPMN) and mucinous cystic neoplasm (MCN) as well as from harmless pseudocysts. STANDARD RADIOLOGICAL METHODS: Cross-sectional imaging with computed tomography (CT) and magnetic resonance imaging (MRI) plays a crucial role in the diagnostics of CPL. METHODICAL INNOVATIONS: An algorithm for the differential diagnostic classification of CPL is presented. PERFORMANCE: The connection to the pancreatic duct is the key diagnostic criterion to differentiate IPMN from all other CPL. An exception to this rule is that pseudocysts can also show a connection to the pancreatic duct. A further classification of CPL with no connection to the pancreatic duct can be made by morphological criteria and correlation of the radiological findings with patient age, sex, history and symptoms. PRACTICAL RECOMMENDATIONS: Depending on the diagnosis and hence the malignant potential the indications for surgery or watch and wait have to be discussed in an interdisciplinary cooperation. Due to its higher soft tissue contrast MRI is often superior to CT for depiction of CPL morphology.

Risk of pancreatic fistula after enucleation of pancreatic tumours.

BACKGROUND: Enucleation is used increasingly for small pancreatic tumours. Data on perioperative outcome after pancreatic enucleation, especially regarding the significance and risk factors associated with postoperative pancreatic fistula (POPF), are limited. This study aimed to assess risk-dependent perioperative outcome after pancreatic enucleation, with a focus on POPF. METHODS: Patients undergoing enucleation for pancreatic lesions between October 2001 and February 2014 were identified from a prospective database. A detailed analysis of morbidity was performed. Risk factors for POPF were assessed by univariable and multivariable analyses. RESULTS: Of 166 enucleations, 94 (56.6 per cent) were performed for cystic and 72 (43.4 per cent) for solid lesions. Morbidity was observed in 91 patients (54.8 per cent). Severe complications occurred in 30 patients (18.1 per cent), and one patient (0.6 per cent) died. Reoperation was necessary in nine patients (5.4 per cent). POPF was the main determinant of outcome and occurred in 68 patients (41.0 per cent): grade A POPF, 34 (20.5 per cent); grade B, ten (6.0 per cent); and grade C, 24 (14.5 per cent). Risk factors independently associated with POPF were: cystic tumour, localization in the pancreatic tail, history of pancreatitis and cardiac co-morbidity. Only cystic morphology was independently associated with clinically relevant POPF (grade B or C), occurring after enucleation in 25 (27 per cent) of 94 patients with cystic tumours versus nine (13 per cent) of 72 patients with solid tumours. Tumour size and distance to the main duct were not associated with risk of POPF. CONCLUSION: Enucleation is a safe procedure in appropriately selected patients with a low rate of severe complications. POPF is the main determinant of outcome and is more frequent after the enucleation of cystic lesions.

Femtosecond time-resolved electronic relaxation dynamics in tetrathiafulvalene.

In the present paper, the ultrafast electronic relaxation of tetrathiafulvalene (TTF) initiated around 4 eV is studied by femtosecond time-resolved velocity-map imaging. The goal is to investigate the broad double structure observed in the absorption spectrum at this energy. By monitoring the transients of the parent cation and its fragments and by varying the pump and the probe wavelengths, two internal conversions and intramolecular vibrational relaxation are detected both on the order of a few hundred of femtoseconds. Photoelectron images permit the assignment of a dark electronic state involved in the relaxation. In addition, the formation of the dimer of TTF has been observed.

[Transoral approach for sublingual-plunging ranula]. / Transoraler Zugang bei sublingualer Tauchranula.

The case of a 7-year-old boy suffering from progressive submental/submandibular swelling is reported. Following clinical and imaging diagnostics (MRI), the suspected diagnosis of a sublingual-plunging ranula was made. Surgery was performed with transoral excision of the sublingual gland in combination with excision of the ranula. Additional submandibular gland excision should be avoided.

Adenosine A(3) receptor-induced proliferation of primary human coronary smooth muscle cells involving the induction of early growth response genes.

In human coronary smooth muscle cells adenosine A(2B) receptors mediate the inhibition of platelet-derived growth factor (PDGF)-induced proliferation via induction of the transcription factor nuclear receptor subfamily 4, group A, member 1 (NR4A1). In the absence of PDGF, adenosine analogues increased proliferation. In the present study we characterised the adenosine receptor mediating the increase in proliferation of these cells and identified involved transcription factors. Cultured human coronary smooth muscle cells were treated with selective A(3) receptor ligands. Effects on proliferation were determined by counting cells and measuring changes in impedance. The induction of transcription factors was assessed by qPCR. The A(3) receptor agonist 2-chloro-IB-MECA (2-chloro-N(6)-(3-iodobenzyl)adenosine-5'-N-methylcarboxamide) enhanced the number of human coronary smooth muscle cells with a half-maximal concentration of only 1 nM. 2-chloro-IB-MECA also increased the expression of the transcription factors early growth response protein (EGR)2 and EGR3, but not of EGR1, NR4A1, NR4A2 and NR4A3. The responses to 2-chloro-IB-MECA were blocked by two A(3) receptor antagonists, MRS1523 (3-propyl-6-ethyl-5[(ethylthio)carbonyl]-2-phenyl-4-propyl-3-pyridine-carboxylate; 10-300 µM) and VUF 5574 (N-(2-methoxyphenyl)-N'-[2-(3-pyridinyl)-4-quinazolinyl]-urea; 1-100 nM, as well as by the phospholipase C-inhibitor U73343 (0.2 µM). Small interfering RNA directed against EGR2 and EGR3 abolished the increases in proliferation induced by 2-chloro-IB-MECA. In summary, this is the first report demonstrating a coupling of smooth muscle adenosine A(3) receptors to increases in proliferation of human coronary smooth cells by the activation of phospholipase C and an induction of the transcriptions factors EGR2 and EGR3. The results facilitate the understanding of the role of adenosine A(3) receptors in the cardiovascular system.

Factor XIII Val34Leu polymorphism is associated with increased factor XIII activation and decreased transcutaneous oxygen readings in patients with diabetic foot ulcers.

AIM: To investigate the relationship between the factor XIII Val34Leu polymorphism, plasma factor XIII activation and transcutaneous oxygen readings in patients with diabetic foot ulcers. Methods Ninety-two consecutive patients with Type diabetes and active foot ulcers were investigated. Plasma factor XIII activation was assessed by ELISA. Genetic polymorphism was detected using the ABI PRISM® SNaPshotTM Multiplex Kit. Results are expressed as median (minimum-maximum). Differences between groups were calculated by Mann-Whitney U-test or χ(2) -test where appropriate. A P-value < .05 was considered significant. RESULTS: e Val34Leu polymorphism was found in 50 subjects (54%) with 42 being heterozygous (Val/Leu) and eight being homozygous (Leu/Leu). The rate of plasma factor XIII activation was increased in patients with Val/Leu or Leu/Leu [Val/Leu or Leu/Leu 152 (55-283) % vs. Val/Val 103 (33-282) %; P < 0.0001]. Transcutaneous oxygen readings were lower in the Val/Leu or Leu/Leu group [Val/Leu or Leu/Leu 16 (0-58) mmHg vs. Val/Val 35 (1-65) mmHg; P = 0.008]. Similarly, plasma factor XIII activation negatively correlated with transcutaneous oxygen readings (r(2) = -0.314; P = 0.014). CONCLUSIONS: Val34Leu polymorphism is associated with increased plasma factor XIII activation and seems to be linked with impaired cutaneous microcirculation in patients with diabetic foot ulcers.

Glitazone-like action of glimepiride and glibenclamide in primary human adipocytes.

AIM: Sulphonylureas (SUs) are among the most widely used oral hypoglycaemic drugs that stimulate insulin secretion. In addition, SUs have pleiotropic effects on other tissues. Conflicting findings have been reported regarding the effects of SUs on adipocytes. We have now investigated the actions of glimepiride and glibenclamide (=glyburide) in primary human adipocytes. METHODS: Primary cultured human white pre-adipocytes were differentiated in vitro according to a standard protocol. Lipid accumulation was assessed by Oil Red O staining and determination of triglyceride content; gene expression was measured by RT PCR and Western blotting. RESULTS: Initially, we characterized the genes regulated during human pre-adipocyte differentiation by performing global microarray analysis. Treatment with glimepiride and glibenclamide caused an increased accumulation of lipid droplets and triglycerides. In addition, genes involved in lipid metabolism were induced and chemokine expression was decreased. Interestingly, the effects of SUs were generally qualitatively and quantitatively similar to those of pioglitazone. In direct comparison, glibenclamide was more potent than glimepiride with respect to the induction of fatty acid binding protein 4 (FABP4) (EC(50) 0.32 vs. 2.8 µM), an important adipocyte marker gene. SU-induced differentiation was virtually completely blocked by the peroxisome proliferator-activated receptor γ (PPARγ)-antagonist T0070907 but not affected by diazoxide, indicating PPARγ activation by SUs. Repaglinide had no effect on adipogenesis, although it causes insulin liberation like SUs. CONCLUSIONS: In primary human pre-adipocytes, glibenclamide and glimepiride strongly induced differentiation, apparently by activating PPARγ. Thus, SUs but not repaglinide may be used to influence insulin resistance beyond their effect on insulin liberation.

Radiological evaluation of pancreatic cancer: What is the significance of arterial encasement >180° after neoadjuvant treatment?

PURPOSE: This study aimed to evaluate contrast-enhanced computed tomography (CE-CT) features for prediction of arterial tumor invasion in pancreatic cancer (PDAC) patients in the event of arterial encasement >180° after neoadjuvant (radio-)chemotherapy (NAT). METHODS: Seventy PDAC patients with seventy-five arteries showing encasement >180° after completion of NAT were analyzed. All patients underwent surgical exploration with either tumor resection including arterial resection, periadventitial dissection (arterial divestment) or confirmation of locally irresectable disease. CE-CT scans were assessed regarding tumor extent and artery-specific imaging features. The results were analyzed on a per-artery basis. Based on the intraoperative and histopathological findings, encased arteries were classified as either invaded or non-invaded. RESULTS: Eighteen radiologically encased arteries were resected; of these, nine had pathologic evidence for tumor invasion. In 42 encased arteries, the tumor could be removed by arterial divestment. In 13 patients with 15 encased arteries, the tumor was deemed technically irresectable. Median tumor size, length of solid soft tissue contact, and degree of circumferential contiguity by solid soft tissue along the artery in CE-CT were significantly lower in the non-invaded than in the invaded artery group (p ≤ 0.017). Imaging features showed moderate accuracies for prediction of arterial invasion (≤72.0 %). The thresholds ≤26 mm for post-NAT solid soft tissue contact and ≤270° for circumferential contiguity by solid soft tissue had high negative predictive values (≥87.5 %). CONCLUSION: Although post-NAT prediction of arterial invasion remains difficult, arteries with ≤270° contiguity by soft tissue and arteries with ≤26 mm length of solid soft tissue contact are unlikely to be invaded, with possible implications for surgical planning.

Assessing the aquatic toxicity and environmental safety of tracer compounds Rhodamine B and Rhodamine WT.

Tracer tests represent a well-established method for delineating key environmental processes in various media and engineered systems. Tracers like Rhodamine B and WT are frequently applied due to their strong fluorescence even at low concentrations.. However, due to a lack of ecotoxicological data, limit values for these tracers cannot be determined. This study fills this critical data gap by providing ecotoxicity data for Rhodamine B and WT using a battery of short-term standardized tests, including growth rate inhibition tests with algae (Raphidocelis subcapitata) and lethality tests using crustaceans (Daphnia magna) and zebrafish (Danio rerio) embryos, and estimating EQS for surface water. For Rhodamine B, the effective and lethal concentration (EC50 and LC50) -causing 50% toxicity were in the range of 14-24 mg/L. For Rhodamine WT, no statistically significant effects were observed (p<0.05) at the tsted concentrations (up to 91, 100 and 200 mg/L for algae, crustaceans and fish embryos, respectively). Thus for all tested organisms, Rhodamine B was more toxic than Rhodamine WT (more than 14 times more toxic for R. subcapitata, 5.6 times for D. magna, 15 times for D. rerio embryos,based on EC10 and LC10 values). These results signify that read-across assessments using ecotoxicity data obtained with Rhodamine B is not advisable for estimating the ecotoxicity of Rhodamine WT. The annual-average quality standard (AA-QS) and maximum allowable concentration quality standard (MAC-QS) for Rhodamine B were found to be 14 and 140 µg/L, respectively. For Rhodamine WT, the corresponding values were estimated to >91 µg/L (AA-QS) and >910 µg/L (MAC-QS). Hence, concentrations below 140 µg/L or 910 µg/L for Rhodamine B and WT, respectively, are not expected to pose a risk to aquatic freshwater life in the case of intermittent discharges, e.g. tracer experiments released in streams.

Nonspecific resistance to bacterial infections. Enhancement by ubiquinone-8.

A lipid fraction from Escherichia coli was extracted with apolar solvents and was found to protect mice from a number of experimental bacterial infections. The benzoquinone, ubiquinone-8, was isolated from this extract by high pressure liquid chromatography and identified as such by nuclear magnetic resonance and mass spectrometry. At a dose of 25 mg/kg this substance was found to provide complete protection against otherwise lethal infections with gram-negative and gram-positive bacteria in mice. Treatment was most effective when given intravenously 24 h before infection. In comparative studies, ubiquinone-8 had a clearly higher activity than ubiquinones-4, Q6, and Q10. A highly significant increase in the clearance rate of bacteria from the blood by the spleen and the liver of treated animals, correlated well with the protective effect of ubiquinone-8. The compound stimulated the ability of mouse macrophages to incorporate sheep erythrocytes and significantly increased the number of antibody-producing cells in spleens of mice.

Atmospheric carbonation reduces bioaccessibility of PAHs in industrially contaminated soil.

Sorptive Bioaccessibility Extraction (SBE) was used to monitor changes in accessibility of polycyclic aromatic hydrocarbons (PAHs) during storage of historically contaminated alkaline soil (Σ US EPA 16 + 2 further PAHs: 2452 ±â€¯69 mg kg-1, n = 3). While total concentrations of PAHs were rather stable during storage for 561 days at 4 °C, PAH accessibility declined by 95% due to atmospheric carbonation. The formation of carbonates was evidenced by an increase of inorganic soil carbon and by carbonate coatings on black soil particles (SEM-EDX) that could be dissolved by providing neutral to acidic soil conditions. Subjecting soil (252 days of storage) to biodegradation at pH 7 resulted in a degraded fraction of PAHs equivalent to the accessible PAH fraction of soil as received (PAHs with log Kow <5). The present study addresses important interactions and relationships between carbonation of soil, aging of PAHs in soil and related changes in PAH accessibility. A main finding was the reversibility of this retention mechanism, a changing environment (e.g. reduction of pH below 8) can result in a rise of accessible PAHs and consequently in an increase of exposure and associated risk.

Actionable perturbations of damage responses by TCL1/ATM and epigenetic lesions form the basis of T-PLL.

T-cell prolymphocytic leukemia (T-PLL) is a rare and poor-prognostic mature T-cell malignancy. Here we integrated large-scale profiling data of alterations in gene expression, allelic copy number (CN), and nucleotide sequences in 111 well-characterized patients. Besides prominent signatures of T-cell activation and prevalent clonal variants, we also identify novel hot-spots for CN variability, fusion molecules, alternative transcripts, and progression-associated dynamics. The overall lesional spectrum of T-PLL is mainly annotated to axes of DNA damage responses, T-cell receptor/cytokine signaling, and histone modulation. We formulate a multi-dimensional model of T-PLL pathogenesis centered around a unique combination of TCL1 overexpression with damaging ATM aberrations as initiating core lesions. The effects imposed by TCL1 cooperate with compromised ATM toward a leukemogenic phenotype of impaired DNA damage processing. Dysfunctional ATM appears inefficient in alleviating elevated redox burdens and telomere attrition and in evoking a p53-dependent apoptotic response to genotoxic insults. As non-genotoxic strategies, synergistic combinations of p53 reactivators and deacetylase inhibitors reinstate such cell death execution.

Phase I trial of oral MAC-321 in subjects with advanced malignant solid tumors.

PURPOSE: MAC-321 is a novel taxane that has demonstrated exceptional activity in human xenograft models when administered intravenously and orally. Preclinical studies of MAC-321 have shown antitumor activity in MDR-expressing and paclitaxel-resistant tumors. This phase I dose escalation study was performed to determine the safety, tolerability, and pharmacokinetic profile of orally administered MAC-321 given once every 21 days. Preliminary antitumor activity of MAC-321 was also examined. METHODS: Key eligibility criteria included adult subjects with refractory solid tumors or solid tumors for which conventional therapy was unsuitable or did not exist, good performance status (ECOG ( 2), and adequate hematologic, hepatic, and renal functions. Plasma pharmacokinetic (PK) sampling was performed during the first cycle of therapy. RESULTS: Five dose levels of MAC-321 ranging from 25 to 75 mg/m(2) were evaluated in 18 subjects (four women and 14 men). MAC-321 was well tolerated at the first three dose levels (25, 37, 50 mg/m(2)). Two subjects developed dose-limiting toxicities (DLTs) at 75 mg/m(2); one subject with grade 3 and one subject with grade 4 neutropenia with fever. Three subjects treated at an intermediate dose level of 60 mg/m(2) had no DLTs. However, the study was terminated prior to completion of the maximal tolerated dose cohort after subjects treated with intravenous MAC-321 in a concurrent study experienced life-threatening toxicities. Other common toxicities included grades 1-2 fatigue and grades 1-2 diarrhea. There was substantial interpatient variability in the PK parameters. MAC-321 was rapidly absorbed with a mean C (max) value of less than 1 h. Mean C (max) and AUC values generally increased in a dose-related manner. The median terminal phase elimination half-life was 45 h (range 20-228 h). Disease stabilization was seen in four subjects with the following tumors: mesothelioma (14 cycles), chondrosarcoma (12 cycles), small cell carcinoma (10 cycles), and prostate carcinoma (6 cycles). CONCLUSIONS: MAC-321 can be safely administered orally once every 21 days up to a dose of 60 mg/m(2). The major DLT was neutropenic fever. Four subjects had disease stabilization.

Towards a situation-and-user-aware multi-modal motorized toilet system to assist older adults with disabilities: A user requirements study.

In the recent decades state of the art technologies appeared in many areas to assist older adults with disabilities. However, one very essential activity of daily life, the toileting remained without any relevant development. The iToilet project of the European Union focuses on the development of an intelligent and motorized toilet system to enable independent toilet use for older adults with disabilities. To begin the development, the user requirements of end-users were assessed by means of focus group interviews and questionnaires. The survey was conducted in Austria and Hungary with the participation of 74 persons in total (41 subjects with movement disorders, 21 caregivers and 12 healthcare managers). From the interviews, the ranking of functions and features based on the number of their mentions was derived. The raw ranking was modulated by the average ratings from the questionnaires that resulted in the final list of priorities. Our results suggest that a safe and intelligent motorized toilet system should have foldable handrails on both sides (especially for wheelchair users), motorized height and tilt adjusting mechanism for the toilet bowl, fixed toilet paper holder on both sides and emergency recognition with call function. Simple operation, storage and retrieval of user specific settings including bowl height, and user identification were also deemed as very important features, while the possibility to control functions with gestures was valued rather low.

Targeting transcription-coupled nucleotide excision repair overcomes resistance in chronic lymphocytic leukemia.

Treatment resistance becomes a challenge at some point in the course of most patients with chronic lymphocytic leukemia (CLL). This applies to fludarabine-based regimens, and is also an increasing concern in the era of more targeted therapies. As cells with low-replicative activity rely on repair that triggers checkpoint-independent noncanonical pathways, we reasoned that targeting the nucleotide excision repair (NER) reaction addresses a vulnerability of CLL and might even synergize with fludarabine, which blocks the NER gap-filling step. We interrogated here especially the replication-independent transcription-coupled-NER ((TC)-NER) in prospective trial patients, primary CLL cultures, cell lines and mice. We screen selected (TC)-NER-targeting compounds as experimental (illudins) or clinically approved (trabectedin) drugs. They inflict transcription-stalling DNA lesions requiring TC-NER either for their removal (illudins) or for generation of lethal strand breaks (trabectedin). Genetically defined systems of NER deficiency confirmed their specificity. They selectively and efficiently induced cell death in CLL, irrespective of high-risk cytogenetics, IGHV status or clinical treatment history, including resistance. The substances induced ATM/p53-independent apoptosis and showed marked synergisms with fludarabine. Trabectedin additionally perturbed stromal-cell protection and showed encouraging antileukemic profiles even in aggressive and transforming murine CLL. This proof-of-principle study established (TC)-NER as a mechanism to be further exploited to resensitize CLL cells.

The fragile X tremor ataxia syndrome in the differential diagnosis of multiple system atrophy: data from the EMSA Study Group.

The recent identification of fragile X-associated tremor ataxia syndrome (FXTAS) associated with premutations in the FMR1 gene and the possibility of clinical overlap with multiple system atrophy (MSA) has raised important questions, such as whether genetic testing for FXTAS should be performed routinely in MSA and whether positive cases might affect the specificity of current MSA diagnostic criteria. We genotyped 507 patients with clinically diagnosed or pathologically proven MSA for FMR1 repeat length. Among the 426 clinically diagnosed cases, we identified four patients carrying FMR1 premutations (0.94%). Within the subgroup of patients with probable MSA-C, three of 76 patients (3.95%) carried premutations. We identified no premutation carriers among 81 patients with pathologically proven MSA and only one carrier among 622 controls (0.16%). Our results suggest that, with proper application of current diagnostic criteria, FXTAS is very unlikely to be confused with MSA. However, slowly progressive disease or predominant tremor are useful red flags and should prompt the consideration of FXTAS. On the basis of our data, the EMSA Study Group does not recommend routine FMR1 genotyping in typical MSA patients.

Solid phase DNA amplification: characterisation of primer attachment and amplification mechanisms.

Different chemical methods used to attach oligonucleotides by their 5'-end on a glass surface were tested in the framework of solid phase PCR where surface-bound instead of freely-diffusing primers are used to amplify DNA. Each method was first evaluated for its capacity to provide a high surface coverage of oligonucleotides essentially attached via a 5'-specific linkage that satisfyingly withstands PCR conditions and leaves the 3'-ends available for DNA polymerase activity. The best results were obtained with 5'-thiol-modified oligonucleotides attached to amino-silanised glass slides using a heterobifunctional cross-linker reagent. It was then demonstrated that the primers bound to the glass surface using the optimal chemistry can be involved in attaching and amplifying DNA molecules present in the reaction mix in the absence of freely-diffusing primers. Two distinct amplification processes called interfacial and surface amplification have been observed and characterised. The newly synthesised DNA can be detected and quantified by radioactive and fluorescent hybridisation assays. These new surface amplification processes are seen as an interesting approach for attachment of DNA molecules by their 5'-end on a solid support and can be used as an alternative route for producing DNA chips for genomic studies.