Optogenetic manipulation of nuclear Dorsal reveals temporal requirements and consequences for transcription.

Morphogen gradients convey essential spatial information during tissue patterning. While both concentration and timing of morphogen exposure are crucial, how cells interpret these graded inputs remains challenging to address. We employed an optogenetic system to acutely and reversibly modulate the nuclear concentration of the morphogen Dorsal (DL), homologue of NF-κB, which orchestrates dorso-ventral patterning in the embryo.

Target gene responses differ when transcription factor levels are acutely decreased by nuclear export versus degradation.

Defining the time of action for morphogens requires tools capable of temporally controlled perturbations. To study how the transcription factor Dorsal affects patterning of the Drosophila embryonic dorsal-ventral axis, we used two light-inducible tags that trigger either nuclear export or degradation of Dorsal under blue light. Nuclear export of Dorsal leads to loss of the high-threshold, ventrally expressed target gene snail (sna), while the low-threshold, laterally expressed target gene short-gastrulation (sog) is retained.

Mechanisms for controlling Dorsal nuclear levels.

Formation of the Dorsal nuclear-cytoplasmic gradient is important for the proper establishment of gene expression patterns along the dorsal-ventral (DV) axis during embryogenesis in . Correct patterning of the DV axis leads to formation of the presumptive mesoderm, neurogenic ectoderm, dorsal ectoderm, and amnioserosa, which are tissues necessary for embryo viability. While Toll signaling is necessary for Dorsal gradient formation, a gradient still forms in the absence of Toll, suggesting there are additional mechanisms required to achieve correct nuclear Dorsal levels.

Target gene responses differ when transcription factor levels are acutely decreased by nuclear export versus degradation.

Defining the time of action for morphogens requires tools capable of temporally controlled perturbations. To study how the transcription factor Dorsal affects patterning of the embryonic dorsal-ventral axis, we used two light-inducible tags that result in either nuclear export or degradation of Dorsal when exposed to blue light. Nuclear export of Dorsal results in loss of expression for the high threshold, ventrally-expressed target gene () but retention of the low threshold, laterally-expressed target gene ().

Two sequential gene expression programs bridged by cell division support long-distance collective cell migration.

The precise assembly of tissues and organs relies on spatiotemporal regulation of gene expression to coordinate the collective behavior of cells. In Drosophila embryos, the midgut musculature is formed through collective migration of caudal visceral mesoderm (CVM) cells, but how gene expression changes as cells migrate is not well understood. Here, we have focused on ten genes expressed in the CVM and the cis-regulatory sequences controlling their expression.

Shifting Mobility Behaviors in Unprecedented Times: A Multigroup MIMIC Model Investigating Intentions to Use On-Demand Ride Services During the COVID-19 Pandemic.

The spread of COVID-19 has been a major disruptive force in people's everyday lives and mobility behavior. The demand for on-demand ride services, such as taxis and ridehailing, has been specifically affected given both restrictions in service operations and users' concerns about virus transmission in shared vehicles. In the early months of the pandemic, demand for these modes decreased by as much as 80%.

A Comparison of Leg Muscle Oxygenation, Cardiorespiratory Responses, and Blood Lactate between Walking and Running at the Same Speed.

It is not known whether different gait modes, or movement patterns, at the same speed elicit differences in muscle oxygen oxygenation, expressed as muscle oxygen saturation (SmO). Thus, the aim of this study was to compare the oxygenation of two leg muscles (vastus lateralis and gastrocnemius medialis), as well as the heart rate, respiratory gases, and blood lactate between two gait modes (walking and running) of the same speed and duration. Ten men walked and ran for 30 min each at 7 km/h in a random, counterbalanced order.

Enhancing brain entry and therapeutic activity of chimeric antigen receptor T cells with intra-arterial NEO100 in a mouse model of CNS lymphoma.

Objective: Malignancies of the CNS are difficult to treat because the blood-brain barrier (BBB) prevents most therapeutics from reaching the intracranial lesions at sufficiently high concentrations. This also applies to chimeric antigen receptor (CAR) T cells, for which systemic delivery is inferior to direct intratumoral or intraventricular injection of the cells. The authors previously reported on a novel approach to safely and reversibly open the BBB of mice by applying intra-arterial (IA) injections of NEO100, a pharmaceutical-grade version of the natural monoterpene perillyl alcohol.

Robots at your doorstep: acceptance of near-future technologies for automated parcel delivery.

The logistics and delivery industry is undergoing a technology-driven transformation, with robotics, drones, and autonomous vehicles expected to play a key role in meeting the growing challenges of last-mile delivery. To understand the public acceptability of automated parcel delivery options, this U.S.

Single-cell transcriptomics illuminates regulatory steps driving anterior-posterior patterning of Drosophila embryonic mesoderm.

Single-cell technologies promise to uncover how transcriptional programs orchestrate complex processes during embryogenesis. Here, we apply a combination of single-cell technology and genetic analysis to investigate the dynamic transcriptional changes associated with Drosophila embryo morphogenesis at gastrulation. Our dataset encompassing the blastoderm-to-gastrula transition provides a comprehensive single-cell map of gene expression across cell lineages validated by genetic analysis.

On the Boredom of Whoredom: Re-Writing the Politics of Sex Work Through Passivity and Femininity.

By elucidating the average everydayness of prostitution, this essay shows-contrary to contemporary conceptions of sex work as either horror or utopia-that whoring is boring. Boredom is a stubborn aspect of modern Western existence. Yet in its philosophical portrayals, it is only described based on masculine parameters, and modeled on male figures such as the .

Inhibition of autophagy and induction of glioblastoma cell death by NEO214, a perillyl alcohol-rolipram conjugate.

Glioblastoma (GBM) is the most aggressive primary brain tumor, exhibiting a high rate of recurrence and poor prognosis. Surgery and chemoradiation with temozolomide (TMZ) represent the standard of care, but, in most cases, the tumor develops resistance to further treatment and the patients succumb to disease. Therefore, there is a great need for the development of well-tolerated, effective drugs that specifically target chemoresistant gliomas.

Nowhere to go? A study of marginalization, social connection, and mental health outcomes among young adults experiencing the COVID-19 pandemic.

Background: The COVID-19 pandemic onset necessitated large-scale closures of third places, potentially exacerbating social barriers experienced by young adults in the United States. To better understand the role of urban form in facilitating socialization, we examine the effects of pandemic-based third place closures on mental health outcomes as mediated by changes in social connection. Because identifying as a racial, gender, or sexual minority can compound baseline disadvantages rooted in systemic inequities, we investigate outcome differences for non-white, woman/nonbinary, and LGBTQ+ young adults to disentangle identity-based nuances of the pandemic experience.

Enhanced brain entry of checkpoint-inhibitory therapeutic antibodies facilitated by intraarterial NEO100 in mouse models of brain-localized malignancies.

Objective: Immune checkpoint-inhibitory therapeutic antibodies have shown striking activity against several types of cancers but are less effective against brain-localized malignancies, in part due to the protective effect of the blood-brain barrier (BBB). The authors hypothesized that intraarterial (IA) delivery of a novel compound, NEO100, has the potential to safely and reversibly open the BBB to enable brain-targeted therapeutic activity of checkpoint-inhibitory antibodies.

Methods: Immunocompetent mice with syngeneic glioblastoma or melanoma cells implanted into their brains were subjected to a single IA injection of NEO100 to open their BBB.

In-person, pick up or delivery? Evolving patterns of household spending behavior through the early reopening phase of the COVID-19 pandemic.

Consumer reactions to COVID-19 pandemic disruptions have been varied, including modifications in spending frequency, amount, product categories and delivery channels. This study analyzes spending data from a sample of 720 U.S.

For whom did telework not work during the Pandemic? understanding the factors impacting telework satisfaction in the US using a multiple indicator multiple cause (MIMIC) model.

The COVID-19 pandemic required employees and businesses across the world to rapidly transition to work from home over extended periods, reaching what is likely the upper bound of telework in many sectors. Past studies have identified both advantages and disadvantages of teleworking. The pandemic experience offers a unique opportunity to examine employees' experiences and perceptions of telework given the broad participation duration and extent.

A Prospective, Cohort Study of SITOIGANAP to Treat Glioblastoma When Given in Combination With Granulocyte-Macrophage Colony-Stimulating Factor/Cyclophosphamide/Bevacizumab/Nivolumab or Granulocyte-Macrophage Colony-Stimulating Factor/Cyclophosphamide/Bevacizumab/Pembrolizumab in Patients Who Failed Prior Treatment With Surgical Resection, Radiation, and Temozolomide.

Background: Glioblastoma (GBM) is the most common primary, malignant brain tumor in adults and has a poor prognosis. The median progression-free survival (mPFS) of newly diagnosed GBM is approximately 6 months. The recurrence rate approaches 100%, and the case-fatality ratio approaches one.

BMP-gated cell-cycle progression drives anoikis during mesenchymal collective migration.

Tissue homeostasis involves the elimination of abnormal cells to avoid compromised patterning and function. Although quality control through cell competition is well studied in epithelial tissues, it is unknown if and how homeostasis is regulated in mesenchymal collectives. Here, we demonstrate that collectively migrating Drosophila muscle precursors utilize both fibroblast growth factor (FGF) and bone morphogenetic protein (BMP) signaling to promote homeostasis via anoikis, a form of cell death in response to substrate de-adhesion.

Barriers to Timely Diagnosis and Treatment of Vector-Borne Diseases in a Changing Climate: A Case Report.

This case study examined current trends in the prevalence of vector-borne diseases and the impact of climate change on disease distribution. Our findings indicate that the dynamics of the Anopheles mosquito population in particular has changed dramatically in the past decade and now poses an increasing threat to human populations previously at low risk for malaria transmission. Given their geographic location and propensity for sustaining vector-borne disease outbreaks, southeastern states are particularly vulnerable to climate-induced changes in vector populations.

brinker levels regulated by a promoter proximal element support germ cell homeostasis.

A limited BMP signaling range in the stem cell niche of the ovary protects against germ cell tumors and promotes germ cell homeostasis. The canonical repressor of BMP signaling in both the Drosophila embryo and wing disc is the transcription factor Brinker (Brk), yet the expression and potential role of Brk in the germarium has not previously been described. Here, we find that brk expression requires a promoter-proximal element (PPE) to support long-distance enhancer action as well as to drive expression in the germarium.

Dueling emergencies: Flood evacuation ridesharing during the COVID-19 pandemic.

Volunteered sharing of resources is often observed in response to disaster events. During evacuations the sharing of resources and vehicles is a crucial mechanism for expanding critical capacity and enabling inclusive disaster response. This paper examines the complexity of rideshare decision-making in the wake of simultaneous emergencies.

NaNuTrap: a technique for in vivo cell nucleus labelling using nanobodies.

In vivo cell labelling is challenging in fast developmental processes because many cell types differentiate more quickly than the maturation time of fluorescent proteins, making visualization of these tissues impossible with standard techniques. Here, we present a nanobody-based method, Nanobody Nuclear Trap (NaNuTrap), which works with the existing Gal4/UAS system in Drosophila and allows for early in vivo cell nuclei labelling independently of the maturation time of the fluorescent protein. This restores the utility of fluorescent proteins that have longer maturation times, such as those used in two-photon imaging, for live imaging of fast or very early developmental processes.

Light-dependent N-end rule-mediated disruption of protein function in Saccharomyces cerevisiae and Drosophila melanogaster.

Here we describe the development and characterization of the photo-N-degron, a peptide tag that can be used in optogenetic studies of protein function in vivo. The photo-N-degron can be expressed as a genetic fusion to the amino termini of other proteins, where it undergoes a blue light-dependent conformational change that exposes a signal for the class of ubiquitin ligases, the N-recognins, which mediate the N-end rule mechanism of proteasomal degradation. We demonstrate that the photo-N-degron can be used to direct light-mediated degradation of proteins in Saccharomyces cerevisiae and Drosophila melanogaster with fine temporal control.

Dynamic patterning by morphogens illuminated by cis-regulatory studies.

Morphogen concentration changes in space as well as over time during development. However, how these dynamics are interpreted by cells to specify fate is not well understood. Here, we focus on two morphogens: the maternal transcription factors Bicoid and Dorsal, which directly regulate target genes to pattern embryos.

Odd-paired is a pioneer-like factor that coordinates with Zelda to control gene expression in embryos.

Pioneer factors such as Zelda (Zld) help initiate zygotic transcription in early embryos, but whether other factors support this dynamic process is unclear. Odd-paired (Opa), a zinc-finger transcription factor expressed at cellularization, controls the transition of genes from pair-rule to segmental patterns along the anterior-posterior axis. Finding that Opa also regulates expression through enhancer along the dorso-ventral axis, we hypothesized Opa's role is more general.