Alu element in the RNA binding motif protein, X-linked 2 (RBMX2) gene found to be linked to bipolar disorder

Objective We have used long-read single molecule, real-time (SMRT) sequencing to fully characterize a similar to 12Mb genomic region on chromosome Xq24-q27, significantly linked to bipolar disorder (BD) in an extended family from a genetic sub-isolate. This family segregates BD in at least four generations with 24 affected individuals. Methods We selected 16 family members for targeted sequencing. The selected individuals either carried the disease haplotype, were non-carriers of the disease haplotype, or served as married-in controls. We designed hybrid capture probes enriching for 5-9Kb fragments spanning the entire 12Mb region that were then sequenced to screen for candidate structural variants (SVs) that could explain the increased risk for BD in this extended family. Results Altogether, 201 variants were detected in the critically linked region. Although most of these represented common variants, three variants emerged that showed near-perfect segregation among all BD type I affected individuals. Two of the SVs were identified in or near genes belonging to the RNA Binding Motif Protein, X-Linked (RBMX) gene family-a 330bp Alu (subfamily AluYa5) deletion in intron 3 of the RBMX2 gene and an intergenic 27bp tandem repeat deletion between the RBMX and G protein-coupled receptor 101 (GPR101) genes. The third SV was a 50bp tandem repeat insertion in intron 1 of the Coagulation Factor IX (F9) gene. Conclusions Among the three genetically linked SVs, additional evidence supported the Alu element deletion in RBMX2 as the leading candidate for contributing directly to the disease development of BD type I in this extended family.Peer reviewe

Screening of Bacteriophage Encoded Toxic Proteins with a Next Generation Sequencing-Based Assay

Bacteriophage vB_EcoM_fHy-Eco03 (fHy-Eco03 for short) was isolated from a sewage sample based on its ability to infect an Escherichia coli clinical blood culture isolate. Altogether, 32 genes encoding hypothetical proteins of unknown function (HPUFs) were identified from the genomic sequence of fHy-Eco03. The HPUFs were screened for toxic properties (toxHPUFs) with a novel, Next Generation Sequencing (NGS)-based approach. This approach identifies toxHPUF-encoding genes through comparison of gene-specific read coverages in DNA from pooled ligation mixtures before electroporation and pooled transformants after electroporation. The performance and reliability of the NGS screening assay was compared with a plating efficiency-based method, and both methods identified the fHy-Eco03 gene g05 product as toxic. While the outcomes of the two screenings were highly similar, the NGS screening assay outperformed the plating efficiency assay in both reliability and efficiency. The NGS screening assay can be used as a high throughput method in the search for new phage-inspired antimicrobial molecules

Cardiogenic shock; focus on ventilatory strategies, the elderly, and biomarker-based risk stratification

Cardiogenic shock (CS) is the most severe form of acute heart failure (HF) characterized by hypotension and systemic hypoperfusion caused by cardiac dysfunction. Prolonged hypotension provokes neurohumoral compensatory mechanisms and systemic inflammatory responses, leading to organ injury followed by multiorgan failure and poor prognosis. CS may be caused by various etiological factors, acute coronary syndromes (ACS) being the most common cause. In addition to prompt recognition of CS, the cause of shock should be treated urgently, as by means of immediate revascularization in the case of ACS-related CS. Although aggressive therapy options, such as mechanical circulatory devices and ventilatory support, may be used, this approach is demanding on the patient, carries risk for complications, and requires additional healthcare resources. Despite advanced therapy options, prognosis in CS is still very poor with a short-term mortality rate up to 40%. Appropriate risk assessment in the early stage of shock is crucial to identify patients most likely to benefit from intensive and costly treatment. The aim of this thesis was to assess the treatment of respiratory failure in CS, focusing on the use of different ventilatory strategies and their impact on outcome; to evaluate the contemporary clinical picture, prognosis, and risk assessment of elderly (≥ 75 years) CS patients; and to investigate prognostic properties of two novel biomarkers in the early phase of CS. The study population consisted of the multinational, observational, prospective CardShock study, which included 219 patients with both ACS and non-ACS etiologies. Study I evaluated the use of different ventilatory strategies in CS. Although most patients (63%) were treated with invasive mechanical ventilation (IMV), a fair number were successfully treated with non-invasive ventilation (NIV) (12%). The intensity of respiratory support required was dependent on the severity of shock; those treated with IMV suffered from more severe shock and had higher 90-day mortality compared with those treated with NIV (49% vs. 27%). However, after balancing the IMV and NIV groups with shock severity and clinical characteristics, the choice of ventilatory strategy itself did not influence the outcome. Study II assessed the key features of elderly (≥ 75 years) CS patients. The elderly constituted a quarter of the population. Despite similar etiology and treatment of shock, they had a higher in-hospital mortality rate compared with younger patients (46% vs. 33%). However, those elderly patients who survived to hospital discharge had a prognosis comparable to that of the younger. The two contemporary risk prediction scores, the CardShock risk score and the IABP-SHOCK II risk score, proved to be useful in the early risk stratification in the elderly patients as well. Furthermore, by incorporating the novel biomarkers into the scores, the risk prediction ability of the scores improved markedly. Study III evaluated concentrations of a novel biomarker, growth differentiation factor 15 (GDF-15), in CS. GDF-15 is a stress-responsive cytokine that is expressed under acute and chronic stressful conditions and it has shown prognostic potential in various diseases. In this study GDF-15 levels were already very high at the beginning of CS and associated with markers of hypoperfusion (high lactate, low pH) and various acute organ dysfunctions (heart, renal, liver) indicating severe circulatory failure. High and increasing levels of GDF-15 were associated with a worse outcome, while low and decreasing levels were indicative of better prognosis. Furthermore, GDF-15 improved the early risk stratification of CS beyond the clinical risk score. Study IV investigated the levels of soluble urokinase-type plasminogen activator receptor (suPAR) in the early stage of CS. suPAR is a novel biomarker secreted in response to inflammatory stimuli and thought to reflect the level of immunoactivation. suPAR has prognostic ability in many acute and chronic diseases, including cardiovascular diseases, cancer, and sepsis. In this study, suPAR levels were clearly elevated during the first days of CS. High levels were associated with both acute and chronic organ dysfunctions. suPAR had independent prognostic potential in CS and improved the risk stratification, especially in the patients with intermediate risk. In conclusion, NIV can be used safely in the treatment of respiratory failure in suitable CS patients. The choice of ventilation strategy did not appear to influence outcome. Elderly patients constitute a considerable portion of CS patients with high mortality. Contemporary risk scores are also useful for early risk prediction in this age group. High levels of the novel biomarkers GDF-15 and suPAR are indicative of severe circulatory failure and end-organ injury, suggesting poor prognosis. These biomarkers may be new prognostic tools in the risk assessment of CS.Sydänperäinen sokki on monitahoinen oireyhtymä, jossa sydämen akuutti toimintahäiriö aiheuttaa sen supistumiskyvyn merkittävän heikentymisen johtaen vaikeaan verenkiertovajaukseen, kudosten hapenpuutteeseen ja monielinvaurioon sekä lopulta hoitamattomana kuolemaan. Oireyhtymän patogeneesissä myös elimistön neurohumoraalisilla vasteilla sekä systeemisen tulehdusreaktion kehittymisellä on oma roolinsa. Mikä tahansa sydämen toimintaa heikentävä sairaus voi olla syynä sydänperäiseen sokkiin, useimmiten kyseessä on laaja sydäninfarkti. Hoidon kulmakivenä on sydämen toimintahäiriön aiheuttaneen syyn tunnistaminen ja välitön korjaaminen. Sokkiin liittyvää verenkierto- ja hengitysvajausta hoidetaan tarvittaessa mekaanisia tukilaitteita avuksi käyttäen tehohoito-olosuhteissa, mikä on kuitenkin potilaille raskasta, altistaa komplikaatioille ja vaatii runsaasti terveydenhuollon resursseja. Kehittyneistä hoitotoimenpiteistä huolimatta kuolleisuus sydänperäiseen sokkiin on edelleen korkea noin 40 %. Hoidossa oleellista on sokin tunnistaminen, raskaista hoidoista hyötyvien korkean riskin potilaiden tunnistaminen ja hoitotoimenpiteiden aloittaminen riittävän varhaisessa vaiheessa ennen peruuttamattomien pääte-elinvaurioiden syntymistä. Tämän väitöskirjan tavoitteena oli selvittää sokkiin liittyvän hengitysvajauksen hoitoa keskittyen eri hengitystukimuotojen käyttöön ja niiden mahdolliseen ennustevaikutukseen sekä tutkia iäkkäiden (≥ 75-vuotiaiden) sokkipotilaiden taudin kliinistä kuvaa ja selvittää ajankohtaisten riskipisteytysmallien käyttökelpoisuutta tämän ikäryhmän ennusteen arvioimisessa. Lisäksi tavoitteena oli arvioida kahden uuden biomarkkerin käyttökelpoisuutta sydänperäisen sokin ennustearviossa. Väitöskirjan tutkimusaineisto on peräisin 219 potilasta käsittävästä monikansallisesta, havainnoivasta CardShock –tutkimuksesta. Ensimmäisessä osatyössä tutkittiin sydänperäiseen sokkiin sairastuneiden potilaiden hengitysvajauksen hoitoa ja eri hengitystukimuotojen käyttöä keskittyen non-invasiiviseen (NIV) hengitystukihoitoon. Tutkimuksessa havaittiin, että sydänperäiseen sokkiin sairastuneiden hengitystuen tarve riippui sokin vaikeusasteesta. Suurin osa potilaista (63%) hoidettiin invasiivisella mekaanisella ventilaatiolla (IMV) keinoilmatien kera, mutta merkittävä osa (12%) pärjäsi pelkästään non-invasiivisella ventilaatiolla (NIV). IMV:lla hoidetut potilaat kärsivät vaikeammasta sokin taudinkuvasta ja heillä oli korkeampi 90 päivän kuolleisuus NIV:llä hoidettuihin verrattuna (49% vs. 27%). Hengitystukistrategian valinnalla ei kuitenkaan ollut vaikutusta ennusteeseen. Toisessa osatyössä selvitettiin iäkkäiden (≥ 75-vuotiaiden) sydänperäiseen sokkiin sairastuneiden potilaiden kliinistä taudinkuvaa, hoitoa ja ennustearviota. Neljäsosa potilaista oli yli 75-vuotiaita. Huolimatta sokin samankaltaisesta etiologiasta ja hoidosta iäkkäiden potilaiden sairaalakuolleisuus oli selvästi nuorempia korkeampi (46% vs. 33%). Toisaalta sokista selvinneiden iäkkäiden ennuste ei eronnut nuorempien ennusteesta. Tutkimuksen mukaan sydänperäiseen sokkiin sairastuneille kehitetyt riskipisteytysmallit toimivat hyvin myös iäkkäillä ja niiden ennustearviota voidaan parantaa yhdistämällä ne biomarkkereiden kanssa. Kolmannessa osatyössä tutkittiin biomerkkiaine GDF-15 pitoisuuksia sydänperäisen sokin alkuvaiheessa sekä niiden yhteyttä ennusteeseen. GDF-15-pitoisuuksien tiedetään nousevan elimistön erilaisissa akuuteissa sekä kroonisissa stressitilanteissa ja GDF-15 omaa ennustearvoa useissa eri sairauksissa. Tutkimuksessa havaittiin, että GDF-15–pitoisuudet olivat hyvin korkeita jo sokin ensivaiheessa. Korkeat pitoisuudet olivat yhteydessä kudosten riittämätöntä verenkiertoa kuvastaviin biomarkkereihin (korkea laktaatti ja matala pH) ja elintoimintahäiriöihin (sydän, maksa, munuaiset) sekä huonompaan ennusteeseen. Lisäksi todettiin, että nouseva GDF-15 –taso oli merkki huonosta ennusteesta, kun taas laskevat pitoisuudet kuvastivat parempaa ennustetta. Yhdistettynä kliiniseen riskipisteytysmalliin GDF-15 paransi sokkipotilaiden ennustearviota huomattavasti. Neljännessä osatyössä määritettiin biomerkkiaine suPAR:n pitoisuuksia sydänperäisen sokin alkuvaiheessa. SuPAR on biomerkkiaine, jonka pitoisuus nousee sekä akuuttien että kroonisten tulehduksellisten tilojen yhteydessä kuvastaen elimistön immunoaktivaatiota. Sen on todettu omaavan ennustearvoa useissa eri sairauksissa, kuten syövissä, sepsiksessä sekä sydän- ja verisuonisairauksissa. Tässä tutkimuksessa suPAR-tasot olivat selvästi koholla sydänperäiseen sokkiin sairastuneilla. Korkeat pitoisuudet olivat yhteydessä eri elintoimintahäiriöihin ja suurempaan kuolemanriskiin. Tutkimuksessa todettiin, että suPAR oli itsenäinen sydänperäisen sokin ennustetekijä ja yhdistettynä kliinisen riskipisteytysmallin kanssa se paransi erityisesti keskiriskin potilaiden ennustearviota. Yhteenvetona voidaan todeta, että sydänperäiseen sokkiin liittyvää hengitysvajausta voidaan hoitaa voidaan turvallisesti myös NIV:lla oikein valikoiduilla potilailla, eikä hengitysvajauksen hoitomuodolla ole merkitystä potilaan ennusteen kannalta. Iäkkäät muodostavat merkittävän osan sydänperäiseen sokkiin sairastuneista potilaista ja sokin ennustearvioon luodut kliiniset riskipisteytysmallit toimivat hyvin myös tässä ikäryhmässä. Tutkitut biomerkkiaineet, GDF-15 ja suPAR, kuvastavat sydänperäiseen sokkiin liittyvää vaikeaa verenkiertovajausta sekä siihen liittyviä elintoimintahäiriöitä ja voivat olla avuksi sokkipotilaiden ennustearviossa

Scientific Opinion on Dietary Reference Values for copper

Following a request from the European Commission, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) derived Dietary Reference Values (DRVs) for copper. Owing to the absence of appropriate biomarkers of copper status and the limitations of available balance studies, the Panel was unable to derive Average Requirements (ARs) and Population Reference Intakes (PRIs). Hence, Adequate Intakes (AIs) were defined based on mean observed intakes in several European Union (EU) countries, given that there is no evidence of overt copper deficiency in the European population. Data from balance studies were used as supportive evidence. For adults, AIs of 1.6 mg/day for men and 1.3 mg/day for women are proposed. For children, AIs are 0.7 mg/day for children aged 1 to < 3 years, 1 mg/day for children aged 3 to < 10 years, and 1.3 and 1.1 mg/day for boys and girls aged 10 to < 18 years, respectively. For infants aged 7–11 months, based on mean observed intakes in four EU countries, an AI of 0.4 mg/day is proposed, which is supported by upwards extrapolation of estimated copper intake in exclusively breast-fed infants. For pregnant women, an increment of 0.2 mg/day is estimated to cover the amount of copper deposited in the fetus and the placenta over the course of pregnancy and in anticipation of the needs for lactation, and for lactating women the same increment is estimated to cover the amount of copper secreted with breast milk. Thus, for pregnant and lactating women, the Panel derived an AI of 1.5 mg/day

National nutrition surveys in Europe: a review on the current status in the 53 countries of the WHO European region

Objectives: The objectives of this study were (1) to determine the coverage of national nutrition surveys in the 53 countries monitored by the World Health Organization (WHO) Regional Office for Europe and identify gaps in provision, (2) to describe relevant survey attributes and (3) to check whether energy and nutrients are reported with a view to providing information for evidence-based nutrition policy planning. Design: Dietary survey information was gathered using three methods: (1) direct email to survey authors and other relevant contacts, (2) systematic review of literature databases and (3) general web-based searches. Survey characteristics relating to time frame, sampling and dietary methodology and nutrients reported were tabled from all relevant surveys found since 1990. Setting: Fifty-three countries of the WHO Regional Office for Europe, which have need for an overview of dietary surveys across the life course. Subjects: European individuals (adults and children) in national diet surveys. Results: A total of 109 nationally representative dietary surveys undertaken post-1990 were found across 34 countries. Of these, 78 surveys from 33 countries were found post-2000, and of these, 48 surveys from 27 countries included children and 60 surveys from 30 countries included adults. No nationally representative surveys were found for 19 of 53 countries, mainly from Central and Eastern Europe. Multiple 24hr recall and food diaries were the most common dietary assessment methods. Only 22 countries reported energy and nutrient intakes from post-2000 surveys; macronutrients were more widely reported than micronutrients. Conclusions: Less than two-thirds of WHO Europe countries have nationally representative diet surveys, mainly collected post-2000. The main availability gaps lie in Central and Eastern European countries, where nutrition policies may therefore lack an appropriate evidence base. Dietary methodological differences may limit the scope for inter-country comparisons