Bordetella pertussis - Vaccination and Strain Variation

Pertussis or whooping cough is a highly contagious vaccine-preventable disease of the human respiratory tract caused by the <i>Bordetella pertussisi> bacteria. In Finland, pertussis vaccinations were started in 1952 leading to a dramatic decrease in the morbidity and mortality. In the late 1990s, the incidence of pertussis increased despite the high vaccination coverage. Strain variation has been connected to the re-emergence of pertussis in countries with long history of pertussis vaccination. In 2005, the pertussis vaccine and the vaccination schedule were changed in Finland. The molecular epidemiology and the strain variation of the B. <i>pertussisi> isolates were examined in Finland and in countries with similar (France) and different (Sweden) vaccination history. Continuous evolution of the B. <i>pertussis i>population in Finland was observed since the 1950s, and the recently circulating isolates were antigenically different from the vaccine strains. Comparison of the circulating isolates from Finland, France and Sweden did not refer to significant differences. Certain type of strains noticed in France already in 1994 mainly caused the recent epidemics in Sweden (1999) and in Finland (2003-4). On several occasions, a new type of strains first appeared in Sweden and some years later in Finland. The B.<i> pertussisi> isolates from the infants were shown to be similar to those from the other age groups. It is suggested that the strains originate from the same reservoir among adolescents and adults. The strain variation does not seem to have a major effect on the morbidity among recently vaccinated individuals, but it might play a role among those who are in the waning phase of immunity. The incidence of pertussis in Finland has remained low since the change of the vaccination programme. This might be related to the epidemic nature of pertussis and the near future will show the real effectiveness of the new vaccination programme. At present, many infants are infected because they are too young to be immunised with the current schedule. New strategies or vaccines are needed to protect those who are the most vulnerable.

Immune Evasion by Borrelia burgdorferi – With Special Reference to CD38-mediated Chemotaxis of Neutrophils and Dendritic Cells

Lyme borreliosis is a tick-transmitted infection caused by the spirochete bacterium <i>Borrelia burgdorferi i> sensu lato. The tick injects bacteria into host skin, where a first line defence, mainly the complement system, neutrophils, dendritic cells and macrophages are ready to attack foreign intruders. However, in the case of Lyme borreliosis, the original immune response in the skin is untypically mild among bacterial infections. A further untypical feature is the ability of <i>B. burgdorferi</i> to disseminate to distant organs, where, in some patients, symptoms appear after years after the original infection. This study aimed at uncovering some of the immune evasion mechanisms utilized by <i>B. burgdorferi</i> against the complement system, neutrophils and dendritic cells. <i>B. burgdorferi</i> was shown to inhibit chemotaxis of human neutrophils towards nformyl- methyl-leucyl-phenylalanine (fMLP). Outer surface protein B (OspB) of <i>B. burgdorferi</i> was shown to promote resistance to the attack of the complement system and neutrophil phagocytosis at low complement concentrations. <i>B. burgdorferi</i> was shown to inhibit migration of dendritic cells in vitro towards CCL19 and CCL21 and also in an in vivo model. This effect was shown to be due to the absence of CD38 on the borrelia-stimulated dendritic cell surface. A defect in p38 mitogen-activated-protein-kinase (p38) signaling was linked to defective CD38 expression. A defect in CD38 expression on <i>B. burgdorferi-i>stimulated neutrophils was also observed. In this study, a number of novel immune evasion strategies utilized by <i>B burgdorferi</i> were chracterized. However, further studies are needed as other immune evasion mechanisms await to be uncovered.

Borrelia burgdorferi evades the effects of ceftriaxone treatment in a mouse model of Lyme borreliosis

<b><i>Borrelia burgdorferi</i> infektoitujen hiirten antibioottihoidon jälkeinen oireilub> Lymen borrelioosi on puutiaisten välittämä monimuotoinen infektiotauti, jonka tunnetuin oire on ns. vaeltava ihottuma eli erythema migrans. Muita tavallisia ilmentymiä ovat erityisesti nivel- ja hermosto-oireet sekä harvemmin sydän- ja silmäoireet. Suurin osa potilaista paranee täysin terveeksi antibioottihoidon avulla, mutta jopa 10 % borrelioosiin sairastuneista oireilee suositusten mukaisesta hoidosta huolimatta. Pitkittyneen oireilun on ajateltu johtuvan mm. infektion laukaisemasta autoimmuunitaudista tai kroonisesta infektiosta, mutta teorioiden tueksi ei ole kyetty esittämään kiistattomia todisteita. Onkin todennäköistä, että antibioottihoidon jälkeisen oireilun takana on useampia mekanismeja eikä yksi teoria selitä kaikkien potilaiden oireilua. Tässä väitöskirjatyössä on tutkittu hoidonjälkeistä borrelioosia hiirimallin avulla. Varhaisvaiheessa (2 viikkoa infektoinnin jälkeen) annettu antibiootti vähensi hiirten nivelturvotusta ja esti B. burgdorferi – bakteerin kasvun kudoksista otetuista näytteissä. Hoidettu¬jen hiirten B. burgdorferi -spesifiset IgG-luokan vasta-aineet pysyivät kuitenkin koholla ja osasta kudosnäytteistä löytyi B. burgdorferi:n DNA:ta PCR-tutkimuksen avulla. Mikäli hiiret hoidettiin myöhäisessä vaiheessa (yli 18 viikkoa infektoinnista) tulokset olivat muuten samanlaiset, mutta keftriaksoni ei vaikuttanut nivelturvotukseen. Näin hiirissä oli aikaansaatu tilanne, joka on hyvin samankaltainen ihmisen hoitoresistentin borrelia-artriitin kanssa: oireet jatkuvat, mutta taudinaiheuttajaa ei saada esiin. Inflammaatiota vaimentavaa anti-TNF-alphaa on käytetty nivelreuman hoidossa menestyksekkäästi huonosti muuhun hoitoon reagoivilla potilailla ja siitä syystä sen ajateltiin voivan vaikuttaa suotuisasti myös <i>B. burgdorferi</i> -infektoitujen hiirten hoidonjälkeiseen nivelturvotukseen. Sillä ei kuitenkaan ollut vaikutusta nivelturvotukseen, mutta yllättäen hoidon jälkeen osa hiirten kudosnäytteistä osoittautui viljelypositiivisiksi. On siis ilmeistä, että hiirimallissamme osa <i>B. burgdorferi</i> spirokeetoista pystyy välttämään keftriaksonihoidon vaikutuksen joko hakeutumalla elimistössä kudokseen, jossa antibiootin pitoisuus ei nouse riittävän korkeaksi, tai ne kykenevät muuntautumaan metabolisesti inaktiiviin tilaan eikä mikrobilääke yhdessä immuunipuolustuksen kanssa onnistu tappamaan niitä. Jatkotutkimuksissa selvitimme <i>B. burgdorferi i>- spirokeetan mahdollista piilopaikkaa tutkimalla antibioottihoidon jälkeen useita eri kudoksia PCR-menetelmällä. Tulosten perusteella spirokeetta näyttää suosivan nivelkudosta tai soluja, joita esiintyy nivelessä runsaasti. On kuitenkin edelleen epäselvää, missä muodossa <i>B. burgdorferi</i> –spirokeetat säilyvät kudoksessa antibioottihoidon jälkeen.

Therapeutic Properties of Superoxide Dismutase 3 and Mesenchymal Stromal Cells in Peripheral Ischemia

The aim of this study was to characterize the cellular mechanisms leading to the beneficial effect of anti-oxidative gene therapy and pro-angiogenic stem cell therapy in acute peripheral ischemia. Post-ischemic events aim to re-establish tissue blood perfusion, to clear cellular debris, and to regenerate lost tissue by differentiation of satellite cells into myoblasts. Although leukocytes have an essential role in clearing cellular debris and promoting angiogenesis, they also contribute to tissue injury through excessive ROS production. First, we investigated the therapeutic properties of extracellular superoxide dismutase (SOD3) gene transfer. SOD3 was shown to reduce oxidative stress, to normalize glucose metabolism, and to enhance cell proliferation in the ischemic muscle. Analysis of the mitogenic Ras-Erk1/2 pathway showed SOD3 mediated induction offering a plausible explanation for enhanced cell proliferation. In addition, SOD3 reduced NF-κB activity by enhancing IκBα expression thus leading to reduced expression of inflammatory cytokines and adhesion molecules with consequent reduction in macrophage infiltration. Secondly, we sought to determine the fate and the effect of locally transplanted mesenchymal stem/stromal cells (MSCs) in acute ischemia. We showed that a vast majority of the transplanted cells are cleared from the injury site within 24 hours after local transplantation. Despite rapid clearance, transplantation was able to temporarily promote angiogenesis and cell proliferation in the muscle. Lack of graft-derived growth factor expression suggests other than secretory function to mediate this observed effect. In conclusion, both SOD3 and MSCs could be utilized to alleviate peripheral ischemia induced tissue injury. We have described a previously unidentified growth regulatory role for SOD3, and suggest a novel mechanism whereby transplanted MSCs enhance the reparative potential of the recipient tissue through physical contacts.

Identifying and Characterizing New Ingredients in vitro for Prebiotic and Synbiotic use

The endogenous microbiota, constituting the microbes that live inside and on humans, is estimated to outnumber human cells by a factor of ten. This commensal microbial population has an important role in many physiological functions, with the densest microbiota population found in the colon. The colonic microbiota is a highly complex and diverse bacterial ecosystem, and a delicate balance exists between the gut microbiota and its host. An imbalance in the microbial ecosystem may lead to severe symptoms in and also beyond the gastrointestinal tract. Due to the important role of the gut microbiota in human health, means of its modification have been introduced in the dietary concepts of pro-, pre- and synbiotics. Prebiotics, which are usually carbohydrates, strive to selectively influence beneficial microbes resident in the colon with the aim of modifying the composition and functionality of the commensal microbial population towards a purportedly healthier one. The study of prebiotic effects on colonic micro-organisms is typically done by using human faecal material, though this provides relatively little information on bacterial populations and metabolic events in different parts of the colon. For this reason, several <i>in vitroi> models have been developed to investigate the gut microbiota. The aim of this doctoral thesis was to screen through some of the promising prebiotic candidates, characterize their effects on the microbiota through the use of two <i>in vitroi> methods (pure microbial cultures and a colon simulator model) and to evaluate their potential as emerging prebiotics or synbiotics when combined with the probiotic <i>Bifidobacterium lactisi> . As a result of the screening work and subsequent colon simulation studies, several compounds with promising features were identified. Xylo-oligosaccharides (XOS), which have previously already shown promise as prebiotic compounds, were well fermented by several probiotic <i>Bifidobacterium lactisi> strains in pure culture studies and in the following simulation studies utilizing the complex microbiota by endogenous <i>B. lactisi> Another promising compound was panose, a trisaccharide belonging to isomalto-oligosaccharides (IMO) that also was also able to modify the microbiota <i>in vitroi> by increasing the number of beneficial microbes investigated. Panose has not been widely studied previously and therefore, this thesis work provided the first data on panose fermentation in mixed colonic microbiota. Galacto-oligosaccharide (GOS) is an established prebiotic, and it was studied here in conjunction with another potential polygosaccharide polydextrose (PDX) and probiotic <i>B. lactisi> Bi-07. In this final study, the synbiotics including GOS were more effective than the constituting pro- or prebiotics alone in modulating the microbiota composition, thus indicating a synergy resulting from the combination. The results obtained in this <i>in vitroi> work can be, and have already been, utilized in product development aimed at the nutritional modification of the human colonic microbiota. Some of the compounds have entered the human clinical intervention phase to nvestigate in more detail the prebiotic and synbiotic properties seen in these <i>in vitroi> studies.

Antimicrobial Resistance in Campylobacter jejuni and Campylobacter coli

<b>Antimicrobial Resistance in <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i>b> Campylobacters are a common cause of bacterial gastroenteritis worldwide, with <i>Campylobacter jejuni</i> and <i>C. coli</i> being the most common species isolated in human infections. If antimicrobial treatment is required, the drugs of choice at the moment are the macrolides and fluoroquinolones. In this thesis, the in vitro resistance profiles of the <i>C. jejuni</i> and <i>C. coli</i> strains were evaluated with emphasis on multidrug resistance. The aim was also to evaluate the different resistance mechanisms against the macrolides. Further, the disk diffusion method was compared to agar dilution method and its repeatability was evaluated, since it has been widely used for the susceptibility testing of campylobacters. The results of the present study showed that resistance to the fluoroquinolones is common in strains isolated from Finnish patients, but resistance to the macrolides is still rare. Multidrug resistance was associated with resistance to both ciprofloxacin and erythromycin. Among the available per oral drugs, least resistance was observed to coamoxiclav There was no resistance to the carbapenems. Sitafloxacin and tigecycline were <i>in vitroi> highly effective towards <i>Campylobacteri> species. A point mutation A2059G of the 23S rRNA gene was the main mechanism behind the macrolide resistance, whereas the efflux pumps did not seem to play an important role when a strain had A2059G mutation. A five amino acids insertion, which has not been described previously, in the ribosomal protein L22 of one highly-resistant C. jejuni strain without mutation in the 23S rRNA gene was also detected. Concerning the disk diffusion method, there was variation in the repeatability In conclusion, macrolides still appear to be the first-choice alternative for suspected <i>Campylobacteri> enteritis. The<i> in vitroi> susceptibilities found suggest that co-amoxiclav might be a candidate for clinical trials on campylobacteriosis, but in life-threatening situations, a carbapenem may be the drug of choice. More studies are needed on whether the disk diffusion test method could be improved or whether all susceptibilities of campylobacters should be done using a MIC based method.

Regulation of cell growth, death, and polarization by ERBB4

<b>Regulation of cell growth, death, and polarization by <i>ERBB4i>b> ErbB4 is a member of the epidermal growth factor receptor (EGFR, ErbB) family. The other members are EGFR, ErbB2 and ErbB3. ErbB receptors are important regulators for example in cardiovascular, neural and breast development but control key cellular functions also in many adult tissues. Abnormal ErbB signaling has been shown to be involved in various illnesses such as cancers and heart diseases. Among the ErbBs, ErbB4 has been shown to have unique signaling characteristics. ErbB4 exists in four alternatively spliced isoforms that are expressed in a tissue-specific manner. Two of the isoforms can be cleaved by membrane proteases, resulting in release of soluble intracellular domains (ICD). Once released into the cytosol, the ICD is capable of translocating into the nucleus and participating in regulation of transcription. The functional differences and the tissue-specific expression patterns suggest isoformspecific roles for ErbB4 isoforms. However, the abilities of ErbB4 isoforms to differently regulate cellular functions were discovered only recently and are not well understood. This study aimed to determine the expression patterns of ErbB4 in normal and diseased tissue, and to define whether the cleavable and non-cleavable isoforms could regulate different target genes and therefore, cellular functions. In this study, a comprehensive ErbB4 expression pattern in several normal tissues, various cancers and non-neoplastic diseases was determined. In addition, the data demonstrated that the cleavable and non-cleavable ErbB4 isoforms could regulate different cellular functions and target genes. Finally, this study defined the cellular responses regulated by ErbB4 during kidney development.

Evolution of Bordetella pertussis post vaccination

<b>Evolution of <i>Bordetella pertussisi> post vaccinationb> Whooping cough or pertussis is caused by the gram-negative bacterium <i>Bordetella pertussisi>. It is a highly contiguous disease in the human respiratory tract. Characteristic of pertussis is a paroxysmal cough with whooping sound during gasps of breath after coughing episodes. It is potentially fatal to unvaccinated infants. The best approach to fight pertussis is to vaccinate. Vaccinations against pertussis have been available from the 1940s. Traditionally vaccines were whole-cell pertussis (wP) preparations as part of the combined diphtheria-tetanus-pertussis (DTP) vaccines. More recently acellular pertussis (aP) vaccines have replaced the wP vaccines in many countries. The aP vaccines are less reactogenic and can also be administered to school children and adults. There are several publications reporting variation in the i>B. pertussisi> virulence factors that are also aP vaccine antigens. This has occurred in the genes coding for pertussis toxin and pertactin about 15 to 30 years after the introduction of pertussis vaccines to immunisation programs. Resurgence of pertussis has also been reported in many countries with high vaccination coverage. In this study the evolution of <i>B. pertussisi> was investigated in Finland, the United Kingdom, Poland, Serbia, China, Senegal and Kenya. These represent countries with a long history of high vaccination coverage with stable vaccines or changes in the vaccine formulation; countries which established high vaccination coverage late; and countries where vaccinations against pertussis were started late. With bacterial cytotoxicity and cytokine measurements, comparative genomic hybridisation, pulsed-field gel electrophoresis (PFGE), genotyping and serotyping it was found that changes in the vaccine composition can postpone the emergence of antigenic variants. It seems that the change in PFGE profiles and the loss of genetic material in the genome of <i>B. pertussisi> are similar in most countries and the vaccine-induced immunity is selecting non-vaccine type strains. However, the differences in the formulation of the vaccines, the vaccination programs and in the coverage of pertussis vaccination have affected the speed and timing of these changes.

Uudenmaan ELY-keskuksen kevyen liikenteen tarveselvitys 2013

Suunnitelmassa on kuvattu Uudenmaan ELY-keskuksen kevyen liikenteen hankkeiden priorisointimenetelmä ja hankekorimenettely. ELY-keskus ylläpitää tietoa maanteiden kevyen liikenteen hanketarpeista sekä niiden kiireellisyydestä ja vaikutuksista hankekorissa. Kevyen liikenteen hankekorissa on tällä hetkellä noin 500 hanketta. Tämän selvityksen laadinnan yhteydessä hankekoriin tehtiin kokonaisvaltainen päivitys, jossa kaikki hankkeet käytiin läpi. Päivitysprosessiin sisältyi myös vuonna 2012 toteutettu laaja kuntakysely, jossa kartoitettiin kuntien näkemyksiä alueensa tärkeimmistä kevyen liikenteen hanketarpeista. Hankkeiden priorisointi perustuu kaikille hankkeille yhteneväisesti määritettyyn tarveindeksiin, jonka pohjalta hankkeet asetetaan asiantuntijatyönä kiireellisyysluokkiin. Tarveindeksi huomioi lukuisia eri tekijöitä, kuten kevyen liikenteen onnettomuuksien määrän, hankkeen pituuden, hankkeen vaikutusalueen väestön ja koululaisten määrän, koulun läheisyyden, sijainnin taajamassa tai haja-asutusalueella, joukkoliikenteen määrän, pitkämatkaisen liikenteen sekä tien tekniset ominaisuudet (muun muassa liikennemäärä, nopeusrajoitus ja pientareen leveys). Tarveindeksi on merkittävä työkalu, mutta se ei yksin määrää hankkeen kiireellisyysluokitusta. Erityisesti hankkeiden käyttäjämäärien arviointiin on viime vuosina panostettu. Kymmenien hankkeiden käyttäjämääriä on laskettu maastossa. Lisäksi ensimmäistä kertaa on hyödynnetty simulointimallin antamia arvioita hankkeiden pyöräilypotentiaalista. Hankkeiden arvioinnissa keskeisinä tekijöinä tarveindeksin ja käyttäjämäärän lisäksi ovat muun muassa kuntien näkemykset hankkeiden kiireellisyydestä, ajoneuvoliikenteen määrä ja sidokset muihin hankkeisiin sekä käytettävissä oleva rahoitus. Työssä käsiteltyjen hankkeiden lopullinen jako kiireellisyysluokkiin tehtiin edellä mainittujen tekijöiden pohjalta iteratiivisena asiantuntijatyönä. Kärkihankejoukkoon nostettiin ne hankkeet, jotka ovat kustannustehokkaita, vähentävät tehokkaimmin henkilövahinkoon johtavia onnettomuuksia, edistävät kestävää liikkumista eli kävelyn ja pyöräilyn kulkumuoto-osuuden kasvattamista sekä tukevat yhdyskuntarakenteen eheyttämistä. Parhaimmat hankkeet nostettiin A-luokkaan ja B-luokkaan jäivät ne maantieverkon hankkeet, jotka eivät tällä hetkellä ole ELY-keskuksen mielestä ajankohtaisia. Kaikkein kiireellisimmät hankkeet ovat A2-luokassa. Näitä hankkeita (9 kpl) Uudenmaan ELY-keskus haluaa edistää yhdessä kuntien kanssa 50/50-periaatteella. Seuraavaksi kiireellisimmät maantieverkon hankkeet ovat A3-luokassa (19 kpl). Näiden edistämiseen ELYkeskus voi osallistua 20 prosentilla. A-luokka käsittää kokonaisuudessaan 28 hanketta, joiden kokonaiskustannusarvion on noin 33 miljoonaa euroa. B-luokan hankkeita ELY-keskus ei rahoita. Uutena luokkana muodostettiin K-luokka. Nämä K-luokan kevyen liikenteen hankkeet ovat hyviä kevyen liikenteen hankkeita, mutta sijaitsevat kuntien asemakaavoittamilla alueilla, joissa maantien tulisi jo nyt olla osa kunnan katuverkkoa. Näiden hankkeiden edistämiseen Uudenmaan ELY-keskus osallistuu ainoastaan silloin kun maantie muutetaan kaduksi kevyen liikenteen hankkeen myötä. K1-luokkaan (12 kpl) on nostettu sellaisia hankkeita, joiden edistämisestä ELY-keskus aikoo neuvotella kuntien kanssa. Kaikkien K-hankkeiden osalta kustannusjako neuvotellaan tapauskohtaisesti. Kevyen liikenteen hankkeet palvelevat eniten paikallista ja lyhytmatkaista liikennettä, joten niiden edistäminen yhteistyössä kuntien kanssa on luontevaa ja nykyisellä ELY-keskuksen rahoitustasolla lähes välttämätöntä. ELY-keskus haluaa kehittää alueiden kävely- ja pyöräilymahdollisuuksia yhdessä kuntien kanssa. Yhdessä hankkeita rahoittamalla on mahdollista edistää hankkeita useammalla alueella. ELY-keskus toivoo, että kunnat ovat aktiivisia ja ottavat yhteyttä mikäli haluavat edistää maanteiden kävely- ja pyöräilyolosuhteita.

De anima separata

Variantti B.

De anima separata

Variantti A.

De anima separata

Variantti C.

Travel changes thought and men : a fresh perspective on the travels of a literary icon

Kirjallisuusarvostelu

Les royaumes de Suede, de Danemarck et de Norwege: divisez en plusieurs provinces, et gouvernemens, etc.

Janamittakaavat: Milles pas geometriques ; Lieues de Suede ; Lieues d'une Heure de Chemin.