La réduction de l’expression de deux gènes codant pour la L-galactono-lactone-1,4-déshydrogénase (GalLDH) et de la GDP-D-mannose-3’,5’-épimérase (GME), enzymes de la voie de biosynthèse de l’ascorbate, a permis de mieux comprendre le rôle physiologique de ces enzymes chez la tomate. D’une part, l’étude de la GalLDH a mis en évidence la régulation complexe du métabolisme de l’ascorbate et la fonction essentielle de cette protéine au sein de la chaîne de transport des électrons au niveau mitochondrial. D’autre part, ce travail a révélé le rôle central de la GME à la fois pour la biosynthèse de l’ascorbate et la biosynthèse des polysaccharides pariétaux, notamment les mannanes et le rhamnogalacturonane II. Chez les plantes sous-exprimant la GME, nous avons pu noter l’incidence de perturbations de la structure pariétale sur les propriétés mécaniques des tiges et des fruits ainsi que sur la fécondation. Ces modifications ont notamment engendré une fragilité accrue des tiges et une stérilité partielle. La GME est donc déterminante pour la qualité nutritionnelle et organoleptique du fruit de tomate. Enfin, dans le cadre d’une approche de biologie intégrative, nos résultats associés aux données issues de plantes sous-exprimant des gènes codant pour des enzymes de la voie de recyclage de l’ascorbate chez la tomate ouvrent des perspectives originales pour l’approfondissement des connaissances sur la régulation et sur l’intégration du métabolisme de l’ascorbate dans le fonctionnement de la cellule. / Down-regulation of two genes encoding the L-galactono-1,4-lactone dehydrogenase (GalLDH) and the GDP-D-mannose-3',5'-epimerase (GME), enzymes of ascorbate biosynthesis pathway, led to a better understanding of the physiological role of these enzymes in tomato plants. On one hand, the study of GalLDH highlighted the complex regulation of ascorbate metabolism and the essential function of this protein in mitochondrial electron transport chain. Moreover, this work revealed the central role of the GME for both the ascorbate biosynthesis and the biosynthesis of cell wall polysaccharides, including mannans and rhamnogalacturonan II. In the GME-silenced plants, we found that modifications of the cell wall structure change the mechanical properties of stems and fruit as well as the fertilization. These changes led to an increase of stem fragility and to an increase of sterility. Therefore, GME plays a crucial role regarding the nutritional and organoleptic quality of tomato fruit. Finally, within the context of a systems biology approach, our results associated to datas obtained with plants silenced for recycling pathway related genes lead to the prospect to unravel the knowledges on the regulation and the integration of ascorbate metabolism in cell functions.

Thesis (Ph. D.)--University of Washington, 2001. / Vita. Includes bibliographical references (leaves 113-137).

京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第22877号 / 理博第4643号 / 新制||理||1667(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)准教授 秋山 秋梅, 教授 沼田 英治, 教授 曽田 貞滋 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM

[pt] A busca constante pela melhoria dos meios de comunicação em banda larga demandou o surgimento de novas tecnologias visando atender a real necessidade de seus usuários. O uso de mobilidade no acesso à internet banda larga como propõem os padrões WiMAX e LTE, impõe a necessidade de se estudar com mais profundidade os parâmetros que caracterizam um canal rádio móvel. Este trabalho objetiva apresentar os resultados experimentais que permitem caracterizar em banda estreita o comportamento do canal de radiopropagação num ambiente urbano. Como resultado das campanhas de medições, são identificados modelos do canal que possibilita ao projetista definir os melhores critérios para a implantação de uma rede móvel de acesso sem fio. Desta forma, são apresentadas duas campanhas de medições, operando nas frequências de 2,5 GHz e 3,5 GHz, destinadas para novos serviços móveis banda larga. / [en] The constant search for improvement of broadband communication systems requires new technologies to attend the increasing needs of the users. The use of mobility in broadband Internet access as proposed in WiMAX and LTE standards, imposes the need to further understand the parameters that characterize a channel mobile radio. This dissertation presents experimental results that allow characterizing the narrow band channel behavior of radio propagation in an urban environment. As a result of measurement campaigns, channel models are identified which allow the designer to define the best criteria to implement a mobile wireless network. The work presents results of two measurement campaigns, at the frequencies of 2.5 GHz and 3.5 GHz, designed for new mobile broadband services.

The discovery of cyclic AMP (cAMP), nearly 50 years ago by Sutherland radically altered the appreciation of metabolic regulation. Since then the presence of cAMP and its tremendous physiological impact has been demonstrated in many prokaryotic systems. In fact, virulence mechanisms of several pathogens known today exploit cAMP dependent pathways. Interestingly the genome of Mycobacterium tuberculosis H37Rv, the causative agent of tuberculosis, encodes as many as 16 adenylyl cyclases (enzymes that convert ATP to 3’, 5’-cAMP) and 10 cyclic-nucleotide binding proteins. Recent reports show that bacterial-derived cAMP manipulates host signaling for bacterial survival, suggesting an important role for cAMP in the pathogenesis of M. tuberculosis. A large number of non-pathogenic species of mycobacteria also share and conserve several of these cAMP metabolism genes, suggesting that cAMP is not only important for pathogenesis but also may play a critical physiological role in the genus. The work carried out in this thesis aims at a better understanding of the role of cAMP by studying the adenylyl cyclases and cyclic AMP receptor proteins (CRPs) from Mycobacterium smegmatis, a non-pathogenic member of the genus. Intracellular cAMP levels in a cell are precisely maintained by modulating the activities of the adenylyl cyclases (cAMP synthesising enzymes), the phosphodiesterases (cAMP hydrolysing enzymes) and the secretion machinery, if any. To assess the role of cAMP in mycobacteria, cAMP levels were measured in M. smegmatis during growth and under various stress conditions. The results show that cAMP levels peak at log phase of growth and decline thereafter. Under acidic conditions or on perturbing the cell-wall, cellular cAMP levels are altered, which indicate a possible role of cAMP in stress adaptation. Earlier work in our laboratory has led to the identification of multiple adenylyl cyclases in the mycobacterial genomes. These cyclases are similar in sequence to the mammalian enzymes and several of them have been shown to be active in vitro displaying a diverse range of biochemical properties. The M. smegmatis genome encodes 10 adenylyl cyclase-like genes. In order to understand the role of cAMP in M. smegmatis, individual cyclases were analysed for their biochemical properties and physiological functions. The work presented in this thesis is concerned with the functional characterization of MSMEG_3578 and MSMEG_3780, two of the several adenylyl cyclases from M. smegmatis. MSMEG_3578 encodes for a protein that comprises two transmembrane domains, an extracellular receptor-like domain, a membrane anchoring HAMP domain and an intracellular cyclase domain. The cyclase domain is closely related to mammalian cyclases but lacks the canonical residues that are critical for the catalysis of class III cyclases. Interestingly, the stop codon of this gene overlaps with the start codon of the downstream gene, MSMEG_3579 (a putative cyclic nucleotide gated mechanosensitive ion channel), suggesting a functional link between the two genes. The conservation of this gene pair across the mycobacterial genus indicates the importance of this putative receptor-effector pair in the physiology of mycobacteria. Additionally, microarray analysis by various groups have shown that this gene pair in Mycobacterium tuberculosis is differentially regulated in conditions that mimic stress the bacteria may experience during infection. In order to ascertain the physiological role of MSMEG_3578, a knock-out M. smegmatis strain was generated and tested for growth and cAMP defects. The knock-out strain showed growth and cAMP profiles similar to the wild-type strain. Over-expression of MSMEG_3578 in M. smegmatis resulted in a significant rise in cAMP levels. Interestingly, over-expression of the MSMEG_3578 adenylyl cyclase in E. coli did not lead to an elevation in cAMP levels indicating that other mycobacterial proteins may be required for the activity of MSMEG_3578 in vivo. In agreement with this, the purified adenylyl cyclase domain of MSMEG_3578 was found to be biochemically inactive in vitro. Additionally, the over-expressing strain has altered colony morphology as compared to the wild type strain. Perturbation of cAMP levels by over-expression of other cyclases also leads to a similar colony morphology phenotype, indicating the phenotype to be controlled by cAMP in general rather than by a specific cyclase in the cell. MSMEG_3780 is a highly conserved, biochemically active adenylyl cyclase, speculated to play a house-keeping function in M. smegmatis. Its orthologs from M. tuberculosis (Rv1647) and M. leprae (ML1399) have been biochemically characterized earlier in our laboratory. To unravel the role of this gene in vivo, the ∆MSMEG_3780 strain was tested for growth and cAMP defects under various conditions. The deletion strain did not show any difference in growth rate or morphology when compared to the wild-type strain. However it showed a reduction in intracellular cAMP levels at the log-phase of growth. Reintroduction of the MSMEG_3780 gene in the deletion strain restored cAMP to wild-type levels, thus indicating a crucial role for this adenylyl cyclase in the maintenance of intracellular cAMP levels during logarithmic growth. In order to investigate the regulation of the MSMEG_3780 gene, its promoter activity was tested under various stress-conditions. Acid-stress specifically resulted in the repression of the MSMEG_3780 promoter activity, a condition which also leads to a decrease in cAMP levels in the cells. Further evidence for the susceptibility of the MSMEG_3780 gene to acid-stress was obtained when the ∆MSMEG_3780 strain failed to reduce intracellular cAMP content upon sustained acid-stress conditions. Since Rv1647 shares similar biochemical properties with MSMEG_3780 and can also heterodimerize with the MSMEG_3780 protein in vitro, it was interesting to test whether the M. tuberculosis ortholog could functionally complement MSMEG_3780. To assess this, a complement strain was generated that contained the Rv1647 gene under the control of MSMEG_3780 promoter, integrated into the genome of ∆MSMEG_3780 strain. Rv1647 protein was able to restore the cAMP phenotype seen on acid stress as well as the cAMP levels in the mutant strain at the log phase of growth. This study indicated the role of cAMP and MSMEG_3780 in acid adaptation and also suggested a non-redundancy of adenylyl cyclases in mycobacteria, where different individual cyclases may have unique functions in the cells. Since Rv1647 could complement the cAMP defective phenotype in ∆MSMEG_3780, this suggests functional parallels between the proteins from the two species. Bacterial adaptation to environmental stress is brought about by a rapid change in its gene expression profile. Cyclic AMP plays an important role by binding to and activating a transcriptional factor, cAMP receptor protein or CRP. We have identified two CRPs from M. smegmatis, viz., MSMEG_0539 and MSMEG_6189 that possess high similarity at the amino acid level (78% overall sequence identity). The CRP ortholog from M. tuberculosis, Rv3676 has been characterized structurally, biochemically and functionally earlier. Western blot and RT-PCR analyses showed that CRPs in M. smegmatis were present during all phases of growth. Both the CRPs were cloned, expressed and shown to bind cAMP. Since the DNA binding domains of Rv3676 and the two M. smegmatis CRPs are nearly identical, the CRPs from M. smegmatis were predicted to bind similar target sequences. Interestingly, a CRP site was identified in the promoter of the MSMEG_3780 gene, suggesting a possible feed-forward or feed-back loop, where the enzymatic product of the adenylyl cyclase now governs its own gene expression. We performed Electrophoretic Mobility Gel Shift Assays (EMSAs) with M. smegmatis lysates to show that CRP binds to the MSMEG_3780 promoter at the CRP site. Subsequent Chromatin Immunoprecipitation (ChIP) assays confirmed that CRP binding to the MSMEG_3780 promoter occurred in vivo. To investigate the role of CRP in the regulation of the MSMEG_3780 gene, luciferase reporter assays with the wild-type and CRP site mutant promoters were carried out. Results suggest that CRP regulates the MSMEG_3780 gene under osmotic stress. However, CRP did not play any role in basal expression of MSMEG_3780 during growth. To assess which CRP of the two is functionally linked to the MSMEG_3780 promoter, we carried out a footprint assay with purified CRPs. It was intriguing to note that both the CRPs were in fact able to bind the promoter albeit under different conditions. Whereas MSMEG_6189 bound the promoter both in the presence and absence of cAMP, MSMEG_0539 bound the promoter only in the presence of cAMP. MSMEG_6189 thus deviates from the accepted CRP paradigm that seeks an absolute requirement of cAMP for specific DNA binding. The present study identifies cAMP as an important stress signal in M. smegmatis. Using MSMEG_3780 as a model gene, the role of cAMP in mycobacteria was studied. The two divergent CRPs that were characterized may function and dictate cAMP-mediated and perhaps independent functions in cells. Taken together, our results provide compelling evidence for the important role of cAMP in the general physiology and stress adaptation in M. smegmatis.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19599号 / 医博第4106号 / 新制||医||1014(附属図書館) / 32635 / 京都大学大学院医学研究科医学専攻 / (主査)教授 柳田 素子, 教授 妹尾 浩, 教授 浅野 雅秀 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No description available.

Différentes translocations génomiques sont fréquemment associées à l'apparition de leucémies myéloïdes aiguës (LMA). Ces translocations génomiques résultent de l’assemblage de deux gènes conduisant à la production d'une protéine de fusion. C'est le cas de la translocation t (3; 5) (q25.1; q34) impliquant le suppresseur tumoral NPM et l'oncogène MLF1 donnant naissance à la protéine de fusion NPM-MLF1. Généralement, les gènes impliqués dans ces translocations contrôlent la croissance cellulaire, la différenciation ou la survie cellulaire. Cependant, pour NPM-MLF1 les causes du gain ou de la perte de fonction associée à la translocation demeurent inconnues car nous ne savons pas comment cette translocation peut favoriser ou participer à l'avènement de la LMA. Le but de ce travail est d’analyser le rôle de NPM-MLF1 dans le cancer et d’examiner comment son activité contribue à la leucémie en faisant des études d’interactions protéine/protéine. En effet, l’étude de la fonction d’une protéine implique souvent de connaître ses partenaires d’interactions. Pour ce faire, la technique de double hybride dans la souche de levure AH109 a été utilisée. Tout d’abord, les ADN complémentaires (ADNc) de MLF1, NPM1 et de NPM-MLF1, MLF1-Like (une partie de MLF1 de l’acide aminé 94 à 157) normaux et mutés du domaine MTG8-Like constitué des acides aminés (a.a.) 151 à 164 de MLF1 (excepté NPM) ont été clonés dans un vecteur d'expression de levure pGBKT7. Les ADNc de GFI-1, mSin3A, PLZF, HDAC1 et HDAC3 ont été clonés dans le plasmide pGADT7 de façon à créer des protéines de fusion synthétiques avec le domaine de liaison à l'ADN et de trans-activation de la protéine GAL4. Le plasmide pGBKT7 possède un gène TRP1 et pGADT7 un gène LEU2 qui permettent la sélection des clones insérés dans la levure. Aussi, le pGBKT7 a un épitope c-myc et pGADT7 un épitope HA qui permet de voir l’expression des protéines par buvardage de type Western. Après la transformation des levures les interactions protéine/protéine ont été observées en vérifiant l’expression des gènes rapporteurs HIS3, LacZ, MEL1, ADE2 de la levure en utilisant des milieux de sélection YPD/-Leu/-Trp, YPD/-Leu/-Trp/-His, YPD/-Leu/-Trp/-His/-Ade, YPD/-Leu/-Trp/+ X-Gal, YPD/-Leu/-Trp/ + X-α-Gal. Ensuite, les interactions trouvées par double-hybride ont été vérifiées dans les cellules érythroleucémiques K562 par immuno-précipitation (IP) de protéines suivies de buvardages Westerns avec les anticorps appropriés. NPM-MLF1, MLF1, MTG8, MLF1-Like surexprimés dans les cellules K562 ont été clonés dans le plasmide pOZ-FH-N. pOZ-FH-N possède un récepteur IL-2 qui permet de sélectionner les cellules qui l’expriment ainsi qu’un tag Flag-HA qui permet de voir l’expression des protéines par buvardage-Western. Les résultats du double-hybride suggèrent une interaction faible de NPM-MLF1 avec HDAC1, HDAC3 et mSin3A ainsi qu’une interaction qui semble plus évidente entre NPM-MLF1 et PLZF, GFI-1. NPM interagit avec GFI-1 et mSin3A. Aussi, MLF1 et MLF1-Like interagissent avec HDAC1, HDAC3, GFI-1, PLZF mais pas avec mSin3A. Les IP suggèrent que NPM-MLF1 interagit avec HDAC1, HDAC3, mSin3A et PLZF. MLF1 et MLF1-Like interagissent avec HDAC1, HDAC3 et mSin3A. L’interaction de NPM-MLF1 avec GFI-1, MLF1 et MLF1-Like avec PLZF et GFI-1 n’a pas encore été vérifiée par IP. Ainsi, nos observations permettent de suggérer que NPM-MF1, MLF1 et NPM pourraient jouer un rôle dans la transcription et la régulation de l’expression de certains gènes importants dans l’hématopoïèse et une variété de processus cellulaires parce qu’ils interagissent avec différents corépresseurs. En déterminant les partenaires protéiques de MLF1, NPM et NPM-MLF1, leurs fonctions et comment NPM-MLF1 influence et modifie le fonctionnement cellulaire normal; il sera possible de renverser le processus de LMA favorisé par la t (3; 5) NPM-MLF1 par la technologie d’interférence à l’ARN. / Abstract Different genomic translocations are frequently associated with the development of acute myeloid leukemia (AML). Genomic translocations can result in the fusion of two genes leading to the formation of a fusion protein. This is the case of the T (3; 5) (q25.1; q34) translocation that implicates the tumour suppressor NPM1 and the oncogene MLF1, giving rise to the fusion protein NPM-MLF1. Generally the genes implicated in these translocations control cell growth, differentiation and survival. However, for NPM-MLF1 the reasons behind the gain or loss of function associated with the translocation are still unknown as we still ignore how this translocation can enhance or take part in the AML development. The goal of my master degree project was to analyse in part the role of NPM-MLF1 in cancer and to examine how its activity can contribute in leukemia through protein/protein interaction assays. The usual study of a protein function implicates the investigation of interacting partners. For this purpose, we used the yeast AH109 to conduct a two-hybrid screen assay. The MLF1, MLF1-Like (amino acid 94 to 157 of MLF1), NPM1 and NPM-MLF1 cDNAs, normal and mutated in the MTG8-Like domain from the amino acid (a.a.) 151 to 164 of MLF1 (with the exception of NPM1) were cloned into the yeast expression vector pGBKT7. The GFI-1, mSin3A, PLZF, HDAC1 and HDA3 cDNAs were cloned into the vector pGADT7. These clones were developed to creat synthetic fusion proteins with the DNA binding or trans-activation domain(s) of the protein Gal4. The pGBKT7 vector contains the TRP1 gene and the pGADT7 the LEU2 gene. These genes were used for the selection of the yeast clones transformed with the plasmids mentioned above. In addition, the pGBKT7 vector has c-myc-tag and the PGADT7 vector the HA-tag allowing the assessment of protein expression through Western Blot analysis. After yeast transformation, the protein/protein interaction were studied while verifying the expression of the reporter genes HIS, LacZ, MEL1, ADE while using the following selective medias YPD/-Leu/-Trp, YPD/-Leu/-Trp/-His, YPD/-Leu/-Trp/-His/-Ade, YPD/-Leu/-Trp/+ X-Gal, YPD/-Leu/-Trp/ + X-α-Gal. The interactions determined by the two-hybrid screening were verified in the erythroleukemic cells K562 using immuno-precipitation (IP) of the proteins followed by western blot using the appropriate antibodies. To achieve this, NPM-MLF1, MLF1, ETO, MLF1-Like cDNAs were cloned into the pOZ-FH-N vector that possess an IL2 receptor, which allows the selection of the positive transformed clones in the cell and a Flag-HA tag that permit the verification of protein expression through Western-blot. The two-hybrid screen results suggest that NPM-MLF1 interacts with HDAC1, HDAC3, PLZF, GFI and mSin3A. NPM interacts with GFI-1 and mSin3A. This has not been yet verified using the IP method. As in the case of MLF1, MLF1-Like interacts with HDAC1, HDAC3, GFI-1 and PLZF. However, no interaction was observed with Sin3A. The IP experiments suggest that NPM-MLF1 interacts with HDAC1, HDAC3, mSin3A and PLZF. MLF1 and MLF1-Like interact with HDAC1, HDAC3 and mSin3A. The interaction of NPM-MLF1 with GFI1 as well as MLF1 and MLF1-Like with PLZF and GFI-1 are not yet verified by IP. Therefore, our observations led to the suggestion that NPM-MLF1, MLF1 and NPM can play a role in the transcription and the regulation of the expression of certain genes that are important for hematopoiesis and a variety through the determination of the protein partners of MLF1, NPM and NPM-MLF1, their functions and how NPM-MLF1 influence/modify the normal cellular function, and by focusing on this study, it might become possible to reverse the AML process that is by the t(3;5) NPM-MLF1 while using the RNA interference technology.

No description available.

Målet med detta projekt har varit att utveckla en monteringsvänlig och kostnadseffektiv ljuddiffuser, en komponent som inte ska påverka ljudnivån och luftflödet för mycket, tillskillnad från den befintliga lösning som idag används i 3nine AB:s oljedimavskiljare. Examensarbetet följer den produktutvecklingsprocess som redogörs i boken Produktutveckling – Konstruktion och design av Karl T. Ulrich och Steven D. Eppinger. Där arbetet har anpassats för tidsramen på 10 veckor och delats upp i fyra faser. Fas1 – Förstudie, Fas 2 – faktainsamling, Fas 3 – Genomförande och Fas 4 – Rapportering. Den lösning som används idag består av fem vikta bitar sträckmetall som har sytts ihop med ståltråd, ljuddiffusern tar lång tid att montera ihop och att montera ned i maskinen. De fem vikta bitarna sträckmetall har vassa kanter efter klippning som försvårar monteringen ytterligare. En ljuddiffuser har en kostnad på 100 kr/st att framställa. För denna lösning togs mätvärden i 3nine AB:s verkstad fram som agerar som referensmätvärden, monteringstid – 333 [s], ljudnivå – 68 [dB], luftflöde – 319 [m ³/h] och DFA – index (mätvärde för monteringsvänlighet) – 5,4 %. Där 100 % ses som optimal monteringsvänlighet och högre DFA-index leder till reducerade kostnader. Då luft strömmar genom maskinen så påverkar detta mätvärdena och möjlig design av ny prototyper, men strömningslära är kunskaper som vi saknar och detta analyser med avseende på detta avgränsades bort. Genom Idéutvecklingsprocesser som Brainstorming, 6-3-5 Brainwritning, Morfologiskmatris, Pughmatris, konceptskisser, Virtuella koncept (3D-CAD) och friformsframställning (3D-utskrivning av prototyp) så togs fem prototyper fram som sedan testades för monteringstid, ljudnivå, luftflöde och DFA-index. Dessa tester resulterade i att det var en prototyp som utmärkte sig med förbättrade resultat jämfört med referensmätvärdena av befintliga ljuddiffusern. Mätvärden för prototypen ”45° väggen”, monteringstid – 16 [s], ljudnivå – 65 [dB], luftflöde – 342 [m ³/h] och DFA – index (mätvärde för monteringsvänlighet) – 93 %. Risk – och FEM-analys genomfördes på prototypen för att identifiera svagheter i konstruktionen, lösningar på dessa rekommenderas i form av små förändringar som t.ex. rundningar vid hörn. Dessa mätvärden redogör att den framtagna lösningen är bättre än dagens lösning och rekommenderas att implementeras och vidareutvecklas av företaget / The aim of this project has been to improve the existing noise diffuser used currently today in the oil-separatingmachines developed by 3nine AB. By reducing noise levels, increasing the air flow, increasing the “ease of assembly” and making it more cost effective. The thesis follows the product development process described in the book “Product Development - Construction and Design” by Karl T. Ulrich and Steven D. Eppinger. The work was adapted for a 10-week timeframe and divided into four phases. Phase 1 - Pre-Study, Phase 2 – Information gathering, Phase 3 - Implementation and Phase 4 - Reporting. The solution used today consists of five folded pieces of stretch metal that have been sewn together with steel wire, the noise diffuser takes a long time to assemble and to fit into the machine. The five folded pieces of stretch metal have sharp edges after cutting, which further complicates the assembly. The production cost for each diffuser is 100 kronor. For the present solution, the measurement values taken at 3nine AB's workshop were set as reference values, assembly time - 333 [s], noise level - 68 [dB], airflow - 319 [m³ / h] and DFA-index (measurement value for ease of assembly) - 5.4%. DFA-index when 100% is seen as the optimal ease of assembly and a higher DFA-index leads to reduced costs. As air flows through the machine, this affects the measured values and possible design of new prototypes, but fluid mechanics is one knowledge we lacked and therefor analysis of this was not possible and delimited. Through Idea Development Processes such as Brainstorming, 6-3-5 Brainwriting, Morphological Matrix, Pugh matrix, Concept Sketches, Virtual Concepts (3D-CAD) and Rapid prototyping (3D-prototype printing), five prototypes were produced, then tested for assembly time, noise level, airflow and DFA -index. These tests resulted in a prototype that featured improved results compared to the reference values of the existing noise diffuser. Measurement values for prototype "45° wall" where assembly time - 16 [s], noise level - 65 [dB], airflow - 342 [m³ / h] and DFA index - 93%. Risk-analysis and FEA was carried out on the same prototype to identify weaknesses in the design. The solutions to these weaknesses are recommended in the form of small design changes such as rounded sharp corners. These measured values state that the solution developed is better than today's solution and is recommended to be implemented and further developed by the company.