Business Plan of D&D fitness studio

Tato práce se zabývá podnikatelským subjektem D&D Fitness studio. Cílem této práce je zjistit, v jakém stavu se studio nachází, poskytnout podklady pro efektivnější řízení, identifikovat podněty ke zlepšení a pomoci se zjištění možností, vedoucích ke zvýšení zisku. Po představení podniku a hlavní podnikatelské náplně, cvičení Bungee workout, se práce zabývá analytickou částí. Nejprve je rozebrána pracovní síla a management, poté finanční stránka a marketingový mix. Vnější prostředí je rozebráno analýzou Porte-rových sil se zvláštním zaměřením na šetření zákazníků a konkurence. Výsledky zjištění shrnuje SWOT analýza. Následně byl vytvořen seznam návrhů, ze kterých bylo vybráno konkrétní řešení. Z něj byly nastaveny SMART cíle a finanční prognózou ověřena jejich relevance. Za své dvouleté působení zaznamenalo fitness studio jen pozvolný vývoj a pohybuje se okolo bodu zvratu. Byly doporučeny změny v přístupu k řízení a zavedení pravidelných kontrol. Dále by vedení mělo podrobněji prozkoumat navržené možnosti, jak zpeněžit studio v nevytížených časech, zajímat se o poskytování licencí partnerským podnikům, začít prodávat zboží a zvážit zvýšení cen.This thesis is concerned with business subject – D&D Fitness studio. The goal of the thesis is to uncover current state of the studio, produce materials to increase effectivity of management, suggest possible improvements and help with creating strategies, leading to increases in profit. After introduction of the studio, and core of the business, the exercise Bungee workout, the thesis transitions to its analytical part. Thesis analysis the workforce, management, finances, and marketing mix. The external environment is explored through Porter’s five forces analysis with emphasis on customers and compe-tition. Outputs are summarized in SWOT analysis. List of possible improvements was created, from which one specific solution was picked. Solution was transformed into SMART goals, that were verified by financial prognosis. During its two years of existence the studio functions on the edge of break-even point. Thesis provides recommenda-tions to change the style of management and to set up regular controlling mecha-nisms. Management should also consider opportunities to monetize less busy times of the studio, granting licences to new partners, selling goods, and increasing prices

Exceptionally inert lanthanide(III) PARACEST MRI contrast agents based on an 18-membered macrocyclic platform

[Abstract] We report a macrocyclic ligand based on a 3,6,10,13-tetraaza-1,8(2,6)-dipyridinacyclotetradecaphane platform containing four hydroxyethyl pendant arms (L1) that forms extraordinary inert complexes with Ln3+ ions. The [EuL1]3+ complex does not undergo dissociation in 1 M HCl over a period of months at room temperature. Furthermore, high concentrations of phosphate and Zn2+ ions at room temperature do not provoke metal-complex dissociation. The X-ray crystal structures of six Ln3+ complexes reveal ten coordination of the ligand to the metal ions through the six nitrogen atoms of the macrocycle and the four oxygen atoms of the hydroxyethyl pendant arms. The analysis of the Yb3+- and Pr3+-induced paramagnetic 1H NMR shifts show that the solid-state structures are retained in aqueous solution. The intensity of the 1H NMR signal of bulk water can be modulated by saturation of the signals of the hydroxy protons of Pr3+, Eu3+, and Yb3+ complexes following chemical-exchange saturation transfer (CEST). The ability of these complexes to provide large CEST effects at 25 and 37 °C and pH 7.4 was confirmed by using CEST magnetic resonance imaging experiments.Ministerio de Ciencia e Innovación; CTQ2011-24487Ministerio de Economía y Competitividad; CTQ2013-43243-

From farm to pharm: taking plant pharmaceuticals from research to production

Affordable production of pharmaceuticals with high potency and low side effects is a major challenge of the 21st century. Peptides are an emerging class of therapeutics that have the potential to marry the specificity and efficacy of protein drugs with the stability and membrane permeability of small molecule drugs. Although many peptides are amenable to chemical synthesis, their cost of production is high, as is the generation of waste products. Peptide production in plants has the potential to be a scalable, cost effective, and a less environmentally taxing alternative. Cyclotides, first discovered in Oldenlandia affinis, are a unique class of backbone cyclic peptides containing three stabilising disulfide bridges that form a knot-like structure. Their stability, and for some variants, the ability to traverse cellular membranes make them ideal candidates for pharmaceutical and agricultural applications. Even though highly constrained sterically, cyclotides are amenable to engineering by replacing native sequences with bioactive epitopes. In this thesis, cyclotide production strategies in plant cell suspensions are examined with a special focus placed on O.\ua0affinis as an archetypical cyclotide producer. Additional species investigated are Clitoria ternatea, Hybanthus enneaspermus, Nicotiana benthamiana, and Petunia hybrida. Insights into how cyclotides and their biosynthetic processing machinery are regulated in suspension plant cells are reported and provide first steps towards an affordable and environmentally friendly peptide production system in plants.Chapter\ua01 introduces the relevant scientific knowledge. The journey starts with a description of proteins and peptides, emphasising cyclotides and their synthesis. Then, current production strategies are compared with plant-based systems. The advantages and disadvantages of bioreactor setups suitable for medium to large scale production of cyclotides in plant cell suspensions are subsequently outlined. Finally, the importance of downstream processing is discussed. At the end of Chapter\ua01, the scope of this thesis is outlined to give an overview of the scientific questions addressed.Chapter\ua02 lays the experimental groundwork upon which the rest of this thesis is built. Protocols to establish C.\ua0ternatea, H.\ua0enneaspermus, N.\ua0benthamiana, O.\ua0affinis, and P.\ua0hybrida suspension cultures were developed and their cyclotide accumulation profiles presented. Cycloviolacin\ua0O2, a cyclotide previously reported only in the Violaceae, was discovered in O.\ua0affinis suspension cells and corroborated in the O.\ua0affinis leaf transcriptome. Two new cyclotides, kalata\ua0B22 and kalata\ua0B23, were characterized from O.\ua0affinis suspensions and were shown to have high sequence similarity to a group of cyclotides found in suspension cells of multiple plant families. Additionally, a strong time dependent effect was observed for cyclotide production in O.\ua0affinis suspension cells and was connected to reduced cyclotide mRNA expression. Cyclotide production was tissue specific in all tested species, an outcome with potential consequences for large scale production.Chapter\ua03 presents the scale-up of plant cell suspension cultures for cyclotide production. C.\ua0ternatea, H.\ua0enneaspermus, and O.\ua0affinis suspensions were cultivated in a rocking motion bioreactor with culture volumes up to 5\ua0L. Growth characteristics and biomass yields of all species were promising; however, contamination, sampling and consequential downregulation of cyclotides in suspension cultures presented both challenges and opportunities which are discussed in depth.Chapter\ua04 investigates several elicitors for boosting cyclotide production in plant cell suspensions. O.\ua0affinis, C.\ua0ternatea, and H.\ua0enneaspermus cultures were either immobilized or treated with methyl jasmonate or sodium chloride. Cyclotide production in C.\ua0ternatea and H.\ua0enneaspermus could not be enhanced, but their base level was already high. Cyclotide production in O.\ua0affinis suspension cells was low but was enhanced by treatment with 3\ua0mM sodium chloride and by cell immobilization.Chapter\ua05 explores Agrobacterium-mediated transformation of plant suspension cells for production of natural and engineered cyclotides. The first reported transformation of O.\ua0affinis was achieved in plant cell packs, which is a stacked filter cake of suspension cells devoid of liquid medium. Several parameters necessary for transformation of plant cell packs were identified, but the observed transformation rates varied greatly. Still, the insights gained will prove useful for research of cyclotide biosynthesis and production in O.\ua0affinis.Chapter\ua06 presents four protocols tested to enable cell cryo-banking of O.\ua0affinis, a feat necessary for large scale production in engineered cell lines. No positive results were observed, but future attempts might find this work a useful staring point.Chapter\ua07 summarizes the results, focused on plant cell suspensions and cyclotide production respectively, whilst they are put in context and possible research paths going forward are discussed. In conclusion, new species were brought into suspension, new cyclotides were discovered, new cyclotide production methods were explored and the first transformation of O.\ua0affinis is reported. The application of knowledge generated in this thesis by analysing cell suspension of cyclotide producing plants could lead to affordable and flexible production of pharmaceutical peptides in plants

Multiphonon-Quenching in near-infrared luminescent lanthanoid cryptates

In dieser Arbeit wurde die Lumineszenzlöschung (Quenching) durch Obertöne hochenergetischer C-H- und C-D-Oszillatoren innerhalb von im nahen Infrarotbereich emittierenden molekularen Lanthanoidkomplexen untersucht. Besonderer Fokus lag dabei auf der quantitativen Untersuchung der C-H- und C-D-Oszillatoren innerhalb von N,N'-dioxidhaltigen Tris(bipyridin)-Kryptaten. Diese Daten wurden im Kontext des Models von Ermolaev und Sveshnikova ausgewertet und ein Ansatz entwickelt, das so genannte Spectral Overlap Integral empirisch anzunähern. Zudem wurde gezeigt, dass das häufig angewendete Energy Gap Law nicht für die quantitative Beschreibung des Quenchings von Lanthanoidenlumineszenz durch hochenergetische Oszillatoren geeignet ist. Darüber hinaus konnte erstmals die radiative Lebenszeit innerhalb von molekularen Yb-Komplexen absichtlich gesenkt werden und durch zusätzliche Deuterierung des Liganden die bis dato höchste Gesamtquantenausbeute für einen molekularen Yb-Komplex erhalten werden