Full-length 16S rRNA gene sequencing using nanopore technology is a fast alternative to conventional short-read 16S rRNA gene sequencing with low initial investment costs that has been used for ...various microbiome studies but has not yet been investigated as an alternative approach for endometrial microbiome analysis. Is in-situ 16S rRNA gene long-read sequencing using portable nanopore sequencing technology feasible and reliable for endometrial microbiome analysis?
A prospective experimental study based on 33 patients seeking infertility treatment between January and October 2019. A 16S rRNA gene long-read nanopore sequencing protocol for analysing endometrial microbiome samples was established, including negative controls for contamination evaluation and positive controls for bias evaluation. Contamination caused by kit and exterior sources was identified and excluded from the analysis. Endometrial samples from 33 infertile patients were sequenced using the optimized long-read nanopore sequencing protocol and compared with conventional short-read sequencing carried out by external laboratories.
Of the 33 endometrial patient samples, 23 successfully amplified (69.7%) and their microbiome was assessed using nanopore sequencing. Of those 23 samples, 14 (60.9%) were Lactobacillus-dominated (>80% of reads mapping to Lactobacillus), with 10 samples resulting in more than 90% Lactobacillus reads. Our long-read nanopore sequencing revealed results similar to two conventional short-read sequencing approaches and to long-read sequencing validation carried out in external laboratories.
In this pilot study, 16S rRNA gene long-read nanopore sequencing was established to analyse the endometrial microbiome in situ that could be widely applied owing to its cost efficiency and portable character.
Therapeutic compensation of deficient bone regeneration is a challenging task and a topic of on-going search for novel treatment strategies. One promising approach for improvement involves non-viral ...gene delivery using the bone morphogenetic protein-2 (BMP-2) gene to provide transient, local and sustained expression of the growth factor. However, since efficiency of non-viral gene delivery is low, this study focused on the improvement of a BMP-2 gene expression system, aiming for compensation of poor transfection efficiency. First, the native BMP-2 gene sequence was modified by codon optimisation and altered by inserting a highly truncated artificial intron (96 bp). Transfection of multiple cell lines and rat adipose-derived mesenchymal stem cells with plasmids harbouring the improved BMP-2 sequence led to a several fold increased expression rate and subsequent osteogenic differentiation. Additionally, comparing expression kinetics of elongation factor 1 alpha (EF1α) promoter with a state of the art CMV promoter revealed significantly higher BMP-2 expression when under the influence of the EF1α promoter. Results obtained by quantification of bone markers as well as osteogenic assays showed reduced sensitivity to promoter silencing effects of the EF1α promoter in rat adipose-derived mesenchymal stem cells. Finally, screening of several protein secretion signals using either luciferase or BMP-2 as reporter protein revealed no superior candidates for potential replacement of the native BMP-2 secretion signal. Taken together, by enhancing the exogenous BMP-2 expression system, low transfection efficiencies in therapeutic applications can be compensated, making safe non-viral systems even more suitable for tissue regeneration approaches.
Lanchester model for three-way combat Kress, Moshe; Caulkins, Jonathan P.; Feichtinger, Gustav ...
European journal of operational research,
01/2018, Letnik:
264, Številka:
1
Journal Article
Recenzirano
•We extend the classic Lanchester model to a three-sided conflict.•The force-allocation between opponents can determine the winner of the battle.•The initial force sizes and attrition rates are ...crucial for the outcome.•By eigenvalue analysis we identify regions in the state space with differing winners.•In some cases an opponent cannot win, but determine the winner by its force allocation.
Lanchester (1960) modeled combat situations between two opponents, where mutual attrition occurs continuously in time, by a pair of simple ordinary (linear) differential equations. The aim of the present paper is to extend the model to a conflict consisting of three parties. In particular, Lanchester’s main result, i.e. his Square Law, is adapted to a triple fight. However, here a central factor – besides the initial strengths of the forces – determining the long run outcome is the allocation of each opponent’s efforts between the other two parties. Depending on initial strengths, (the) solution paths are calculated and visualized in appropriate phase portraits. We are able identify regions in the state space where, independent of the force allocation of the opponents, always the same combatant wins, regions, where a combatant can win if its force allocation is wisely chosen, and regions where a combatant cannot win itself but determine the winner by its forces allocation. As such, the present model can be seen as a forerunner of a dynamic game between three opponents.
This study aimed to define the genotypic and phenotypic spectrum of reversible acute liver failure (ALF) of infancy resulting from biallelic pathogenic TRMU variants and determine the role of ...cysteine supplementation in its treatment.
Individuals with biallelic (likely) pathogenic variants in TRMU were studied within an international retrospective collection of de-identified patient data.
In 62 individuals, including 30 previously unreported cases, we described 47 (likely) pathogenic TRMU variants, of which 17 were novel, and 1 intragenic deletion. Of these 62 individuals, 42 were alive at a median age of 6.8 (0.6-22) years after a median follow-up of 3.6 (0.1-22) years. The most frequent finding, occurring in all but 2 individuals, was liver involvement. ALF occurred only in the first year of life and was reported in 43 of 62 individuals; 11 of whom received liver transplantation. Loss-of-function TRMU variants were associated with poor survival. Supplementation with at least 1 cysteine source, typically N-acetylcysteine, improved survival significantly. Neurodevelopmental delay was observed in 11 individuals and persisted in 4 of the survivors, but we were unable to determine whether this was a primary or a secondary consequence of TRMU deficiency.
In most patients, TRMU-associated ALF was a transient, reversible disease and cysteine supplementation improved survival.
Summary
This study investigated the implication of a preceding high-trauma fracture on subsequent high- and low-trauma fractures at different skeletal sites in postmenopausal women and similarly aged ...men at an age range of 54 to 70 years. A preceding high-trauma fracture increases the risk of future low-trauma non-vertebral fractures including hip.
Introduction
Little is known about the impact of the skeletal fracture site in conjunction with the severity of a past fracture (high- or low-trauma preceding fracture) and its effect on future fracture risk.
Methods
Patients with de novo high- and low-trauma fractures admitted to seven large trauma centers across Austria between 2000 and 2012 were stratified into sex and different age groups. Kaplan-Meier estimates, Cox proportional hazards regression models (HR), and likelihood calculations estimated effects of age, sex, and the anatomic region on the probability of a subsequent fracture in the same patient.
Results
Included in the study were 433,499 female and male patients at an age range of 0 to 100 years with 575,772 de novo high- and low-trauma fractures. In the age range of 54–70 years, subsequent fractures were observed in 16% of females and 12.1% of males. A preceding high-trauma fracture was associated with 12.9% of subsequent fractures, thereof 6.5% of high- and 6.4% of low-trauma in origin, usually at the hip, humerus, or pelvis. The highest effect sizes were observed for femur, humerus, and thorax fractures with hazard ratios (HR) of 1.26, 1.18, and 1.14. After splitting into high-trauma preceding and subsequent low-trauma fractures, the femoral neck (HR = 1.59), the female sex (HR = 2.02), and age (HR = 1.03) were discriminators for increased future fracture risk.
Conclusions
Preceding high-trauma fractures increase the risk of future low-trauma non-vertebral fractures including hip. For each patient with a fracture, regardless of the severity of the trauma, osteoporosis should be taken into clinical consideration.
•Innovative marketing model involving experience quality is proposed.•Optimal marketing mix is determined.•Guidelines for subsidizing quality are derived.•History dependent equilibria (indifference ...threshold, Skiba point) are identified.
For certain goods or services, the quality of the product can be assessed by customers only after consumption. We determine the optimal time paths for pricing, advertising and quality for a profit-maximizing firm facing demand that is influenced both by this experience quality as well as by advertising. In particular, there may exist two optimal trajectories separated by an indifference threshold in which the firm has the same utility of converging to either of the two long-run steady states. Or, to put it in a nutshell: the optimal mix of marketing instruments may lead to history-dependence. One implication is that there may be a market failure such that a government subsidy could help reach the steady state that is best for the Economy in the sense of having greater sales and a higher quality product.
Mitochondrial disorders are clinically and genetically diverse, with isolated complex III (CIII) deficiency being relatively rare. Here, we describe two affected cousins, presenting with recurrent ...episodes of severe lactic acidosis, hyperammonaemia, hypoglycaemia and encephalopathy. Genetic investigations in both cases identified a homozygous deletion of exons 2 and 3 of UQCRH, which encodes a structural complex III (CIII) subunit. We generated a mouse model with the equivalent homozygous Uqcrh deletion (Uqcrh−/−), which also presented with lactic acidosis and hyperammonaemia, but had a more severe, non‐episodic phenotype, resulting in failure to thrive and early death. The biochemical phenotypes observed in patient and Uqcrh−/− mouse tissues were remarkably similar, displaying impaired CIII activity, decreased molecular weight of fully assembled holoenzyme and an increase of an unexpected large supercomplex (SXL), comprising mostly of one complex I (CI) dimer and one CIII dimer. This phenotypic similarity along with lentiviral rescue experiments in patient fibroblasts verifies the pathogenicity of the shared genetic defect, demonstrating that the Uqcrh−/− mouse is a valuable model for future studies of human CIII deficiency.
SYNOPSIS
This work describes the first confirmed pathogenic variant in UQCRH in two children presenting with recurrent episodes of metabolic crisis. A mouse model harbouring the equivalent variant in Uqcrh shared similar biochemical phenotypes of impaired CIII activity and abnormal OXPHOS supercomplex structures, but with more severe manifestation.
A 2.2 kb homozygous deletion of exons 2 and 3 of UQCRH (chr1:g.46,774,245‐46,776,461) was identified through WES in two cousins that presented with episodes of lactic acidosis, hyperammonaemia and impaired glucose homeostasis.
A viable mouse model carrying the equivalent two‐exon deletion in Uqcrh was generated and systematically phenotyped, displaying important characteristics of human CIII disorders.
CIII activity and assembly in both patient fibroblasts and Uqcrh−/− mice were impaired by Uqcrh/UQCRH absence. Its lack of expression, however, still allows dimerisation.
Aberrant assemblies of respiratory supercomplexes including an increase of a large supercomplex (SXL) were revealed by complexome profiling in both patient fibroblasts and Uqcrh−/− mouse tissue.
This work describes the first confirmed pathogenic variant in UQCRH in two children presenting with recurrent episodes of metabolic crisis. A mouse model harbouring the equivalent variant in Uqcrh shared similar biochemical phenotypes of impaired CIII activity and abnormal OXPHOS supercomplex structures, but with more severe manifestation.
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