Identification and expression of the 20 kd structural protein gene of colitis bacteriophage

The 2.3 kb BamHI fragment from the colitis bacteriophage DNA was transcribed and translated into a 20 kd structural protein P6, in a coupled transcription-translation system derived from Escherichia coli. This protein was expressed in vivo by the 2.3 kb DNA cloned in pBR322. The gene with the regulatory elements for this protein was located on the 680 bp AvaII fragment of the insert DNA. It hybridized with two RNAs of sizes 520 and 1630 nucleotides indicating that both are messengers for the 20 kd protein. Dot-blot hybridization showed that the transcripts for P6 reached a maximum level at 12 min after phage infection.

Väinö Kantele lähetystyöntekijänä kommunistien valtaannousun aikana Kiinassa 1948—1951

Tämä tutkimus tarkastelee pastori Väinö Kanteleen työtä lähetystyöntekijänä Kiinassa hänen kolmannella työkaudellaan 1948 - 1951. Tutkimuksen keskeisimmät lähteet ovat Kanteleen perheelleen osoittamat päiväkirjamaiset kirjeet sekä Suomen Lähetysseuran (SLS) johtokunnan kokousten ja lähettien kokousten pöytäkirjat. Tutkimus on mikrohistoriallinen sivuten henkilö- ja lähetyshistoriaa. Kantele saapui Yungšunin asemalle maaliskuussa 1948. Asemalle oli kaivattu kokenutta mieslähettiä. Työkauden alku sujui hyvin Kiinan poliittisen tilanteen levottomuudesta huolimatta. Tilanne kuitenkin kiristyi kommunistien saadessa lisää kannatusta. SLS kutsui lähetit pois työalueelta keväällä 1949. Kantele päätti jäädä Kiinaan. Yungšunin seurakunta oli toivonut hänen jäämistään. Lisäksi mahdollisuudet lähetystyön tekemiseen olivat yhä olemassa huolimatta Kiinan kansantasavallan perustamisesta lokakuussa 1949. Olosuhteet alkoivat kuitenkin asteittain kiristyä. Hajanaisten sotilasjoukkojen sekä kommunistiarmeijan liikkuminen ympäri Kiinaa häiritsivät lähetystyötä. Sotilaat muun muassa valtasivat lähetysasemia käyttöönsä. Ongelmana olivat myös inflaatio ja postin kulun epävarmuus. Henkilökohtaisesti Kanteletta vaivasi myös tunne oman uskonsa heikkoudesta. Vuonna 1950 Kiinan poliittiset uudistukset rajoittivat lähetystyötä entisestään. Heinäkuussa julkaistu Kirkkojen manifesti vaati kirkkojen pikaista irrottautumista ulkomaisista yhteyksistään, käytännössä lähetysjärjestöistä. Samana vuonna aloitettiin maareformi. Seurakuntiin asetettiin kokoontumiskielto reformin toteuttamisen ajaksi. Yungšunissa kieltoa ei kumottu enää Kanteleen työkauden aikana. Uudistusten myötä Kanteleen liikkumisvapautta rajoitettiin. Viranomaiset hakivat hänet välillä moneksi päiväksi kuulusteluihin. Lopulta hän eli käytännössä vankina asemallaan. Tammikuussa 1951 hän koki, ettei lähetystyön tekemiseen ole mahdollisuuksia, ja anoi maastapoistumislupaa viranomaisilta. Alkoi pitkä ja epävarma odotus. Kantele alkoi laulaa kiinankielisiä virsiä kovaan ääneen asemalla koska koki, että se oli ainoa jäljellä oleva keino levittää kristinuskon sanomaa. Tapansa vuoksi hänet tunnettiin laulavana evankelistana. Tie kotiin avautui lopulta heinäkuussa 1951. Kantele koki työnsä lähetyssaarnaajana tärkeäksi ja rakkaaksi. Hän oli valmis työskentelemään hankalissa olosuhteissa. Hän oli hyvin ihmisläheinen ja kuvaili kirjeissään paljon tapaamiaan ihmisiä. Kantele oli käytännön ihminen eikä hänen kirjoituksissaan ole havaittavissa taipumusta opillisiin pohdintoihin.

Leskien ja orpojen muuttuva asema luo- ja kisii-yhteisöissä Länsi-Keniassa

Tutkielma käsittelee naisleskien ja orpojen elämää luo- ja kisii-yhteisöissä Länsi-Kenian maaseudulla. Tarkoituksena on selvittää, kuinka yhteiskunnassa tapahtuvat taloudelliset ja sosiaaliset muutokset ovat vaikuttaneet orpojen ja leskien asemaan patriarkaalisissa yhteisöissä. Tutkielma pyrkii osoittamaan paikallistason muutosten kulttuurisidonnaisuuden erilaisia muutosteemoja tarkastelemalla. Erityisesti on tarkasteltu aids-epidemian ja maapulan vaikutuksia leskien ja orpojen lähisuhteisiin, maankäyttöoikeuksiin ja luojen leviraattisuhteeseen. Tutkielman aineisto koostuu kahden kuukauden kenttätyöjakson aikana tehdyistä haastatteluista ja osallistuvasta havainnoinnista. Kenttätyö on tehty Suomen Luterilaisen Evankeliumiyhdistyksen ja Kenian Evangelic Lutheran Churchin orpotyöprojektin yhteydessä syksyllä 2007. Aids-epidemian myötä huollettavien määrä maaseutuyhteisöissä on kasvanut. Samanaikaisesti on havaittu, että sukulaisten muodostama turvaverkko on heikentynyt. Turvaverkon heikentyminen liittyy resurssipulaan ja arvojen muutokseen. Lesket ja orvot ovat vaarassa jäädä ilman tarvitsemaansa tukea ja apua. Kisiiden ja luojen sukulaisuusjärjestelmässä naisilla ei ole siskon tai tyttären rooliin liittyviä oikeuksia. Tutkielmasta käy ilmi, että leskien ja orpojen häätäminen kotitiloilta on yleistä, vaikka perinteisesti heidän asemansa maankäyttäjänä on ollut turvattu. Luo-leski saatetaan häätää myös hänen kieltäytyessään leviraattisuhteesta. Häätöjen keskeinen motiivi on saada lesken käytössä ollut maapala miesvainajan sukulaisten haltuun. Häädetyt lesket rinnastuvat Thomas Håkanssonin tutkimiin avoliitosta häädettyihin kisii-naisiin, joita hän nimittää suvuttomiksi naisiksi. Pekka Seppälän tutkimusta luhyoiden sukupolvenvaihdoksista käytetään maankäyttöoikeuksiin liittyvän analyysin pohjana. Luojen leviraattisuhteessa on tapahtunut merkittäviä muutoksia aids-epidemian aikana. Länsi-Kenian aids-tilanne on muuta maata huonompi. Alueen asukkailla on valistuskampanjoiden ansiosta hyvät tiedot sairaudesta, mutta he toimivat usein tämän tiedon vastaisesti. Tutkielman mukaan keskeisin syy näennäiselle välinpitämättömyydelle on leimautumisen pelko. Aidsiin liittyvä stigma on voimakasta ja sen vuoksi tauti pyritään salaamaan mahdollisimman pitkään. Sukulaismiehet kieltäytyvät usein leskenperijän roolista hiv-tartunnan pelossa. Tämä on luonut kysyntää ammattiperijöille, jotka suorittavat seksuaalisia rituaaleja leskille korvausta vastaan. Tutkielmassa esitellään rituaalisen seksin merkitystä luoille ja todetaan sen olevan tärkein syy leviraattikäytännön jatkumiselle. Luo-leskien perinteistä asemaa tarkastellaan Betty Potashin tutkimusten valossa.

Drug epidemiology, interactions and pharmacogenetics : A post-mortem database study

Use of adverse drug combinations, abuse of medicinal drugs and substance abuse are considerable social problems that are difficult to study. Prescription database studies might fail to incorporate factors like use of over-the-counter drugs and patient compliance, and spontaneous reporting databases suffer from underreporting. Substance abuse and smoking studies might be impeded by poor participation activity and reliability. The Forensic Toxicology Unit at the University of Helsinki is the only laboratory in Finland that performs forensic toxicology related to cause-of-death investigations comprising the analysis of over 6000 medico-legal cases yearly. The analysis repertoire covers most commonly used drugs and drugs of abuse, and the ensuing database contains also background information and information extracted from the final death certificate. In this thesis, the data stored in this comprehensive post-mortem toxicology database was combined with additional metabolite and genotype analyses that were performed to complete the profile of selected cases. The incidence of drug combinations possessing serious adverse drug interactions was generally low (0.71%), but it was notable for the two individually studied drugs, a common anticoagulant warfarin (33%) and a new generation antidepressant venlafaxine (46%). Serotonin toxicity and adverse cardiovascular effects were the most prominent possible adverse outcomes. However, the specific role of the suspected adverse drug combinations was rarely recognized in the death certificates. The frequency of bleeds was observed to be elevated when paracetamol and warfarin were used concomitantly. Pharmacogenetic factors did not play a major role in fatalities related to venlafaxine, but the presence of interacting drugs was more common in cases showing high venlafaxine concentrations. Nicotine findings in deceased young adults were roughly three times more prevalent than the smoking frequency estimation of living population. Contrary to previous studies, no difference in the proportion of suicides was observed between nicotine users and non-nicotine users. However, findings of abused substances, including abused prescription drugs, were more common in the nicotine users group than in the non-nicotine users group. The results of the thesis are important for forensic and clinical medicine, as well as for public health. The possibility of drug interactions and pharmacogenetic issues should be taken into account in cause-of-death investigations, especially in unclear cases, medical malpractice suspicions and cases where toxicological findings are scarce. Post-mortem toxicological epidemiology is a new field of research that can help to reveal problems in drug use and prescription practises.

Maturation of a transient outward potassium current in mouse fetal hypothalamic neurons in culture

The whole-cell voltage clamp technique was used to record potassium currents in mouse fetal hypothalamic neurons developing in culture medium from days 1 to 17. The neurons were derived from fetuses of IOPS/OF1 mice on the 14th day of gestation. The mature neurons (>six days in culture) showed both a transient potassium current and a non-inactivating delayed rectifier potassium current. These were identified pharmacologically by using the potassium channel blockers tetraethyl ammonium chloride and 4-aminopyridine, and on the basis of their kinetics and voltage sensitivities. The delayed rectifier potassium current had a threshold of −20 mV, a slow time-course of activation, and was sustained during the voltage pulse. The 4-aminopyridine-sensitive current was transient, and was activated from a holding potential more negative (−80 mV) than that required for evoking the delayed rectifier potassium current (−40 mV). The delayed rectifier potassium current was detectable from day 1 onwards, while the transient potassium current showed a distinct developmental trend. The time-constant of inactivation became faster with age in culture. The half steady-state inactivation potential showed a shift towards less negative membrane potentials with age, and the relationship was best described by a logarithmic regression equation.The developmental trend of the transient potassium current may relate functionally to the progressive morphological changes, and the appearance of synaptic connections during ontogenesis.

Sotalapsuus, äitiys ja isoäitiys : Kertomuksia sotalapsuuden varjossa

War children were sent away to shelter without their parents to other Nordic countries, mainly to Sweden. The phenomenon was remarkable. During the Second World War nearly 80,000 children were sent from their homes by trains or boats. These children travelled to foster homes where they were placed with new parents looking after them. After the conclusion of the peace, for some months or sometimes years later, orders were given to send the children back to their families in Finland. Returning back to Finland and to their biological parents and families was not always easy. Deep bonds between the children and their foster families were created and leaving caused grief to those small travellers once again. In some cases, distances were created in the relations between Mothers and their daughters. Many had forgotten their Finnish, and returning to school proved difficult. Some of the war children felt rootlessness, a result of being torn away from their family and culture. The aim of this study is to describe how former war children became mothers by themselves, and later on grandmothers. The study also explores how they describe the meaning of the war and their childhood in their own parenthood and what were their experiences of time in foster homes. Seven former war children and three daughters were interviewed for this study. Interviews were biographical. A narrative approach and thematic reading (by Riessman 2008) has guided the analysis of the texts. According to the results of this study, the importance of having your own home , family and security in childhood relationships is significant. Caring and having responsibility for disadvantaged others was important for former war children. What come from the detailed experiences of the 'war childhood' most of all were the difficulties they found on returning to Finland. Some of them had become very attached to their foster parents. There were varying degrees of language problems among the returnees. Some of the interviewees had completely forgotten their native language. Given that, starting the school at home was difficult. They also remembered continuous travelling.When asked on the outcome of their relationship with their biological mother, most interviewees were happy, with a few experiencing some distance in this relationship. Security and being available to protect their children were important in their own motherhood and grand motherhood. In difficult family situations like divorce, they wanted to give their time and support for helping with grandchildren. Another important aspect in family life is interaction between all its members. Talking things through in families and also in War Child Associations was highly valued. However, talking of war childhood had been silenced in some families. In conclusion, the experiences of former war children should take in consideration when difficult situations between parents and children or children s positions in war zones are resolved. War children also have a lot to give for further educational study.

Immunochemical analysis of prolamins in gluten-free foods

People with coeliac disease have to maintain a gluten-free diet, which means excluding wheat, barley and rye prolamin proteins from their diet. Immunochemical methods are used to analyse the harmful proteins and to control the purity of gluten-free foods. In this thesis, the behaviour of prolamins in immunological gluten assays and with different prolamin-specific antibodies was examined. The immunoassays were also used to detect residual rye prolamins in sourdough systems after enzymatic hydrolysis and wheat prolamins after deamidation. The aim was to characterize the ability of the gluten analysis assays to quantify different prolamins in varying matrices in order to improve the accuracy of the assays. Prolamin groups of cereals consist of a complex mixture of proteins that vary in their size and amino acid sequences. Two common characteristics distinguish prolamins from other cereal proteins. Firstly, they are soluble in aqueous alcohols, and secondly, most of the prolamins are mainly formed from repetitive amino acid sequences containing high amounts of proline and glutamine. The diversity among prolamin proteins sets high requirements for their quantification. In the present study, prolamin contents were evaluated using enzyme-linked immunosorbent assays based on ω- and R5 antibodies. In addition, assays based on A1 and G12 antibodies were used to examine the effect of deamidation on prolamin proteins. The prolamin compositions and the cross-reactivity of antibodies with prolamin groups were evaluated with electrophoretic separation and Western blotting. The results of this thesis research demonstrate that the currently used gluten analysis methods are not able to accurately quantify barley prolamins, especially when hydrolysed or mixed in oats. However, more precise results can be obtained when the standard more closely matches the sample proteins, as demonstrated with barley prolamin standards. The study also revealed that all of the harmful prolamins, i.e. wheat, barley and rye prolamins, are most efficiently extracted with 40% 1-propanol containing 1% dithiothreitol at 50 °C. The extractability of barley and rye prolamins was considerably higher with 40% 1-propanol than with 60% ethanol, which is typically used for prolamin extraction. The prolamin levels of rye were lowered by 99.5% from the original levels when an enzyme-active rye-malt sourdough system was used for prolamin degradation. Such extensive degradation of rye prolamins suggest the use of sourdough as a part of gluten-free baking. Deamidation increases the diversity of prolamins and improves their solubility and ability to form structures such as emulsions and foams. Deamidation changes the protein structure, which has consequences for antibody recognition in gluten analysis. According to the resuts of the present work, the analysis methods were not able to quantify wheat gluten after deamidation except at very high concentrations. Consequently, deamidated gluten peptides can exist in food products and remain undetected, and thus cause a risk for people with gluten intolerance. The results of this thesis demonstrate that current gluten analysis methods cannot accurately quantify prolamins in all food matrices. New information on the prolamins of rye and barley in addition to wheat prolamins is also provided in this thesis, which is essential for improving gluten analysis methods so that they can more accurately quantify prolamins from harmful cereals.