Deletion of the three distal S1 motifs of Saccharomyces cerevisiae Rrp5p abolishes pre-rRNA processing at site A(2) without reducing the production of functional 40S subunits

Yeast Rrp5p, one of the few trans-acting proteins required for the biogenesis of both ribosomal subunits, has a remarkable two-domain structure. Its C-terminal region consists of seven tetratricopeptide motifs, several of which are crucial for cleavages at sites

Are torso asymmetry and torso displacements in a computer brace model associated with initial in-brace correction in adolescent idiopathic scoliosis?

Background: Lack of initial in-brace correction is strongly predictive for brace treatment failure in adolescent idiopathic scoliosis (AIS) patients. Computer-aided design (CAD) technology could be useful in quantifying the trunk in 3D and brace characteristics in order to further investigate the effect of brace modifications on initial in-brace correction and subsequently long-term brace treatment success. The purpose of this pilot study was to identify parameters obtained from 3D surface scans which influence the initial in-brace correction (IBC) in a Boston brace in patients with AIS.Methods: Twenty-fiveAIS patients receiving a CAD-based Boston brace were included in this pilot study consisting of 11 patients with Lenke classification type 1 and 14 with type 5 curves. The degree of torso asymmetry and segmental peak positive and negative torso displacements were analyzed with the use of patients’ 3D surface scans and brace models for potential correlations with IBC. Results: The mean IBC of the major curve on AP view was 15.9% (SD = 9.1%) for the Lenke type 1 curves, and 20.1% (SD = 13.9%) for the type 5 curves. The degree of torso asymmetry was weakly correlated with patient’s pre-brace major curve Cobb angle and negligible correlated with major curve IBC. Mostly weak or negligible correlations were observed between IBC and the twelve segmental peak displacements for both Lenke type 1 and 5 curves.Conclusion: Based on the results of this pilot study, the degree of torso asymmetry and segmental peak torso displacements in the brace model alone are not clearly associated with IBC.</p

Are torso asymmetry and torso displacements in a computer brace model associated with initial in-brace correction in adolescent idiopathic scoliosis?

Background: Lack of initial in-brace correction is strongly predictive for brace treatment failure in adolescent idiopathic scoliosis (AIS) patients. Computer-aided design (CAD) technology could be useful in quantifying the trunk in 3D and brace characteristics in order to further investigate the effect of brace modifications on initial in-brace correction and subsequently long-term brace treatment success. The purpose of this pilot study was to identify parameters obtained from 3D surface scans which influence the initial in-brace correction (IBC) in a Boston brace in patients with AIS.Methods: Twenty-fiveAIS patients receiving a CAD-based Boston brace were included in this pilot study consisting of 11 patients with Lenke classification type 1 and 14 with type 5 curves. The degree of torso asymmetry and segmental peak positive and negative torso displacements were analyzed with the use of patients’ 3D surface scans and brace models for potential correlations with IBC. Results: The mean IBC of the major curve on AP view was 15.9% (SD = 9.1%) for the Lenke type 1 curves, and 20.1% (SD = 13.9%) for the type 5 curves. The degree of torso asymmetry was weakly correlated with patient’s pre-brace major curve Cobb angle and negligible correlated with major curve IBC. Mostly weak or negligible correlations were observed between IBC and the twelve segmental peak displacements for both Lenke type 1 and 5 curves.Conclusion: Based on the results of this pilot study, the degree of torso asymmetry and segmental peak torso displacements in the brace model alone are not clearly associated with IBC.</p

Foreign gene expression in Hansenula polymorpha. A system for the synthesis of small functional peptides

We describe the synthesis and purification of two functional peptides, namely human insulin-like growth factor II (IGF-II) and Xenopus laevis magainin II in Hansenula polymorpha after their synthesis as hybrid proteins fused to the C terminus of endogenous amine oxidase. The hybrid genes, placed under control of the H. polymorpha alcohol oxidase promoter (PAOX), were integrated into the genomic alcohol oxidase locus, yielding stable production strains. High-level synthesis of the fusion proteins, exceeding 20% of total cellular protein, was obtained when the transformed strains were grown in methanol-limited chemostat cultures; when expressed by itself, i.e. in the absence of the amine oxidase gene, IGF-II could not be recovered from crude cell extracts, probably as a result of rapid proteolytic degradation. Accumulation in peroxisomes did not significantly affect the IGF-II protein stability when expressed in the absence of the carrier protein. Apparently, fusion to the large (±78 kDa) amine oxidase carrier particularly stabilizes the peptides and prevents them from proteolysis. After partial purification, the fusion partners were readily separated by factor Xa treatment

Hansenula polymorpha Swi1p and Snf2p are essential for methanol utilisation

We have cloned the Hansenula polymorpha SWI1 and SNF2 genes by functional complementation of mutants that are defective in methanol utilisation. These genes encode proteins similar to Saccharomyces cerevisiae Swi1p and Snf2p, which are subunits of the SWI/SNF complex. This complex belongs to the family of nucleosome-remodeling complexes that play a role in transcriptional control of gene expression. Analysis of the phenotypes of constructed H. polymorpha SWI1 and SNF2 disruption strains indicated that these genes are not necessary for growth of cells on glucose, sucrose, or various organic nitrogen sources which involve the activity of peroxisomal oxidases. Both disruption strains showed a moderate growth defect on glycerol and ethanol, but were fully blocked in methanol utilisation. In methanol-induced cells of both disruption strains, two peroxisomal enzymes involved in methanol metabolism, alcohol oxidase and dihydroxyacetone synthase, were hardly detectable, whereas in wild-type cells these proteins were present at very high levels. We show that the reduction in alcohol oxidase protein levels in H. polymorpha SWI1 and SNF2 disruption strains is due to strongly reduced expression of the alcohol oxidase gene. The level of Pex5p, the receptor involved in import of alcohol oxidase and dihydroxyacetone synthase into peroxisomes, was also reduced in both disruption strains compared to that in wild-type cells.

Heavy menstrual bleeding on direct factor Xa inhibitors: Rationale and design of the MEDEA study

Background: In premenopausal women, treatment with direct oral factor Xa inhibitors is associated with an increased risk of heavy menstrual bleeding (HMB) compared with vitamin K antagonists (VKA). Treatment with the direct oral thrombin inhibitor dabigatran appears to be associated with a reduced risk of HMB compared with VKA. These findings come from small observational studies or post hoc analyses of trials in which HMB was not a primary outcome. Use of tranexamic acid during the menstrual period may be effective in patients with HMB, but prospective data regarding efficacy and safety in patients on anticoagulant treatment are lacking. Rationale and Design: A direct comparison of a factor Xa inhibitor and a thrombin inhibitor with HMB as primary outcome, as well as an evaluation of the effects of adding tranexamic acid in women with anticoagulant-associated HMB is highly relevant for clinical practice. The MEDEA study is a randomized, open-label, pragmatic clinical trial to evaluate management strategies in premenopausal women with HMB associated with factor Xa inhibitor therapy. Outcomes: Women using factor Xa inhibitors with proven HMB, as assessed by a pictorial blood loss assessment chart (PBAC) score of >150, will be randomized to one of three study arms: (i) switch to dabigatran; (ii) continue factor Xa inhibitor with addition of tranexamic acid during the menstrual period; or (iii) continue factor Xa inhibitor without intervention. The primary outcome is the difference in PBAC score before and after randomization. Here, we present the rationale and highlight several unique features in the design of the study

Identification of regulatory variants associated with genetic susceptibility to meningococcal disease

Non-coding genetic variants play an important role in driving susceptibility to complex diseases but their characterization remains challenging. Here, we employed a novel approach to interrogate the genetic risk of such polymorphisms in a more systematic way by targeting specific regulatory regions relevant for the phenotype studied. We applied this method to meningococcal disease susceptibility, using the DNA binding pattern of RELA - a NF-kB subunit, master regulator of the response to infection - under bacterial stimuli in nasopharyngeal epithelial cells. We designed a custom panel to cover these RELA binding sites and used it for targeted sequencing in cases and controls. Variant calling and association analysis were performed followed by validation of candidate polymorphisms by genotyping in three independent cohorts. We identified two new polymorphisms, rs4823231 and rs11913168, showing signs of association with meningococcal disease susceptibility. In addition, using our genomic data as well as publicly available resources, we found evidences for these SNPs to have potential regulatory effects on ATXN10 and LIF genes respectively. The variants and related candidate genes are relevant for infectious diseases and may have important contribution for meningococcal disease pathology. Finally, we described a novel genetic association approach that could be applied to other phenotypes