Degenerative and regenerative pathways underlying Duchenne muscular dystrophy revealed by single-nucleus RNA sequencing

Duchenne muscular dystrophy (DMD) is a fatal muscle disorder characterized by cycles of degeneration and regeneration of multinucleated myofibers and pathological activation of a variety of other muscle-associated cell types. The extent to which different nuclei within the shared cytoplasm of a myofiber may display transcriptional diversity and whether individual nuclei within a multinucleated myofiber might respond differentially to DMD pathogenesis is unknown. Similarly, the potential transcriptional diversity among nonmuscle cell types within dystrophic muscle has not been explored. Here, we describe the creation of a mouse model of DMD caused by deletion of exon 51 of the dystrophin gene, which represents a prevalent disease-causing mutation in humans. To understand the transcriptional abnormalities and heterogeneity associated with myofiber nuclei, as well as other mononucleated cell types that contribute to the muscle pathology associated with DMD, we performed single-nucleus transcriptomics of skeletal muscle of mice with dystrophin exon 51 deletion. Our results reveal distinctive and previously unrecognized myonuclear subtypes within dystrophic myofibers and uncover degenerative and regenerative transcriptional pathways underlying DMD pathogenesis. Our findings provide insights into the molecular underpinnings of DMD, controlled by the transcriptional activity of different types of muscle and nonmuscle nuclei

Navigating education in the context of COVID-19 lockdowns and school closures : challenges and resilience among adolescent girls and young women in South Africa

Gender related vulnerabilities and inequalities place female learners at high risk of school disengagement due to COVID-19 disruptions. Understanding the impacts of school closures and educational disruptions on female learners in South Africa is critical to inform appropriate, gender-sensitive policies, and programs, to mitigate further exacerbation of educational inequalities. We examined the effects that COVID-19 and lockdowns have had on the educational experiences of adolescent girls and young women (AGYW) aged 15–24, in six districts of South Africa characterized by high rates of HIV, teenage pregnancy and socio-economic hardship. Following a concurrent triangulation mixed-methods approach, we conducted a cross-sectional survey with 515 AGYW, and qualitative interviews with 50 AGYW. More than half of survey participants enrolled in education had been unable to continue with their studies. Factors associated with educational disruption included low socio-economic status, lack of cell phone access and household food insecurity. Qualitative narratives included challenges with online learning and studying at home in resource restricted settings, and insufficient support from schools and teachers. However, despite multiple barriers to continuing education, some AGYW demonstrated educational resilience, enabled by psychosocial and structural support, and resource access. Our findings lend support to an emerging evidence base showing that the closure of schools and tertiary institutions, combined with challenging home environments, and a lack of access to appropriate technology, has disproportionately impacted the most vulnerable AGYW, exacerbating pre-existing educational inequalities within the South African education system. Addressing structural barriers to educational equity, particularly in the pandemic context, including access of technology and the internet, is urgent

Adaptation and resilience : lessons learned from implementing a combination health and education intervention for adolescent girls and young women in South Africa during the COVID-19 pandemic

The COVID-19 pandemic has been associated with reduced access to health services and worsening health outcomes for HIV and sexual and reproductive health (SRH). Through the analysis of data from an evaluation study of a combination intervention for adolescent girls and young women (AGYW) in South Africa, we sought to examine the way in which implementation and service provision were impacted by the COVID-19 pandemic and related restrictions, describing the adaptation implementers made to respond to this context. The intervention was implemented from 2019 in South African districts identified as high priority, given the high rates of HIV and teenage pregnancy amongst AGYW. The South African government introduced the first COVID-19 lockdown in March 2020. We conducted in-depth interviews with 38 intervention implementers in the period from November 2020 to March 2021. Respondents described various ways in which the COVID-19 pandemic and related restrictions had limited their ability to implement the intervention and provide services as planned. As a result, AGYW intervention beneficiary access to SRH and psychosocial services was disrupted. Implementers described several ways in which they attempted to adapt to the pandemic context, such as offering services remotely or door-to-door. Despite attempts to respond to the context and adapt services, overall COVID-19 negatively affected implementation and service provision, and heightened issues around community acceptability of the programs. Our findings can help to inform efforts to reduce health service disruption, increase health system resilience, and ensure continuous SRH service provision to AGYW in times of pandemics and other crises

Autobiography as unconventional history: Constructing the author

The experience of historians as autobiographers has led them to reconsider the nature of historical knowledge and the function of the historian as an intermediary between the past and present. In the new theoretical context of the social sciences and historiography, we can take this proposal further and consider autobiography as a valid form of history—or, at least, as ‘unconventional history’, understood as negotiations with history that transcend or subvert traditional chronological monographs, posit the ‘subjective’ as a useful form of knowledge, and engage the constructed nature of the text. Taking this hypothesis as a starting point, this article reads historians' autobiographical texts to explore if we can/should continue to defend the classic distinction between subject and object, historian scientist and historian author. In this article I compare the work of several historian autobiographers that permit us to identify different methodologies in approaching the story of the self that also reflects different theoretical conceptions of history. I argue that historians that may be considered ‘constructionist’, such as Fernand Braudel, Annie Kriegel, George Duby, and Eric Hobsbawm, design their autobiographies in the same way they articulate their historical texts: by foregrounding objectivity and establishing critical distance between the subject—the historian who narrates the story—and the object—one's own life. Unconventional or experimental approaches, such as those espoused by Robert Rosenstone, Dominick LaCapra, or Clifford Geertz, result in more self-conscious autobiographies, which are, paradoxically, often more realistic and more revealing of the epistemological nature of life writing. ----------------- La experiencia de los historiadores como autobiógrafos les ha llevado a reconsiderar la naturaleza del conocimiento histórico y la función del historiador como un intermediario entre el pasado y el presente. En el nuevo contexto teórico de las ciencias sociales y la historiografía podemos tomar esta propuesta más allá y considerar la autobiografía como una forma válida de historia-o, al menos, de historia ‘poco convencional’-, entendida como negociaciones con la historia que trascienden o subvierten las tradicionales monografías cronológicas, plantean lo "subjetivo" como una forma útil de conocimiento y participan de la naturaleza construida del texto. Tomando esta hipótesis como punto de partida, este artículo lee los textos autobiográficos de los historiadores para explorar si se puede / debe seguir defendiendo la clásica distinción entre sujeto y objeto, historiador científico e historiador escritor. En este artículo comparo el trabajo de varios historiadores autobiógrafos que nos permiten identificar las diferentes metodologías para acercarse a la historia del yo y que también reflejan las diferentes concepciones teóricas de la historia. Sostengo que los historiadores que pueden considerarse "constructivistas", como Fernand Braudel, Annie Kriegel, George Duby y Eric Hobsbawm, diseñan sus autobiografías de la misma forma que articulan sus textos históricos: poniendo en primer plano la objetividad y estableciendo una distancia crítica entre el sujeto -el historiador que narra la historia-y el objeto- la vida de cada uno. Enfoques no convencionales o experimentales, como los expuestos por Robert Rosenstone, Dominick LaCapra, o Clifford Geertz, resultan autobiografías más autoconscientes, que son, paradójicamente, a menudo más realistas y más reveladoras de la naturaleza epistemológica de la escritura de la vida

In Vivo Analysis of MEF2 Transcription Factors in Synapse Regulation and Neuronal Survival

MEF2 (A–D) transcription factors govern development, differentiation and maintenance of various cell types including neurons. The role of MEF2 isoforms in the brain has been studied using in vitro manipulations with only MEF2C examined in vivo. In order to understand specific as well as redundant roles of the MEF2 isoforms, we generated brain-specific deletion of MEF2A and found that Mef2aKO mice show normal behavior in a range of paradigms including learning and memory. We next generated Mef2a and Mef2d brain-specific double KO (Mef2a/dDKO) mice and observed deficits in motor coordination and enhanced hippocampal short-term synaptic plasticity, however there were no alterations in learning and memory, Schaffer collateral pathway long-term potentiation, or the number of dendritic spines. Since previous work has established a critical role for MEF2C in hippocampal plasticity, we generated a Mef2a, Mef2c and Mef2d brain-specific triple KO (Mef2a/c/dTKO). Mef2a/c/d TKO mice have early postnatal lethality with increased neuronal apoptosis, indicative of a redundant role for the MEF2 factors in neuronal survival. We examined synaptic plasticity in the intact neurons in the Mef2a/c/d TKO mice and found significant impairments in short-term synaptic plasticity suggesting that MEF2C is the major isoform involved in hippocampal synaptic function. Collectively, these data highlight the key in vivo role of MEF2C isoform in the brain and suggest that MEF2A and MEF2D have only subtle roles in regulating hippocampal synaptic function

Myogenin Regulates Exercise Capacity and Skeletal Muscle Metabolism in the Adult Mouse

Although skeletal muscle metabolism is a well-studied physiological process, little is known about how it is regulated at the transcriptional level. The myogenic transcription factor myogenin is required for skeletal muscle development during embryonic and fetal life, but myogenin's role in adult skeletal muscle is unclear. We sought to determine myogenin's function in adult muscle metabolism. A Myog conditional allele and Cre-ER transgene were used to delete Myog in adult mice. Mice were analyzed for exercise capacity by involuntary treadmill running. To assess oxidative and glycolytic metabolism, we performed indirect calorimetry, monitored blood glucose and lactate levels, and performed histochemical analyses on muscle fibers. Surprisingly, we found that Myog-deleted mice performed significantly better than controls in high- and low-intensity treadmill running. This enhanced exercise capacity was due to more efficient oxidative metabolism during low- and high-intensity exercise and more efficient glycolytic metabolism during high-intensity exercise. Furthermore, Myog-deleted mice had an enhanced response to long-term voluntary exercise training on running wheels. We identified several candidate genes whose expression was altered in exercise-stressed muscle of mice lacking myogenin. The results suggest that myogenin plays a critical role as a high-level transcriptional regulator to control the energy balance between aerobic and anaerobic metabolism in adult skeletal muscle

Absence of RIP140 Reveals a Pathway Regulating glut4-Dependent Glucose Uptake in Oxidative Skeletal Muscle through UCP1-Mediated Activation of AMPK

Skeletal muscle constitutes the major site of glucose uptake leading to increased removal of glucose from the circulation in response to insulin. Type 2 diabetes and obesity are often associated with insulin resistance that can be counteracted by exercise or the use of drugs increasing the relative proportion of oxidative fibers. RIP140 is a transcriptional coregulator with a central role in metabolic tissues and we tested the effect of modulating its level of expression on muscle glucose and lipid metabolism in two mice models. Here, we show that although RIP140 protein is expressed at the same level in both oxidative and glycolytic muscles, it inhibits both fatty acid and glucose utilization in a fiber-type dependent manner. In RIP140-null mice, fatty acid utilization increases in the extensor digitorum longus and this is associated with elevated expression of genes implicated in fatty acid binding and transport. In the RIP140-null soleus, depletion of RIP140 leads to increased GLUT4 trafficking and glucose uptake with no change in Akt activity. AMPK phosphorylation/activity is inhibited in the soleus of RIP140 transgenic mice and increased in RIP140-null soleus. This is associated with increased UCP1 expression and mitochondrial uncoupling revealing the existence of a signaling pathway controlling insulin-independent glucose uptake in the soleus of RIP140-null mice. In conclusion, our findings reinforce the participation of RIP140 in the maintenance of energy homeostasis by acting as an inhibitor of energy production and particularly point to RIP140 as a promising therapeutic target in the treatment of insulin resistance

Microgenomic Analysis in Skeletal Muscle: Expression Signatures of Individual Fast and Slow Myofibers

BACKGROUND: Skeletal muscle is a complex, versatile tissue composed of a variety of functionally diverse fiber types. Although the biochemical, structural and functional properties of myofibers have been the subject of intense investigation for the last decades, understanding molecular processes regulating fiber type diversity is still complicated by the heterogeneity of cell types present in the whole muscle organ. METHODOLOGY/PRINCIPAL FINDINGS: We have produced a first catalogue of genes expressed in mouse slow-oxidative (type 1) and fast-glycolytic (type 2B) fibers through transcriptome analysis at the single fiber level (microgenomics). Individual fibers were obtained from murine soleus and EDL muscles and initially classified by myosin heavy chain isoform content. Gene expression profiling on high density DNA oligonucleotide microarrays showed that both qualitative and quantitative improvements were achieved, compared to results with standard muscle homogenate. First, myofiber profiles were virtually free from non-muscle transcriptional activity. Second, thousands of muscle-specific genes were identified, leading to a better definition of gene signatures in the two fiber types as well as the detection of metabolic and signaling pathways that are differentially activated in specific fiber types. Several regulatory proteins showed preferential expression in slow myofibers. Discriminant analysis revealed novel genes that could be useful for fiber type functional classification. CONCLUSIONS/SIGNIFICANCE: As gene expression analyses at the single fiber level significantly increased the resolution power, this innovative approach would allow a better understanding of the adaptive transcriptomic transitions occurring in myofibers under physiological and pathological condition

Management practices for control of ragwort species

The ragwort species common or tansy ragwort (Jacobaea vulgaris, formerly Senecio jacobaea), marsh ragwort (S. aquaticus), Oxford ragwort (S. squalidus) and hoary ragwort (S. erucifolius) are native in Europe, but invaded North America, Australia and New Zealand as weeds. The abundance of ragwort species is increasing in west-and central Europe. Ragwort species contain different groups of secondary plant compounds defending them against generalist herbivores, contributing to their success as weeds. They are mainly known for containing pyrrolizidine alkaloids, which are toxic to grazing cattle and other livestock causing considerable losses to agricultural revenue. Consequently, control of ragwort is obligatory by law in the UK, Ireland and Australia. Commonly used management practices to control ragwort include mechanical removal, grazing, pasture management, biological control and chemical control. In this review the biology of ragwort species is shortly described and the different management practices are discussed

Eccentric Exercise Activates Novel Transcriptional Regulation of Hypertrophic Signaling Pathways Not Affected by Hormone Changes

Unaccustomed eccentric exercise damages skeletal muscle tissue, activating mechanisms of recovery and remodeling that may be influenced by the female sex hormone 17β-estradiol (E2). Using high density oligonucleotide based microarrays, we screened for differences in mRNA expression caused by E2 and eccentric exercise. After random assignment to 8 days of either placebo (CON) or E2 (EXP), eighteen men performed 150 single-leg eccentric contractions. Muscle biopsies were collected at baseline (BL), following supplementation (PS), +3 hours (3H) and +48 hours (48H) after exercise. Serum E2 concentrations increased significantly with supplementation (P<0.001) but did not affect microarray results. Exercise led to early transcriptional changes in striated muscle activator of Rho signaling (STARS), Rho family GTPase 3 (RND3), mitogen activated protein kinase (MAPK) regulation and the downstream transcription factor FOS. Targeted RT-PCR analysis identified concurrent induction of negative regulators of calcineurin signaling RCAN (P<0.001) and HMOX1 (P = 0.009). Protein contents were elevated for RND3 at 3H (P = 0.02) and FOS at 48H (P<0.05). These findings indicate that early RhoA and NFAT signaling and regulation are altered following exercise for muscle remodeling and repair, but are not affected by E2