Finding a (Pine) Needle in a Haystack: Chloroplast Genome Sequence Divergence in Rare and Widespread Pines

Critical to conservation efforts and other investigations at low taxonomic levels, DNA sequence data offer important insights into the distinctiveness, biogeographic partitioning and evolutionary histories of species. The resolving power of DNA sequences is often limited by insufficient variability at the intraspecific level. This is particularly true of studies involving plant organelles, as the conservative mutation rate of chloroplasts and mitochondria makes it difficult to detect polymorphisms necessary to track genealogical relationships among individuals, populations and closely related taxa, through space and time. Massively parallel sequencing (MPS) makes it possible to acquire entire organelle genome sequences to identify cryptic variation that would be difficult to detect otherwise. We are using MPS to evaluate intraspecific chloroplast-level divergence across biogeographic boundaries in narrowly endemic and widespread species of Pinus. We focus on one of the world\u27s rarest pines - Torrey pine (Pinus torreyana) - due to its conservation interest and because it provides a marked contrast to more widespread pine species. Detailed analysis of nearly 90% ( approximately 105 000 bp each) of these chloroplast genomes shows that mainland and island populations of Torrey pine differ at five sites in their plastome, with the differences fixed between populations. This is an exceptionally low level of divergence (1 polymorphism/ approximately 21 kb), yet it is comparable to intraspecific divergence present in widespread pine species and species complexes. Population-level organelle genome sequencing offers new vistas into the timing and magnitude of divergence within species, and is certain to provide greater insight into pollen dispersal, migration patterns and evolutionary dynamics in plants

[Prevalence of claw disorders in swiss cattle farms].

INTRODUCTION The project «Healthy claws - the foundation for the future» aims to establish a Swiss national claw health monitoring based on digital recordings by claw trimmers during claw trimming. To assess claw health on the participating farms, between-herd prevalence, within-herd prevalence and cow prevalence of all claw disorders based on the «ICAR Claw Health Atlas» were calculated during this study. Claw trimmers underwent an intensive training and examination in order to ensure data quality. To guarantee the representativity of the prevalences, only farm claw trimmings were considered, where ≥ 80 % of the cows in a herd were trimmed. The calculations were based on 7108 cows and 403 heifers from 238 farms, during the period from February 2020 to February 2021. At least one claw disorder was present in 99,2 % of the farms, with 49,6 % of the heifers and 77,7 % of the cows having at least one claw disorder. The high prevalence is seen as a result of all ICAR claw disorders being considered, whereas not all of them are painful and consequently not all of them cause lameness. The absence of lameness assessment limits the evaluation of existing herd problems. High between-herd and cow prevalences were observed for the following claw disorders: heel horn erosion (92,9 %/64,7 %), digital dermatitis (55,9 %/20,7 %), white line disease (81,5 %/17,7 %) and sole hemorrhage (66,4 %/11,6 %). Asymmetric claws, corkscrew claws, scissor claws, horn fissure, interdigital phlegmon, swelling of the coronet and/or bulb and toe necrosis had low prevalences. The proportion of cows treated with a hoof block (0,5 %) was comparatively small in regard of the cows suffering from ulcers (5,6 %) and white line abscesses (2,5 %). The median within-herd prevalence of digital dermatitis was 5,6 %, with a maximal within-herd prevalence of 87,5 %. Despite the contagious nature of digital dermatitis, no increase of between-herd and cow prevalence has been observed in the past ten years throughout Switzerland. Based on this data, the Swiss claw health situation can be monitored, compared over time and improved in the future

[Risk factors for infectious foot disorders on two Swiss cattle mountain pastures].

In the present study, risk groups for infectious foot disorders were identified on two large Swiss cattle mountain pastures by analyzing animal and treatment data of a total of 3256 animals of the bovine species. Both mountain pastures were part of the Federal Office for Agriculture (FOAG) consultancy project «Healthy animals, attractive herdsmen positions and less medication on large cattle mountain pastures». The project was launched in 2020 following the increased incidence of lameness on these mountain pastures. Bacteriological and histological analyses were to provide information as to whether the most common foot disorder was interdigital phlegmon (IP) or whether digital dermatitis also occurred. Further, the temporal distribution of cases over the mountain pasture season and the influence of mountain pasture and year were investigated and interpreted for the project years 2020 to 2022, and treatment incidences were compared between years. Multiple treatment cycles in the same individual were classified into persistent infections and new infections. Nineteen of 394 first-treated cattle were clinically examined, 12 of them were additionally sampled for bacteriological and histological analyses. All cases examined showed, both clinically and following laboratory analyses, typical characteristics for IP. In contrast, there was no specific evidence for the presence of digital dermatitis. No persistent infections occurred during treatment with benzylpenicillin. Multivariate logistic regression analyses identified > 365-730-day-old cattle (odds ratio OR 8,29), as well as inseminated (OR 5,30) and non-inseminated (OR 7,85) heifers as risk groups for the disease studied (p < 0,05). Association with the oestrus activity of non-inseminated heifers and a generally higher locomotor activity in heifers compared to cows - with a correspondingly increased risk of injury - is conceivable. Meat breeds had a reduced risk compared to dairy breeds (OR 0,29). Breed differences in behavior and/or the effectiveness of the local immune response might have an impact. Knowing about these risk groups can be put to use in the future when selecting animals to be taken to the mountain pastures and/or when planning pasture management in order to reduce the prevalence of infectious foot disorders and thereby the use of antibiotics

Intrinsic MYH7 expression regulation contributes to tissue level allelic imbalance in hypertrophic cardiomyopathy

HCM, the most common inherited cardiac disease, is mainly caused by mutations in sarcomeric genes. More than a third of the patients are heterozygous for mutations in the MYH7 gene encoding for the β-myosin heavy chain. In HCM-patients, expression of the mutant and the wildtype allele can be unequal, thus leading to fractions of mutant and wildtype mRNA and protein which deviate from 1:1. This so-called allelic imbalance was detected in whole tissue samples but also in individual cells. There is evidence that the severity of HCM not only depends on the functional effect of the mutation itself, but also on the fraction of mutant protein in the myocardial tissue. Allelic imbalance has been shown to occur in a broad range of genes. Therefore, we aimed to examine whether the MYH7-alleles are intrinsically expressed imbalanced or whether the allelic imbalance is solely associated with the disease. We compared the expression of MYH7-alleles in non-HCM donors and in HCM-patients with different MYH7-missense mutations. In the HCM-patients, we identified imbalanced as well as equal expression of both alleles. Also at the protein level, allelic imbalance was determined. Most interestingly, we also discovered allelic imbalance and balance in non-HCM donors. Our findings therefore strongly indicate that apart from mutation-specific mechanisms, also non-HCM associated allelic-mRNA expression regulation may account for the allelic imbalance of the MYH7 gene in HCM-patients. Since the relative amount of mutant mRNA and protein or the extent of allelic imbalance has been associated with the severity of HCM, individual analysis of the MYH7-allelic expression may provide valuable information for the prognosis of each patient

Conformance checking using activity and trace embeddings

Conformance checking describes process mining techniques used to compare an event log and a corresponding process model. In this paper, we propose an entirely new approach to conformance checking based on neural network-based embeddings. These embeddings are vector representations of every activity/task present in the model and log, obtained via act2vec, a Word2vec based model. Our novel conformance checking approach applies the Word Mover’s Distance to the activity embeddings of traces in order to measure fitness and precision. In addition, we investigate a more efficiently calculated lower bound of the former metric, i.e. the Iterative Constrained Transfers measure. An alternative method using trace2vec, a Doc2vec based model, to train and compare vector representations of the process instances themselves is also introduced. These methods are tested in different settings and compared to other conformance checking techniques, showing promising results

Targeted Capture Sequencing in Whitebark Pine Reveals Range-Wide Demographic and Adaptive Patterns Despite Challenges of a Large, Repetitive Genome

Whitebark pine (Pinus albicaulis) inhabits an expansive range in western North America, and it is a keystone species of subalpine environments. Whitebark is susceptible to multiple threats – climate change, white pine blister rust, mountain pine beetle, and fire exclusion – and it is suffering significant mortality range-wide, prompting the tree to be listed as ‘globally endangered’ by the International Union for Conservation of Nature and ‘endangered’ by the Canadian government. Conservation collections (in situ and ex situ) are being initiated to preserve the genetic legacy of the species. Reliable, transferrable, and highly variable genetic markers are essential for quantifying the genetic profiles of seed collections relative to natural stands, and ensuring the completeness of conservation collections. We evaluated the use of hybridization-based target capture to enrich specific genomic regions from the 27 GB genome of whitebark pine, and to evaluate genetic variation across loci, trees, and geography. Probes were designed to capture 7,849 distinct genes, and screening was performed on 48 trees. Despite the inclusion of repetitive elements in the probe pool, the resulting dataset provided information on 4,452 genes and 32% of targeted positions (528,873 bp), and we were able to identify 12,390 segregating sites from 47 trees. Variations reveal strong geographic trends in heterozygosity and allelic richness, with trees from the southern Cascade and Sierra Range showing the greatest distinctiveness and differentiation. Our results show that even under non-optimal conditions (low enrichment efficiency; inclusion of repetitive elements in baits), targeted enrichment produces high quality, codominant genotypes from large genomes. The resulting data can be readily integrated into management and gene conservation activities for whitebark pine, and have the potential to be applied to other members of 5-needle pine group (Pinus subsect. Quinquefolia) due to their limited genetic divergence

Wafer scale transfer route for top down III nitride nanowire LED arrays based on the femtosecond laser lift off technique

The integration of gallium nitride GaN nanowire light emitting diodes nanoLEDs on flexible substrates offers opportunities for applications beyond rigid solid state lighting e.g., for wearable optoelectronics and bendable inorganic displays . Here, we report on a fast physical transfer route based on femtosecond laser lift off fs LLO to realize wafer scale top down GaN nanoLED arrays on unconventional platforms. Combined with photolithography and hybrid etching processes, we successfully transferred GaN blue nanoLEDs from a full two inch sapphire substrate onto a flexible copper Cu foil with a high nanowire density 107 wires cm2 , transfer yield 99.5 , and reproducibility. Various nanoanalytical measurements were conducted to evaluate the performance and limitations of the fs LLO technique as well as to gain insights into physical material properties such as strain relaxation and assess the maturity of the transfer process. This work could enable the easy recycling of native growth substrates and inspire the development of large scale hybrid GaN nanowire optoelectronic devices by solely employing standard epitaxial LED wafers i.e., customized LED wafers with additional embedded sacrificial materials and a complicated growth process are not require

A pipeline to quantify serum and cerebrospinal fluid microRNAs for diagnosis and detection of relapse in paediatric malignant germ-cell tumours

Background:The current biomarkers alpha-fetoprotein and human chorionic gonadotropin have limited sensitivity and specificity for diagnosing malignant germ-cell tumours (GCTs). MicroRNAs (miRNAs) from the miR-371-373 and miR-302/367 clusters are overexpressed in all malignant GCTs, and some of these miRNAs show elevated serum levels at diagnosis. Here, we developed a robust technical pipeline to quantify these miRNAs in the serum and cerebrospinal fluid (CSF). The pipeline was used in samples from a cohort of exclusively paediatric patients with gonadal and extragonadal malignant GCTs, compared with appropriate tumour and non-tumour control groups.Methods:We developed a method for miRNA quantification that enabled sample adequacy assessment and reliable data normalisation. We performed qRT-PCR profiling for miR-371-373 and miR-302/367 cluster miRNAs in a total of 45 serum and CSF samples, obtained from 25 paediatric patients.Results:The exogenous non-human spike-in cel-miR-39-3p and the endogenous housekeeper miR-30b-5p were optimal for obtaining robust serum and CSF qRT-PCR quantification. A four-serum miRNA panel (miR-371a-3p, miR-372-3p, miR-373-3p and miR-367-3p): (i) showed high sensitivity/specificity for diagnosing paediatric extracranial malignant GCT; (ii) allowed early detection of relapse of a testicular mixed malignant GCT; and (iii) distinguished intracranial malignant GCT from intracranial non-GCT tumours at diagnosis, using CSF and serum samples.Conclusions:The pipeline we have developed is robust, scalable and transferable. It potentially promises to improve clinical management of paediatric (and adult) malignant GCTs

Inference of reticulate evolutionary histories by maximum likelihood: the performance of information criteria

Background: Maximum likelihood has been widely used for over three decades to infer phylogenetic trees from molecular data. When reticulate evolutionary events occur, several genomic regions may have conflicting evolutionary histories, and a phylogenetic network may provide a more adequate model for representing the evolutionary history of the genomes or species. A maximum likelihood (ML) model has been proposed for this case and accounts for both mutation within a genomic region and reticulation across the regions. However, the performance of this model in terms of inferring information about reticulate evolution and properties that affect this performance have not been studied. Results: In this paper, we study the effect of the evolutionary diameter and height of a reticulation event on its identifiability under ML. We find both of them, particularly the diameter, have a significant effect. Further, we find that the number of genes (which can be generalized to the concept of "non-recombining genomic regions") that are transferred across a reticulation edge affects its detectability. Last but not least, a fundamental challenge with phylogenetic networks is that they allow an arbitrary level of complexity, giving rise to the model selection problem. We investigate the performance of two information criteria, the Akaike Information Criterion (AIC) and the Bayesian Information Criterion (BIC), for addressing this problem. We find that BIC performs well in general for controlling the model complexity and preventing ML from grossly overestimating the number of reticulation events. Conclusion: Our results demonstrate that BIC provides a good framework for inferring reticulate evolutionary histories. Nevertheless, the results call for caution when interpreting the accuracy of the inference particularly for data sets with particular evolutionary features