88 research outputs found

    PCR inhibition in stool samples in relation to age of infants

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    Background: PCR is rapidly replacing traditional methods in diagnostic virus laboratories. PCR inhibitors,which are often present in clinical samples, may lead to false negative test results.Objectives: The aim was to study the presence of PCR inhibitors in stool samples collected from 3- to24-month old children.Study design: Total RNA fraction extracted from stool samples was spiked with a standardized amount ofSemliki Forest Virus RNA and amplified using specific PCR primers. The presence of PCR inhibitors wasdetected by a decrease in amplification rate compared to spiked water samples. Inhibition in differentage groups and dietary origin of PCR inhibitors were analyzed by comparing the samples taken duringexclusive and non-exclusive breastfeeding periods. The inactivation of PCR inhibitors was also assessed.Results: Complete inhibition was seen in 12% (13/108) and partial inhibition in 19% (21/108) of the samples.Inhibition was seen in none of the stool samples (0/31) taken from infants younger than 6 monthscompared to 17% of samples (13/77) taken from6 to 24 months old infants (p more common in younger age group. Addition of bovine serum albumin (BSA) into the reaction mixtureseliminated the effect of inhibitors leading to all samples being positive.Conclusions: PCR inhibitors are frequent in stool samples. They may originate from dietary componentsand can lead to false negative PCR results. The addition of BSA to the cDNA and PCR reactions proved tobe an easy and effective method for eliminating the inhibitory effect of these compounds

    Enrichment of rare variants in population isolates : single AICDA mutation responsible for hyper-IgM syndrome type 2 in Finland

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    Antibody class-switch recombination and somatic hypermutation critically depend on the function of activation-induced cytidine deaminase (AID). Rare variants in its gene AICDA have been reported to cause autosomal recessive AID deficiency (autosomal recessive hyper-IgM syndrome type 2 (HIGM2)). Exome sequencing of a multicase Finnish family with an HIGM2 phenotype identified a rare, homozygous, variant (c.416T > C, p.(Met139Thr)) in the AICDA gene, found to be significantly enriched in the Finnish population compared with other populations of European origin (38.56-fold, P <0.001). The population history of Finland, characterized by a restricted number of founders, isolation and several population bottlenecks, has caused enrichment of certain rare disease-causing variants and losses of others, as part of a phenomenon called the Finnish Disease Heritage. Accordingly, rare founder mutations cause the majority of observed Finnish cases in these mostly autosomal recessive disorders that consequently are more frequent in Finland than elsewhere. Screening of all currently known Finnish patients with an HIGM2 phenotype showed them to be homozygous for p.(Met139Thr). All the Finnish p.(Met139Thr) carriers with available data on their geographic descent originated from the eastern and northeastern parts of Finland. They were observed to share more of their genome identity by descent (IBD) than Finns in general (P <0.001), and they all carried a 207.5-kb ancestral haplotype containing the variant. In conclusion, the identified p.(Met139Thr) variant is significantly enriched in Finns and explains all thus far found AID deficiencies in Finland.Peer reviewe

    Oxygen influenced intergranular crack propagation: analysing microstructure and chemistry in the crack tip region

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    Ni-base superalloys have for decades been studied with regard to environmentally influenced intergranular crack propagation. For high temperature fatigue frequencies <0.1 Hz, it has been shown that an oxygen-rich environment promotes time-dependent crack growth while at >0.1 Hz and/or in inert environments (e.g. vacuum) crack growth is cycle dependent. Oxygen interaction at, or ahead of, the crack tip has been pointed out as the reason for the degraded mechanical properties. While many aspects of this type of crack growth have been previously investigated there is still no consensus about the detailed mechanisms, mainly due to the lack of in-detail investigations of the crack-tip region. Here, crack tip regions in the Ni-base superalloy Alloy 718 were studied. Specimens were subjected to 90 s hold-times at 550 degrees C and 650 degrees C. Crack growth was arrested before final fracture, allowing cross-sectional analyses of the crack-tip region using scanning electron microscopy (SEM). Detailed studies of the crack-tip region were performed using transmission electron microscopy (TEM) and atom probe tomography (APT). For both APT and TEM samples, site-specific focussed ion beam (FIB) sample preparation was performed in a combined FIB-SEM system. The methodology of accessing and analysing the crack tip region is shown. Initial results on oxidation, oxygen penetration and plastic deformation are shown and discussed

    Norway spruce and spruce shoot aphid as indicators of traffic pollution

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