An improved automated immunoassay for C-reactive protein on the Dimension® clinical chemistry system

Recent clinical data indicate that the measurement of the concentration of C-reactive protein (CRP) requires a higher sensitivity and wider dynamic range than most of the current methods can offer. Our goal was to develop a totally automated and highly sensitive CRP assay with an extended range on the Dimension® clinical chemistry system based on particle-enhanced turbidimetric-immunoassay (PETIA) technology. The improved method was optimized and compared to the Binding Site's radial immunodiffusion assay using disease state specimens to minimize interference. Assay performance was assessed on the Dimension® system in a 12-instrument inter-laboratory comparison study. A split-sample comparison (n = 622) was performed between the improved CRP method on the Dimension® system and the N Latex CRP mono method on the Behring Nephelometer, using a number of reagent and calibrator lots on multiple instruments. The method was also referenced to the standard material, CRM470, provided by the International Federation of Clinical Chemistry (IFCC). The improved CRP method was linear to 265.1mg/l with a detection limit between 0.2 and 0.5mg/l. The method detects antigen excess from the upper assay limit to 2000mg/l, thereby allowing users to retest the sample with dilution. Calibration was stable for 60 days. The within-run reproducibility (CV) was less than 5.1% and total reproducibility ranged from 1.1 to 6.7% between 3.3 and 265.4mg/l CRP. Linear regression analysis of the results on the improved Dimension® method (DM) versus the Behring Nephelometer (BN) yielded the following equation: DM = 0.99 × BN − 0.37; r = 0.992. Minimal interference was observed from sera of patients with elevated IgM, IgG and IgA. The recovery of the IFCC standard was within 100 ± 7 % across multiple lots of reagent and calibrator. The improved CRP method provided a sensitive, accurate and rapid approach to quantify CRP in serum and plasma on the Dimension® clinical chemistry system. The ability to detect antigen excess eliminated reporting falsely low results caused by the ‘prozone effect’

Whole-genome sequencing illuminates multifaceted targets of selection to humic substances in Eurasian perch

Extreme environments are inhospitable to the majority of species, but some organisms are able to survive in such hostile conditions due to evolutionary adaptations. For example, modern bony fishes have colonized various aquatic environments, including perpetually dark, hypoxic, hypersaline and toxic habitats. Eurasian perch (Perca fluviatilis) is among the few fish species of northern latitudes that is able to live in very acidic humic lakes. Such lakes represent almost "nocturnal" environments; they contain high levels of dissolved organic matter, which in addition to creating a challenging visual environment, also affects a large number of other habitat parameters and biotic interactions. To reveal the genomic targets of humic-associated selection, we performed whole-genome sequencing of perch originating from 16 humic and 16 clear-water lakes in northern Europe. We identified over 800,000 single nucleotide polymorphisms, of which >10,000 were identified as potential candidates under selection (associated with >3000 genes) using multiple outlier approaches. Our findings suggest that adaptation to the humic environment may involve hundreds of regions scattered across the genome. Putative signals of adaptation were detected in genes and gene families with diverse functions, including organism development and ion transportation. The observed excess of variants under selection in regulatory regions highlights the importance of adaptive evolution via regulatory elements, rather than via protein sequence modification. Our study demonstrates the power of whole-genome analysis to illuminate the multifaceted nature of humic adaptation and provides the foundation for further investigation of causal mutations underlying phenotypic traits of ecological and evolutionary importance

Ferredoxin containing bacteriocins suggest a novel mechanism of iron uptake in <i>Pectobacterium spp</i>

In order to kill competing strains of the same or closely related bacterial species, many bacteria produce potent narrow-spectrum protein antibiotics known as bacteriocins. Two sequenced strains of the phytopathogenic bacterium &lt;i&gt;Pectobacterium carotovorum&lt;/i&gt; carry genes encoding putative bacteriocins which have seemingly evolved through a recombination event to encode proteins containing an N-terminal domain with extensive similarity to a [2Fe-2S] plant ferredoxin and a C-terminal colicin M-like catalytic domain. In this work, we show that these genes encode active bacteriocins, pectocin M1 and M2, which target strains of &lt;i&gt;Pectobacterium carotovorum&lt;/i&gt; and &lt;i&gt;Pectobacterium atrosepticum&lt;/i&gt; with increased potency under iron limiting conditions. The activity of pectocin M1 and M2 can be inhibited by the addition of spinach ferredoxin, indicating that the ferredoxin domain of these proteins acts as a receptor binding domain. This effect is not observed with the mammalian ferredoxin protein adrenodoxin, indicating that &lt;i&gt;Pectobacterium spp.&lt;/i&gt; carries a specific receptor for plant ferredoxins and that these plant pathogens may acquire iron from the host through the uptake of ferredoxin. In further support of this hypothesis we show that the growth of strains of &lt;i&gt;Pectobacterium carotovorum&lt;/i&gt; and &lt;i&gt;atrosepticum&lt;/i&gt; that are not sensitive to the cytotoxic effects of pectocin M1 is enhanced in the presence of pectocin M1 and M2 under iron limiting conditions. A similar growth enhancement under iron limiting conditions is observed with spinach ferrodoxin, but not with adrenodoxin. Our data indicate that pectocin M1 and M2 have evolved to parasitise an existing iron uptake pathway by using a ferredoxin-containing receptor binding domain as a Trojan horse to gain entry into susceptible cells

Cohort Profile: COVIDMENT: COVID-19 cohorts on mental health across six nations

Why were the cohorts set up? With more than 218 million cases and 4.5 million deaths worldwide (Worldometers, 31 August 2021), the COVID-19 pandemic has had an unprecedented influence on the global economy and population health. As a potent global disaster, it is likely to significantly affect the incidence of adverse mental health symptoms and psychiatric disorders, particularly in vulnerable and highly affected populations. The World Health Organization and leading scientific journals have alerted concerning the potential adverse mental health impact of COVID-19 and emphasized the need for multinational research in this area, which additionally provides new insights into disease mechanisms

Tilausliikenne ja matkanjärjestäjä: luvat ja arvonlisäverotus, case: Tilausliikenne Pekka Palmgren Ky

Opinnäytetyön aiheena oli selvittää, mitä kaikkea tilausliikenneyrityksen pitää osata ottaa huomioon niin tilausliikenteen osalta kuin myös matkanjärjestäjän osalta kotimaassa ja ulkomailla. Opinnäytetyössä tutkittaviksi kohdiksi määritettiin tilausliikenteen yleiset luvat kotimaassa, tilausliikenteeltä vaaditut luvat ulkomailla, arvonlisäverotus kotimaassa, eri maiden arvonlisäverokohtelu ulkomailla sekä matkanjärjestäjältä vaaditut toimenpiteet kotimaassa ja ulkomailla. Opinnäytetyön tarkoituksena oli toimia manuaalina tilausliikenneyrityksille, jotka harkitsevat omien matkojen järjestämistä. Opinnäytetyön teoreettisessa osassa käsiteltiin ensin Suomessa tilausliikenteeltä ja matkanjärjestäjältä vaaditut lainmukaiset luvat sekä muut vaatimukset, jonka jälkeen siirryttiin tarkastelemaan Euroopan Unionin jäsenmaissa vallitsevia säännöksiä lähinnä arvonlisäveron kannalta. Euroopan Unionin aluetta tarkasteltiin ensin yleisesti, ja sitten muutamien maiden käytäntöä selvitettiin vähän tarkemmin: Ruotsi, Viro, Latvia, Liettua, Saksa, Tanska ja Belgia. Lopuksi käsiteltiin Euroopan Unionin ulkopuolisten maiden vaatimuksia ja sääntöjä. Tarkemmin tutustuttiin Venäjän ja Norjan käytäntöihin. Opinnäytetyössä oli haasteellista löytää tietoa varsinkin ulkomaiden arvonlisäverotuksesta. Ongelmaksi muodostui myös Suomen lain osalta matkanjärjestäjän puutteellinen määrittäminen, johon saatiin selvennystä uuden valmismatkalain ansiosta. Empiirisessä osiossa tarkasteltiin kohdeyrityksen nykyistä käytäntöä matkanjärjestäjänä ja sekä pohdittiin tulevaisuudessa mahdollisten matkojen lisäyksien vaatimien toimenpiteiden kannattavuutta. Kuluttajavirastolle annettavan vakuuden suuruutta määritettäessä laskettiin erilaisten ideaalisten matkojen määrä, ja arvioitiin keskimääräiset matkustajamäärät erilaisilla matkoilla. Erilaisten matkojen ennakkolaskutuksen tarvetta pohdittiin, ja kaikkien matkojen ennakkolaskutusta verrattiin pidempien matkojen ennakkolaskutukseen. Opinnäytetyössä todettiin, että rekisteröitymiset arvonlisäverovelvolliseksi eri yhteisömaihin eivät ole kannattavia pienille yrityksille, kun matkoja on vain muutama vuosittain. Matkailualan arvonlisäverotus on syystäkin monimutkainen, joten eri maiden vaatimukset kannattaa tarkistaa etukäteen ennen matkaa.The subject of this thesis was to examine what a charter-traffic company and a travel agent both in one's home country and abroad have to take into consideration. The main subjects in this thesis were determined to be common licenses of charter traffic, required charter-traffic permits abroad, value-added tax in the home country, value-added tax practices in different countries and what actions travel agents have to do in their home country and abroad. The purpose of this thesis was to create a manual for charter-traffic companies, who consider arranging their own trips. In the theoretical section of this thesis I first examined legal licenses and other requirements from charter-traffic companies and travel agents in Finland, and then moved on to the viewpoint of current legal provisions, especially on value-added tax, in member countries of the European Union. At first the European Union was studied generally, and then the practices of a number of countries were studied a little closer, namely those of Sweden, Estonia, Latvia, Lithuania, Germany, Denmark and Belgium. Lastly I studied requirements and rules in countries outside of the European Union. The practices of Russia and Norway were examined in more detail. It was challenging to find information for this thesis especially about value-added taxation in foreign countries. The undefined concept of the travel agent in Finnish law was also problematic, but the new Package Travel Act was able to clarify the situation. In the empirical section I examined existing practices of the case company as a travel agent and pondered whether it would be profitable to increase the number of trips and what kind of operations it would require. To determine the amount of the pledge required by the Consumer Agency I calculated the ideal amount of different travels and evaluated the average number of passengers on different trips. The need of advance invoicing of different trips was pondered, and the advance invoicing of all trips was compared to the advance invoicing of longer trips. In this thesis I came to the conclusion that value-added tax registrations in different EU countries aren't profitable to small companies when there are only a few trips annually. The value-added taxation in tourism is complex for a reason; therefore the requirements of different countries should be checked in advance before the trip