Особливості планування і реалізації проектів ресторанного бізнесу

Ресторанний бізнес є однією із найбільш значущих складових індустрії гостинності. Водночас, ресторанний бізнес, з одного боку, є одним із засобів високоліквідного використання капіталу, а з іншого − середовищем із високим ступенем конкурентності. У всьому світі він є одним із найбільш розповсюджених видів малого бізнесу, тому заклади та підприємства ведуть між собою постійну боротьбу за сегментацію ринку, за пошук нових та за утримання постійних споживачів їхньої продукції та послуг. Всі заклади та підприємства повинні мати високий рівень конкурентоспроможності та мати свою унікальність

Chaperone upregulation reduces aggregation analysed by immunohistochemistry and SDS PAGE.

<p>A) Confocal microscope images showing the reduction in aggregation of all TDP isoforms in the adult motor neuron cell bodies in the 10-day-old brain upon co-expression with CG14207 compared to flies not expressing CG14207, 10 days after induction of expression. TDP isoforms are detected by anti-HA staining (red) and nuclei by TOTO-3 staining (blue). Scale bars = 10 µm. B) Western blot detection of RIPA soluble and urea soluble fractions of TDP-43 from fly head homogenates in the presence or absence of the co-expression of CG14207 under reducing conditions, using an anti-HA antibody. β-actin was used as a loading control.</p

Neurotoxicity of TDP-43 expression in <i>Drosophila melanogaster</i>.

<p>A) Scheme of constructs used for expression. Both constructs have an HA-tagged N-terminus. The nuclear localisation sequence (NLS) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031899#pone.0031899-Winton1" target="_blank">[39]</a> is indicated by two black rectangles between residues 82 and 98, the RNA-recognition motifs (RRM) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031899#pone.0031899-Winton1" target="_blank">[39]</a> or RNA-binding domains (RRM 1, between residues 106 and 175, and RRM2, between 191 and 262) are indicated by two gray rectangles, and the nuclear export sequence (NES) is represented by the black rectangle between residues 239 and 250 within RRM 2. Arrowheads indicate caspase cleavage sites at residues 89 and 219 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031899#pone.0031899-Buratti1" target="_blank">[40]</a>. B) Optical and scanning electron microscope images demonstrating the effects of the TDP-43 and TDP-25 constructs when expression is driven in the photoreceptor neurons with <i>gmr-GAL4</i>. Colour pictures represent optical micrographs, taken at 7.5× magnification, of fly eyes and flies of the same genotype imaged using a scanning electron microscope at 200× magnification are shown in the adjacent picture. Scale bars represent 20 µm. C) Low and high magnification confocal microscopy images of imaginal eye discs of third instar larvae expressing TDP-43 or TDP-25 in the photoreceptors under the control of <i>gmr-GAL4</i>. TDP-43 and TDP-25 distribution is shown by anti-HA (red) staining, anti-ubiquitin (green) reveals ubiquitinated species, and TOTO-3 (blue) reveals nuclei. White arrowheads indicate aggregates, white arrows indicate the location of the morphogenetic furrow, and white boxes represent the area shown at higher magnification. Low magnification scale bars = 100 µm and high magnification scale bars = 10 µm.</p

Genetic upregulation of an endogenous <i>Drosophila</i> chaperone reduces toxicity of TDP-43 and its disease associated fragment.

<p>A) Light and scanning electron microscope images demonstrating the effects of the TDP-43 and TDP-25 constructs when expression is driven by <i>gmr-GAL4</i>, in the presence and absence of co-expression of the chaperone CG14207. Light micrographs taken at 7.5× magnification, of fly eyes and inset are flies of the same genotype imaged using a scanning electron microscope at 200× magnification. Adjacent are low and high magnification confocal microscopy images of imaginal eye discs of third instar larvae. TDP-43 and TDP-25 distribution is shown by anti-HA (red) staining. TOTO-3 (blue) detects nuclei. White arrowheads indicate aggregates. White arrows indicate the location of the morphogenetic furrows and white boxes represent the areas shown at higher magnification. Low magnification scale bars = 100 µm and high magnification scale bars = 10 µm. B) Western blot detecting soluble (RIPA soluble) and insoluble (9 M urea soluble) fractions of TDP-43 and TDP-25 from fly brain homogenates in the presence or absence of the co-expression of CG14207, under reducing conditions. β-actin was used as a loading control. C) Western blot detecting soluble (RIPA soluble) fractions under non-reducing conditions, using an anti-HA antibody. β-actin was used as a loading control. D) Light microscope images demonstrating the effects of the TDP-25 construct when expression is driven by a different <i>gmr-GAL4</i> driver, which generates expression to a greater extent than that used previously, in the presence and absence of the co-expression of CG14207. Light micrographs of <i>Drosophila</i> eyes taken at 7× magnification.</p

Chaperone upregulation has differential effects on neuronal dysfunction when TDP-43 is expressed in the adult motor neurons.

<p>A) Plot showing the Kaplan Meier survival data of TDP-43 and TDP-25 when expressed in adult motor neurons using a temperature sensitive Gal80 <i>D42</i>-<i>GAL4</i> driver compared to non-transgenic flies (WT). Lifespans were analysed by Kaplan-Meier statistics, n = 100 grouped into 10 tubes of 10 flies per genotype, differences between genotypes were analysed by Mann-Whitney U tests. B) Graph representing differences in motor function, analysed by climbing assay, when expressing TDP-43 and TDP-25 with and without CG14207 in adult motor neurons using the <i>Gal80-D42-GAL4</i> driver.</p

Selenium-Enhanced Electron Microscopic Imaging of Different Aggregate Forms of a Segment of the Amyloid β Peptide in Cells

The aggregation of misfolded proteins is a common feature underlying a wide range of age-related degenerative disorders, including Alzheimer’s and Parkinson’s diseases. A key aspect of understanding the molecular origins of these conditions is to define the manner in which specific types of protein aggregates influence disease pathogenesis through their interactions with cells. We demonstrate how selenium-enhanced electron microscopy (SE-EM), combined with tomographic reconstruction methods, can be used to image, here at a resolution of 5–10 nm, the interaction with human macrophage cells of amyloid aggregates formed from Aβ_25–36, a fragment of the Aβ peptide whose self-assembly is associated with Alzheimer’s disease. We find that prefibrillar aggregates and mature fibrils are distributed into distinct subcellular compartments and undergo varying degrees of morphological change over time, observations that shed new light on the origins of their differential toxicity and the mechanisms of their clearance. In addition, the results show that SE-EM provides a powerful and potentially widely applicable means to define the nature and location of protein assemblies <i>in situ</i> and to provide detailed and specific information about their partitioning and processing