| 1. |
- Klionsky, Daniel J., et al.
(author)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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In: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Research review (peer-reviewed)abstract
- In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
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| 2. |
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| 3. |
- Danese, E., et al.
(author)
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Impact of the CYP4F2 p.V433M Polymorphism on Coumarin Dose Requirement: Systematic Review and Meta-Analysis
- 2012
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In: Clinical Pharmacology and Therapeutics. - : Springer Science and Business Media LLC. - 1532-6535 .- 0009-9236. ; 92:6, s. 746-756
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Research review (peer-reviewed)abstract
- A systematic review and a meta-analysis were performed to quantify the accumulated information from genetic association studies investigating the impact of the CYP4F2 rs2108622 (p.V433M) polymorphism on coumarin dose requirement. An additional aim was to explore the contribution of the CYP4F2 variant in comparison with, as well as after stratification for, the VKORC1 and CYP2C9 variants. Thirty studies involving 9,470 participants met prespecified inclusion criteria. As compared with CC-homozygotes, T-allele carriers required an 8.3% (95% confidence interval (CI): 5.6-11.1%; P < 0.0001) higher mean daily coumarin dose than CC homozygotes to reach a stable international normalized ratio (INR). There was no evidence of publication bias. Heterogeneity among studies was present (I-2 = 43%). Our results show that the CYP4F2 p.V433M polymorphism is associated with interindividual variability in response to coumarin drugs, but with a low effect size that is confirmed to be lower than those contributed by VKORC1 and CYP2C9 polymorphisms.
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| 4. |
- Anders, S., et al.
(author)
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European roadmap on superconductive electronics - Status and perspectives
- 2010
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In: Physica C: Superconductivity and its Applications. - : Elsevier BV. - 0921-4534. ; 470:23-24, s. 2079-2126
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Journal article (peer-reviewed)abstract
- For four decades semiconductor electronics has followed Moore's law: with each generation of integration the circuit features became smaller, more complex and faster. This development is now reaching a wall so that smaller is no longer any faster. The clock rate has saturated at about 3-5 GHz and the parallel processor approach will soon reach its limit. The prime reason for the limitation the semiconductor electronics experiences is not the switching speed of the individual transistor, but its power dissipation and thus heat. Digital superconductive electronics is a circuit- and device-technology that is inherently faster at much less power dissipation than semiconductor electronics. It makes use of superconductors and Josephson junctions as circuit elements, which can provide extremely fast digital devices in a frequency range - dependent on the material - of hundreds of GHz: for example a flip-flop has been demonstrated that operated at 750 GHz. This digital technique is scalable and follows similar design rules as semiconductor devices. Its very low power dissipation of only 0.1 mu W per gate at 100 GHz opens the possibility of three-dimensional integration. Circuits like microprocessors and analogue-to-digital converters for commercial and military applications have been demonstrated. In contrast to semiconductor circuits, the operation of superconducting circuits is based on naturally standardized digital pulses the area of which is exactly the flux quantum Phi(0). The flux quantum is also the natural quantization unit for digital-to-analogue and analogue-to-digital converters. The latter application is so precise, that it is being used as voltage standard and that the physical unit 'Volt' is defined by means of this standard. Apart from its outstanding features for digital electronics, superconductive electronics provides also the most sensitive sensor for magnetic fields: the Superconducting Quantum Interference Device (SQUID). Amongst many other applications SQUIDs are used as sensors for magnetic heart and brain signals in medical applications, as sensor for geological surveying and food-processing and for non-destructive testing. As amplifiers of electrical signals. SQUIDs can nearly reach the theoretical limit given by Quantum Mechanics. A further important field of application is the detection of very weak signals by 'transition-edge' bolo-meters, superconducting nanowire single-photon detectors, and superconductive tunnel junctions. Their application as radiation detectors in a wide frequency range, from microwaves to X-rays is now standard. The very low losses of superconductors have led to commercial microwave filter designs that are now widely used in the USA in base stations for cellular phones and in military communication applications. The number of demonstrated applications is continuously increasing and there is no area in professional electronics, in which superconductive electronics cannot be applied and surpasses the performance of classical devices. Superconductive electronics has to be cooled to very low temperatures. Whereas this was a bottleneck in the past, cooling techniques have made a huge step forward in recent years: very compact systems with high reliability and a wide range of cooling power are available commercially, from microcoolers of match-box size with milli-Watt cooling power to high-reliability coolers of many Watts of cooling power for satellite applications. Superconductive electronics will not replace semiconductor electronics and similar room-temperature techniques in standard applications, but for those applications which require very high speed, low-power consumption, extreme sensitivity or extremely high precision, superconductive electronics is superior to all other available techniques. To strengthen the European competitiveness in superconductor electronics research projects have to be set-up in the following field: - Ultra-sensitive sensing and imaging. - Quantum measurement instrumentation. - Advanced analogue-to-digital converters. - Superconductive electronics technology.
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| 5. |
- Arpaia, Riccardo, 1985, et al.
(author)
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Highly homogeneous YBCO/LSMO nanowires for photoresponse experiments
- 2014
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In: Superconductor Science and Technology. - : IOP Publishing. - 0953-2048 .- 1361-6668. ; 27:4
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Journal article (peer-reviewed)abstract
- By using nanolithography and a soft etching procedure, we have realized YBa2Cu3O7-x/La0.7Sr0.3MnO3 (YBCO/LSMO) nanowires, with cross sections down to 100 x 50 nm(2) that ensure the cover age of areas up to 10 x 30 mu m(2). The LSMO layer acts as a capping for YBCO, minimizing the degradation of the superconducting properties taking place during the patterning; moreover, as a ferromagnetic manganite, it is expected to accelerate the relaxation dynamics of quasiparticles in YBCO, making such a system potentially attractive for applications in superconducting ultrafast optoelectronics. The reproducibility of the values of the critical current densities measured in different devices with the same geometry makes our nanowires ideal candidates for photoresponse experiments. First measurements have shown a satisfactory photoresponse from YBCO/LSMO devices.
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| 6. |
- Bressan, E., et al.
(author)
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Experimental and computational investigation of Morse taper conometric system reliability for the definition of fixed connections between dental implants and prostheses
- 2014
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In: Proceedings of the Institution of Mechanical Engineers Part H-Journal of Engineering in Medicine. - : SAGE Publications. - 0954-4119 .- 2041-3033. ; 228:7, s. 674-681
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Journal article (peer-reviewed)abstract
- Nowadays, dental implantology is a reliable technique for treatment of partially and completely edentulous patients. The achievement of stable dentition is ensured by implant-supported fixed dental prostheses. Morse taper conometric system may provide fixed retention between implants and dental prostheses. The aim of this study was to investigate retentive performance and mechanical strength of a Morse taper conometric system used as implant-supported fixed dental prostheses retention. Experimental and finite element investigations were performed. Experimental tests were achieved on a specific abutment-coping system, accounting for both cemented and non-cemented situations. The results from the experimental activities were processed to identify the mechanical behavior of the coping-abutment interface. Finally, the achieved information was applied to develop reliable finite element models of different abutment-coping systems. The analyses were developed accounting for different geometrical conformations of the abutment-coping system, such as different taper angle. The results showed that activation process, occurred through a suitable insertion force, could provide retentive performances equal to a cemented system without compromising the mechanical functionality of the system. These findings suggest that Morse taper conometrical system can provide a fixed connection between implants and dental prostheses if proper insertion force is applied. Activation process does not compromise the mechanical functionality of the system.
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| 7. |
- Farina, R., et al.
(author)
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The bleeding site: a multi-level analysis of associated factors
- 2013
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In: Journal of Clinical Periodontology. - : Wiley. - 0303-6979. ; 40:8, s. 735-742
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Journal article (peer-reviewed)abstract
- Aim To evaluate the association between the probability of a sulcus/pocket to bleed on probing (BoP) and patient related as well as site-specific characteristics. Methods Data from 88960 sites were retrospectively derived from the clinical record charts of 601 adult patients. BoP (positive/negative) had been recorded at the initial periodontal visit after probing pocket depth (PPD) assessment. To analyse the influence of patient-, tooth- and site-related factors on the probability for a site to be BoP+, a logistic, 3-level model was built with BoP as the binary outcome variable. Results (i) The mean probability to be BoP+ for a site with PPD=3mm was 18%, and the log odds increased by 0.69 for each 1mm increment in PPD; (ii) a significantly higher risk for BoP+ was observed for inter-proximal versus approximal surfaces, posterior teeth versus anterior teeth, females versus males, while a significantly lower risk was observed for smokers versus non-smokers; (iii) when controlling for the significant covariates, different BoP+ probabilities could still be observed among sites in patients with a different susceptibility to BoP. Conclusions The probability of a site to be BoP+ was associated with either site-specific (i.e. PPD, tooth aspect, tooth type) or patient-related factors (i.e. gender, smoking status).
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| 8. |
- Gertow, Karl, et al.
(author)
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Identification of the BCAR1-CFDP1-TMEM170A Locus as a Determinant of Carotid Intima-Media Thickness and Coronary Artery Disease Risk
- 2012
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In: Circulation: Cardiovascular Genetics. - 1942-325X .- 1942-3268. ; 5:6, s. 656-665
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Journal article (peer-reviewed)abstract
- Background-Carotid intima-media thickness (cIMT) is a widely accepted marker of subclinical atherosclerosis. To date, large-scale investigations of genetic determinants of cIMT are sparse. Methods and Results-To identify cIMT-associated genes and genetic variants, a discovery analysis using the Illumina 200K CardioMetabochip was conducted in 3430 subjects with detailed ultrasonographic determinations of cIMT from the IMPROVE (Carotid Intima Media Thickness [IMT] and IMT-Progression as Predictors of Vascular Events in a High Risk European Population) study. Segment-specific IMT measurements of common carotid, bifurcation, and internal carotid arteries, and composite IMT variables considering the whole carotid tree (IMTmean, IMTmax, and IMTmean-max), were analyzed. A replication stage investigating 42 single-nucleotide polymorphisms for association with common carotid IMT was undertaken in 5 independent European cohorts (total n=11 590). A locus on chromosome 16 (lead single-nucleotide polymorphism rs4888378, intronic in CFDP1) was associated with cIMT at significance levels passing multiple testing correction at both stages (array-wide significant discovery P=6.75x10(-7) for IMTmax; replication P=7.24x10(-6) for common cIMT; adjustments for sex, age, and population substructure where applicable; minor allele frequency 0.43 and 0.41, respectively). The protective minor allele was associated with lower carotid plaque score in a replication cohort (P=0.04, n=2120) and lower coronary artery disease risk in 2 case-control studies of subjects with European ancestry (odds ratio [95% confidence interval] 0.83 [0.77-0.90], P=6.53x10(-6), n=13 591; and 0.95 [0.92-0.98], P=1.83x10(-4), n= 82 297, respectively). Queries of human biobank data sets revealed associations of rs4888378 with nearby gene expression in vascular tissues (n=126-138). Conclusions-This study identified rs4888378 in the BCAR1-CFDP1-TMEM170A locus as a novel genetic determinant of cIMT and coronary artery disease risk in individuals of European descent. (Circ Cardiovasc Genet. 2012;5:656-665.)
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| 9. |
- Gibaud, Thomas, et al.
(author)
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New routes to food gels and glasses
- 2012
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In: Faraday Discussions. - : Royal Society of Chemistry (RSC). - 1364-5498 .- 1359-6640. ; 158, s. 267-284
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Journal article (peer-reviewed)abstract
- We describe the possibility to create solid-like protein samples whose structural and mechanical properties can be varied and tailored over an extremely large range in a very controlled way through an arrested spinodal decomposition process. We use aqueous lysozyme solutions as a model globular protein system. A combination of video microscopy, small-angle neutron and X-ray scattering and reverse Monte Carlo modeling is used to characterize the structure of the bicontinuous network with two coexisting phases of a dilute protein solution and a glassy or arrested dense protein backbone at all relevant length scales. Rheological measurements are then used to determine the complex mechanical response of these protein gels as a function of protein concentration and quench temperature. While in particular the origin of the dependence of the mechanical properties on quench depth and concentration is not well understood currently, it seems ultimately connected to the particular bicontinuous structure of the arrested spinodal network created by the interplay between the early stage of a spinodal decomposition and the position of the glass line. We then generalize this behavior and discuss how this could open up new routes to prepare gel-like food systems with adjustable structural and mechanical properties. We present results from a first feasibility study where we use a depletion interaction caused by the addition of small non-adsorbing polymers to suspensions of casein micelles in order to create food gels with tunable structural and mechanical properties through an arrested spinodal decomposition process.
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| 10. |
- Kaspersen, Jørn D., et al.
(author)
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Generic structures of cytotoxic liprotides : nano-sized complexes with oleic acid cores and shells of disordered proteins
- 2014
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In: ChemBioChem (Print). - : Wiley-VCH Verlagsgesellschaft. - 1439-4227 .- 1439-7633. ; 15:18, s. 2693-2702
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Journal article (peer-reviewed)abstract
- The cytotoxic complex formed between alpha-lactalbumin and oleic acid (OA) has inspired many studies on protein-fatty acid complexes, but structural insight remains sparse. After having used small-angle X-ray scattering (SAXS) to obtain structural information, we present a new, generic structural model of cytotoxic protein-oleic acid complexes, which we have termed liprotides (lipids and partially denatured proteins). Twelve liprotides formed from seven structurally unrelated proteins and prepared by different procedures all displayed core-shell structures, each with a micellar OA core and a shell consisting of flexible, partially unfolded protein, which stabilizes the OA micelle. The common structure explains similar effects exerted on cells by different liprotides and is consistent with a cargo off-loading of the OA into cell membranes.
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