A Genomic and Genetic Analysis of Doublesex Targets and Function in Drosophila Sexual Dimorphism

Sex determination pathways are diverse throughout the animal kingdom, but converge upon conserved genes that encode products that regulate sexual dimorphism. One such downstream factor across many diverged sex determination pathways is the Drosophila doublesex (dsx) gene. The role of doublesex is highly conserved in different insects and dsx homologs (dsx, mab-3 related transcription factors, DMRTs) play roles in sexual differentiation in a diverse array of metazoans. In Drosophila, nearly all manifestations of sexual dimorphism between males and females are regulated by doublesex, yet there are only three known direct targets of DSX, which cannot account for the differences in regulation by DSX in sexually dimorphic tissues. To gain a comprehensive understanding of DSX targets, we undertook multiple experimental approaches that allowed us to identify genes that were bound by DSX, genes whose expression changed in response to DSX perturbation, and genes that function in dsx-expressing cells. DSX protein binding was assayed by ChIP-seq and DamID-seq on S2 cells expressing tagged DSX-M or DSX-F. We also examined DSX occupancy in adult fat body and gonads using DamID-seq or DamID-chip. These experiments identified 3,717 genes bound by DSX in at least one occupancy dataset. Strikingly, we found that genes with the highest levels of DSX occupancy were bound by DSX in all occupancy data sets. This suggests one main mechanism of DSX action would be binding to potential targets in all tissues/contexts rather than having context-dependent targets. In this model of DSX action, additional inputs (such as segmental identity) would be needed to enact transcriptional regulation of bound genes in the appropriate context. Further strengthening this model, although 2,668 genes are bound by DSX in our adult 
fat body occupancy data, less than 1% of these occupied genes show large and robust transcriptional changes in response to acute changes in DSX isoform. We found that predicted DSX targets are significantly enriched in genes that yield phenotypes in sexually dimorphic tissues after RNAi knockdown in dsx-expressing cells (p=0.002). 41 (70.7%) of high probability DSX targets had phenotypes in at least one sexual dimorphic tissue compared to 7 (31.8%) of low probability targets. Altogether, the occupancy, transcriptional profiling, and functional testing have provided a detailed description of how dsx regulates sexual development. New dsx-interacting genes include genes involved in insect hormone signaling. We have identified the Ecdysone receptor gene as a target of DSX. Since the Drosophila gonad represents an excellent model to dissect how DSX acts on a particular time and place to promote development of a sexually dimorphic tissue, we examined the Ecdysone receptor gene, which is involved in ecdysteroid signaling, for roles in gonad sexual development. My data supports the hypothesis that the steroid hormone ecdysone elicits a different response in the male vs. female gonad and that this difference is regulated by DSX and may be important for proper formation of the ovary vs. the testis. Rather than being strictly a genetic process, results from our experiments may demonstrate that sexual differentiation in the gonad occurs through a combination of signals that include sex specific hormone signaling. Since the formation of the gonad may represent processes that are conserved from flies to man, this research will provide insight into conserved genes that regulate developmentally similar pathways whose outcome generates major differences observed between the sexes

Exited Prostitution Survivor Policy Platform

Survivors of prostitution propose a policy reform platform including three main pillars of priority: criminal justice reforms, fair employment, and standards of care. The sexual exploitation of prostituted individuals has lasting effects which can carry over into many aspects of life. In order to remedy these effects and give survivors the opportunity to live a full and free life, we must use a survivor-centered approach to each of these pillars to create change. First, reform is necessary in the criminal justice system to recognize survivors as victims of crime and not perpetrators, while holding those who exploited them fully responsible. Second, reform is necessary to assist survivors in finding fair employment by offering vocational training, financial counseling, and educational scholarships, as well as offering employment opportunities that utilize survivors’ vast array of skills and interests. Finally, standards of care for survivors exiting prostitution should focus on supporting survivors in our journeys and support short- and long-term resources that empower us. These systemic changes are necessary to recognize survivors as the valuable human beings we are and to support survivors in fulfilling our vast potential

The importance of sesquiterpene oxidation products for secondary organic aerosol formation in a springtime hemiboreal forest

Secondary organic aerosols (SOAs) formed from biogenic volatile organic compounds (BVOCs) constitute a significant fraction of atmospheric particulate matter and have been recognized to significantly affect the climate and air quality. Atmospheric SOA particulate mass yields and chemical composition result from a complex mixture of oxidation products originating from a diversity of BVOCs. Many laboratory and field experiments have studied SOA particle formation and growth in the recent years. However, a large uncertainty still remains regarding the contribution of BVOCs to SOA. In particular, organic compounds formed from sesquiterpenes have not been thoroughly investigated, and their contribution to SOA remains poorly characterized. In this study, a Filter Inlet for Gases and Aerosols (FI-GAERO) combined with a high-resolution time-of-flight chemical ionization mass spectrometer (CIMS), with iodide ionization, was used for the simultaneous measurement of gas-phase and particle-phase oxygenated compounds. The aim of the study was to evaluate the relative contribution of sesquiterpene oxidation products to SOA in a springtime hemiboreal forest environment. Our results revealed that monoterpene and sesquiterpene oxidation products were the main contributors to SOA particles. The chemical composition of SOA particles was compared for times when either monoterpene or sesquiterpene oxidation products were dominant and possible key oxidation products for SOA particle formation were identified for both situations. Surprisingly, sesquiterpene oxidation products were the predominant fraction in the particle phase in some periods, while their gas-phase concentrations remained much lower than those of monoterpene products. This can be explained by favorable and effective partitioning of sesquiterpene products into the particle phase. The SOA particle volatility determined from measured thermograms increased when the concentration of sesquiterpene oxidation products in SOA particles was higher than that of monoterpenes. Overall, this study demonstrates that sesquiterpenes may have an important role in atmospheric SOA formation and oxidation chemistry, in particular during the spring recovery period.Peer reviewe

The proadhesive phenotype of systemic sclerosis skin promotes myeloid cell adhesion via ICAM-1 and VCAM-1

Objective. SSc is characterized by microvascular abnormalities and leucocyte infiltration. Previous studies have suggested a proadhesive phenotype in SSc skin, but the functional consequences of this phenotype are not fully understood. Molecules known to mediate leucocyte adhesion include those present at intracellular junctions, such as junctional adhesion molecule-B (JAM-B), JAM-C and CD99, as well as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). The aim of this study was to examine adhesive interactions in SSc skin. Methods. The expression of JAM-B, JAM-C, CD99, ICAM-1 and VCAM-1 in SSc skin was determined by immunohistology and cell surface ELISA. Myeloid U937 cell–SSc dermal fibroblast adhesion assays or in situ adhesion assays to SSc skin were performed. Results. JAM-C and CD99 expression on endothelial cells (ECs) in SSc skin was decreased compared with expression on normal ECs. CD99 was overexpressed on mononuclear cells in SSc skin and on SSc dermal fibroblasts. Neutralizing ICAM-1 inhibited the binding of U937 cells to SSc dermal fibroblasts. In addition, blocking both ICAM-1 and VCAM-1 inhibited U937 cell adhesion to either proximal (less involved) or distal (more involved) SSc skin. Conclusions. These studies show that JAM-C and CD99 are aberrantly expressed in SSc skin. However, these adhesion molecules do not mediate myeloid cell–SSc skin adhesion. In contrast, we demonstrate an important role for ICAM-1 and VCAM-1 in the retention of myeloid cells in SSc skin, suggesting that targeting these molecules may be useful SSc therapies.NIH (grants AI-40987, AR-48267 and AR-19616)Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/77484/1/Rheumatology 48; 734-740, 2009.pdf-

The proadhesive phenotype of systemic sclerosis skin promotes myeloid cell adhesion via ICAM-1 and VCAM-1

Objective. SSc is characterized by microvascular abnormalities and leucocyte infiltration. Previous studies have suggested a proadhesive phenotype in SSc skin, but the functional consequences of this phenotype are not fully understood. Molecules known to mediate leucocyte adhesion include those present at intracellular junctions, such as junctional adhesion molecule-B (JAM-B), JAM-C and CD99, as well as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). The aim of this study was to examine adhesive interactions in SSc skin. Methods. The expression of JAM-B, JAM-C, CD99, ICAM-1 and VCAM-1 in SSc skin was determined by immunohistology and cell surface ELISA. Myeloid U937 cell-SSc dermal fibroblast adhesion assays or in situ adhesion assays to SSc skin were performed. Results. JAM-C and CD99 expression on endothelial cells (ECs) in SSc skin was decreased compared with expression on normal ECs. CD99 was overexpressed on mononuclear cells in SSc skin and on SSc dermal fibroblasts. Neutralizing ICAM-1 inhibited the binding of U937 cells to SSc dermal fibroblasts. In addition, blocking both ICAM-1 and VCAM-1 inhibited U937 cell adhesion to either proximal (less involved) or distal (more involved) SSc skin. Conclusions. These studies show that JAM-C and CD99 are aberrantly expressed in SSc skin. However, these adhesion molecules do not mediate myeloid cell-SSc skin adhesion. In contrast, we demonstrate an important role for ICAM-1 and VCAM-1 in the retention of myeloid cells in SSc skin, suggesting that targeting these molecules may be useful SSc therapies

The proadhesive phenotype of systemic sclerosis skin promotes myeloid cell adhesion via ICAM-1 and VCAM-1

Objective. SSc is characterized by microvascular abnormalities and leucocyte infiltration. Previous studies have suggested a proadhesive phenotype in SSc skin, but the functional consequences of this phenotype are not fully understood. Molecules known to mediate leucocyte adhesion include those present at intracellular junctions, such as junctional adhesion molecule-B (JAM-B), JAM-C and CD99, as well as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). The aim of this study was to examine adhesive interactions in SSc skin. Methods. The expression of JAM-B, JAM-C, CD99, ICAM-1 and VCAM-1 in SSc skin was determined by immunohistology and cell surface ELISA. Myeloid U937 cell-SSc dermal fibroblast adhesion assays or in situ adhesion assays to SSc skin were performed. Results. JAM-C and CD99 expression on endothelial cells (ECs) in SSc skin was decreased compared with expression on normal ECs. CD99 was overexpressed on mononuclear cells in SSc skin and on SSc dermal fibroblasts. Neutralizing ICAM-1 inhibited the binding of U937 cells to SSc dermal fibroblasts. In addition, blocking both ICAM-1 and VCAM-1 inhibited U937 cell adhesion to either proximal (less involved) or distal (more involved) SSc skin. Conclusions. These studies show that JAM-C and CD99 are aberrantly expressed in SSc skin. However, these adhesion molecules do not mediate myeloid cell-SSc skin adhesion. In contrast, we demonstrate an important role for ICAM-1 and VCAM-1 in the retention of myeloid cells in SSc skin, suggesting that targeting these molecules may be useful SSc therapies

The NRG1 exon 11 missense variant is not associated with autism in the Central Valley of Costa Rica

<p>Abstract</p> <p>Background</p> <p>We are conducting a genetic study of autism in the isolated population of the Central Valley of Costa Rica (CVCR). A novel Neuregulin 1 (NRG1) missense variant (exon 11 G>T) was recently associated with psychosis and schizophrenia (SCZ) in the same population isolate.</p> <p>Methods</p> <p>We genotyped the NRG1 exon 11 missense variant in 146 cases with autism, or autism spectrum disorder, with CVCR ancestry, and both parents when available (N = 267 parents) from 143 independent families. Additional microsatellites were genotyped to examine haplotypes bearing the exon 11 variant.</p> <p>Results</p> <p>The NRG1 exon 11 G>T variant was found in 4/146 cases including one de novo occurrence. The frequency of the variant in case chromosomes was 0.014 and 0.045 in the parental non-transmitted chromosomes. At least 6 haplotypes extending 0.229 Mb were associated with the T allele. Three independent individuals, with no personal or family history of psychiatric disorder, shared at least a 1 megabase haplotype 5' to the T allele.</p> <p>Conclusion</p> <p>The NRG1 exon 11 missense variant is not associated with autism in the CVCR.</p

Briefing Two: Justice for All and the Economic Crisis

The world faces its biggest economic crisis in almost 100 years. COVID-19's economic impacts are sure to last longer than the public health emergency and will trigger a massive increase in justice problems. Unemployment is rising, people are increasingly threatened by eviction, many companies are fighting to stave off bankruptcy. In our briefing on Justice for All and the Economic Crisis we present strategies for how justice systems can help, not hinder economic recovery, and how justice leaders can take action to reshape justice systems and support more inclusive, sustainable, and resilient patterns of growth.This report was commisioned by Pathfinders for Peaceful, Just and Inclusive Societie

Briefing One: Justice for All and the Public Health Emergency

The COVID-19 pandemic is an unprecedented global emergency. It is not only a health crisis but also a human rights crisis. Justice actors face daunting responsibilities as they design, implement, and enforce new measures to prevent the spread of infection. Measures that heighten the risk of human rights abuses can undermine trust, at a time when the justice system most needs to maintain the public’s confidence. For better or for worse, justice systems and justice workers are on the frontline of this pandemic. If we get our response right, societies will be better able to confront the pandemic effectively and fairly. That will build the foundations for reset and recovery. If we get it wrong, it is no exaggeration to say that people will die unnecessarily. In the Justice for All report released last year, the Task Force on Justice noted that 1.5 billion people had a justice problem that they could not resolve. Now as well as before the pandemic, marginalized communities – already poorly served by justice systems – face the highest risks, as do vulnerable groups. The pandemic is widening the justice gap, with a sharp increase in the problems that many people face and the ability of justice actors to respond declining. This briefing – Justice for All and the Public Health Emergency – discusses the most pressing priorities that the public health emergency poses for justice leaders and proposes seven areas for urgent action as the tide of infections continues to rise. It will soon be followed by a second briefing to cover the role justice plays in the economic crisis and recovery, and in building cohesion and hope for a better world. In the health sector we are seeing a massive global effort, with people coming together in response to the pandemic. This includes unprecedented international cooperation, a global drive to find treatment and a vaccine, and intensive international sharing and learning among health professionals as they battle the pandemic. This briefing too has been a collective effort, but it is only the beginning. It is also a call to action for the justice community to rally to help countries under extraordinary pressure to get it right. We call on everyone working for justice – globally, nationally, locally; in government, civil society, community organizations or the private sector – to pull together to resolve the justice problems the pandemic is creating, to prevent injustices from occurring, and to use justice as a platform for people to play the fullest possible role in their economies and societies.This report was commisioned by Pathfinders for Peaceful, Just and Inclusive Societie

ISPD gene mutations are a common cause of congenital and limb-girdle muscular dystrophies

Dystroglycanopathies are a clinically and genetically diverse group of recessively inherited conditions ranging from the most severe of the congenital muscular dystrophies, Walker-Warburg syndrome, to mild forms of adult-onset limb-girdle muscular dystrophy. Their hallmark is a reduction in the functional glycosylation of α-dystroglycan, which can be detected in muscle biopsies. An important part of this glycosylation is a unique O-mannosylation, essential for the interaction of α-dystroglycan with extracellular matrix proteins such as laminin-α2. Mutations in eight genes coding for proteins in the glycosylation pathway are responsible for ∼50% of dystroglycanopathy cases. Despite multiple efforts using traditional positional cloning, the causative genes for unsolved dystroglycanopathy cases have escaped discovery for several years. In a recent collaborative study, we discovered that loss-of-function recessive mutations in a novel gene, called isoprenoid synthase domain containing (ISPD), are a relatively common cause of Walker-Warburg syndrome. In this article, we report the involvement of the ISPD gene in milder dystroglycanopathy phenotypes ranging from congenital muscular dystrophy to limb-girdle muscular dystrophy and identified allelic ISPD variants in nine cases belonging to seven families. In two ambulant cases, there was evidence of structural brain involvement, whereas in seven, the clinical manifestation was restricted to a dystrophic skeletal muscle phenotype. Although the function of ISPD in mammals is not yet known, mutations in this gene clearly lead to a reduction in the functional glycosylation of α-dystroglycan, which not only causes the severe Walker-Warburg syndrome but is also a common cause of the milder forms of dystroglycanopathy