Evaluating the Advantages of Implementing a CRM solution

CRM-järjestelmän avulla pyritään yleisesti tehostamaan liiketoimintaprosesseja. Yrityksestä, kuin myös toimialasta riippuen hyödyt sekä tavoitteet vaihtelevat jonkin verran. Usein CRM-järjestelmän avulla saavutettavien hyötyjen mittaaminen ja arviointi organisaatiossa koetaan hankalaksi ja täten todellinen hyötyjen arviointi jää suppeaksi. Tämän tutkimuksen tavoitteena oli kartoittaa etuja, joita CRM-järjestelmän käyttöönotto on todellisuudessa tuonut yrityksille. Tutkimus kulminoituu laskentamalliin, jonka avulla CRM käytön hyödyt konvertoidaan konkreettiseksi rahasummaksi. Tutkimuksessa pyritään vastaamaan seuraaviin tutkimuskysymyksiin: 1. Kun yritys suunnitteli CRM-sovelluksen käyttöönottoa, mitkä olivat järjestelmälle asetetut tavoitteet? 2. Onko yritys saavuttanut CRM-järjestelmälle asetetut tavoitteet? 3. Onko yritys saavuttanut odottamattomia hyötyjä CRM-järjestelmän käytön avulla? 4. Mitkä ovat CRM-järjestelmän tärkeimmät kehityskohteet käyttökokemusten perusteella? Tutkimus perustuu CRM-kirjallisuuden avulla luotuun teoriaviitekehykseen. Teoriaviitekehyksen pohjalta luotiin ideaalimalli ja mallin pohjalta web-survey -kysely. Kyselyn tulosten perusteella toteutettiin haastatteluja, joilla pyrittiin saamaan tutkimustuloksiin syvyyttä.

Suluntekolaitteen toteutusvaihtoehdot: Kannattavuustarkastelu ja valinta

Diplomityön toimeksiantajayritys on 20 henkilöä työllistävä konepaja, jonka yksi liiketoiminta-alueista on koneenrakennus. Työn tutkimusongelmana oli selvittää, mikä on kannattavin kolmesta tarkasteltavasta teknisestä vaihtoehdosta lujitemuoviputken suluntekolaitteen toteuttamiseksi. Kyseessä on tapaustutkimus, jossa on käytetty esimerkkiasiakkaan, muovituotteita teollisuudelle valmistavan yrityksen, lähtötietoja. Kannattavuustekijöiksi määriteltiin kannattavuus asiakkaan näkökulmasta, kannattavuus toimittajan näkökulmasta sekä vaihtoehtoihin liittyvä epävarmuus. Asiakkaan näkökulmasta kannattavuutta on arvioitu investoinnin edullisuutena, toimittajan näkökulmasta projektin kannattavuutena taloudellisesta näkökulmasta ja vaihtoehtoihin liittyvää epävarmuutta muun muassa herkkyysanalyyseillä. Kannattavuustarkastelu on toteutettu Excel -mallilla, joka sisältää pisteytysjärjestelmän kannattavimman vaihtoehdon valitsemiseksi. Osana tutkimusta laadittiin sulunteon laatuvirheitä koskeva laatukustannusselvitys ja hinnoiteltiin laitteet. Kannattavuustarkastelun tuloksena todettiin, että sulunteko kannattaa koneellistaa ja että kannattavin tarkasteltavista vaihtoehdoista on konstruktio, jossa kudokset kelataan muotille rinnakkain. Todettiin, että laatukustannukset ovat merkittävä kustannuserä. Laadittua Excel-mallia voidaan käyttää myös muiden tuotteistusprojektien kannattavuustarkasteluun. Toimeksiantajayritys voi käyttää työssä saavutettuja tuloksia myyntiargumenttina.

Ferromagnetic properties of pyrolytic nanocarbon

The work reported in this thesis is dedicated to irreversible magnetic properties in pyrolytic nanocarbon samples. Based on atomic force microscope images, the samples consist of carbon clusters with radius 30..120 nm. These are treated as single-domain nanoparticles. Magnetic hysteresis, field cooled, zero field cooled and thermoremanent magnetization measurements were performed using an RF SQUID magnetometer and ferromagnetic behaviour was observed. Analysis suggests that the ferromagnetic ordering is associated with defects in a thin surface layer, whose thickness is independent of particle size. Critical radius for single-domain particles, critical radius for coherent rotation, magnetic layer thickness, distance between elementary magnetic moments, saturation magnetization, exchange stiffness constant and anisotropy energy density are also presented.

COMPARISON BETWEEN WOOD DRYING DEFECT SCORES: SPECIMEN TESTING X ANALYSIS OF KILN-DRIED BOARDS

It is important to develop drying technologies for Eucalyptus grandis lumber, which is one of the most planted species of this genus in Brazil and plays an important role as raw material for the wood industry. The general aim of this work was to assess the conventional kiln drying of juvenile wood of three clones of Eucalyptus grandis. The specific aims were to compare the behavior between: i) drying defects indicated by tests with wood specimens and conventional kiln-dried boards; and ii) physical properties and the drying quality. Five 11-year-old trees of each clone were felled, and only flatsawn boards of the first log were used. Basic density and total shrinkage were determined, and the drying test with wood specimens at 100 °C was carried out. Kiln drying of boards was performed, and initial and final moisture content, moisture gradient in thickness, drying stresses and drying defects were assessed. The defect scoring method was used to verify the behavior between the defects detected by specimen testing and the defects detected in kiln-dried boards. As main results, the drying schedule was too severe for the wood, resulting in a high level of boards with defects. The behavior between the defects in the drying test with specimens and the defects of kiln-dried boards was different, there was no correspondence, according to the defect scoring method.

Tuotannonohjausjärjestelmän kehittäminen laivanrakennustelakalla

Diplomityön tavoitteena oli selvittää tuotannon vallitseva toimintatapa, niin organisaation rakenteen, kuin myös sen toimintojen osalta. Vallitsevan toimintatavan epäkohtien perusteella pyrittiin luomaan uusi toimintatapa, jonka mukaan tuotannon tulisi toimia. Työn laajempana tarkoituksena oli edesauttaa tuotannonohjausjärjestelmän kehitystä ja käyttöönottoa STX Europen Turun telakalla. Työssä tutkittiin runkotuotannon tuotanto-organisaation prosesseja ja tuotannon toimintatapaa uuden tuotannonohjausjärjestelmän aikana. Tutkimuksessa selvitettiin tuotannon prosessien toimintaa ja havaintojen perusteella kuvattiin toimintojen epäkohdat. Tuotanto-organisaation toiminta kuvattiin yleisellä tasolla, jolloin epäkohtien vaikutus muuhun organisaatioon ja sen toimintaan ilmenivät. Havaintojen perusteella kävi ilmi, että tuotanto-organisaation toiminta ei ollut tuotannonohjausjärjestelmän vaatimalla tasolla. Tehtävienkuvauksia ei oltu laadittu tarvittavalla tarkkuudella. Asianomaiset henkilöt eivät olleet tietoisia heidän vastuualueistaan ja velvollisuuksistaan toimiessaan uuden tuotannonohjausjärjestelmän kanssa. Kuvauksen perusteella luotiin uusi ehdotus tuotanto-organisaation toimintatavalle. Työn johtopäätöksenä todettiin tarve uudelle tuotantoa tukevalle prosessille sekä prosessikuvauksien luomiselle koko tuotanto-organisaation toiminnasta. Johtopäätöksissä todettiin tuotannonohjausjärjestelmän olevan käyttöönottovalmiudessa, mutta mahdollisiin kehityskohteisiin tulisi ohjata resursseja.

The spindle assembly checkpoint as a drug target - Novel small-molecule inhibitors of Aurora kinases

Cell division (mitosis) is a fundamental process in the life cycle of a cell. Equal distribution of chromosomes between the daughter cells is essential for the viability and well-being of an organism: loss of fidelity of cell division is a contributing factor in human cancer and also gives rise to miscarriages and genetic birth defects. For maintaining the proper chromosome number, a cell must carefully monitor cell division in order to detect and correct mistakes before they are translated into chromosomal imbalance. For this purpose an evolutionarily conserved mechanism termed the spindle assembly checkpoint (SAC) has evolved. The SAC comprises a complex network of proteins that relay and amplify mitosis-regulating signals created by assemblages called kinetochores (KTs). Importantly, minor defects in SAC signaling can cause loss or gain of individual chromosomes (aneuploidy) which promotes tumorigenesis while complete failure of SAC results in cell death. The latter event has raised interest in discovery of low molecular weight (LMW) compounds targeting the SAC that could be developed into new anti-cancer therapeutics. In this study, we performed a cell-based, phenotypic high-throughput screen (HTS) to identify novel LMW compounds that inhibit SAC function and result in loss of cancer cell viability. Altogether, we screened 65 000 compounds and identified eight that forced the cells prematurely out of mitosis. The flavonoids fisetin and eupatorin, as well as the synthetic compounds termed SACi2 and SACi4, were characterized in more detail utilizing versatile cell-based and biochemical assays. To identify the molecular targets of these SAC-suppressing compounds, we investigated the conditions in which SAC activity became abrogated. Eupatorin, SACi2 and SACi4 preferentially abolished the tensionsensitive arm of the SAC, whereas fisetin lowered also the SAC activity evoked by lack of attachments between microtubules (MTs) and KTs. Consistent with the abrogation of SAC in response to low tension, our data indicate that all four compounds inhibited the activity of Aurora B kinase. This essential mitotic protein is required for correction of erratic MT-KT attachments, normal SAC signaling and execution of cytokinesis. Furthermore, eupatorin, SACi2 and SACi4 also inhibited Aurora A kinase that controls the centrosome maturation and separation and formation of the mitotic spindle apparatus. In line with the established profound mitotic roles of Aurora kinases, these small compounds perturbed SAC function, caused spindle abnormalities, such as multi- and monopolarity and fragmentation of centrosomes, and resulted in polyploidy due to defects in cytokinesis. Moreover, the compounds dramatically reduced viability of cancer cells. Taken together, using a cell-based HTS we were able to identify new LMW compounds targeting the SAC. We demonstrated for the first time a novel function for flavonoids as cellular inhibitors of Aurora kinases. Collectively, our data support the concept that loss of mitotic fidelity due to a non-functional SAC can reduce the viability of cancer cells, a phenomenon that may possess therapeutic value and fuel development of new anti-cancer drugs.

Regulation of epidermal tight junctions by calcium ATPases and p38

The epidermis is the upper layer of the skin and keratinocytes are its most abundant cells. Tight junctions are cell junctions located in the granular layer of the epidermis. They maintain the polarity of the cells and regulate the movement of water-soluble molecules. Epidermal tight junctions may lose their integrity when there are defects in intercellular calcium regulation. Hailey-Hailey and Darier´s disease are dominantly inherited, blistering skin diseases. Hailey-Hailey disease is caused by mutations in the ATP2C1 gene encoding a calcium/manganese ATPase SPCA1 of the Golgi apparatus. Darier´s disease is caused by mutations in the ATP2A2 gene encoding a calcium ATPase SERCA2 of the endoplasmic reticulum. p38 regulates the differentiation of keratinocytes. The overall regulation of epidermal tight junctions is not well understood. The present study examined the regulation of tight junctions in the human epidermis with a focus on calcium ATPases and p38. Skin from Hailey-Hailey and Darier´s disease patients was studied by using immunofluorescence labeling which targeted intercellular junction proteins. Transepidermal water loss was also measured. ATP2C1 gene expression was silenced in cultured keratinocytes, by siRNA, which modeled Hailey-Hailey disease. Expression of intercellular junction proteins was studied at the mRNA and protein levels. Squamous cell carcinoma and normal human keratinocytes were used as a model for impaired and normal keratinocyte differentiation, and the role of p38 isoforms alpha and delta in the regulation of intercellular junction proteins was studied. Both p38 isoforms were silenced by adenovirus cell transduction, chemical inhibitors or siRNA and keratinocyte differentiation was assessed. The results of this thesis revealed that: i.) intercellular junction proteins are expressed normally in acantholytic skin areas of patients with Hailey-Hailey or Darier´s disease but the localization of ZO-1 expanded to the stratum spinosum; ii.) tight junction proteins, claudin-1 and -4, are regulated by ATP2C1 in non-differentiating keratinocytes; and iii.) p38 delta regulates the expression of tight junction protein ZO-1 in proliferating keratinocytes and in squamous cell carcinoma derived cells. ZO-1 silencing, however, did not affect the expression of other tight junction proteins, suggesting that they are differently regulated. This thesis introduces new mechanisms involved in the regulation of tight junctions revealing new interactions. It provides novel evidence linking intracellular calcium regulation and tight junctions.

Toimituskyvyn kehittäminen prosessin materiaalipuutteita hallitsemalla

Toimituskyky on yrityksen suorituskykyä kuvaava tekijä, jolla on merkittävä vaikutus asiakastyytyväisyyteen, erityisesti valmistusteollisuudessa. Toimituskyky muodostuu materiaalin saatavuudesta ja logistisen järjestelmän toimitusvarmuudesta, joten hyvä toimituskyky edellyttää materiaalipuutteiden hallintaa. Tämän diplomityön tavoitteena on esittää, miten toimituskykyä voidaan kehittää tilaus-toimitusprosessin materiaalipuutteita hallitsemalla. Tutkimus toteutettiin konstruktiivisella tutkimusotteella case-tutkimuksena havainnoimalla toimintaa case-yrityksessä sekä analysoimalla case-yrityksen kirjallista materiaalia ja arkistoja. Yrityksessä havaittuja materiaalipuutteisiin liittyviä ongelmia tarkasteltiin tilaus-toimitusprosessin näkökulmasta prosessijohtamisen ja systemaattisen ongelmanratkaisun teorioiden avulla. Tutkimuksen tuloksena laadittiin kolme käytännönläheistä ratkaisuehdotusta havaittuihin ongelmiin; (1) materiaalipuutteiden syy-seuraussuhteita kuvaavat ongelma-syy-seurausketjut, (2) materiaalipuutteiden ongelmanratkaisumalli systemaattisen ongelmanratkaisun tueksi sekä (3) visuaalinen tilaus-toimitusprosessimalli, joka painottaa osaprosessien yhteyttä koko prosessin toimituskykyyn ja toimitusvarmuuteen. Tulosten mukaan materiaalipuutteet tulisi käsittää prosessin laatuvirheinä, jotka antavat arvokasta tietoa siitä, että prosessissa on laatuongelmia. Tulosten perusteella yrityksen toimituskykyä voidaan kehittää havainnoimalla tilaus-toimitusprosessin laatuvirheitä, selvittämällä laatuvirheiden syy-seuraussuhteet systemaattisesti ongelmanratkaisumallia hyödyntäen sekä toimimalla prosessiajattelun mukaisesti tilaus-toimitusprosessin toimituskyvyn jatkuvaa parantamista tavoitellen. Tutkimusongelman tarkastelutapaa ja työn tuloksia voidaan soveltaa samankaltaisiin tapauksiin, joissa tilaus-toimitusprosessin laatuvirheet, esimerkiksi materiaalipuutteet, paljastavat kehittämistä vaativia epäkohtia prosessin toimintatavoissa. Tilaus-toimitusprosessin toimituskykyä voidaan kehittää vain, jos panostetaan ajan hallintaan ja kykyyn toimia asiakaslupausten ja sopimusten mukaisesti.

Usability comparison of open source content management systems

The purpose of this thesis is to study, investigate and compare usability of open source cms. The thesis examines and compares usability aspect of some open source cms. The research is divided into two complementary parts –theoretical part and analytical part. The theoretical part mainly describes open source web content management systems, usability and the evaluation methods. The analytical part is to compare and analyze the results found from the empirical research. Heuristic evaluation method was used to measure usability problems in the interfaces. The study is fairly limited in scope; six tasks were designed and implemented in each interface for discovering defects in the interfaces. Usability problems were rated according to their level of severity. Time it took by each task, level of problem’s severity and type of heuristics violated will be recorded, analyzed and compared. The results of this study indicate that the comparing systems provide usable interfaces, and WordPress is recognized as the most usable system.

Tukin mittaus ja optimointi sahalinjassa

Tukin mittaus ennen sahausta ja sahausasetteen optimointi on kehittynyt paljon viimeisen 10 vuoden aikana. Sahauksen kannattavuuden huonontuessa raaka-aineen tehokas hyödyntäminen on muodostunut tärkeäksi osaksi prosessia. Mittalaitteiden tekniikan kehityttyä on ollut mahdollista mitata tukin muoto ja halkaisijat eri kohdista entistä tarkemmin. Sahausasetteen optimoinnilla pyritään raaka-aineen mahdollisimman tehokkaaseen käyttöön eli saamaan mahdollisimman hyvä saanto jokaisesta yksittäisestä sahatusta tukista. Mittaustarkkuus on suoraan kytköksissä sahausasetteen optimointi tuloksen onnistumiseen. Yleisesti tukin mittaus ennen sahausta ja sahausasetteen optimointi tulevat samalta toimittajalta. Työssä tarkasteltiin kahden eri toimittajan tukkimittareita sekä optimoinnin onnistumista sen perusteella. Käytössä oli lasikuituinen mallitukki, jota mitattiin kummankin toimittajan mittareilla. Näin voitiin suoraan vertailla mittauksen ja optimoinnin onnistumista ja verrata sitä optimaalisiin tuloksiin. Työssä käytettiin kandidaatintyössä luomaani toimintamallia tukkimittarin tarkkuuden toteamiseksi. Mittaus- ja optimointivirheistä pystyttiin laskemaan, kuinka paljon tappiota sahalaitokselle aiheutui verrattuna optimaaliseen mittaus- ja optimointitulokseen. Jo pienetkin virheet optimoinnissa ja mittauksessa vaikuttavat sahauksen kannattavuuteen, kun tarkastellaan sahalaitosta jossa sahataan 8000 – 10 000 tukkia yhden työvuoron aikana. Tulosten perusteella mittarit mittaavat hieman virheellisesti, ja kummankin mittarin mittausten perusteella saatiin eri sahausasete optimointitulokseksi. Mittavirheen takia voitiin todeta, että parantamalla mittaustarkkuutta voidaan sahauksen kannattavuutta parantaa.

Neuronal and endothelial nitric oxide synthase gene knockout mice

Targeted disruption of the neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) genes has led to knockout mice that lack these isoforms. These animal models have been useful to study the roles of nitric oxide (NO) in physiologic processes. nNOS knockout mice have enlarged stomachs and defects in the inhibitory junction potential involved in gastrointestinal motility. eNOS knockout mice are hypertensive and lack endothelium-derived relaxing factor activity. When these animals are subjected to models of focal ischemia, the nNOS mutant mice develop smaller infarcts, consistent with a role for nNOS in neurotoxicity following cerebral ischemia. In contrast, eNOS mutant mice develop larger infarcts, and show a more pronounced hemodynamic effect of vascular occlusion. The knockout mice also show that nNOS and eNOS isoforms differentially modulate the release of neurotransmitters in various regions of the brain. eNOS knockout mice respond to vessel injury with greater neointimal proliferation, confirming that reduced NO levels seen in endothelial dysfunction change the vessel response to injury. Furthermore, eNOS mutant mice still show a protective effect of female gender, indicating that the mechanism of this protection cannot be limited to upregulation of eNOS expression. The eNOS mutant mice also prove that eNOS modulates the cardiac contractile response to ß-adrenergic agonists and baseline diastolic relaxation. Atrial natriuretic peptide, upregulated in the hearts of eNOS mutant mice, normalizes cGMP levels and restores normal diastolic relaxation.

ß-Spectrin São PauloII, a novel frameshift mutation of the ß-spectrin gene associated with hereditary spherocytosis and instability of the mutant mRNA

Hereditary spherocytosis (HS) is a common inherited anemia characterized by the presence of spherocytic red cells. Defects in several membrane protein genes have been involved in the pathogenesis of HS. ß-Spectrin-related HS seems to be common. We report here a new mutation in the ß-spectrin gene coding region in a patient with hereditary spherocytosis. The patient presented acanthocytosis and spectrin deficiency and, at the DNA level, a novel frameshift mutation leading to HS, i.e., a C deletion at codon 1392 (ß-spectrin São PauloII), exon 20. The mRNA encoding ß-spectrin São PauloII was very unstable and the mutant protein was not detected in the membrane or in other cellular compartments. It is interesting to note that frameshift mutations of the ß-spectrin gene at the 3' end allow the insertion of the mutant protein in the red cell membrane, leading to a defect in the auto-association of the spectrin dimers and consequent elliptocytosis. On the other hand, ß-spectrin São PauloII protein was absent in the red cell membrane, leading to spectrin deficiency, HS and the presence of acanthocytes.

Spectroscopic Studies of III-V Semiconductor Materials for Improved Devices

Defects in semiconductor crystals and at their interfaces usually impair the properties and the performance of devices. These defects include, for example, vacancies (i.e., missing crystal atoms), interstitials (i.e., extra atoms between the host crystal sites), and impurities such as oxygen atoms. The defects can decrease (i) the rate of the radiative electron transition from the conduction band to the valence band, (ii) the amount of charge carriers, and (iii) the mobility of the electrons in the conduction band. It is a common situation that the presence of crystal defects can be readily concluded as a decrease in the luminescence intensity or in the current flow for example. However, the identification of the harmful defects is not straightforward at all because it is challenging to characterize local defects with atomic resolution and identification. Such atomic-scale knowledge is however essential to find methods for reducing the amount of defects in energy-efficient semiconductor devices. The defects formed in thin interface layers of semiconductors are particularly difficult to characterize due to their buried and amorphous structures. Characterization methods which are sensitive to defects often require well-defined samples with long range order. Photoelectron spectroscopy (PES) combined with photoluminescence (PL) or electrical measurements is a potential approach to elucidate the structure and defects of the interface. It is essential to combine the PES with complementary measurements of similar samples to relate the PES changes to changes in the interface defect density. Understanding of the nature of defects related to III-V materials is relevant to developing for example field-effect transistors which include a III-V channel, but research is still far from complete. In this thesis, PES measurements are utilized in studies of various III-V compound semiconductor materials. PES is combined with photoluminescence measurements to study the SiO2/GaAs, SiNx/GaAs and BaO/GaAs interfaces. Also the formation of novel materials InN and photoluminescent GaAs nanoparticles are studied. Finally, the formation of Ga interstitial defects in GaAsN is elucidated by combining calculational results with PES measurements.

To Divide or Not to Divide; MicroRNAs and Small Compounds as Modulators of Mitosis

Mitosis is under the stringent quality control of the spindle assembly checkpoint (SAC). However, in cancer cells this control can fail, leading to excessive cellular proliferation and ultimately to the formation of a tumor. Novel cancer cell selective therapies are needed to stop the uncontrolled cell proliferation and tumor growth. The aim of the research presented in this thesis was to identify microRNAs (miRNAs) that could play a role in cancer cell proliferation as well as low molecular weight (LMW) compounds that could interfere with cell division. The findings could be used to develop better cancer diagnostics and therapies in the future. First, a high-throughput screen (HTS) was performed to identify LMW compounds that possess a similar chemical interaction field as rigosertib, an anti-cancer compound undergoing clinical trials. A compound termed Centmitor-1 was discovered that phenocopied the cellular impact of rigosertib by affecting the microtubule dynamics. Next, another HTS aimed at identifying compounds that would target the Hec1 protein, which mediates the interaction between spindle microtubules and chromosomes. Perturbation of this connection should prevent cell division and induce cell death. A compound termed VTT-006 was discovered that abrogated mitosis in several cell line models and exhibited binding to Hec1 in vitro. Lastly, using a cell-based HTS two miRNAs were identified that affected cancer cell proliferation via Aurora B kinase, which is an important mitotic regulator. MiR-378a-5p was found to indirectly suppress the production of the kinase whereas let-7b showed direct binding to the 3’UTR of Aurora B mRNA and repressed its translation. The miRNA-mediated perturbation of Aurora B induced defects in mitosis leading to abnormal chromosome segregation and induction of aneuploidy. The results of this thesis provide new information on miRNA signaling in cancer, which could be utilized for diagnostic purposes. Moreover, the thesis introduces two small compounds that may benefit future drug research.

The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease

Chronic granulomatous disease (CGD) is an inherited disorder of the innate immune system characterized by a defective oxidative burst of phagocytes and subsequent impairment of their microbicidal activity. Mutations in one of the NADPH-oxidase components affect gene expression or function of this system, leading to the phenotype of CGD. Defects in gp91-phox lead to X-linked CGD, responsible for approximately 70% of CGD cases. Investigation of the highly heterogeneous genotype of CGD patients includes mutation analysis, Northern blot or Western blot assays according to the particular case. The aim of the present study was to use reverse transcription (RT)-PCR for the analysis of molecular defects responsible for X-linked CGD in eight Brazilian patients and to assess its potential for broader application to molecular screening in CGD. Total RNA was prepared from Epstein B virus-transformed B-lymphocytes and reverse transcribed using random hexamers. The resulting cDNA was PCR-amplified by specific and overlapping pairs of primers designed to amplify three regions of the gp91-phox gene: exons 1-5, 3-9, and 7-13. This strategy detected defective gp91-phox expression in seven patients. The RT-PCR results matched clinical history, biochemical data (nitroblue tetrazolium or superoxide release assay) and available mutation analysis in four cases. In three additional cases, RT-PCR results matched clinical history and biochemical data. In another case, RT-PCR was normal despite a clinical history compatible with CGD and defective respiratory burst. We conclude that this new application of RT-PCR analysis - a simple, economical and rapid method - was appropriate for screening molecular defects in 7 of 8 X-linked CGD patients.