Development of the Acoustic Emission Based Diagnostics Concept for the Rotary Lime Kiln

Reaaliaikainen, ennakoiva kunnonvalvonta on erittäin tärkeä osa modernin tehtaan tai tuotantolinjan toimintaa. Diplomityön teettäjä haluaa edelleen kehittää akustiseen emissioon perustuvaa kunnonvalvonta järjestelmäänsä, jotta siitä olisi enemmän hyötyä asiakkaalle. Diplomityö sisältää johdannonakustiseen emissioon ja akustisiin emissio sensoreihin. Työn tavoitteena oli kehittää päätöksentekojärjestelmä, jota käytettäisiin työn teettäjän valmistamien sensoreiden antaman tiedon automatisoituun analysointiin. Työssä on vertailtu kolmea eri ohjelmistotoimittajaa ja heidän ohjelmiaan, ja tehty ehdotus hankittavasta ohjelmistosta. Lisäksi työssä on kehitetty ohjeita, joiden avulla ohjelmisto ohjelmoidaan tuottamaan reaaliaikaista tietoa ja huolto-ohjeita sen käyttäjille. Lisäksi työssä annetaan ehdotuksia kunnonvalvonta- ja päätöksentekojärjestelmän edelleen kehittämiseen.

Service product structure supported by remote diagnostics

Työn tavoitteena oli määrittää etähuoltokonseptin yleinen rakenne sekä selvittää asiakkaille tarjottavia modulaarisia palvelutuotteita. Aluksi selvitettiin huoltokonseptin rakennetta kirjallisuuden ja asiantuntijoiden haastattelujen avulla. Asiantuntijoiden haastattelut toteutettiin vapaamuotoisesti kysymyslistaa apuna käyttäen. Tämän lisäksi työssä pohditaan KM, CRM ja PDM rooleja etähuoltokonseptin kannalta sekä tutkaillaan etähuollon tulevaisuuden näkymiä.Diplomityössä käsitellään modulaarisuutta yleisellä tasolla. Moduularisuus on moniulotteinen termi. Usein sillä tarkoitetaan yrityksen sisäistä tuotekehityksen hallintaa. Toisaalta se voidaan nähdä myös asiakkaalle tarjottavina tuotteina ja tässä diplomityössä on keskitytty tähän puoleen. Loppuosa diplomityöstä käsittelee palvelutuotteiden tuotteistamisprosessia.Diplomityön tuloksena hahmoteltiin etähuolto konseptia ja tutkittiin mahdollisia palvelutuotteita, joita voidaan sisällyttää etähuoltokonseptiin. Tämän lisäksi toteutettiin osa tuotteistamisprosessista. Etähuollon kehittäminen on haasteellista. Haasteeksi jää tuotekehitystoiminnan ja palvelupuolen toiminnan tehostaminen, jotta tuotteita voitaisiin jo suunnittelupuolella kehittää etähuollon tarpeisiin. Kehittäminen vaatii molempien puolien aktiivista osallistumista.

Genetic Basis and Diagnostics of Extended-Spectrum Beta-Lactamases among Enterobacteriaceae in Finland

Since the introduction of antibiotic agents, the amount and prevalence of Beta-lactam resistant enterobacteria has become an increasing problem. Many enterobacteria are opportunistic pathogens that easily acquire resistance mechanisms and genes, which make the situation menacing. These bacteria have acquired resistance and can hydrolyse extended spectrum cephalosporins and penicillins by producing enzymes called extended-spectrum Beta-lactamases (ESBLs). ESBL-producing bacteria are most commonly found in the gastro-intestinal tract of colonised patients. These resistant strains can be found in both health-care associated and community-acquired isolates. The detection and treatment of infections caused by bacteria producing ESBLs are problematic. This study investigated the genetic basis of extended-spectrum Beta-lactamases in Enterobacteriaceae, especially in Escherichia coli and Klebsiella pneumoniae isolates. A total of 994 Finnish Enterobacteriaceae strains, collected at 26 hospital laboratories, during 2000 and 2007 were analysed. For the genetic basis studies, PCR, sequencing and pyrosequencing methods were optimised. In addition, international standard methods, the agar dilution and disk diffusion methods were performed for the resistance studies, and the susceptibility of these strains was tested for antimicrobial agents that are used for treating patients. The genetic analysis showed that bla_CTX-M was the most prevalent gene among the E. coli isolates, while blaS_HV-12 was the most common Beta-lactamase gene in K. pneumoniae. The susceptibility testing results showed that about 60% of the strains were multidrug resistant. The prevalence of ESBL-producing isolates in Finland has been increasing since 2000. However, the situation in Finland is still much better than in many other European countries.

Free Prostate-specific Antigen Forms and Kallikrein-related Peptidase 2: Tools for Prostate Cancer Diagnostics

Prostate-specific antigen (PSA) is a marker that is commonly used in estimating prostate cancer risk. Prostate cancer is usually a slowly progressing disease, which might not cause any symptoms whatsoever. Nevertheless, some cases of cancer are aggressive and need to be treated before they become life-threatening. However, the blood PSA concentration may rise also in benign prostate diseases and using a single total PSA (tPSA) measurement to guide the decision on further examinations leads to many unnecessary biopsies, over-detection, and overtreatment of indolent cancers which would not require treatment. Therefore, there is a need for markers that would better separate cancer from benign disorders, and would also predict cancer aggressiveness. The aim of this study was to evaluate whether intact and nicked forms of free PSA (fPSA-I and fPSA-N) or human kallikrein-related peptidase 2 (hK2) could serve as new tools in estimating prostate cancer risk. First, the immunoassays for fPSA-I and free and total hK2 were optimized so that they would be less prone to assay interference caused by interfering factors present in some blood samples. The optimized assays were shown to work well and were used to study the marker concentrations in the clinical sample panels. The marker levels were measured from preoperative blood samples of prostate cancer patients scheduled for radical prostatectomy. The association of the markers with the cancer stage and grade was studied. It was found that among all tested markers and their combinations especially the ratio of fPSA-N to tPSA and ratio of free PSA (fPSA) to tPSA were associated with both cancer stage and grade. They might be useful in predicting the cancer aggressiveness, but further follow-up studies are necessary to fully evaluate the significance of the markers in this clinical setting. The markers tPSA, fPSA, fPSA-I and hK2 were combined in a statistical model which was previously shown to be able to reduce unnecessary biopsies when applied to large screening cohorts of men with elevated tPSA. The discriminative accuracy of this model was compared to models based on established clinical predictors in reference to biopsy outcome. The kallikrein model and the calculated fPSA-N concentrations (fPSA minus fPSA-I) correlated with the prostate volume and the model, when compared to the clinical models, predicted prostate cancer in biopsy equally well. Hence, the measurement of kallikreins in a blood sample could be used to replace the volume measurement which is time-consuming, needs instrumentation and skilled personnel and is an uncomfortable procedure. Overall, the model could simplify the estimation of prostate cancer risk. Finally, as the fPSA-N seems to be an interesting new marker, a direct immunoassay for measuring fPSA-N concentrations was developed. The analytical performance was acceptable, but the rather complicated assay protocol needs to be improved until it can be used for measuring large sample panels.

Molecular diagnostics of rare inherited SYNDROMES with a view on diagnostic test development

CHARGE syndrome, Sotos syndrome and 3p deletion syndrome are examples of rare inherited syndromes that have been recognized for decades but for which the molecular diagnostics only have been made possible by recent advances in genomic research. Despite these advances, development of diagnostic tests for rare syndromes has been hindered by diagnostic laboratories having limited funds for test development, and their prioritization of tests for which a (relatively) high demand can be expected. In this study, the molecular diagnostic tests for CHARGE syndrome and Sotos syndrome were developed, resulting in their successful translation into routine diagnostic testing in the laboratory of Medical Genetics (UTUlab). In the CHARGE syndrome group, mutation was identified in 40.5% of the patients and in the Sotos syndrome group, in 34%, reflecting the use of the tests in routine diagnostics in differential diagnostics. In CHARGE syndrome, the low prevalence of structural aberrations was also confirmed. In 3p deletion syndrome, it was shown that small terminal deletions are not causative for the syndrome, and that testing with arraybased analysis provides a reliable estimate of the deletion size but benign copy number variants complicate result interpretation. During the development of the tests, it was discovered that finding an optimal molecular diagnostic strategy for a given syndrome is always a compromise between the sensitivity, specificity and feasibility of applying a new method. In addition, the clinical utility of the test should be considered prior to test development: sometimes a test performing well in a laboratory has limited utility for the patient, whereas a test performing poorly in the laboratory may have a great impact on the patient and their family. At present, the development of next generation sequencing methods is changing the concept of molecular diagnostics of rare diseases from single tests towards whole-genome analysis.

Switchable lanthanide fluorescence probes in homogenous molecular diagnostics

The number of molecular diagnostic assays has increased tremendously in recent years.Nucleic acid diagnostic assays have been developed, especially for the detection of human pathogenic microbes and genetic markers predisposing to certain diseases. Closed-tube methods are preferred because they are usually faster and easier to perform than heterogenous methods and in addition, target nucleic acids are commonly amplified leading to risk of contamination of the following reactions by the amplification product if the reactions are opened. The present study introduces a new closed-tube switchable complementation probes based PCR assay concept where two non-fluorescent probes form a fluorescent lanthanide chelate complex in the presence of the target DNA. In this dual-probe PCR assay method one oligonucleotide probe carries a non-fluorescent lanthanide chelate and another probe a light absorbing antenna ligand. The fluorescent lanthanide chelate complex is formed only when the non-fluorescent probes are hybridized to adjacent positions into the target DNA bringing the reporter moieties in close proximity. The complex is formed by self-assembled lanthanide chelate complementation where the antenna ligand is coordinated to the lanthanide ion captured in the chelate. The complementation probes based assays with time-resolved fluorescence measurement showed low background signal level and hence, relatively high nucleic acid detection sensitivity (low picomolar target concentration). Different lanthanide chelate structures were explored and a new cyclic seven dentate lanthanide chelate was found suitable for complementation probe method. It was also found to resist relatively high PCR reaction temperatures, which was essential for the PCR assay applications. A seven-dentate chelate with two unoccupied coordination sites must be used instead of a more stable eight- or nine-dentate chelate because the antenna ligand needs to be coordinated to the free coordination sites of the lanthanide ion. The previously used linear seven-dentate lanthanide chelate was found to be unstable in PCR conditions and hence, the new cyclic chelate was needed. The complementation probe PCR assay method showed high signal-to-background ratio up to 300 due to a low background fluorescence level and the results (threshold cycles) in real-time PCR were reached approximately 6 amplification cycles earlier compared to the commonly used FRET-based closed-tube PCR method. The suitability of the complementation probe method for different nucleic acid assay applications was studied. 1) A duplex complementation probe C. trachomatis PCR assay with a simple 10-minute urine sample preparation was developed to study suitability of the method for clinical diagnostics. The performance of the C. trachomatis assay was equal to the commercial C. trachomatis nucleic acid amplification assay containing more complex sample preparation based on DNA extraction. 2) A PCR assay for the detection of HLA-DQA1*05 allele, that is used to predict the risk of type 1 diabetes, was developed to study the performance of the method in genotyping. A simple blood sample preparation was used where the nucleic acids were released from dried blood sample punches using high temperature and alkaline reaction conditions. The complementation probe HLA-DQA1*05 PCR assay showed good genotyping performance correlating 100% with the routinely used heterogenous reference assay. 3) To study the suitability of the complementation probe method for direct measurement of the target organism, e.g., in the culture media, the complementation probes were applied to amplificationfree closed-tube bacteriophage quantification by measuring M13 bacteriophage ssDNA. A low picomolar bacteriophage concentration was detected in a rapid 20- minute assay. The assay provides a quick and reliable alternative to the commonly used and relatively unreliable UV-photometry and time-consuming culture based bacteriophage detection methods and indicates that the method could also be used for direct measurement of other micro-organisms. The complementation probe PCR method has a low background signal level leading to a high signal-to-background ratio and relatively sensitive nucleic acid detection. The method is compatible with simple sample preparation and it was shown to tolerate residues of urine, blood, bacteria and bacterial culture media. The common trend in nucleic acid diagnostics is to create easy-to-use assays suitable for rapid near patient analysis. The complementation probe PCR assays with a brief sample preparation should be relatively easy to automate and hence, would allow the development of highperformance nucleic acid amplification assays with a short overall assay time.

Upconverting diagnostics: Multiplex assays utilizing upconversion luminescence technology

Conventional diagnostics tests and technologies typically allow only a single analysis and result per test. The aim of this study was to propose robust and multiplex array-inwell test platforms based on oligonucleotide and protein arrays combining the advantages of simple instrumentation and upconverting phosphor (UCP) reporter technology. The UCPs are luminescent lanthanide-doped crystals that have a unique capability to convert infrared radiation into visible light. No autofluorescence is produced from the sample under infrared excitation enabling the development of highly sensitive assays. In this study, an oligonucleotide array-in-well hybridization assay was developed for the detection and genotyping of human adenoviruses. The study provided a verification of the advantages and potential of the UCP-based reporter technology in multiplex assays as well as anti-Stokes photoluminescence detection with a new anti- Stokes photoluminescence imager. The developed assay was technically improved and used to detect and genotype adenovirus types from clinical specimens. Based on the results of the epidemiological study, an outbreak of adenovirus type B03 was observed in the autumn of 2010. A quantitative array-in-well immunoassay was developed for three target analytes (prostate specific antigen, thyroid stimulating hormone, and luteinizing hormone). In this study, quantitative results were obtained for each analyte and the analytical sensitivities in buffer were in clinically relevant range. Another protein-based array-inwell assay was developed for multiplex serodiagnostics. The developed assay was able to detect parvovirus B19 IgG and adenovirus IgG antibodies simultaneously from serum samples according to reference assays. The study demonstrated that the UCPtechnology is a robust detection method for diverse multiplex imaging-based array-inwell assays.

Serpulina-suvun bakteereiden diagnostiikka sekä sioista vuosina 1993-1995 eristettyjen Serpulina-suvun bakteereiden tarkastelua

Summary: The diagnostics of Serpulina bacteria and a review on Finnish swine colonic spirochaetes isolated during 1993-1995

Naudan alkiodiagnostiikka

Summary: Bovine embryo diagnostics

BSE taudin kartoitus, nopea diagnostiikka ja ennaltaehkäisy

Summary: Survey, diagnostics and prevention of BSE

HIV-pikatestien testaus ja tulosten vertaaminen rutiinidiagnostiikan testituloksiin

Teimme opinnäytetyömme Helsingin ja Uudenmaan sairaanhoitopiirin laboratorion eli HUSLABin virologian osastolla hiv- hepatiittityöpisteessä. Työn tavoitteena oli selvittää kuinka luotettavia immunokromatografiset HIV-pikatestit ovat. Testasimme kolmea eri HIV-pikatestiä ja vertasimme niistä saatuja tuloksia virologian osaston rutiinidiagnostiikan tuloksiin. Pikatestit olivat Determine HIV - 1/2 (Abbott), CORE HIV 1&2 (Core Diagnostics) ja IMMUNOFLOW HIV 1-HIV2 (Core Diagnostics). Testasimme 100 seeruminäytettä jokaisella kolmella HIV-pikatestillä. Näytteistä vahvoja positiivisia oli 20, tuoreita tartuntoja 20, vääriä reaktiivisia 20 ja negatiivisia 40. Tulosten käsittelyssä käytimme SPSS ja Excel -ohjelmia. Tuloksiemme perusteella kaikkien HIV-pikatestien spesifisyys oli parempi kuin niiden sensitiivisyys. HIV-pikatestit eivät antaneet täysin luotettavia tuloksia. HIV-pikatestit eivät välttämättä havaitse tuoreita infektioita ja tämän takia saattaisi tulla vääriä diagnooseja. HIV-pikatestit saattavat myös antaa vääriä positiivisia tuloksia. Testin luotettavuus on hyvä, jos testataan pitkälle edenneitä HIV-infektioita, joissa vasta-aineet ovat korkeita. Tulosten perusteella rutiinidiagnostiikan testeillä HIV-infektio saadaan paremmin diagnosoitua kuin HIV-pikatesteillä. Suomessa ei mielestämme ole tarpeellista käyttää HIV-pikatestejä, sillä laboratoriotekniikka on hyvin kehittynyttä ja välimatkat lyhyitä. Suuren HIV-prevalenssin maissa HIV-pikatestit olisivat mielestämme hyödyllisiä, jotta mahdollisimman moni saataisiin testattua.

Upgrading of thethird generation of the Remote Diagnostic Systems for ABB Marine Propulsion Systems

Remote diagnostics has become very popular in marine industry. Diagnostic systems are improved all the time and the newest monitoring systems are constantly taken into operation. Most vessels are, however, rigged up by outmoded facilities, which should be updated. In this work the principles of operating of such a remote diagnostic system as the ABB's third generation remote diagnostic system (RDS) and the newest Propulsion Condition Management System (PCMS) are studied. As a result of the thesis the ways of upgrading the old systems of RDS are presented. Specifically, the work focuses on the establishment of the connection between Advant controller AC-110 from the old system and PCMS server, and in other words how to get Modbus data by OPC server.

Sähkökoneiden etädiagnostiikan kenttätason tiedonsiirtotarpeen arviointi ja instrumentoinnin kehittäminen

Tässä diplomityössä tarkastellaan sähkökoneiden tyypillisiä vikoja ja joitain näiden havaitsemiseen käytettäviä mittauksia ja analyysejä; mittausten tuottamaa tietomäärää arvioidaan. Muutamia teollisuuslaitoksissa käytettyjä väyliä ja tiedonsiirtoprotokollia esitellään, ja analysoidaan kunnonvalvonnan mittauksien siirtämisen näillä väylillä vaatimaa väylän datansiirtonopeutta. Valvottavien sähkökoneiden määrän ylärajaa arvioidaan kunkin väylän/protokollan tapauksessa. Työssä esitellään ratkaisuja sähkökoneiden etädiagnostiikan tiedonsiirron toteuttamiseksi ja arvioidaan valvottavien sähkökoneiden määrän ylärajaa kussakin tapauksessa. Lisäksi työssä suunnitellaan ja toteutetaan kunnonvalvonnan mitta-antureiden kanssa käytettävä mittaustiedon keräily-yksikkö. Keräily-yksikön sisältävä kunnonvalvonnan ja etädiagnostiikan tiedonkeruu- ja tiedonsiirtojärjestelmä asennetaan kahteen pilot-kohteeseen: sellutehtaalle ja pienvesivoimalaan.

Lyhyen kantaman radiolähettimien soveltuvuus sähkökäyttöjen kunnon-valvonnan ja etädiagnostiikan tiedonsiirtotarpeisiin

Lyhyen kantaman radiolähettimien käyttö on kasvanut muutaman viime vuoden aikana huomattavasti. Osaltaan tähän on vaikuttanut uusien taajuusalueiden allokointi lupavapaiden radiolähettimien käyttöön, standardoitujen tekniikoiden yleistyminen ja laitteiden parempi saatavuus. Tässä työssä tarkastellaan lyhyen kantaman radiolähettimien soveltuvuutta sähkökäyttöjen kunnonvalvonnan ja etädiagnostiikan tiedonsiirtotarpeisiin. Työssä luodaan katsaus teollisuusympäristön vaikutuksiin langattomassa tiedonsiirrossa ja tarkastellaan sähkökäyttöjen kunnonvalvonnan ja etädiagnostiikan vaatimuksia tiedonsiirrolle ja tiedonsiirtojärjestelmälle. Työssä käydään läpi lyhyen kantaman radiolähettimiin liittyvät säädökset ja luodaan katsaus standardoituihin sekä valmistajakohtaisiin lyhyen kantaman radiotekniikoihin. Tiedonsiirtojärjestelmän suunnittelun kannalta tarkastellaan tiedonsiirtojärjestelmän topologiaa, protokollaa ja saatavilla olevia tekniikoita ja niiden implementoimista järjestelmään. Valmistajakohtaisia lyhyen kantaman radiomoduulien toimintaa ja Bluetoothin sisältävää älypuhelinta tutkitaan laboratoriomittauksin. Lopuksi kootaan yhteen langattoman tiedonsiirron ja eri tekniikoiden soveltuvuus sähkökäyttöjen kunnonvalvonnan ja etädiagnostiikan sovelluksiin.