Depth-Sensing Indentation as a Micro- and Nanomechanical Approach to Characterisation of Mechanical Properties of Soft, Biological, and Biomimetic Materials

Classical methods of material testing become extremely complicated or impossible at micro-/nanoscale. At the same time, depth-sensing indentation (DSI) can be applied without much change at various length scales. However, interpretation of the DSI data needs to be done carefully, as length-scale dependent effects, such as adhesion, should be taken into account. This review paper is focused on different DSI approaches and factors that can lead to erroneous results, if conventional DSI methods are used for micro-/nanomechanical testing, or testing soft materials. We also review our recent advances in the development of a method that intrinsically takes adhesion effects in DSI into account: the Borodich-Galanov (BG) method, and its extended variant (eBG). The BG/eBG methods can be considered a framework made of the experimental part (DSI by means of spherical indenters), and the data processing part (data fitting based on the mathematical model of the experiment), with such distinctive features as intrinsic model-based account of adhesion, the ability to simultaneously estimate elastic and adhesive properties of materials, and non-destructive nature

Stress Alters Rates and Types of Loss of Heterozygosity in Candida albicans

Genetic diversity is often generated during adaptation to stress, and in eukaryotes some of this diversity is thought to arise via recombination and reassortment of alleles during meiosis. Candida albicans, the most prevalent pathogen of humans, has no known meiotic cycle, and yet it is a heterozygous diploid that undergoes mitotic recombination during somatic growth. It has been shown that clinical isolates as well as strains passaged once through a mammalian host undergo increased levels of recombination. Here, we tested the hypothesis that stress conditions increase rates of mitotic recombination in C. albicans, which is measured as loss of heterozygosity (LOH) at specific loci. We show that LOH rates are elevated during in vitro exposure to oxidative stress, heat stress, and antifungal drugs. In addition, an increase in stress severity correlated well with increased LOH rates. LOH events can arise through local recombination, through homozygosis of longer tracts of chromosome arms, or by whole-chromosome homozygosis. Chromosome arm homozygosis was most prevalent in cultures grown under conventional lab conditions. Importantly, exposure to different stress conditions affected the levels of different types of LOH events, with oxidative stress causing increased recombination, while fluconazole and high temperature caused increases in events involving whole chromosomes. Thus, C. albicans generates increased amounts and different types of genetic diversity in response to a range of stress conditions, a process that we term “stress-induced LOH” that arises either by elevating rates of recombination and/or by increasing rates of chromosome missegregation

Excitable Oil Droplets ‐ FRET Across a Liquid‐Liquid Phase Boundary

YesFRET forms the basis for energy transfer in biological systems and organisms and it has become an investigative tool in the analysis of protein‐protein interactions and in the study of semiconductors (SC). Until now, FRET has been restricted to the simultaneous presence of both components in the same phase. Here, we report on the first successful prototype demonstrating interfacial FRET. This innovative FRET between inorganic SC‐nanoparticles and illuminating protein chimeras takes place across an oil/water interface. As a ′proof of concept′ oil droplets were stabilized by hydrophobin‐derivatives in aqueous solution. These proteins possess the ability to attach fused functional domains close to an interface. Moreover, an optically active nanostructure directly docks to the hydrophobin at the oil/water interface. Due to its modular design, this signal amplification array has the potential to be exploited in numerous fields ranging from biosensors, biotechnology to medical applications

In middle-aged and old obese patients, training intervention reduces leptin level: A meta-analysis

BACKGROUND: Leptin is one of the major adipokines in obesity that indicates the severity of fat accumulation. It is also an important etiological factor of consequent cardiometabolic and autoimmune disorders. Aging has been demonstrated to aggravate obesity and to induce leptin resistance and hyperleptinemia. Hyperleptinemia, on the other hand, may promote the development of age-related abnormalities. While major weight loss has been demonstrated to ameliorate hyperleptinemia, obese people show a poor tendency to achieve lasting success in this field. The question arises whether training intervention per se is able to reduce the level of this adipokine. OBJECTIVES: We aimed to review the literature on the effects of training intervention on peripheral leptin level in obesity during aging, in order to evaluate the independent efficacy of this method. In the studies that were included in our analysis, changes of adiponectin levels (when present) were also evaluated. DATA SOURCES: 3481 records were identified through searching of PubMed, Embase and Cochrane Library Database. Altogether 19 articles were suitable for analyses. STUDY ELIGIBILITY CRITERIA: Empirical research papers were eligible provided that they reported data of middle-aged or older (above 45 years of age) overweight or obese (body mass index above 25) individuals and included physical training intervention or at least fitness status of groups together with corresponding blood leptin values. STATISTICAL METHODS: We used random effect models in each of the meta-analyses calculating with the DerSimonian and Laird weighting methods. I-squared indicator and Q test were performed to assess heterogeneity. To assess publication bias Egger's test was applied. In case of significant publication bias, the Duval and Tweedie's trim and fill algorithm was used. RESULTS: Training intervention leads to a decrease in leptin level of middle-aged or older, overweight or obese male and female groups, even without major weight loss, indicated by unchanged serum adiponectin levels. Resistance training appears to be more efficient in reducing blood leptin level than aerobic training alone. CONCLUSIONS: Physical training, especially resistance training successfully reduces hyperleptinemia even without diet or major weight loss

Ploidy variation in fungi: Polyploidy, aneuploidy, and genome evolution

The ability of an organism to replicate and segregate its genome with high fidelity is vital to its survival and for the production of future generations. Errors in either of these steps (replication or segregation) can lead to a change in ploidy or chromosome number. While these drastic genome changes can be detrimental to the organism, resulting in decreased fitness, they can also provide increased fitness during periods of stress. A change in ploidy or chromosome number can fundamentally change how a cell senses and responds to its environment. Here, we discuss current ideas in fungal biology that illuminate how eukaryotic genome size variation can impact the organism at a cellular and evolutionary level. One of the most fascinating observations from the past 2 decades of research is that some fungi have evolved the ability to tolerate large genome size changes and generate vast genomic heterogeneity without undergoing canonical meiosis

Neocentromeres Form Efficiently at Multiple Possible Loci in Candida albicans

Centromeres are critically important for chromosome stability and integrity. Most eukaryotes have regional centromeres that include long tracts of repetitive DNA packaged into pericentric heterochromatin. Neocentromeres, new sites of functional kinetochore assembly, can form at ectopic loci because no DNA sequence is strictly required for assembly of a functional kinetochore. In humans, neocentromeres often arise in cells with gross chromosome rearrangements that rescue an acentric chromosome. Here, we studied the properties of centromeres in Candida albicans, the most prevalent fungal pathogen of humans, which has small regional centromeres that lack pericentric heterochromatin. We functionally delimited centromere DNA on Chromosome 5 (CEN5) and then replaced the entire region with the counter-selectable URA3 gene or other marker genes. All of the resulting cen5Δ::URA3 transformants stably retained both copies of Chr5, indicating that a functional neocentromere had assembled efficiently on the homolog lacking CEN5 DNA. Strains selected to maintain only the cen5Δ::URA3 homolog and no wild-type Chr5 homolog also grew well, indicating that neocentromere function is independent of the presence of any wild-type CEN5 DNA. Two classes of neocentromere (neoCEN) strains were distinguishable: “proximal neoCEN” and “distal neoCEN” strains. Neocentromeres in the distal neoCEN strains formed at loci about 200–450 kb from cen5Δ::URA3 on either chromosome arm, as detected by massively parallel sequencing of DNA isolated by CENP-ACse4p chromatin immunoprecipitation (ChIP). In the proximal neoCEN strains, the neocentromeres formed directly adjacent to cen5Δ::URA3 and moved onto the URA3 DNA, resulting in silencing of its expression. Functional neocentromeres form efficiently at several possible loci that share properties of low gene density and flanking repeated DNA sequences. Subsequently, neocentromeres can move locally, which can be detected by silencing of an adjacent URA3 gene, or can relocate to entirely different regions of the chromosome. The ability to select for neocentromere formation and movement in C. albicans permits mechanistic analysis of the assembly and maintenance of a regional centromere

CNS targets of adipokines

This is the author accepted manuscript. The final version is available from American Physiological Society via the DOI in this record.Our understanding of adipose tissue as an endocrine organ has been transformed over the last twenty years. During this time a number of adipocyte-derived factors or adipokines have been identified. This paper will review evidence for how adipokines acting via the central nervous system (CNS) regulate normal physiology and disease pathology. The reported CNS-mediated effects of adipokines are varied and include the regulation of energy homeostasis, autonomic nervous system activity, the reproductive axis, neurodevelopment, cardiovascular function, and cognition. Due to the wealth of information available and the diversity of their known functions, the archetypal adipokines leptin and adiponectin will be the focused on extensively. Other adipokines with established CNS actions will also be discussed. Due to the difficulties associated with studying CNS function on a molecular level in humans, the majority of our knowledge, and as such the studies described in this paper, comes from work in experimental animal models; however, where possible the relevant data from human studies are also highlighted

New nano-hybrid material - built from CdSe/ZnS quantum dots and bacteriorhodopsin - with optoelectrical and biological applications : development and optical and structural characterisation

Ce travail est consacré à l’élaboration et à la caractérisation d’un nouveau matériau hybride présentant des propriétés de transfert d’énergie, à base de la protéine photochromique bactériorhodopsine (bR) acceptrice d’énergie et de boites quantiques (QDs) de CdSe/ZnS fluorescentes donneuses d’énergie. Le but est d’essayer d’améliorer et d’utiliser les propriétés photochromiques et photochimiques de la bR en utilisant un transfert d’énergie par résonancede Förster (FRET) qui peut exister avec des QDs semiconductrices. Ce nouveau matériau hybride pourrait servir dans divers domaines, tels que des commutateurs optiques ou lescellules photovoltaïques.Les propriétés physiques d’un tel composé et son efficacité dépendant fortement de sa structure, ce travail à consister à élaborer le complexe bR/QD et à le caractériser par microscopie à force atomique (AFM) et par spectrométries optiques (fluorescence,absorption) afin de mettre en évidence un éventuel effet FRET entre bR/QDs. Nous avons montré qu’en modifiant la charge de surface des QDs nous pouvions optimiser l’efficacité duFRET. Cet effet provient d’une modification des forces électrostatiques entraînant un arrangement des QDs différent à la surface des membranes en fonction de leur charge de surface. En renforçant la liaison bR/QD à l’aide de biotine et de streptavidine, nous avons atteint une efficacité maximale de FRET de 80 %.Lors de nombreuses expériences que nous avons menées, nous avons montré parAFM et dichroïsme circulaire que la présence de QDs accélère fortement le processus de monomérisation de la bR par du détergent ce qui pourrait avoir des applications en biologie.In this work, we built and characterized a new hybrid material with energy transfer properties made from photochromic protein Bacteriorhodopsin (bR) which is the energy acceptor and CdSe/ZnS fluorescent quantum dots (QDs) which are the energy donor. Our aim was to improve the photochromic and photo-physical bR properties by using a Förster resonance energy transfer (FRET) which should exist in presence of semiconductor QDs. This new hybrid material should have applications in many fields such as optical switching or photovoltaics.Since optical properties and efficiency of such a material are highly dependent of its structure, our work was to develop and characterize bR/QD complex by atomic force microscopy (AFM) and optical spectrometries (fluorescence, absorption) in order to demonstrate FRET between bR and QDs. By tuning QDs surface charges, we proved that we were able to optimize FRET, by optimization of the electrostatic interactions between bR and QDs. This effect was due to a better organization of QDs on the membrane surface when interactions were optimized. We also showed that when QD and bR are linked by biotin streptavidin link they revealed the maximum FRET efficiency of 80 %.During all these experiments, we showed that QDs could induce a quicker detergent monomerization of bR. This is supported by AFM images and circular dichroïsm measurements. This effect should find applications in biology