Miracle-dz/UniFAR-FTUs huggingface dataset:
dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression

Abstract Distinct epigenomic profiles of histone marks have been associated with gene expression, but questions regarding the causal relationship remain. Here we investigated the activity of a broad collection of genomically targeted epigenetic regulators that could write epigenetic marks associated with a repressed chromatin state (G9A, SUV39H1, Krüppel-associated box (KRAB), DNMT3A as well as the first targetable versions of Ezh2 and Friend of GATA-1 (FOG1)). dCas9 fusions produced target gene repression over a range of 0- to 10-fold that varied by locus and cell type. dCpf1 fusions were unable to repress gene expression. The most persistent gene repression required the action of several effector domains; however, KRAB-dCas9 did not contribute to persistence in contrast to previous reports. A ‘direct tethering’ strategy attaching the Ezh2 methyltransferase enzyme to dCas9, as well as a ‘recruitment’ strategy attaching the N-terminal 45 residues of FOG1 to dCas9 to recruit the endogenous nucleosome remodeling and deacetylase complex, were both successful in targeted deposition of H3K27me3. Surprisingly, however, repression was not correlated with deposition of either H3K9me3 or H3K27me3. Our results suggest that so-called repressive histone modifications are not sufficient for gene repression. The easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation.
Overhydration, underhydration, and total body sodium: A tricky “ménage a trois” in dialysis patients

Overhydration is a frequent complication in dialysis patients. It has been linked with hypertension, left ventricular hypertrophy, arterial stiffness, atherosclerosis uremic cardiomyopathy, and all‐cause mortality or cardiovascular morbidity. In addition, predialysis underhydration is also associated with increased risk of death in ESRD patients. In this context, the optimal evaluation of hydration status is a must. However, this mission is not easy or accurate. In the last 10 years, several new methods have been tested in dialysis patients, particularly bioimpedance and lung ultrasonography. The precise clinical value of these techniques in the daily care of hemodialysis patients is not obvious yet. Sodium is also an important piece of this puzzle. Salt intake and/or removal of sodium during dialysis are essential determinants of optimal hydration status. Recent studies have revealed that salt and water homeostasis is also dependent of tissue sodium storage—increased in hemodialysis patients. However, the significance of increased sodium tissue storage as a cardiovascular risk factor and the relationship between tissue sodium content and hard CV endpoint have not yet been elucidated yet.
Baculovirus expression of the nucleoprotein gene of measles virus and utility of the recombinant protein in diagnostic enzyme immunoassays

A recombinant baculovirus that expresses the nucleoprotein gene of measles virus (Edmonston vaccine strain) under the transcriptional control of the polyhedrin promoter was generated. The expressed protein (B-MVN) comigrated with the authentic viral nucleoprotein as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it was phosphorylated. The B-MVN protein proved to be reactive with monoclonal antibodies in radioimmunoprecipitations, and it was immunogenic, eliciting in mice antisera that recognized the native nucleoprotein. In addition, the B-MVN protein was evaluated as a replacement source of antigen for whole virus in enzyme immunoassays (EIAs) for detection of measles virus-specific immunoglobulin M (IgM) and IgG antibodies. A capture IgM EIA with the B-MVN protein as antigen detected specific IgM antibodies in 18 (72%) acute- and all convalescent-phase specimens from 25 clinical measles cases and exceeded 99% specificity with 120 control specimens. An indirect IgG EIA with the B-MVN protein detected specific IgG antibodies in 129 of 131 (98%) serum specimens with antibodies to measles virus, and results obtained from testing 268 additional serum specimens were better correlated with measles virus-neutralizing antibodies than those obtained with a commercial EIA.
EARLY LIFE HISTORIES, OCEAN CURRENTS, AND THE POPULATION GENETICS OF CARIBBEAN REEF FISHES

Tropical reef fishes, along with many benthic invertebrates, have a life cycle that includes a sedentary, bottom‐dwelling reproductive phase and a planktonic stage that occurs early in development. The adult benthic populations occupy disjunct, patchy habitats; the extent of gene flow due to dispersal of the planktonic life stage is generally unknown.
Impaired sympathetic baroreflexes in prehypertensive Dahl hypertension-sensitive rats.

Baroreflex responsiveness was assessed in Dahl rats that had been fed a low-salt diet to retard subsequent development of hypertension. After five weeks there were no differences in baseline pressures, but all responses elicited reflexly by elevating blood pressure with phenylephrine were less pronounced in hypertension-sensitive (DS) than in hypertension-resistant (DR) rats. Diminished responses included the following: acceleration in afferent aortic nerve activity, bradycardia, and inhibition of splanchnic and renal sympathetic nerve activity. By contrast, opposite responses elicited by lowering blood pressure with sodium nitroprusside did not differ significantly. Selective baroreflex attenuation (i.e. which was demonstrable only when blood pressure was elevated but not when it was lowered) in DS rats may have been due to differences either in pharmacology of the two drugs, or in sinoaortic vessel wall geometry. The baroreflex impairment shown here could represent a genetic predisposition that eventually elevates blood pressure in DS rats by increasing sympathetic vasomotor tone in the renal and splanchnic vascular beds.
Students' Satisfaction in Higher Education

Students’ satisfaction can be defined as a short-term attitude resulting from an evaluation of students’ educational experience, services and facilities. Earlier it was measured by common satisfaction frameworks but later higher education specify satisfaction models were developed. The objective of this review is to render all available constructive literature about students’ satisfaction with a sound theoretical and empirical background. Data were collected from refereed journals and conference papers, and are constructively analyzed from different point of views to filter a sound background for future studies. The first section of the paper discuss students’ satisfaction, satisfaction models and frameworks used by previous researchers around the world and second section explain the empirical findings of previous studies in real world context.
Exhaled nitric oxide corresponds with office evaluation of asthma control

Exhaled NO (ENO) has been studied as a noninvasive marker of airway inflammation, and has been shown to be elevated in asthma patients. The aim of this study was to investigate whether ENO measurements differ significantly between groups of asthmatic children with different disease control and to compare ENO measurements with the clinical assessment of asthma control. Seventy‐three children between 5–18 years old with a diagnosis of asthma were recruited. ENO was measured online during a slow vital capacity maneuver. The mean of three plateau NO levels was used for analysis. Baseline and postbronchodilator spirometry were performed. The assessment of disease control was based on the frequency of use of beta2‐agonists, occurrence of day‐ and nighttime asthma symptoms, and spirometry results. Twenty‐one children (group 1) had good asthma control. In 31 patients (group 2), asthma control was acceptable. In 21 patients (group 3), asthma was insufficiently controlled. ENO levels were (median (quartiles)): group 1, 11 ppb (9–21); group 2, 15 ppb (11–26); and group 3, 28 ppb (19–33). Measurements were significantly different between all three groups (P = 0.009, Kruskal‐Wallis), between groups 1 and 3 (P = 0.01, Mann‐Whitney U test), and between groups 2 and 3 (P = 0.01, Mann‐Whitney‐U test). The same was true for reversibility testing. We found significantly different ENO levels between a group of pediatric asthma patients with insufficient and good/sufficient control, as defined by clinical assessment. These results suggest that ENO measurements may be useful for monitoring asthma patients. Pediatr Pulmonol. 2003; 36:283–289. © 2003 Wiley‐Liss, Inc.
Bcl‐2 protein expression in carcinomas originating from the follicular epithelium of the thyroid gland

The bcl‐2 product has been related to the block of programmed cell death (apoptosis) both in lymphoid and in epithelial cells. The pathological expression of bcl‐2 has been investigated mainly in haematological malignancies. Here we have investigated bcl‐2 expression in a model of epithelial tumours represented by the spectrum of carcinomas arising from the follicular epithelium of the human thyroid gland. The analysis was carried out by immunocytochemistry on archival material using monoclonal antibodies against bcl‐2 and thyroglobulin (Tg) on consecutive sections of 94 well‐differentiated carcinomas (WDCs), 19 poorly differentiated carcinomas (PDCs), and 22 undifferentiated carcinomas (UCs) of the thyroid gland. In a subset of 5 cases of UC showing a differentiated component (UC‐D), the expression of p53 protein was also investigated. As controls, fetal and adult normal thyroid glands and adenomas were analysed. bcl‐2 expression was detected in 74 of 94 cases (78·7 per cent) of WDC, 16 of 19 cases (84·2 per cent) of PDC, and 3 of 22 cases (13·6 per cent) of UC. Simultaneous expression of bcl‐2 protein and Tg was observed in 74 of 94 cases (78·7 per cent) of WDC, 13 of 19 cases (68·4 per cent) of PDC, and in no case of UC. bcl‐2 and Tg immunostaining was detected in all fetal and normal thyroid glands as well as in the adenoma specimens examined. In the subset of UC‐D, mutual exclusion of bcl‐2 and p53 expression was observed in the undifferentiated and differentiated components (p53 but not bcl‐2 expressed in the former). Our data indicate that within the spectrum of thyroid carcinomas bcl‐2 expression, similarly to Tg, is strictly correlated with cell differentiation and that its abrogation is restricted to UCs, in which alterations of the p53 gene represent a late genetic event related to tumour progression.
Validity of the Dyslexia Screener

Optometrists see many patients who complain of reading difficulty. The Dyslexia Screener (TDS) is an instrument which has been developed to determine, within 5 min, whether or not a patient shows evidence of having one of the three basic types of dyslexia (a coding problem in reading and spelling). Results using TDS were compared with the Dyslexia Determination Test (DDT). TDS rating scores were shown to contain much of the same clinical information as the DDT rating scores
Cell density downregulates DNA synthesis and proliferation during osteogenesis in vitro

Abstract. The classical models of in vitro cell culture comprise fibroblasts and epithelial cells. Osteogenic cells represent another interesting cell model; however, it is not known whether during osteogenesis cell density regulates cell growth as seen in cultures of fibroblasts and epithelial cells. We selected MC3T3‐E1 cells for study because they are an osteogenic cell line that, when subcultured, grow to confluence and form multilayers of cells in conventional cultures by continued proliferation, as do fibroblasts. Once maximum cell density is obtained, proliferation is down regulated resulting in a mixed population of quiescent and dividing cells. We used this model to determine whether downregulation of proliferation as expressed by cell number and DNA synthesis is cell density‐dependent. MC3T3‐E1 cells were cultured over a period of 34 days to determine their kinetics, viability, ability to synthesize DNA, distribution within phases of the cell cycle and cell number‐response relationships. Our results show that (1) viability ranged between 92% and 96% and the cell number 2.5 x 105 per cm2 once cultures reached steady state, (2) most cells entered the G0/G1 phase of the cell cycle on day 7, (3) there was no correlation between the proportion of cells in S phase and downregulation of DNA synthesis, (4) a direct relationship exists between cell density and downregulation of DNA synthesis on day 8, (5) the minimum time for cells to be cultured before downregulation of DNA synthesis begins is independent of cell number, and (6) downregulation of DNA synthesis is reversible. These results suggest that density‐dependent downregulation of DNA synthesis may be a mechanism of growth control for osteogenic cells in vitro that operates more like density‐dependent growth control in cultures of fibroblasts rather than epithelial cells.
High power density Z-source resonant wireless charger with line frequency sinusoidal charging

Wireless charger for Electric Vehicles (EVs) is an off-line application and it needs power factor correction (PFC) function, which usually consists of a front-end boost PFC and a cascaded DC/DC converter. Z-source resonant converter (ZSRC), a single-stage solution with low cost and high efficiency, was proposed for EV wireless charger lately. The Z-source capacitors in the ZSRC are designed to absorb the double line frequency ripple in this single-phase application. Sinusoidal charging, which allows the double line frequency ripple to propagate to the output, is another solution aiming at reducing the bulky capacitors. Recently, a comparison of low frequency (120 Hz) sinusoidal charging and DC charging shows negligible impact on Li-ion batteries' performance. Therefore, ZSRC's capacitor, the biggest component, can be reduced dramatically from millifarad to several microfarads with sinusoidal charging technique, which features the ZSRC high power density. Also, it keeps the Z-source's benefit of boost ability and being immune to shoot-through problems. In this paper, the sinusoidal charging behavior for the ZSRC is modeled, and a control scheme that has both PFC function and load regulation for sinusoidal charging is proposed. It can pass IEC61000-3-2 Class A criterion. Experimental results based on a 1-kW prototype with 20-cm air gap between the primary and secondary side are presented to illustrate the proposed control scheme.
Eu Law as Private International Law? Reconceptualising the Country-of-Origin Principle as Vested-Rights Theory

If private international law is widely considered too technical to stir passions in the wider population, such considerations may have to be rethought. Recently, people all over Europe took to the streets to protest against a proposed norm of private international law – the “country-of-origin” principle stated in Article 16 of the proposed services directive.2 Had the proposal become law, providers of October 2006 Journal of Private International Law 195
Brain Acetylcholinesterase Activity in Alzheimer Disease Measured by Positron Emission Tomography

Brain acetylcholinesterase activity was measured in 14 patients with Alzheimer disease and 14 age-matched control subjects by positron emission tomography with a radioactive acetylcholine analogue. Kinetic analysis was performed to calculate k 3, an index of acetylcholinesterase activity. The k3 values were significantly reduced in the neocortex, hippocampus, and amygdala of all patients with Alzheimer disease, suggesting a loss of cholinergic innervation from the basal forebrain. Most profound reductions of k3 values were observed in the temporal (−30%) and parietal cortices (−31%), although reductions of k3 values were relatively uniform in the cerebral neocortex. This technique may be a powerful tool for early diagnosis of Alzheimer disease and also for therapeutic monitoring of acetylcholinesterase inhibitors in Alzheimer disease.
Myofibroblast Upregulators are Elevated in Joint Capsules in Posttraumatic Contractures

We hypothesized specific growth factors are increased in the elbow capsules of patients with post traumatic elbow contractures. A model of surgically induced joint contracture in rabbit knees was developed to study the growth factor expression in joint contractures. This study demonstrates this model mimics the human condition and analyzes how the growth factor levels decrease with time in rabbit knees with contractures. Reverse transcription polymerase chain reaction was used to measure mRNA levels of transforming growth factor-β1, connective tissue growth factor, ED-A of fibronectin, and α-smooth muscle actin normalized to a housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase. In the joint capsules of patients with elbow contractures, mRNA levels were increased for transforming growth factor-β1, connective tissue growth factor, and α-smooth muscle actin. In the joint capsules of rabbit knees with contractures, mRNA levels were increased for transforming growth factor-β1, connective tissue growth factor, ED-A of fibronectin, and α-smooth muscle actin. The mRNA levels for transforming growth factor-β1, connective tissue growth factor, and α-smooth muscle actin decreased with time in rabbit knees. The elevated levels of these myofibroblast upregulators and fibrogenic growth factors could explain the previously reported increase in myofibroblasts and collagen mRNA levels. The rabbit knee model correlated well with the human post traumatic elbow contractures.Level of Evidence: Level II, prospective comparative study. See the Guidelines for Authors for a complete description of levels of evidence.
Effects of capsule endoscopy on cardiac pacemakers.

BACKGROUND AND STUDY AIMS Capsule endoscopy is a new, noninvasive diagnostic technique which enables visualization of the mucosa of the small intestine in physiological conditions and without the need to subject the patient to external radiation. Wireless capsule video endoscopy is contraindicated in patients with a cardiac pacemaker. However, on the basis of the characteristics of the radiofrequency band used by the endoscopic capsule, together with a series of other factors, it is possible that the patient's use of a pacemaker should not be a contraindication to capsule endoscopy. PATIENTS AND METHODS This work had two phases: an in vitro study, using an interference detector, and an in vivo study involving 20 patients with cardiac pacemakers who showed symptoms or signs that justified a capsule endoscopy investigation. RESULTS No interference was observed during the first, in vitro, phase of the study. In the second phase, patients wore a Holter recorder for a mean time of 10 hours, during which the average number of recorded QRS complexes exceeded 30,000. All the pacemakers functioned normally. Interference due only to myopotentials was detected in a single patient in whom modification of the pacemaker programming was necessary. In the other 19 patients, neither sensing nor stimulation failures were observed. CONCLUSIONS No interference between the pacemaker and the endoscopic capsule was observed. All the pacemakers functioned normally, and no increased incidence of adverse effects was observed. Neither was any pacemaker-induced interference observed on the capsule endoscopy images.
Oligopeptide transport and regulation of extracellular proteolysis are required for growth of Aspergillus fumigatus on complex substrates but not for virulence

Moulds are characterized by their saprophytic lifestyle that is based on osmotrophy. Among them, Aspergillus fumigatus has emerged as the leading cause of fungal infections in the presence of an underlying immunodeficiency. To assess the role of its nutritional versatility for virulence, transcriptional profiling studies in the presence of varying sources of nitrogen were carried out and revealed an extensive reprogramming of the fungal transcriptome when shifting to a proteinaceous growth substrate. Transcripts encoding metabolic activities were predominantly upregulated, as were proteinases and transport activities. To probe whether fundamental aspects of its osmotrophic lifestyle, that is, extracellular proteolysis and uptake of oligopeptides, are required for A. fumigatus pathogenicity, serial gene replacements were carried out, which eventually yielded an octuple deletion mutant ablated for the opt gene family. This strain displayed no growth defect on various substrates, but supplementary reduction of extracellular proteolytic activity by additional deletion of the prtT gene revealed a synthetic phenotype on porcine lung tissue agar. Virulence studies in a murine model of pulmonary aspergillosis did not disclose any attenuation in virulence of these deletants. Our data emphasize a high degree of redundancy encoded by the A. fumigatus genome that secures nutrient supply for growth and, therefore, virulence.
Introduction of a heterologous editing site into the tobacco plastid genome: the lack of RNA editing leads to a mutant phenotype.

The psbF mRNA is edited in spinach plastids by a C to U conversion, changing a serine to a conserved phenylalanine codon. In tobacco at this position a phenylalanine codon is present at the DNA level, and the psbF mRNA here is not edited. To test if the psbF editing capacity is evolutionarily conserved, the tobacco psbF gene was modified to match the corresponding spinach sequence. The endogenous tobacco gene was replaced with the modified copy using biolistic transformation. We report here that the heterologous editing site remains unmodified in transplastomic tobacco plants. The lack of editing is associated with slower growth, lowered chlorophyll content and high chlorophyll fluorescence, a phenotype characteristic of photosynthetic mutants. This finding confirms that the editing of the psbF mRNA is an essential processing step for protein function and thus provides direct proof for the biological significance of plant organellar RNA editing. Given that a mutant phenotype is associated with the lack of editing, it seems likely that the evolutionary loss of the site‐specific capacity for psbF editing was preceded by the mutation that eliminated the editing requirement.
Young park users’ attitudes and behaviour to sun protection

The increase in skin cancer prevalence globally has prompted a range of health promotion sun safety initiatives. An area where evidence has been lacking is on the long-term impact of some of these initiatives on the attitudes and sun protection behaviour of young adults and of the sun protection measures used by people using city parks. This article disseminates a study that examined the knowledge, attitude and behaviour of 18- to 28-year-old Caucasian park users. An interview questionnaire was used with behaviour validation incorporated to corroborate the results and reduce recall bias. A cross comparison of answers and placement into pre-coded responses were made at regular intervals to ensure consistency of data collection. Knowledge of risks associated with sun exposure and knowledge of sun protection methods was high. The most common sources of knowledge on skin cancer prevention were parents and family, followed by television, then magazines and newspapers. Surprisingly, the citing of school sun safety health promotion initiatives as a source of knowledge was low. The vast majority of females and males felt that a suntan had aesthetic qualities and made them look more attractive and healthy. Only a small number of the participants’ sun protection behaviour in the park corresponded with their reported normal sun protection behaviour. Males in this study use sunscreen less than females. Females also used sunscreen with a higher sun protection factor. Seeking a tan is intentional behaviour undertaken by the majority of the participants, although females were more likely to seek a tan in comparison to males. The majority of participants had experienced sunburn in the summer period with some reporting severe sunburn. Recommendations are made for a gender specific health promotion approach, which targets familial education with a supportive environment in the school or public domain.