CO2-laserin soveltaminen WFU-paperin reunan leikkaamiseen

Tämän diplomityön tarkoitus oli selvittää CO2-laserin soveltuvuutta paperin reunan leikkaamiseen ennen liimapuristinta nopealle modernilleWFU-paperikoneelle. Merkittävimmät leikkauskokeet tehtiin leikkaamalla paperirataa todenmukaisissa olosuhteissa koekoneella. Koeleikkauksissa tutkittiin myös päällystettyjen paperilajien CO2-laserleikkausta. Paperin reunan koeleikkauksia suoritettiin leikkauspöytää vasten sekä telaa vasten. Leikkauspöytänä käytettiin korkeapainevesileikkaimelle suunniteltua leikkauspöytää, jota paranneltiin tämän työn yhteydessä paremmin laserleikkaukselle sopivaksi. Telaa vasten suoritetuilla leikkauksilla lasertehon tarve oli suurempi verrattuna pöytää vasten leikkaukseen samalla leikkausnopeudella ja samalla paperilajilla. Laserlaitteiston mitoittaminen oikein pelkkien arkkileikkauskokeiden perusteella ei ollut mahdollista. Myös rullilla tapahtuvia leikkauskokeita tarvittiin. Pöytää vasten tapahtuvaan leikkaamiseen tarvitaan 1000 W laserlaite paperin molemmille reunoille. Tämän tehoiset laserlaitteet riittävät kaikille Changshun PK 1:n paperilajeille niiden lajikohtaiseen maksimiajonopeuteen. Testatuista kolmesta polttovälistä antoi 5" polttoväli parhaat leikkaustulokset. Polttopisteen paikka voi vaihdella ± 0.75 mm paperin pinnasta vaikuttamatta silti merkittävästi lasertehon tarpeeseen. Laserleikatun paperin reuna oli hyvälaatuinen, eikä laserleikkaus aiheuttanut paperille epätoivottuja asioita kuten mustumista.

Kartonkikoneen tuotannonsuunnittelu jalostuslähtöisesti

Työn tarkoituksena oli selvittää monipope-menetelmän taustaa, menetelmän karkea tekninen toteutus, sovellettavuus kartonkikone KA1:n tuotantoon ja menetelmän mukanaan tuomat edut ja haitat, sekä ratkaisumallit mahdollisten tuotannollisten häiriötilanteiden ratkaisemista varten. Työssä selvitettiin myös tuotannon, tuotannon-suunnittelun, varastoinnin, asiakaspalvelun ja tuottavuuden tehostamista monipope-menetelmän avulla, sekä tuotannonsuunnittelun jalostuslähtöisyyden korostamista. Työn kirjallisuusosassa käsitellään lyhyesti toiminnanohjauksen, tuotannonsuunnittelun ja materiaalinhallinnan perusteiden lisäksi tuotannon-suunnittelussa hyödynnettävän tietojärjestelmän toimintaa ja kartonkitehtaan tuotantoprosessi kartonkikoneiden ja muovipäällystyskoneiden osalta. Kokeellisessa osassa perehdyttiin tuotannon ja tuotannonsuunnittelun tämän hetkiseen tilaan, selvitettiin monipope-menetelmän idea ja sen karkea tekninen toteutus ja tutkittiin monipopemenetelmän sovellettavuutta pohjautuen toteutuneeseen tuotantoon ja nykyisiin tuotannon rajoitteisiin. Välivarastointikapasiteetin rajoittaminen johti päällystyskapasiteetin uudelleen organisoimisen ja nykyisen kapasiteetin riittävyyden selvittämiseen koetrimmitysten avulla. Lisäksi selvitettiin uuden menetelmän avulla saavutettavaa tuotannon nousua erityisesti jatkojalostuksen osalta. Monipope-menetelmän sovellettavuustutkimuksissa havaittiin uuden menetelmän tuovan mukanaan lukuisia etuja ja säästöjä muun muassa materiaalikulujen alentumisen ja vähentyneiden työvaiheiden kautta. Kartonkikone KA1:n tuotteiden jatkojalostuksen tuotannon nousuksi määritettiin noin 2 %:ia ja tämän lisäksi uusi menetelmä yksinkertaistaisi ja helpottaisi koko kartonkitehtaan rullaliikennettä ja varastojen hallintaa. Menetelmä toisi uusia toimintamalleja KA1:n tuotannon häiriötilanteiden varalle, lisäisi tuotannonsuunnittelun joustavuutta ja helpottaisi myös asiakaspalvelun ja laaduntarkkailun kehittämistä. Tutkimuksen tuloksena voidaan todeta, että monipope-menetelmän tuomat edut ovat kiistattomat. Menetelmän soveltamisen mahdollisuuksia tulisikin miettiä jatkossa investointeja suunnitellessa, jotta menetelmän toteuttaminen kohtuullisin investointikustannuksin nykyisten tuotantoedellytysten lisäksi olisi mahdollista.

Sahalaitosten häiriömittauskäytännön yhtenäistäminen

Työn tavoitteena oli selvittää kahdeksan sahalaitoksen häiriöseurannan nykytila sekä selvittämisen jälkeen ehdottaa sahalaitoksille otettavaksi käyttöön yhtenäinen samoihin häiriön luokitus- ja kirjaamisperiaatteisiin perustuva mittauskäytäntö. Yhtenäisen käytännön myötä sahalaitoksilta saatavat häiriötiedot ovat sahalaitosten kesken vertailukelpoisia. Mittauskäytännön yhtenäistämisellä pyritään nostamaan tuotantolinjojen käytettävyyttä. Sahalaitosten häiriömittauskäytännöissä oletettiin olevan merkittäviä eroja. Työ suoritettiin lähettämällä kysymykset sähköpostilla etukäteen sahalaitoksille, jonka jälkeen käytiin sahalaitoksissa haastattelemassa häiriöasioiden kanssa tekemisissä olevia henkilöitä. Haastattelujen jälkeen sahalaitoksilta saatuja tietoja täydennettiin sähköpostin välityksellä. Näiden toimintatapojen lisäksi tutkittiin sahalaitoksilta saatuja häiriöraportteja ja syykoodilistoja sekä verrattiin tutkimuksessa satuja tuloksia aihetta käsittelevään kirjallisuuteen. Tutkimustulokset vahvistavat työn aloitusvaiheessa tehdyn oletuksen, jonka mukaan sahalaitosten häiriömittauskäytännöissä on merkittäviä eroja. Tutkimusta hyödyntäen asiantuntijatyöryhmä yhtenäisti sahalinjojen ja tasaamoiden pien-häiriöiden ja varsinaisten häiriöiden laukaisurajat sekä häiriöiden mittauspaikat. Myös syykoodilistat yhtenäistettiin. Tämän lisäksi muodostettiin yhtenäinen vertailuraportti, jotta sahalaitosten häiriötietojen vertaileminen on mahdollista.

Alumiinipainevalujen purseen- ja jäysteenpoiston kehitysmahdollisuudet

Valupurseiden ja jäysteiden poistaminen on osa alumiinipainevalujen tuotantoprosessia. Työssä on tutkittu käytössä olevien ja uusien menetelmien mahdollisuuksia taloudellisempaan tuotantoon. Purseiden ja jäysteiden poistamiseen käytettävien menetelmien lisäksi tutkimuskohteita ja ideoita on haettu muista metallien työstömenetelmistä. Valupurseiden ja jäysteiden määritelmiä, muodostumista ja luokittelua on esitelty laajasti. Menetelmien tutkimus on painottunut valupurseiden poistamiseen ja valun jälkeistä leikkaamista on tutkittu erityisesti sisäpuolisten muotojen työstämiseen käytettyjen pistintyökalujen kautta. Muotin ulostyöntötapin purseen poistaminen on ollut tärkeä asia menetelmien tutkimuksissa. Valupurseiden, leikkaus- ja koneistusjäysteiden poistamiseksi lastuavista työstömenetelmistä tutkittuja ovat koneistaminen koneistuskeskuksella, aventaminen, hiertopuhallus, suihkuhiertäminen, vesisuihkuleikkaus, ultraäänityöstö, harjaus, painehiertäminen, hiominen kohdistetuilla ja kohdistamattomilla menetelmillä. Myös terminen jäysteenpoistomenetelmä (TEM), kemiallinen työstö (ECM) ja laserleikkaus on otettu esiin tutkimuksessa. Työn tuloksena on näkemys tutkittujen menetelmien jatkokehitystarpeesta ja mahdollisuudesta soveltaa niitä sarjatuotantoon.

Characterisation of Human Neutral and Lysosomal alpha-Mannosidases

Neutral alpha-mannosidase and lysosomal MAN2B1 alpha-mannosidase belong to glycoside hydrolase family 38, which contains essential enzymes required for the modification and catabolism of asparagine-linked glycans on proteins. MAN2B1 catalyses lysosomal glycan degradation, while neutral α-mannosidase is most likely involved in the catabolism of cytosolic free oligosaccharides. These mannose containing saccharides are generated during glycosylation or released from misfolded glycoproteins, which are detected by quality control in the endoplasmic reticulum. To characterise the biological function of human neutral α-mannosidase, I cloned the alpha-mannosidase cDNA and recombinantly expressed the enzyme. The purified enzyme trimmed the putative natural substrate Man9GlcNAc to Man5GlcNAc, whereas the reducing end GlcNAc2 limited trimming to Man8GlcNAc2. Neutral α-mannosidase showed highest enzyme activity at neutral pH and was activated by the cations Fe₂+, Co2+ and Mn₂+, Cu2+ in turn had a strong inhibitory effect on alpha-mannosidase activity. Analysis of its intracellular localisation revealed that neutral alpha-mannosidase is cytosolic and colocalises with proteasomes. Further work showed that the overexpression of neutral alpha-mannosidase affected the cytosolic free oligosaccharide content and led to enhanced endoplasmic reticulum associated degradation and underglycosylation of secreted proteins. The second part of the study focused on MAN2B1 and the inherited lysosomal storage disorder α-mannosidosis. In this disorder, deficient MAN2B1 activity is associated with mutations in the MAN2B1 gene. The thesis reports the molecular consequences of 35 alpha-mannosidosis associated mutations, including 29 novel missense mutations. According to experimental analyses, the mutations fall into four groups: Mutations, which prevent transport to lysosomes are accompanied with a lack of proteolytic processing of the enzyme (groups 1 and 3). Although the rest of the mutations (groups 2 and 4) allow transport to lysosomes, the mutated proteins are less efficiently processed to their mature form than is wild type MAN2B1. Analysis of the effect of the mutations on the model structure of human lysosomal alpha-mannosidase provides insights on their structural consequences. Mutations, which affect amino acids important for folding (prolines, glycines, cysteines) or domain interface interactions (arginines), arrest the enzyme in the endoplasmic reticulum. Surface mutations and changes, which do not drastically alter residue volume, are tolerated better. Descriptions of the mutations and clinical data are compiled in an α-mannosidosis database, which will be available for the scientific community. This thesis provides a detailed insight into two ubiquitous human alpha-mannosidases. It demonstrates that neutral alpha-mannosidase is involved in the degradation of cytosolic oligosaccharides and suggests that the regulation of this α-mannosidase is important for maintaining the cellular homeostasis of N-glycosylation and glycan degradation. The study on alpha-mannosidosis associated mutations identifies multiple mechanisms for how these mutations are detrimental for MAN2B1 activity. The α-mannosidosis database will benefit both clinicians and scientific research on lysosomal alpha‑mannosidosis.

Aging based maintenance and reinvestment scheduling of electric distribution

The maintenance of electric distribution network is a topical question for distribution system operators because of increasing significance of failure costs. In this dissertation the maintenance practices of the distribution system operators are analyzed and a theory for scheduling maintenance activities and reinvestment of distribution components is created. The scheduling is based on the deterioration of components and the increasing failure rates due to aging. The dynamic programming algorithm is used as a solving method to maintenance problem which is caused by the increasing failure rates of the network. The other impacts of network maintenance like environmental and regulation reasons are not included to the scope of this thesis. Further the tree trimming of the corridors and the major disturbance of the network are not included to the problem optimized in this thesis. For optimizing, four dynamic programming models are presented and the models are tested. Programming is made in VBA-language to the computer. For testing two different kinds of test networks are used. Because electric distribution system operators want to operate with bigger component groups, optimal timing for component groups is also analyzed. A maintenance software package is created to apply the presented theories in practice. An overview of the program is presented.

Methods of genetic diversity creation and functional display for directed evolution experiments

Protein engineering aims to improve the properties of enzymes and affinity reagents by genetic changes. Typical engineered properties are affinity, specificity, stability, expression, and solubility. Because proteins are complex biomolecules, the effects of specific genetic changes are seldom predictable. Consequently, a popular strategy in protein engineering is to create a library of genetic variants of the target molecule, and render the population in a selection process to sort the variants by the desired property. This technique, called directed evolution, is a central tool for trimming protein-based products used in a wide range of applications from laundry detergents to anti-cancer drugs. New methods are continuously needed to generate larger gene repertoires and compatible selection platforms to shorten the development timeline for new biochemicals. In the first study of this thesis, primer extension mutagenesis was revisited to establish higher quality gene variant libraries in Escherichia coli cells. In the second study, recombination was explored as a method to expand the number of screenable enzyme variants. A selection platform was developed to improve antigen binding fragment (Fab) display on filamentous phages in the third article and, in the fourth study, novel design concepts were tested by two differentially randomized recombinant antibody libraries. Finally, in the last study, the performance of the same antibody repertoire was compared in phage display selections as a genetic fusion to different phage capsid proteins and in different antibody formats, Fab vs. single chain variable fragment (ScFv), in order to find out the most suitable display platform for the library at hand. As a result of the studies, a novel gene library construction method, termed selective rolling circle amplification (sRCA), was developed. The method increases mutagenesis frequency close to 100% in the final library and the number of transformants over 100-fold compared to traditional primer extension mutagenesis. In the second study, Cre/loxP recombination was found to be an appropriate tool to resolve the DNA concatemer resulting from error-prone RCA (epRCA) mutagenesis into monomeric circular DNA units for higher efficiency transformation into E. coli. Library selections against antigens of various size in the fourth study demonstrated that diversity placed closer to the antigen binding site of antibodies supports generation of antibodies against haptens and peptides, whereas diversity at more peripheral locations is better suited for targeting proteins. The conclusion from a comparison of the display formats was that truncated capsid protein three (p3Δ) of filamentous phage was superior to the full-length p3 and protein nine (p9) in obtaining a high number of uniquely specific clones. Especially for digoxigenin, a difficult hapten target, the antibody repertoire as ScFv-p3Δ provided the clones with the highest affinity for binding. This thesis on the construction, design, and selection of gene variant libraries contributes to the practical know-how in directed evolution and contains useful information for scientists in the field to support their undertakings.

Feasibility of industrial implementation of laser cutting into paper making machine

Laser cutting implementation possibilities into paper making machine was studied as the main objective of the work. Laser cutting technology application was considered as a replacement tool for conventional cutting methods used in paper making machines for longitudinal cutting such as edge trimming at different paper making process and tambour roll slitting. Laser cutting of paper was tested in 70’s for the first time. Since then, laser cutting and processing has been applied for paper materials with different level of success in industry. Laser cutting can be employed for longitudinal cutting of paper web in machine direction. The most common conventional cutting methods include water jet cutting and rotating slitting blades applied in paper making machines. Cutting with CO2 laser fulfils basic requirements for cutting quality, applicability to material and cutting speeds in all locations where longitudinal cutting is needed. Literature review provided description of advantages, disadvantages and challenges of laser technology when it was applied for cutting of paper material with particular attention to cutting of moving paper web. Based on studied laser cutting capabilities and problem definition of conventional cutting technologies, preliminary selection of the most promising application area was carried out. Laser cutting (trimming) of paper web edges in wet end was estimated to be the most promising area where it can be implemented. This assumption was made on the basis of rate of web breaks occurrence. It was found that up to 64 % of total number of web breaks occurred in wet end, particularly in location of so called open draws where paper web was transferred unsupported by wire or felt. Distribution of web breaks in machine cross direction revealed that defects of paper web edge was the main reason of tearing initiation and consequent web break. The assumption was made that laser cutting was capable of improvement of laser cut edge tensile strength due to high cutting quality and sealing effect of the edge after laser cutting. Studies of laser ablation of cellulose supported this claim. Linear energy needed for cutting was calculated with regard to paper web properties in intended laser cutting location. Calculated linear cutting energy was verified with series of laser cutting. Practically obtained laser energy needed for cutting deviated from calculated values. This could be explained by difference in heat transfer via radiation in laser cutting and different absorption characteristics of dry and moist paper material. Laser cut samples (both dry and moist (dry matter content about 25-40%)) were tested for strength properties. It was shown that tensile strength and strain break of laser cut samples are similar to corresponding values of non-laser cut samples. Chosen method, however, did not address tensile strength of laser cut edge in particular. Thus, the assumption of improving strength properties with laser cutting was not fully proved. Laser cutting effect on possible pollution of mill broke (recycling of trimmed edge) was carried out. Laser cut samples (both dry and moist) were tested on the content of dirt particles. The tests revealed that accumulation of dust particles on the surface of moist samples can take place. This has to be taken into account to prevent contamination of pulp suspension when trim waste is recycled. Material loss due to evaporation during laser cutting and amount of solid residues after cutting were evaluated. Edge trimming with laser would result in 0.25 kg/h of solid residues and 2.5 kg/h of lost material due to evaporation. Schemes of laser cutting implementation and needed laser equipment were discussed. Generally, laser cutting system would require two laser sources (one laser source for each cutting zone), set of beam transfer and focusing optics and cutting heads. In order to increase reliability of system, it was suggested that each laser source would have double capacity. That would allow to perform cutting employing one laser source working at full capacity for both cutting zones. Laser technology is in required level at the moment and do not require additional development. Moreover, capacity of speed increase is high due to availability high power laser sources what can support the tendency of speed increase of paper making machines. Laser cutting system would require special roll to maintain cutting. The scheme of such roll was proposed as well as roll integration into paper making machine. Laser cutting can be done in location of central roll in press section, before so-called open draw where many web breaks occur, where it has potential to improve runability of a paper making machine. Economic performance of laser cutting was done as comparison of laser cutting system and water jet cutting working in the same conditions. It was revealed that laser cutting would still be about two times more expensive compared to water jet cutting. This is mainly due to high investment cost of laser equipment and poor energy efficiency of CO2 lasers. Another factor is that laser cutting causes material loss due to evaporation whereas water jet cutting almost does not cause material loss. Despite difficulties of laser cutting implementation in paper making machine, its implementation can be beneficial. The crucial role in that is possibility to improve cut edge strength properties and consequently reduce number of web breaks. Capacity of laser cutting to maintain cutting speeds which exceed current speeds of paper making machines what is another argument to consider laser cutting technology in design of new high speed paper making machines.

De novo transcriptome assembly and annotation of the isopod Idotea balthica

The Baltic Sea is a unique environment that contains unique genetic populations. In order to study these populations on a genetic level basic molecular research is needed. The aim of this thesis was to provide a basic genetic resource for population genomic studies by de novo assembling a transcriptome for the Baltic Sea isopod Idotea balthica. RNA was extracted from a whole single adult male isopod and sequenced using Illumina (125bp PE) RNA-Seq. The reads were preprocessed using FASTQC for quality control, TRIMMOMATIC for trimming, and RCORRECTOR for error correction. The preprocessed reads were then assembled with TRINITY, a de Bruijn graph-based assembler, using different k-mer sizes. The different assemblies were combined and clustered using CD-HIT. The assemblies were evaluated using TRANSRATE for quality and filtering, BUSCO for completeness, and TRANSDECODER for annotation potential. The 25-mer assembly was annotated using PANNZER (protein annotation with z-score) and BLASTX. The 25-mer assembly represents the best first draft assembly since it contains the most information. However, this assembly shows high levels of polymorphism, which currently cannot be differentiated as paralogs or allelic variants. Furthermore, this assembly is incomplete, which could be improved by sampling additional developmental stages.