Active Directoryn Group Policyjen parhaat käytännöt : case Palmia

Opinnäytetyön toimeksiantajana oli Helsingin kaupungin Palmia liikelaitos. Tämä opinnäytetyö tehtiin, koska Palmialla otettiin käyttöön Windows 8 -käyttöjärjestelmä tablet-tietokoneissa ja uusi käyttöjärjestelmä tarvitsi myös ryhmäkäytäntöasetusten määrittämisen. Ryhmäkäytännöt oli määritelty myös sekavasti. Työssä selvitetään aluksi ryhmäkäytäntöjen toimintaperiaatetta ja esitetään yleisiä parhaita käytäntöjä, joiden mukaan ryhmäkäytäntöjä tulisi määritellä. Tietolähteenä on käytetty pääasiallisesti Microsoft-ympäristöihin perehtyneiden asiantuntijoiden Internet-sivuja ja blogeja. Työn aluksi tehtiin nykytilan kartoitus, josta saatiin yleiskäsitys, mitä on tehty hyvin ja mitkä osa-alueet vaativat muutoksia. Tämän kartoituksen pohjalta laadittiin kehitysehdotuksia, joista osa päätettiin toteuttaa. Windows 8 -käyttöjärjestelmän asetuksiin ja optimointiin määriteltiin asetukset linjassa olemassa olevan ympäristön asetusten kanssa. Opinnäytetyön tuloksena syntyi Palmialle yksinkertainen ryhmäkäytäntöjen ympäristö, jota on helppo ylläpitää. Ryhmäkäytäntöobjektit on suunniteltu niin, että ne painottavat vähäistä vaikutusta loppukäyttäjään. Lisäksi havaittiin, että ryhmäkäytäntöjä ei tule jättää määrittelyn jälkeen vaille huomiota, koska tietokoneissa oleva käyttöjärjestelmä ja käytetyt sovellukset päivittyvät jatkuvasti ja ympäristön asetuksien tulee olla koko ajan ajantasaiset.This thesis was commissioned by the City of Helsinki Public Utility Palmia as they introduced the Windows 8 operating system in their tablet computers. The purpose of the thesis was to define a proper Group Policy configuration as Group Policies were poorly defined. First, the principle of Group Policies is described and what the general best practices are on how Group Policies should be set. The data source is mainly Internet sites and blogs of experts specialized in Microsoft environments. The thesis was started by studying the current state of Group Policies, which gave an overview of what has been done well and what areas require changes. On the basis of this survey development proposals were drawn up and some of those were decided to implement. As a result of this study Palmia got a simple Group Policy environment that is easy to maintain. Group Policies were designed so that the impact on the end user is minimized. In addition, it was found out that the Group Policies should not be left ignored after they have once been defined and set. That is because the operating systems and applications of computers are continuously updated and the environment settings must be constantly up-to-date

Rapid diagnostic tests for resource-poor areas

Rapid diagnostic tests, such as lateral flow immunoassays may enable diagnostics in resource poor areas, thus improving health outcomes by helping to control and eliminate infectious diseases. This thesis focuses on the development of sensitive tools for the rapid diagnosis of three infectious diseases prevalent in resource poor areas: hepatitis C, pertussis and malaria. For malaria and hepatitis C, rapid diagnostic tests with improved sensitivity could help in ongoing efforts to eliminate the diseases. For pertussis (whooping cough), a field-usable rapid serodiagnostic test would help in disease surveillance and control. New methods to enhance the sensitivity of lateral flow immunoassays include preconcentrating the sample before the immunoassay takes place, controlling the flow of reagents in the lateral flow strip to allow more complex assays and using high-sensitivity luminescent instrument-read labels. All the above methods involve trade-offs between the sensitivity, complexity, and affordability of the test. In publication I of this thesis, immunoassays for anti-HCV antibodies were developed using a single multiepitope protein antigen and a luminescent europium-chelate label. In II and III, quantitative lateral flow immunoassays utilizing luminescent europium nanoparticle labels were developed and assessed. The results obtained with patient samples using the lateral flow immunoassay for anti-pertussis antibodies (II) correlated well with a traditional enzyme immunoassay. In IV, an higly sensitive lateral flow immunoassay utilizing up-converting nanophosphor labels was developed for Plasmodium falciparum infection i.e. malaria, and the test performance was evaluated with P. falciparum culture samples. The analytical sensitivity of P. falciparum detection was improved up to 250-fold as compared to a standard lateral flow test. The results of the publications included in this thesis show that the use of intrumentread luminescent labels in rapid lateral flow immunoassays allows the development of highly sensitive and quantitative point-of-care tests, which could be used in-resource poor areas. Particularly, an ultrasensitive test for the detection of P. falciparum could detect asymptomatic carriers of the malaria parasite and thus support malaria elimination efforts.Pikadiagnostiikka-testit, kuten lateraalivirtaus-immunomääritykset mahdollistavat diagnostiikan käytön köyhillä ja eristyneillä alueilla, mikä auttaa kontrolloimaan ja hävittämään infektiotauteja. Tämä väitöskirja käsittelee kolmen infektiotaudin pikadiagnostiikkaa: hepatiitti C , hinkuyskä ja malaria. Herkemmät pikadiagnostiikkatestit erityisesti malarialoiselle ja hepatiitti C –infektioille voisivat auttaa yrityksissä hävittää nämä taudit. Lisäksi vieritestaukseen soveltuva pikatesti hinkuyskälle auttaisi taudin valvonnassa ja helpottaisi taudin leviämisen estämistä. Lateraalivirtaus-immunomääritysten herkkyyden parantamiseksi on kehitetty useita menetelmiä. Näihin kuuluvat näytteen konsentrointi ennen testin suorittamista, määrityksen komponenttien virtauksen kontrollointi ja luminoivien mittalaitteilla luettavien leimojen käyttö. Kaikki nämä metodit edellyttävät kompromisseja kehitetyn testin herkkyyden, helppokäyttöisyyden ja edullisuuden välillä. Väitöskirjatyön ensimmäisessä julkaisussa kehitettiin immunomäärityksiä HCV vastaaineille käyttäen multiepitooppiproteiini-antigeenia ja luminoivaa europiumkelaattileimaa. Toisessa ja kolmannessa julkaisussa kehitettiin luminoivia europiumnanopartikkeleita käyttäviä kvantitatiivisia lateraalivirtaus-immunomäärityksiä. Toisessa julkaisussa kehitetty hinkuyskän vasta-aineita tunnistavaa lateraalivirtausimmunomääritys korreloi hyvin potilasnäytteillä perinteisen entsyymi-immunomäärityksen kanssa. Neljännessä julkaisussa kehitettiin herkkä käänteisviritteisiä nanopartikkelileimoja käyttävä lateraalivirtaus-immunomääritys Plasmodium falciparum malarialoisen havaitsemiseksi verestä. Kehitetty testi oli noin 250 kertaa herkempi havaitsemaan malarialoisia kuin aikaisemmin käytetyt lateraalivirtaustestit. Tämän väitöskirjatyön tulokset osoittavat, että käyttämällä mittalaitteilla luettavia leimoja lateraalivirtaus-immunomäärityksissä voidaan kehittää erittäin herkkiä kvantitatiivisia pikatestejä resurssiköyhille alueille. Erittäin herkkää malaria-pikatestiä voitaisiin erityisesti käyttää havaitsemaan myös malarialoisen oireettomat kantajat ja täten tukemaan malarian hävittämisohjelmia

Generation Green – A holistic approach to implementation of green principles and practices in educational programmes in pharmaceutical and medical sciences at the University of Helsinki

Solving the environmental and sustainability challenges associated with drug development, manufacturing, distribution, use, and end -of -life requires comprehensive change in the mindset of healthcare professionals on all fronts. Besides current professionals, the faculty teachers and students have a critical role in facilitating the implementation of green principles and practices in educational programs, but no change occurs unless the need and the tools are properly established and their impact understood. This article describes the evolution of green pharmacy practice in the Faculty of Pharmacy at the University of Helsinki, following a previously published framework for change management. Furthermore, the article describes the dissemination of the principles and good practices into medical education.Peer reviewe

High-sensitivity lateral flow immunoassay with a fluorescent lanthanide nanoparticle label

Lateral flow (LF) immunoassays are commonly used for point-of-care testing and typically incorporate visually read reporters, such as gold particles. To improve sensitivity and develop quantitative LF immunoassays, visual reporters can be replaced by fluorescent reporters detected by an instrument. In this study, we used fluorescent europium(III) chelate doped nanoparticle (Eu-np) reporters to develop a quantitative high-sensitivity LF immunoassay for free prostate specific antigen (fPSA). Furthermore, we tested different simplified formats of the assay and the effect of different modifiable parameters on the detection limit of the assay: dynamic range, assay duration and number of assay steps. The molar detection limits of the different assay formats were compared with published detection limits of LF immunoassays with different reporters. The cutoff was calculated from 11 female serum samples. The detection limit of the sensitivity optimized fPSA assay with fPSA spiked into pooled female serum was 0.01 ng/ml, which is approximately 100-fold lower than the most sensitive gold particle LF assays and 10-fold lower than other Eu-np and carbon nanoparticle based LF immunoassays. Thus, Eu-np reporters can be used to develop highly sensitive and quantitative LF immunoassays.</p

Escherichia coli–expressed near full length HIV-1 envelope glycoprotein is a highly sensitive and specific diagnostic antigen

Background: The Human Immunodeficiency Virus type 1 (HIV-1) envelope glycoprotein gp160, useful in detecting anti-HIV-1 antibodies, is difficult to express in heterologous hosts. The major hurdles are its signal sequence, strong hydrophobic regions and heavy glycosylation. While it has not been possible to express full length recombinant (r)-gp160 in E. coli, it can be expressed in insect and mammalian cells, but at relatively higher cost. In this work, we report E. coli-based over-expression of r-gp160 variant and evaluate its performance in diagnostic immunoassays for the detection of anti-HIV-1 antibodies. Methods: A deletion variant of r-gp160 lacking hydrophobic regions of the parent full length molecule was expressed in E. coli and purified to near homogeneity using single-step Ni (II)-affinity chromatography. Biotinylated and europium (III) chelate-labeled versions of this antigen were used to set up one- and two-step time-resolved fluorometric double antigen sandwich assays. The performance of these assays was evaluated against a collection of well-characterized human sera (n = 131), that included an in-house panel and four commercially procured panels. Results: In-frame deletion of three hydrophobic regions, spanning amino acid residues 1–43, 519–538 and 676–706, of full length HIV-1 gp160 resulted in its expression in E. coli. Both the one- and two-step assays manifested high sensitivity unambiguously identifying 75/77 and 77/77 HIV-1 positive sera, respectively. Both assays also identified all 52 HIV-seronegative sera correctly. Between the two assays, the mean signal-to-cutoff value of the two-step assay was an order of magnitude greater than that of the one-step assay. Both assays were highly specific manifesting no cross-reactivity towards antibodies specific to other viruses like hepatitis B, C and human T cell leukemia viruses. Conclusions: This study has demonstrated the expression of r-gp160 variant in E. coli, by deletion of hydrophobic regions, and its purification in reasonable yields. This underscores the potential for cost saving in antigen production. Evaluation of this antigen in a double antigen sandwich two-step assay showed it to be a highly sensitive and specific HIV-1 diagnostic reagent. The amenability of this assay to the one-step format suggests its potential utility in developing a rapid point-of-care HIV-1 diagnostic test

Ultrasensitive and Robust Point-of-Care Immunoassay for the Detection of Plasmodium falciparum Malaria.

Plasmodium falciparum malaria is widespread in the tropical and subtropical regions of the world. There is ongoing effort to eliminate malaria from endemic regions, and sensitive point-of-care (POC) diagnostic tests are required to support this effort. However, current POC tests are not sufficiently sensitive to detect P. falciparum in asymptomatic individuals. After extensive optimization, we have developed a highly sensitive and robust POC test for the detection of P. falciparum infection. The test is based on upconverting nanophosphor-based lateral flow (UCNP-LF) immunoassay. The developed UCNP-LF test was validated using whole blood reference panels containing samples at different parasite densities covering eight strains of P. falciparum from different geographical areas. The limit of detection was compared to a WHO-prequalified rapid diagnostic test (RDT). The UCNP-LF achieved a detection limit of 0.2-2 parasites/μL, depending on the strain, which is 50- to 250-fold improvement in analytical sensitivity over the conventional RDTs. The developed UCNP-LF is highly stable even at 40 °C for at least 5 months. The extensively optimized UCNP-LF assay is as simple as the conventional malaria RDTs and requires 5 μL of whole blood as sample. Results can be read after 20 min from sample addition, with a simple photoluminescence reader. In the absence of a reader device at the testing site, the strips after running the test can be transported and read at a central location with access to a reader. We have found that the test and control line signals are stable for at least 10 months after running the test. The UCNP-LF has potential for diagnostic testing of both symptomatic and asymptomatic individuals

Microparticle - based platform for point-of-care immunoassays

There is a need for quantitative and sensitive, yet simple point-of-care immunoassays: We have developed a point-of-care microparticle-based immunoassay platform which combines the performance of a microliter well based assay with the usability of a rapid assay. The platform contained a separate reaction and detection chambers and microparticles for the solid-phase. Photoluminescent up-converting nanoparticles (UCNPs) were used as labels. The platform was tested with a cardiac troponin I assay, and a limit of detection of 19.7 ng/L was obtained. This study demonstrates the feasibility of developing point-of-care assays on the new platform for various analytes of interests

Generation Green – Incorporation of environmental aspects into the curricula of pharmaceutical and medical education

Peer reviewe

Double-Antigen Lateral Flow Immunoassay for the Detection of Anti-HIV-1 and -2 Antibodies Using Upconverting Nanoparticle Reporters

Rapid diagnostic tests (RDTs) are often used for the detection of anti-human immunodeficiency virus (HIV) antibodies in remote locations in low- and middle-income countries (LMIC) with low or limited access to central laboratories. The typical format of an RDT is a lateral flow assay (LFA) with visual interpretation prone to subjectivity. This risk of misinterpretation can be overcome with luminescent upconverting nanoparticle reporters (UCNPs) measured with a miniaturized easy-to-use reader instrument. An LFA with UCNPs for anti-HIV-1/2 antibodies was developed and the assay performance was evaluated extensively with challenging patient sample panels. Sensitivity (n = 145) of the UCNP-LFA was 96.6% (95% CI: 92.1–98.8%) and specificity (n = 309) was 98.7% (95% CI: 96.7–99.7%). Another set of samples (n = 200) was used for a comparison between the UCNP-LFA and a conventional visual RDT. In this comparison, the sensitivities for HIV-1 were 96.4% (95% CI: 89.8–99.3%) and 97.6% (95% CI: 91.6–99.7%), for the UCNP-LFA and conventional RDT, respectively. The specificity was 100% (95% CI: 96.4–100%) for both assays. The developed UCNP-LFA demonstrates the applicability of UCNPs for the detection of anti-HIV antibodies. The signal measurement is done by a reader instrument, which may facilitate automated result interpretation, archiving and transfer of data from de-centralized locations.</p