The Creation: Haydn\u27s Drama

This paper focuses on Haydn’s use of drama in regard to his oratorio The Creation. To understand Haydn’s use of drama, we considered three main aspects. The first aspect is the means by which Haydn came about The Creation text and the meaning it held for him. The second aspect is an analysis of the work, particularly looking at text painting, comedic elements, and orchestration. Finally, we will discuss the reception of Haydn’s work and the importance it held in relation to the aesthetics of the time. The Creation was a staple of Haydn’s work, and would prove to be a large influence on future composers, as well as writers and artists

Understanding early organogenesis using a simplified in situ hybridization protocol in Xenopus

Organogenesis is the study of how organs are specified and then acquire their specific shape and functions during development. The Xenopuslaevis embryo is very useful for studying organogenesis because their large size makes them very suitable for identifying organs at the earliest steps in organogenesis. At this time, the primary method used for identifying a specific organ or primordium is whole mount in situ hybridization with labeled antisense RNA probes specific to a gene that is expressed in the organ of interest. In addition, it is relatively easy to manipulate genes or signaling pathways in Xenopus and in situ hybridization allows one to then assay for changes in the presence or morphology of a target organ. Whole mount in situ hybridization is a multi-day protocol with many steps involved. Here we provide a simplified protocol with reduced numbers of steps and reagents used that works well for routine assays. In situ hybridization robots have greatly facilitated the process and we detail how and when we utilize that technology in the process. Once an in situ hybridization is complete, capturing the best image of the result can be frustrating. We provide advice on how to optimize imaging of in situ hybridization results. Although the protocol describes assessing organogenesis in Xenopus laevis, the same basic protocol can almost certainly be adapted to Xenopus tropicalis and other model systems

Developmental origins for kidney disease due to Shroom3 deficiency

CKD is a significant health concern with an underlying genetic component. Multiple genome-wide association studies (GWASs) strongly associated CKD with the shroomfamilymember 3 (SHROOM3) gene, which encodes an actin-associated protein important in epithelial morphogenesis. However, the role of SHROOM3 in kidney development and function is virtually unknown. Studies in zebrafish and rat showed that alterations in Shroom3 can result in glomerular dysfunction. Furthermore, human SHROOM3 variants can induce impaired kidney function in animal models. Here, we examined the temporal and spatial expression of Shroom3 in the mammalian kidney. We detected Shroom3 expression in the condensing mesenchyme, Bowman\u27s capsule, and developing and mature podocytes in mice. Shroom3 null (Shroom3Gt/Gt) mice showed marked glomerular abnormalities, including cystic and collapsing/degenerating glomeruli, and marked disruptions in podocyte arrangement and morphology. These podocyte-specific abnormalities are associated with altered Rho-kinase/myosin II signaling and loss of apically distributed actin. Additionally, Shroom3 heterozygous (Shroom3Gt/+) mice showed developmental irregularities that manifested as adult-onset glomerulosclerosis and proteinuria. Taken together, our results establish the significance of Shroom3 in mammalian kidney development and progression of kidney disease. Specifically, Shroom3 maintains normal podocyte architecture in mice via modulation of the actomyosin network, which is essential for podocyte function. Furthermore, our findings strongly support the GWASs that suggest a role for SHROOM3 in human kidney disease

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Semaphorin3f as a Spatial Regulator of Embryogenesis

During embryogenesis, cells integrate both spatial and temporal information from their surroundings to influence proliferation, migration, differentiation and physiological functions. Understanding the molecular mechanisms which confer spatial identity is essential to our understanding of tissue development and human disease. In this thesis I explore multiple roles for the secreted chemotactic ligand Semaphorin3f (Sema3f) in different biological contexts. Using zebrafish (Danio rerio) as a model I take advantage of the duplicated genome to study loss of function of both orthologs, Sema3fa and Sema3fb, in discrete contexts due to their differential expression. First, I show that in the eye Sema3fa produced by progenitors is necessary for the generation of amacrine cells within the temporal retina and the spatially-organized transcriptome of stem cells in the ciliary marginal zone (CMZ). Second, I define an endogenous role of Sema3fa to maintain the avascularity of the neural retina and refine the branch pattern of intraocular vessels. Loss of Sema3fa results in the pathologic angiogenesis of leaky blood vessels into the neural retina. Last, I unveil a role for Sema3fb produced by cardiomyocyte progenitors in the differentiation of the ventricle of the developing heart. Overall, my work provides the first evidence of a Sema3 involved in retinal progenitor cell and cardiomyocyte differentiation, and elucidates the endogenous role of Sema3fa as a negative regulator of retinal blood vessels in the embryo and adult. My data exemplifies the necessity of spatial information conferred by a single chemotactic molecule, Sema3f, to impact differentiation and cellular biology

Shroom3 deficient mice show congenital heart defects

Congenital heart defects (CHDs) are associated with a number of genetic and environmental risk factors affecting approximately 1% of newborns. Shroom3 is an actin binding and microtubule organizing protein essential for neural tube closure in mouse, Xenopus and chick. In Xenopus shroom3 expression is found within the forming heart and loss of activity results in malformed hearts. In addition, SHROOM3 has recently been associated with heterotaxy in a human patient. Mice homozygous for the Shroom3 gene trap die at birth due to exencephaly and here, I provide evidence that the majority of these mice have CHDs, including septal defects, semilunar valve abnormalities, decreased ventricle wall thickness and functional deficits. I fully describe the expression pattern of Shroom3 in heart development, demonstrating it is widely expressed throughout the myocardium. My study is an initial step in characterizing the cell activities that drive cardiac morphogenesis and that result in CHDs when disrupted

Application of game theory in Swedish raw material market : Investigating the pulpwood market

Studien går ut på att analysera marknadsstrukturen för två industriföretag(Holmen och SCA) under antagandet att båda konkurrerar mot varandragenom att köpa rå material samt genom att sälja förädlade produkter.Produktmarknaden som undersöks är pappersmarknaden och antas varakoncentrerad. Rå materialmknaden som undersöks ärmassavedmarknaden och antas karaktäriseras som en duopsony. Detvisade sig att Holmen och SCA köper massaved från en stor mängdskogsägare. Varje företag skapar varje månad en prislista där de bestämmerbud priset föassaved. Priset varierar beroende på region. Både SCA ochHolmen väljer mellan två strategiska beslut, antigen att buda högt pris ellerlågt pris. Genom spelteori så visade det sig att båda industriföretagenanvänder mixade strategier då de i vissa tillfällen budar högt och i andratillfällen budar lågt. Nash jämviktslägen för mixade strategier räknades utmatematiskt och analyserades genom dynamisk spelteori.Marknadskoncentrationen för pappersmarknaden undersöktes viaHerfindahl-Hirschman index (HHI). Porters femkraftsmodell användes föratt analysera industri konkurrensen. Resultatet visade attproduktmarknaden är koncentrerad då HHI testerna gav höga indexvärdenmellan 3100 och 1700. Det existerade dessutom ett Nash jämviktsläge fö mixade strategier som gav SCA förväntad lönsamhet 1651 miljoner kronoroch Holmen 1295 miljoner kronor. Dynamisk spelteori visade att SCA ochHolmens budgivning följer ett mönster och att högt/lågt bud beror påavvikelser från Nash jämviktslägets sannolikhetsdistribution. Nashjämviktslägets råder ifall sannolikhetsdistributionerna vid låg budgivningär 68,6 procent för SCA och 66,7 procent för Holmen. Detta gav indikatore för icke samarbetsvilliga spel. Slutsatsen är att om två spelare (kvarnar) nårThe research aims to analyze the market structure of two companies in th forest industry (Holmen and SCA) with the assumption that thes companies compete at buying raw materials and selling products. Theproduct market in this study is the paper market under the assumption thatboth companies operate in a concentrated product market. The rawmatial market that one investigates in this study is the pulpwood marketunder the assumption that it is a duopsony. What this study has concludedis that Holmen and SCA buy pulpwood from lots of different self-managingforest owners. Each company creates a monthly pricelist where they decidethe bid price of pulpwood. The amount varies depending on the region. Bot SCA and Holmen chooses between two strategic decisions, either to bid highor to bid low. Through game theory, it has been clear that each company usesmixed strategies as they sometimes give high bids and sometimes give lowbids. The Nash equilibrium for mixed strategies have been calculatedmathematically and analyzed through the dynamics of game theory. As fore market concentration, the product market has been investigatedthrough the Herfindahl-Hirschman index (HHI). Porter's five-force modelwas used to analyze the industry competition. The results showed that theproduct market is concentrated as the HHI tests gave High index scoresbetween 3100 and 1700. In addition, there existed a Nash equilibrium in amixed strategy that gave SCA expected payoff 1651 million SEK and Holmen1295 million SEK. The dynamic game theory showed that SCA and Holmen'sbidding follows a repeating trajectory and that the high/low bidding is dueto deviations from Nash equilibrium probability distribution. The Nashequilibrium situation prevails if the probability distribution at low biddingis 68.6 percent for SCA and 66,7 percent for Holmen. This providedindicators for a non-cooperative game. The conclusion is that if two player

Semaphorin3f as a cardiomyocyte derived regulator of heart chamber development

Abstract Background During development a pool of precursors form a heart with atrial and ventricular chambers that exhibit distinct transcriptional and electrophysiological properties. Normal development of these chambers is essential for full term survival of the fetus, and deviations result in congenital heart defects. The large number of genes that may cause congenital heart defects when mutated, and the genetic variability and penetrance of the ensuing phenotypes, reveals a need to understand the molecular mechanisms that allow for the formation of chamber-specific cardiomyocyte differentiation. Methods We used in situ hybridization, immunohistochemistry and functional analyses to identify the consequences of the loss of the secreted semaphorin, Sema3fb, in the development of the zebrafish heart by using two sema3fb CRISPR mutant alleles. Results We find that in the developing zebrafish heart sema3fb mRNA is expressed by all cardiomyocytes, whereas mRNA for a known receptor Plexina3 (Plxna3) is expressed preferentially by ventricular cardiomyocytes. In sema3fb CRISPR zebrafish mutants, heart chamber development is impaired; the atria and ventricles of mutants are smaller in size than their wild type siblings, apparently because of differences in cell size and not cell numbers. Analysis of chamber differentiation indicates defects in chamber specific gene expression at the border between the ventricular and atrial chambers, with spillage of ventricular chamber genes into the atrium, and vice versa, and a failure to restrict specialized cardiomyocyte markers to the atrioventricular canal (AVC). The hypoplastic heart chambers are associated with decreased cardiac output and heart edema. Conclusions Based on our data we propose a model whereby cardiomyocytes secrete a Sema cue that, because of spatially restricted expression of the receptor, signals in a ventricular chamber-specific manner to establish a distinct border between atrial and ventricular chambers that is important to produce a fully functional heart. Video abstrac

Polarity and morphogenesis of the eye epithelium requires the adhesion junction associated adaptor protein Traf4

<p>During development, neuroepithelial progenitors acquire apico-basal polarity and adhere to one another via apically located tight and adherens junction complexes. This polarized neuroepithelium must continue to integrate cells arising through cell divisions and intercalation, and allow for cell movements, at the same time as undergoing morphogenesis. Cell proliferation, migration and intercalation all occur in the morphing embryonic eye. To understand how eye development might depend on dynamic epithelial adhesion, we investigated the function of a known regulator of junctional plasticity, Tumour necrosis factor receptor-associated factor 4 (Traf4). <i>traf4a</i> mRNA is expressed in the developing eye vesicle over the period of optic cup morphogenesis, and Traf4a loss leads to disrupted evagination and elongation of the eye vesicles, and aberrant organization and apico-basal polarity of the eye epithelium. We propose a model whereby Traf4a regulates apical junction plasticity in nascent eye epithelium, allowing for its polarization and morphogenesis.</p> <p><b>Symbols and Abbreviations</b>: AB: apico-basal; aPKC: atypical protein kinase-C; CRISPR: clustered regularly-interspaced short palindromic repeats; GFP: green fluorescent protein; hpf: hours post-fertilization; MO: antisense morpholino oligonucleotide; pHH3: phospho histone H3; ss: somite stage; Traf4: Tumour necrosis factor receptor-associated factor 4; ZO-1: zona occludens-1</p