Production of transgenic strawberries by temporary immersion bioreactor system and verification by TAIL-PCR

BACKGROUND: Strawberry (Fragaria × ananassa) is an economically important soft fruit crop with polyploid genome which complicates the breeding of new cultivars. For certain traits, genetic engineering offers a potential alternative to traditional breeding. However, many strawberry varieties are quite recalcitrant for Agrobacterium-mediated transformation, and a method allowing easy handling of large amounts of starting material is needed. Also the genotyping of putative transformants is challenging since the isolation of DNA for Southern analysis is difficult due to the high amount of phenolic compounds and polysaccharides that complicate efficient extraction of digestable DNA. There is thus a need to apply a screening method that is sensitive and unambiguous in identifying the different transformation events. RESULTS: Hygromycin-resistant strawberries were developed in temporary immersion bioreactors by Agrobacterium-mediated gene transfer. Putative transformants were screened by TAIL-PCR to verify T-DNA integration and to distinguish between the individual transformation events. Several different types of border sequence arrangements were detected. CONCLUSION: This study demonstrates that temporary immersion bioreactor system suits well for the regeneration of transgenic strawberry plants as a labour-efficient technique. Small amount of DNA required by TAIL-PCR is easily recovered even from a small transformant, which allows rapid verification of T-DNA integration and detection of separate gene transfer events. These techniques combined clearly facilitate the generation of transgenic strawberries but should be applicable to other plants as well

No association in maternal serum levels of TMAO and its precursors in pre-eclampsia and in non-complicated pregnancies

Only a few studies have explored the role of microbiota-dependent metabolite trimethylamine N-oxide (TMAO) in non-complicated pregnancy and in pre-eclampsia (PE). We enrolled 139 PE and 29 healthy pregnant women in a nested case control study. We hypothesized that elevated levels of circulating TMAO and its precursors choline and glycine betaine in the late second or in third trimester might contribute to the PE and are associated with the onset of the disease and clinical features such as elevated blood pressure. The association with a few available lifestyle factors (use of fish and physical activity) was also evaluated. In contrast with the previous findings, there was no difference in TMAO concentration between PE and healthy women. In addition, TMAO concentration was not associated with any of the PE related clinical features, angiogenic or inflammatory markers. In future, it is crucial to obtain longitudinal data on TMAO in both non-complicated and in PE pregnancies before we could have more detailed understanding of TMAO.Peer reviewe

Diet- and microbiota-related metabolite, 5-aminovaleric acid betaine (5-AVAB), in health and disease

5-Aminovaleric acid betaine (5-AVAB) is a trimethylated compound associated with the gut microbiota, potentially produced endogenously, and related to the dietary intake of certain foods such as whole grains. 5-AVAB accumulates within the metabolically active tissues and has been typically found in higher concentrations in the heart, muscle, and brown adipose tissue. Furthermore, 5-AVAB has been associated with positive health effects such as fetal brain development, insulin secretion, and reduced cancer risk. However, it also has been linked with some negative health outcomes such as cardiovascular disease and fatty liver disease. At the cellular level, 5-AVAB can influence cellular energy metabolism by reducing β-oxidation of fatty acids. This review will focus on the metabolic role of 5-AVAB with respect to both physiology and pathology. Moreover, the analytics and origin of 5-AVAB and related compounds will be reviewed

Fearful dogs have increased plasma glutamine and gamma-glutamyl glutamine

Anxiety-related disorders, including fearfulness are common and leading welfare problems among the worldwide dog population. The etiology of anxieties is complex and affected by genetic and environmental factors. Thus, there is a need for more comprehensive approaches, such as metabolomics, to understand the causes of anxiety and to identify anxiety-related biomarkers for more efficient diagnostic and treatment options. To study metabolic alterations related to canine fearfulness, a non-targeted plasma metabolite profiling was performed in a cohort of 20 fearful and 21 non-fearful dogs. The results showed that nine metabolic features were significantly associated with fearfulness. The most prominent change included increased plasma glutamine and gamma-glutamyl glutamine (gamma-Glu Gln) in fearful dogs across breeds. Alterations in glutamine metabolism have previously been associated with several psychiatric disorders, indicating the relevance of this finding also in dogs. In addition, we describe a novel breed-specific association between renal biomarker symmetric dimethylarginine (SDMA) and canine fearfulness. These observed metabolic alterations may result from high levels of prolonged psychological stress in fearful dogs.Peer reviewe

Serum metabolites associated with wholegrain consumption using nontargeted metabolic profiling: a discovery and reproducibility study

Purpose: To identify fasting serum metabolites associated with WG intake in a free-living population adjusted for potential confounders. Methods: We selected fasting serum samples at baseline from a subset (n = 364) of the prospective population-based Kuopio Ischaemic Heart Disease Risk Factor Study (KIHD) cohort. The samples were analyzed using nontargeted metabolomics with liquid chromatography coupled with mass spectrometry (LC–MS). Association with WG intake was investigated using both random forest followed by linear regression adjusted for age, BMI, smoking, physical activity, energy and alcohol consumption, and partial Spearman correlation adjusted for the same covariates. Features selected by any of these models were shortlisted for annotation. We then checked if we could replicate the findings in an independent subset from the same cohort (n = 200). Results: Direct associations were observed between WG intake and pipecolic acid betaine, tetradecanedioic acid, four glucuronidated alkylresorcinols (ARs), and an unknown metabolite both in discovery and replication cohorts. The associations remained significant (FDR<0.05) even after adjustment for the confounders in both cohorts. Sinapyl alcohol was positively correlated with WG intake in both cohorts after adjustment for the confounders but not in linear models in the replication cohort. Some microbial metabolites, such as indolepropionic acid, were positively correlated with WG intake in the discovery cohort, but the correlations were not replicated in the replication cohort. Conclusions: The identified associations between WG intake and the seven metabolites after adjusting for confounders in both discovery and replication cohorts suggest the potential of these metabolites as robust biomarkers of WG consumption

Kestävän kehityksen projekti peruskoulun 6. luokalla

Tiivistelmä. Kestävä kehitys on käsitteenä laaja ja siitä on monia eri tulkintoja. Ensimmäisen kerran kestävää kehitystä käsiteltiin vuonna 1987 Gro Harlem Brundtlandin johtamassa Ympäristön ja kehityksen maailmankomissiossa. Komission raportissa Our Common Future kestävä kehitys määritellään seuraavasti: ”Kestävä kehitys on kehitystä, joka tyydyttää nykyhetken tarpeet viemättä tulevilta sukupolvilta mahdollisuutta tyydyttää omat tarpeensa.” Kestävää kehitystä määritellään kolmen ulottuvuuden, ekologisen, taloudellisen sekä sosiaalisen ja kulttuurisen ulottuvuuden avulla. Kestävän kehityksen käsittelyssä ympäristökasvatus on tärkeässä asemassa. Ympäristökasvatuksen tulisi suuntautua viidelle eri osa-alueelle: tietoisuuden, tiedon, asenteiden, taitojen ja osallistumisen alueille. Ympäristökasvatuksen tavoitteena on auttaa oppijaa tiedostamaan ympäristö kokonaisuutena ja herkistymään sen ongelmille, auttaa hankkimaan tietoa ympäristöstä, auttaa selkiyttämään arvoja ja tunteita suhteessa ympäristöön, auttaa hankkimaan taitoja ympäristöongelmien ratkaisemiseen sekä tarjota mahdollisuus aktiivisen osallistumiseen ympäristöongelmien ratkaisemiseksi. Tutkimukselle asetettiin kaksi tutkimustehtävää, joista ensimmäinen kuvaa projektin suunnittelu-, toteutus- ja arviointiprosessin. Toisen tutkimustehtävän avulla selvitetään projektin vaikutusta oppilaiden tietoihin kestävästä kehityksestä kirjoitelmia analysoiden. Aineistona tässä tutkimuksessa on projektin suunnittelu- ja toteutusvaiheessa kerätyt tutkijoiden muistiinpanot, jakso- ja tuntisuunnitelmat, tutkijoiden kirjoittama tutkimuspäiväkirja sekä oppilailta projektin alussa ja lopussa kerätyt kirjoitelmat. Tämä tutkimus on luonteeltaan kvalitatiivinen ja siinä sovelletaan toimintatutkimuksen strategiaa. Helmikuussa 2013 toteutettiin kestävän kehityksen projekti oululaisessa perusopetuksen koulun 6. luokalla. Oppilaita luokassa oli 20. Projekti toteutettiin luokanopettajaopintoihin sisältyvän teemaharjoittelun yhteydessä. Projektin tavoitteena oli edistää kestävää kehitystä opetuksen ja kasvatuksen avulla. Projektin avulla haluttiin lisätä oppilaiden tietoisuutta kestävästä kehityksestä ja rohkaista oppilaita löytämään tapoja, jotka mahdollistavat kestävän elämäntavan. Tutkimuksen tuloksissa todettiin, että oppilaat kirjoittivat projektin jälkeen niistä asioista, joita toteutuksessa korostettiin. Oppilaat kirjoittivat, että on tärkeää huomioida tulevat sukupolvet ja mainitsivat kestävän kehityksen kolme ulottuvuutta kirjoitelmissaan. Ekologisesti kestävä kehitys painottui sekä projektin alussa että lopussa kerätyissä kirjoitelmissa. Erot ensimmäisten ja toisten kirjoitelmien välillä olivat merkittäviä. Projektin lopussa kerätyt kirjoitelmat olivat tekstimäärältään huomattavasti pidempiä. Kirjoitelmissa kestävää kehitystä kuvailtiin monipuolisesti ja oppilaiden tietomäärä aiheeseen liittyen oli selvästi lisääntynyt. Kestävän kehityksen projekti sisälsi vain yhden suunnittelun, toiminnan ja arvioinnin kierroksen. Syklien määrä ei kuitenkaan ole mielekäs arviointikriteeri toimintatutkimukselle, sillä jo yhteen sykliin voi sisältyä sekä jatkuvuus että spiraalimainen kehittäminen. Tärkeämpää kuin syklien määrä on se, että toimintatutkimuksen perusidea eli suunnittelu, toiminta ja arviointi toteutuivat. Tutkimus on osoitus siitä, että hyvin suunniteltu ja toteutettu projekti antaa mahdollisuuden kestävän tulevaisuuden kannalta tärkeiden arvojen tuomiseksi esiin opetuksessa. Kestävän kehityksen projektista oli käytännön hyötyä sekä oppilaille että tutkijoille itselleen oman opettajuuden kehittymisessä. Opettamalla kestävää kehitystä alakoulussa luodaan perusta maapallon säilymiselle hyvänä myös tuleville sukupolville. Opettajina oli ilahduttavaa huomata, että lähes kaikki oppilaat sisäistivät tämän asian projektin aikana. Opettajan tehtävä on tarjota erilaisia näkökulmia maailmaan — erilaisiin tapoihin elää ja ajatella

Maternal microbiota-derived metabolic profile in fetal murine intestine, brain and placenta

Background The maternal microbiota affects the development of the offspring by microbial metabolites translocating to the fetus. To reveal the spectrum of these molecular mediators of the earliest host-microbe interactions, we compared placenta, fetal intestine and brain from germ-free (GF) and specific pathogen free (SPF) mouse dams by non-targeted metabolic profiling. Results One hundred one annotated metabolites and altogether 3680 molecular features were present in significantly different amounts in the placenta and/or fetal organs of GF and SPF mice. More than half of these were more abundant in the SPF organs, suggesting their microbial origin or a metabolic response of the host to the presence of microbes. The clearest separation was observed in the placenta, but most of the molecular features showed significantly different levels also in the fetal intestine and/or brain. Metabolites that were detected in lower amounts in the GF fetal organs included 5-aminovaleric acid betaine, trimethylamine N-oxide, catechol-O-sulphate, hippuric and pipecolic acid. Derivatives of the amino acid tryptophan, such as kynurenine, 3-indolepropionic acid and hydroxyindoleacetic acid, were also less abundant in the absence of microbiota. Ninety-nine molecular features were detected only in the SPF mice. We also observed several molecular features which were more abundant in the GF mice, possibly representing precursors of microbial metabolites or indicators of a metabolic response to the absence of microbiota. Conclusions The maternal microbiota has a profound impact on the fetal metabolome. Our observations suggest the existence of a multitude of yet unidentified microbially modified metabolites which pass through the placenta into the fetus and potentially influence fetal development.Peer reviewe

Side-stream products of malting: a neglected source of phytochemicals

Whole grain consumption reduces the risk of several chronic diseases. A major contributor to the effect is the synergistic and additive effect of phytochemicals. Malting is an important technological method to process whole grains; the main product, malted grain, is used mainly for brewing, but the process also yields high amounts of side-stream products, such as rootlet. In this study, we comprehensively determined the phytochemical profile of barley, oats, rye, and wheat in different stages of malting and the subsequent extraction phases to assess the potential of malted products and side-streams as a dietary source of bioactive compounds. Utilizing semi-quantitative LC–MS metabolomics, we annotated 285 phytochemicals from the samples, belonging to more than 13 chemical classes. Malting significantly altered the levels of the compounds, many of which were highly increased in the rootlet. Whole grain cereals and the malting products were found to be a diverse and rich source of phytochemicals, highlighting the value of these whole foods as a staple. The characterization of phytochemicals from the 24 different sample types revealed previously unknown existence of some of the compound classes in certain species. The rootlet deserves more attention in human nutrition, rather than its current use mainly as feed, to benefit from its high content of bioactive components