Neurological manifestations and molecular genetics of acute intermittent porphyria in North Western Russia

Acute intermittent porphyria (AIP, MIM #176000) is an inherited metabolic disease due to a partial deficiency of the third enzyme, hydroxymethylbilane synthase (HMBS, EC: 4.3.1.8), in the haem biosynthesis. Neurological symptoms during an acute attack, which is the major manifestation of AIP, are variable and relatively rare, but may endanger a patient's life. In the present study, 12 Russian and two Finnish AIP patients with severe neurological manifestations during an acute attack were studied prospectively from 1995 to 2006. Autonomic neuropathy manifested as abdominal pain (88%), tachycardia (94%), hypertension (75%) and constipation (88%). The most common neurological sign was acute motor peripheral neuropathy (PNP, 81%) often associated with neuropathic sensory loss (54%) and CNS involvement (85%). Despite heterogeneity of the neurological manifestations in our patients with acute porphyria, the major pattern of PNP associated with abdominal pain, dysautonomia, CNS involvement and mild hepatopathy could be demonstrated. If more strict inclusion criteria for biochemical abnormalities (>10-fold increase in excretion of urinary PBG) are applied, neurological manifestations in an acute attack are probably more homogeneous than described previously, which suggests that some of the neurological patients described previously may not have acute porphyria but rather secondary porphyrinuria. Screening for acute porphyria using urinary PBG is useful in a selected group of neurological patients with acute PNP or encephalopathy and seizures associated with pain and dysautonomia. Clinical manifestations and the outcome of acute attacks were used as a basis for developing a 30-score scale of the severity of an acute attack. This scale can easily be used in clinical practice and to standardise the outcome of an attack. Degree of muscle weakness scored by MRC, prolonged mechanical ventilation, bulbar paralysis, impairment of consciousness and hyponatraemia were important signs of a poor prognosis. Arrhythmia was less important and autonomic dysfunction, severity of pain and mental symptoms did not affect the outcome. The delay in the diagnosis and repeated administrations of precipitating factors were the main cause of proceeding of an acute attack into pareses and severe CNS involvement and a fatal outcome in two patients. Nerve conduction studies and needle EMG were performed in eleven AIP patients during an acute attack and/or in remission. Nine patients had severe PNP and two patients had an acute encephalopathy but no clinically evident PNP. In addition to axonopathy, features suggestive of demyelination could be demonstrated in patients with severe PNP during an acute attack. PNP with a moderate muscle weakness was mainly pure axonal. Sensory involvement was common in acute PNP and could be subclinical. Decreased conduction velocities with normal amplitudes of evoked potentials during acute attacks with no clinically evident PNP indicated subclinical polyneuropathy. Reversible symmetrical lesions comparable with posterior reversible encephalopathy syndrome (PRES) were revealed in two patients' brain CT or MRI during an acute attack. In other five patients brain MRI during or soon after the symptoms was normal. The frequency of reversible brain oedema in AIP is probably under-estimated since it may be short-lasting and often indistinguishable on CT or MRI. In the present study, nine different mutations were identified in the HMBS gene in 11 unrelated Russian AIP patients from North Western Russia and their 32 relatives. AIP was diagnosed in nine symptom-free relatives. The majority of the mutations were family-specific and confirmed allelic heterogeneity also among Russian AIP patients. Three mutations, c.825+5G>C, c.825+3_825+6del and c.770T>C, were novel. Six mutations, c.77G>A (p.R26H), c.517C>T (p.R173W), c.583C>T (p.R195C), c.673C>T (p.R225X), c.739T>C (p.C247R) and c.748G>C (p.E250A), have previously been identified in AIP patients from Western and other Eastern European populations. The effects of novel mutations were studied by amplification and sequencing of the reverse-transcribed total RNA obtained from the patients' lymphoblastoid or fibroblast cell lines. The mutations c.825+5G>C and c.770T>C resulted in varyable amounts of abnormal transcripts, r.822_825del (p.C275fsX2) and [r.770u>c, r.652_771del, r.613_771del (p.L257P, p.G218_L257del, p.I205_L257del)]. All mutations demonstrated low residual activities (0.1-1.3 %) when expressed in COS-1 cells confirming the causality of the mutations and the enzymatic defect of the disease. The clinical outcome, prognosis and correlation between the HMBS genotype and phenotype were studied in 143 Finnish and Russian AIP patients with ten mutations (c.33G>T, c.97delA, InsAlu333, p.R149X, p.R167W, p.R173W, p.R173Q, p.R225G, p.R225X, c.1073delA) and more than six patients in each group. The patients were selected from the pool of 287 Finnish AIP patients presented in a Finnish Porphyria Register (1966-2003) and 23 Russian AIP patients (diagnosed 1995-2003). Patients with the p.R167W and p.R225G mutations showed lower penetrance (19% and 11%) and the recurrence rate (33% and 0%) in comparison to the patients with other mutations (range 36 to 67% and 0 to 66%, respectively), as well as milder biochemical abnormalities [urinary porphobilinogen 47±10 vs. 163±21 mol/L, p<0.001; uroporphyrin 130±40 vs. 942±183 nmol/L, p<0.001] suggesting a milder form of AIP in these patients. Erythrocyte HMBS activity did not correlate with the porphobilinogen excretion in remission or the clinical of the disease. In all AIP severity patients, normal PBG excretion predicted freedom from acute attacks. Urinary PBG excretion together with gender, age at the time of diagnosis and mutation type could predict the likelihood of acute attacks in AIP patients.

Fatigue fractures in military conscripts : A study on risk factors, diagnostics and long-term consequences

Fatigue fracture is an overuse injury commonly encountered in military and sports medicine, and known to relate to intensive or recently intensified physical activity. Bone responds to increased stress by enhanced remodeling. If physical stress exceeds bone s capability to remodel, accumulation of microfractures can lead to bone fatigue and stress fracture. Clinical diagnosis of stress fractures is complex and based on patient s anamnesis and radiological imaging. Bone stress fractures are mostly low-risk injuries, healing well after non-operative management, yet, occurring in high-risk areas, stress fractures can progress to displacement, often necessitating surgical treatment and resulting in prolonged morbidity. In the current study, the role of vitamin D as a predisposing factor for fatigue fractures was assessed using serum 25OHD level as the index. The average serum 25OHD concentration was significantly lower in conscripts with fatigue fracture than in controls. Evaluating TRACP-5b bone resorption marker as indicator of fatigue fractures, patients with elevated serum TRACP-5b levels had eight times higher probability of sustaining a stress fracture than controls. Among the 154 patients with exercise induced anterior lower leg pain and no previous findings on plain radiography, MRI revealed a total of 143 bone stress injuries in 86 patients. In 99% of the cases, injuries were in the tibia, 57% in the distal third of the tibial shaft. In patients with injury, forty-nine (57%) patients exhibited bilateral stress injuries. In a 20-year follow-up, the incidence of femoral neck fatigue fractures prior to the Finnish Defence Forces new regimen in 1986 addressing prevention of these fractures was 20.8/100,000, but rose to 53.2/100,000 afterwards, a significant 2.6-fold increase. In nineteen subjects with displaced femoral neck fatigue fractures, ten early local complications (in first postoperative year) were evident, and after the first postoperative year, osteonecrosis of the femoral head in six and osteoarthritis of the hip in thirteen patients were found. It seems likely that low vitamin D levels are related to fatigue fractures, and that an increasing trend exists between TRACP-5b bone resorption marker elevation and fatigue fracture incidence. Though seldom detected by plain radiography, fatigue fractures often underlie unclear lower leg stress-related pain occurring in the distal parts of the tibia. Femoral neck fatigue fractures, when displaced, lead to long-term morbidity in a high percentage of patients, whereas, when non-displaced, they do not predispose patients to subsequent adverse complications. Importantly, an educational intervention can diminish the incidence of fracture displacement by enhancing awareness and providing instructions for earlier diagnosis of fatigue fractures.

Detection of renal dysfunction during and after anaesthesia and surgery - evaluation of the influence of inorganic fluoride, ketorolac and clonidine

Drugs and surgical techniques may have harmful renal effects during the perioperative period. Traditional biomarkers are often insensitive to minor renal changes, but novel biomarkers may more accurately detect disturbances in glomerular and tubular function and integrity. The purpose of this study was first, to evaluate the renal effects of ketorolac and clonidine during inhalation anesthesia with sevoflurane and isoflurane, and secondly, to evaluate the effect of tobacco smoking on the production of inorganic fluoride (F-) following enflurane and sevoflurane anesthesia as well as to determine the effect of F- on renal function and cellular integrity in surgical patients. A total of 143 patients undergoing either conventional (n = 75) or endoscopic (n = 68) inpatient surgery were enrolled in four studies. The ketorolac and clonidine studies were prospective, randomized, placebo controlled and double-blinded, while the cigarette smoking studies were prospective cohort studies with two parallel groups. As a sign of proximal tubular deterioration, a similar transient increase in urine N-acetyl-beta-D-glucosaminidase/creatinine (U-NAG/crea) was noted in both the ketorolac group and in the controls (baseline vs. at two hours of anesthesia, p = 0.015) with a 3.3 minimum alveolar concentration hour sevoflurane anesthesia. Uncorrelated U-NAG increased above the maximum concentration measured from healthy volunteers (6.1 units/l) in 5/15 patients with ketorolac and in none of the controls (p = 0.042). As a sign of proximal tubular deterioration, U-glutathione transferase-alpha/crea (U-GST-alpha/crea) increased in both groups at two hours after anesthesia but a more significant increase was noted in the patients with ketorolac. U-GST-alpha/crea increased above the maximum ratio measured from healthy volunteers in 7/15 patients with ketorolac and in 3/15 controls. Clonidine diminished the activation of the renin-angiotensin aldosterone system during pneumoperitoneum; urine output was better preserved in the patients treated with clonidine (1/15 patients developed oliguria) than in the controls (8/15 developed oliguria (p=0.005)). Most patients with pneumoperitoneum and isoflurane anesthesia developed a transient proximal tubular deterioration, as U-NAG increased above 6.1 units/L in 11/15 patients with clonidine and in 7/15 controls. In the patients receiving clonidine treatment, the median of U-NAG/crea was higher than in the controls at 60 minutes of pneumoperitoneum (p = 0.01), suggesting that clonidine seems to worsen proximal tubular deterioration. Smoking induced the metabolism of enflurane, but the renal function remained intact in both the smokers and the non-smokers with enflurane anesthesia. On the contrary, smoking did not induce sevoflurane metabolism, but glomerular function decreased in 4/25 non-smokers and in 7/25 smokers with sevoflurane anesthesia. All five patients with S-F- ≥ 40 micromol/L, but only 6/45 with S-F- less than 40 micromol/L (p = 0.001), developed a S-tumor associated trypsin inhibitor concentration above 3 nmol/L as a sign of glomerular dysfunction. As a sign of proximal tubulus deterioration, U-beta 2-microglobulin increased in 2/5 patients with S-F- over 40 micromol/L compared to 2/45 patients with the highest S-F- less than 40 micromol/L (p = 0.005). To conclude, sevoflurane anesthesia may cause a transient proximal tubular deterioration which may be worsened by a co-administration of ketorolac. Clonidine premedication prevents the activation of the renin-angiotensin aldosterone system and preserves normal urine output, but may be harmful for proximal tubules during pneumoperitoneum. Smoking induces the metabolism of enflurane but not that of sevoflurane. Serum F- of 40 micromol/L or higher may induce glomerular dysfunction and proximal tubulus deterioration in patients with sevoflurane anesthesia. The novel renal biomarkers warrant further studies in order to establish reference values for surgical patients having inhalation anesthesia.

The Structure and Functions of Hantavirus Nucleocapsid Protein

Hantaviruses, members of the genus Hantavirus in the Bunyaviridae family, are enveloped single-stranded RNA viruses with tri-segmented genome of negative polarity. In humans, hantaviruses cause two diseases, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS), which vary in severity depending on the causative agent. Each hantavirus is carried by a specific rodent host and is transmitted to humans through excreta of infected rodents. The genome of hantaviruses encodes four structural proteins: the nucleocapsid protein (N), the glycoproteins (Gn and Gc), and the polymerase (L) and also the nonstructural protein (NSs). This thesis deals with the functional characterization of hantavirus N protein with regard to its structure. Structural studies of the N protein have progressed slowly and the crystal structure of the whole protein is still not available, therefore biochemical assays coupled with bioinformatical modeling proved essential for studying N protein structure and functions. Presumably, during RNA encapsidation, the N protein first forms intermediate trimers and then oligomers. First, we investigated the role of N-terminal domain in the N protein oligomerization. The results suggested that the N-terminal region of the N protein forms a coiled-coil, in which two antiparallel alpha helices interact via their hydrophobic seams. Hydrophobic residues L4, I11, L18, L25 and V32 in the first helix and L44, V51, L58 and L65 in the second helix were crucial for stabilizing the structure. The results were consistent with the head-to-head, tail-to-tail model for hantavirus N protein trimerization. We demonstrated that an intact coiled-coil structure of the N terminus is crucial for the oligomerization capacity of the N protein. We also added new details to the head-to-head, tail-to-tail model of trimerization by suggesting that the initial step is based on interaction(s) between intact intra-molecular coiled-coils of the monomers. We further analyzed the importance of charged aa residues located within the coiled-coil for the N protein oligomerization. To predict the interacting surfaces of the monomers we used an upgraded in silico model of the coiled-coil domain that was docked into a trimer. Next the predicted target residues were mutated. The results obtained using the mammalian two-hybrid assay suggested that conserved charged aa residues within the coiled-coil make a substantial contribution to the N protein oligomerization. This contribution probably involves the formation of interacting surfaces of the N monomers and also stabilization of the coiled-coil via intramolecular ionic bridging. We proposed that the tips of the coiled-coils are the first to come into direct contact and thus initiate tight packing of the three monomers into a compact structure. This was in agreement with the previous results showing that an increase in ionic strength abolished the interaction between N protein molecules. We also showed that residues having the strongest effect on the N protein oligomerization are not scattered randomly throughout the coiled-coil 3D model structure, but form clusters. Next we found evidence for the hantaviral N protein interaction with the cytoplasmic tail of the glycoprotein Gn. In order to study this interaction we used the GST pull-down assay in combination with mutagenesis technique. The results demonstrated that intact, properly folded zinc fingers of the Gn protein cytoplasmic tail as well as the middle domain of the N protein (that includes aa residues 80 248 and supposedly carries the RNA-binding domain) are essential for the interaction. Since hantaviruses do not have a matrix protein that mediates the packaging of the viral RNA in other negatve stranded viruses (NSRV), hantaviral RNPs should be involved in a direct interaction with the intraviral domains of the envelope-embedded glycoproteins. By showing the N-Gn interaction we provided the evidence for one of the crucial steps in the virus replication at which RNPs are directed to the site of the virus assembly. Finally we started analysis of the N protein RNA-binding region, which is supposedly located in the middle domain of the N protein molecule. We developed a model for the initial step of RNA-binding by the hantaviral N protein. We hypothesized that the hantaviral N protein possesses two secondary structure elements that initiate the RNA encapsidation. The results suggest that amino acid residues (172-176) presumably act as a hook to catch vRNA and that the positively charged interaction surface (aa residues 144-160) enhances the initial N-RNA interacation. In conclusion, we elucidated new functions of hantavirus N protein. Using in silico modeling we predicted the domain structure of the protein and using experimental techniques showed that each domain is responsible for executing certain function(s). We showed that intact N terminal coiled-coil domain is crucial for oligomerization and charged residues located on its surface form a interaction surface for the N monomers. The middle domain is essential for interaction with the cytoplasmic tail of the Gn protein and RNA binding.

TOTEM early measurements

The status of the TOTEM experiment is described as well as the prospects for the measurements in the early LHC runs. The primary goal of TOTEM is the measurement of the total p-p cross section, using a method independent of the luminosity. A final accuracy of 1% is ex- pected with dedicated β∗ = 1540 m runs, while at the beginning a 5% resolution is achievable with a β∗ = 90 m optics. Accordingly to the running scenarios TOTEM will be able to measure the elastic scattering in a wide range of t and to study the cross-sections and the topologies of diffractive events. In a later stage, physics studies will be extended to low-x and forward physics collaborating with CMS as a whole experimental apparatus.

Valkoinen läiskä kartalla : Perusvärinnimet ja niiden merkityskentät unkarin- ja suomenkielisissä idiomeissa

Työni tavoitteena on vertailla unkarilaisia ja suomalaisia idiomeja, joissa esiintyy jokin perusvärinnimi. Perusvärinnimiä ovat fehér (valkoinen), fekete (musta), szürke (harmaa), piros tai vörös (punainen), zöld (vihreä), sárga (keltainen), kék (sininen) ja barna (ruskea). Erityisesti pyrin ottamaan selvää käytetäänkö samaa värinnimeä molemmissa kielissä samasta asiasta vai onko värinnimien käytössä tarkasteltujen kielten välillä eroja. Vertailen unkarilaisia idiomeja suomalaisiin ja käytän vertailussa Tamás Forgácsin esittämää mallia ekvivalenssin lajeista. Aineisto on kerätty unkarin- ja suomenkielisistä fraasi- ja idiomisanakirjoista sekä yleissanakirjoista. Idiomi on yksi fraseologismien alaryhmistä. Idiomi koostuu vähintään kahdesta lekseemistä, jotka voidaan kirjoittaa joko yhteen tai erikseen. Idiomi on puhtaimmillaan opaakki, kiteytynyt sanaliitto, mutta idiomi voi olla myös osittain sananmukaisesti tulkittavissa. Idiomaattisuuden ja metaforisuuden aste on jatkumo, jonka toisessa päässä ovat täysin läpinäkymättömät ja toisessa transparentit sanaliitot. Idiomit ovat puhtaimmillaan tiettyyn kieleen sidottuja. Idiomit voivat olla kuitenkin laajalevikkisiä, koska niitä on lainattu eri kieliin suoraan kääntämällä esimerkiksi Raamatusta tai antiikin teksteistä. Viime vuosisatojen ja vuosikymmenten idiomit ovat hypoteettisesti useimmiten käännöslainoja jostain Euroopan valtakielestä. Värinnimitutkimuksen tunnetuimpia teorioita on Berlinin ja Kayn teoria kielessä esiintyvien värinnimien järjestyksestä. Heidän mukaansa musta, valkoinen ja punainen esiintyvät maailman kielissä kaikista yleisimmin, sininen, ruskea ja harmaa puolestaan esiintyvät vain korkeakulttuureissa, eivät esimerkiksi luonnonkansojen kielissä. Hypoteesini on, että aineistossa esiintyvien värinnimien lukumäärä korreloi jossain määrin Berlinin ja Kayn esittämän värinnimien järjestyksen kanssa. Täysin identtisiä idiomeja sanastoltaan ja merkitykseltään löytyy aineistosta suhteellisen paljon, useimmat näistä ovat valkoisen, mustan tai harmaan värin sisältäviä idiomeja. Täysin toisiaan vastaavista idiomeista suurin osa on tullut unkariin ja suomeen Raamatusta tai lainattu sellaisenaan englannista, saksasta tai ranskasta. Osittaisessa vastaavuussuhteessa ja funktionaalisessa vastaavuussuhteessa löytyy värinnimien kannalta mielenkiintoisimmat idiomit: keltainen ja vihreä esiintyvät parissa idiomissa ristikkäin. Pääasiassa idiomeissa esiintyvät värinnimet ovat unkarin ja suomen välillä identtisiä, mikäli unkarilaiselle idiomille löytyy vastine suomen kielestä. Idiomit ilman vastinetta ovat aineiston suurin ja samalla myös kirjavin ryhmä analysoitavaksi, pienin ryhmä puolestaan ovat leksikaalisen vastaavuuden ryhmään kuuluvat idiomit. Suurin osa unkarilaisista idiomeista on työssä analysoitu, mutta suomenkieliset vastineettomat idiomit jäävät vähäiselle huomiolle tutkimusongelman näkökulmasta johtuen. Aineiston 143 unkarilaisesta idiomista noin puolet sijoittuu nollaekvivalenssin eli vastineettomien idiomien ryhmään. Täydellisen ekvivalenssin ryhmä on toiseksi suurin, leksikaalisen vastaavuuden ryhmä pienin. Idiomeissa esiintyvistä väreistä yleisimpiä ovat musta, valkoinen, punainen ja vihreä. Ruskeaa väriä ei esiinny unkarilaisissa idiomeissa lainkaan, suomalaisissa idiomeissa viidessä. Unkarin ja suomen värien merkityskentät vastaavat pitkälti toisiaan. Ainoastaan keltainen ja vihreä esiintyivät selkeästi ristikkäin: unkariksi kateus on keltaista ja suomeksi vihreää, tulokkaaseen liitetään unkarissa vihreä, suomessa keltainen väri.

The Influence of World Factors on Finnish Stock Market Volatility

This paper investigates to what extent the volatility of Finnish stock portfolios is transmitted through the "world volatility". We operationalize the volatility processes of Finnish leverage, industry, and size portfolio returns by asymmetric GARCH specifications according to Glosten et al. (1993). We use daily return data for January, 2, 1987 to December 30, 1998. We find that the world shock significantly enters the domestic models, and that the impact has increased over time. This applies also for the variance ratios, and the correlations to the world. The larger the firm, the larger is the world impact. The conditional variance is higher during recessions. The asymmetry parameter is surprisingly non-significant, and the leverage hypothesis cannot be verified. The return generating process of the domestic portfolio returns does usually not include the world information set, thus indicating that the returns are generated by a segmented conditional asset pricing model.

Seerumin kreatiinikinaasi- ja aspartaattiaminotransferaasiaktiivisuuden vaikutus kalkkunoiden (Meleagris gallopavo) käyttäytymiseen erikorkuisissa kuljetushäkeissä

Kalkkunoiden teuraskuljetuksissa lintuja pidetään kuljetuspäällyksissä, joiden korkeuden on esitetty olevan riittämätön suurimpien kalkkunakukkojen kuljetukseen, mutta tieteellisiä tutkimuksia aiheesta on vähän. Jalostuksella saavutetut nopeakasvuiset ja suuret lihakset voivat lisäksi altistaa kalkkunat mahdollisesti kivuliaille lihassairauksille. Tutkielman tavoitteena oli selvittää lihasentsyymiarvojen vaikutusta kalkkunoiden käyttäytymiseen erikorkuisissa kuljetuspäällyksissä. Tutkimuksessa analysoitiin kreatiinikinaasin (CK) ja aspartaattiaminotransferaasin (ASAT) aktiivisuutta kalkkunoiden seerumissa, sillä näiden solunsisäisten entsyymien yhtäaikainen esiintyminen seerumissa on todettu olevan merkki lihasvauriosta ja täten kuvaavan eläimen heikentynyttä hyvinvointia. Tutkimuksessa käytettiin 36 lihantuotantoon jalostettua kalkkunakukkoa. Kutakin lintua testattiin kahtena eri päivänä noin viikon välein. Testattavien lintujen paino oli keskimäärin 16,5 ± 0,2 kiloa. Linnut olivat eri testikerroilla satunnaistetusti erikorkuisissa häkeissä. Häkkikorkeudet olivat 40, 55 ja 90 cm. Linnut olivat paikallaan olevissa häkeissä kuusi tuntia, jonka ajan niiden käyttäytymistä videoitiin. Kustakin linnusta otettiin verinäyte yhdellä testauskerralla kuvaamisen päätyttyä ja seerumista analysoitiin CK ja ASAT. Kalkkunoiden CK-aktiivisuus oli 25450,5 ±10402,6 IU/l ja ASAT-aktiivisuus 625,0 ±143,7 IU/l. Häkkikorkeudella ei ollut tilastollisesti merkitsevää korrelaatiota CK- eikä ASAT-aktiivisuuden kanssa (p = 0,86 ja p = 0,68), mutta CK- ja ASAT-arvot korreloivat positiivisesti keskenään (p < 0,001). Sekä CK- että ASAT-aktiivisuuksien ollessa korkeat linnut makasivat vähemmän 55 cm ja 90 cm korkeissa häkeissä testijakson ensimmäisinä tunteina. Paino ja CK-aktiivisuus olivat positiivisesti korreloituneet (p = 0,001). Kalkkunoiden lihasentsyymiarvoista on saatavilla heikosti tietoa, mutta verrattuna moniin muihin eläinlajeihin kalkkunoiden seerumin CK- ja ASAT-arvot ovat huomattavan korkeat. 40 cm korkeissa häkeissä linnut eivät voineet tilan ahtauden vuoksi seistä normaalissa asennossa jalat ojennettuina kuten muissa häkkikorkeuksissa. Linnuilla kivun liittymistä lihassairauksiin ei ole kuvattu, mutta kipu on varsin todennäköistä akuutissa vaiheessa. Kalkkunat siis saattavat mahdollisuuksien mukaan välttää makaamista kivun vuoksi. Jalostuksella saatu rintalihaksen nopea kasvu suhteettoman suureksi on todettu voivan aiheuttaa lihasvaurioita. On mahdollista, että jo suuri koko itsessään saattaa aiheuttaa kalkkunoille kipua. Tekijät, jotka kohottavat seerumin lihasentsyymiarvoja, aiheuttavat joka tapauksessa myös hyvinvointiongelmia. Nykyisin käytössä olevat matalat, 40 cm korkeat, kuljetuspäällykset voivat heikentää etenkin suurikokoisten kalkkunakukkojen kuljetuksen aikaista hyvinvointia, koska ne estävät lintuja seisomasta luonnollisessa asennossa ja niissä liikkuminen on muutenkin hyvin rajoitettua. Jotta jalostettujen kalkkunoiden lihasentsyymiarvoista voitaisiin tehdä tarkempia johtopäätöksiä, tarvitaan lisää tutkimuksia lihasentsyymiarvojen perustason määrittämiseksi. Lisätutkimuksia tarvitaan myös kalkkunoiden mahdollisesti kokeman kivun ja lihasentsyymiarvojen välisestä yhteydestä.

Kelan asumistukitilasto 2010

Kelan asumistukitilasto 2010 sisältää keskeiset tiedot Kelan maksamista eläkkeensaajan asumistuista, yleisistä asumistuista sekä opintotuen asumislisistä. Julkaisu tarjoaa tilastoja näiden etuuksien saajista, asumisolosuhteista sekä maksetuista euromääristä. Eläkkeensaajan asumistuki, yleinen asumistuki ja opintotuen asumislisä ovat yksin asuville ja perheille myönnettäviä, suoraan asumismenoja pienentäviä asumistukia, joiden toimeenpano kuuluu Kelalle. Kela maksaa asumisen tukemiseen myös sotilasavustuksen asumisavustusta, mutta toimeentulotuen tyyppinen asumisavustus ei sisälly julkaisun tilastotaulukoihin.

The challenge of LC/MS/MS multi-mycotoxin analysis - Heracles battling the Hydra?

Mycotoxins are secondary metabolites of filamentous fungi. They pose a health risk to humans and animals due to their harmful biological properties and common occurrence in food and feed. Liquid chromatography/mass spectrometry (LC/MS) has gained popularity in the trace analysis of food contaminants. In this study, the applicability of the technique was evaluated in multi-residue methods of mycotoxins aiming at simultaneous detection of chemically diverse compounds. Methods were developed for rapid determination of toxins produced by fungal genera of Aspergillus, Fusarium, Penicillium and Claviceps from cheese, cereal based agar matrices and grains. Analytes were extracted from these matrices with organic solvents. Minimal sample clean-up was carried out before the analysis of the mycotoxins with reversed phase LC coupled to tandem MS (MS/MS). The methods were validated and applied for investigating mycotoxins in cheese and ergot alkaloid occurrence in Finnish grains. Additionally, the toxin production of two Fusarium species predominant in northern Europe was studied. Nine mycotoxins could be determined from cheese with the method developed. The limits of quantification (LOQ) allowed the quantification at concentrations varying from 0.6 to 5.0 µg/kg. The recoveries ranged between 96 and 143 %, and the within-day repeatability (as relative standard deviation, RSDr) between 2.3 and 12.1 %. Roquefortine C and mycophenolic acid could be detected at levels of 300 up to 12000 µg/kg in the mould cheese samples analysed. A total of 29 or 31 toxins could be analysed with the method developed for agar matrices and grains, with the LOQs ranging overall from 0.1 to 1250 µg/kg. The recoveries ranged generally between 44 and 139 %, and the RSDr between 2.0 and 38 %. Type-A trichothecenes and beauvericin were determined from the cereal based agar and grain cultures of F. sporotrichioides and F. langsethiae. T-2 toxin was the main metabolite, the average levels reaching 22000 µg/kg in the grain cultures after 28 days of incubation. The method developed for ten ergot alkaloids from grains allowed their quantification at levels varying from 0.01 to 10 µg/kg. The recoveries ranged from 51 to 139 %, and the RSDr from 0.6 to 13.9 %. Ergot alkaloids were measured in barley and rye at average levels of 59 and 720 µg/kg, respectively. The two most prevalent alkaloids were ergocornine and ergocristine. The LC/MS methods developed enabled rapid detection of mycotoxins in such applications where several toxins co-occurred. Generally, the performance of the methods was good, allowing reliable analysis of the mycotoxins of interest with sufficiently low quantification limits. However, the variation in validation results highlighted the challenges related to optimising this type of multi-residue methods. New data was obtained about the occurrence of mycotoxins in mould cheeses and of ergot alkaloids in Finnish grains. In addition, the study revealed the high mycotoxin-producing potential of two common fungi in Finnish crops. The information can be useful when risks related to fungal and mycotoxin contamination will be assessed.

Effects of gender, and SLCO1B1 and CYP7A1 polymorphisms on plasma bile acids in humans and the pharmacokinetics of therapeutic ursodeoxycholic acid

Bile acids are important steroid-derived molecules essential for fat absorption in the small intestine. They are produced in the liver and secreted into the bile. Bile acids are transported by bile flow to the small intestine, where they aid the digestion of lipids. Most bile acids are reabsorbed in the small intestine and return to the liver through the portal vein. The whole recycling process is referred to as the enterohepatic circulation, during which only a small amount of bile acids are removed from the body via faeces. The enterohepatic circulation of bile acids involves the delicate coordination of a number of bile acid transporters expressed in the liver and the small intestine. Organic anion transporting polypeptide 1B1 (OATP1B1), encoded by the solute carrier organic anion transporter family, member 1B1 (SLCO1B1) gene, mediates the sodium independent hepatocellular uptake of bile acids. Two common SNPs in the SLCO1B1 gene are well known to affect the transport activity of OATP1B1. Moreover, bile acid synthesis is an important elimination route for cholesterol. Cholesterol 7α-hydroxylase (CYP7A1) is the rate-limiting enzyme of bile acid production. The aim of this thesis was to investigate the effects of SLCO1B1 polymorphism on the fasting plasma levels of individual endogenous bile acids and a bile acid synthesis marker, and the pharmacokinetics of exogenously administered ursodeoxycholic acid (UDCA). Furthermore, the effects of CYP7A1 genetic polymorphism and gender on the fasting plasma concentrations of individual endogenous bile acids and the bile acid synthesis marker were evaluated. Firstly, a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of bile acids was developed (Study I). A retrospective study examined the effects of SLCO1B1 genetic polymorphism on the fasting plasma concentrations of individual bile acids and a bile acid synthesis marker in 65 healthy subjects (Study II). In another retrospective study with 143 healthy individuals, the effects of CYP7A1 genetic polymorphism and gender as well as SLCO1B1 polymorphism on the fasting plasma levels of individual bile acids and the bile acid synthesis marker were investigated (Study III). The effects of SLCO1B1 polymorphism on the pharmacokinetics of exogenously administered UDCA were evaluated in a prospective genotype panel study including 27 healthy volunteers (Study IV). A robust, sensitive and simple HPLC-MS/MS method was developed for the simultaneous determination of 16 individual bile acids in human plasma. The method validation parameters for all the analytes met the requirements of the FDA (Food and Drug Administration) bioanalytical guidelines. This HPLC-MS/MS method was applied in Studies II-IV. In Study II, the fasting plasma concentrations of several bile acids and the bile acid synthesis marker seemed to be affected by SLCO1B1 genetic polymorphism, but these findings were not replicated in Study III with a larger sample size. Moreover, SLCO1B1 polymorphism had no effect on the pharmacokinetic parameters of exogenously administered UDCA. Furthermore, no consistent association was observed between CYP7A1 genetic polymorphism and the fasting plasma concentrations of individual bile acids or the bile acid synthesis marker. In contrast, gender had a major effect on the fasting plasma concentrations of several bile acids and also total bile acids. In conclusion, gender, but not SLCO1B1 or CYP7A1 polymorphisms, has a major effect on the fasting plasma concentrations of individual bile acids. Moreover, the common genetic polymorphism of CYP7A1 is unlikely to influence the activity of CYP7A1 under normal physiological conditions. OATP1B1 does not play an important role in the in vivo disposition of exogenously administered UDCA.