RhoA GEF Mcf2lb regulates rosette integrity during collective cell migration.

Multicellular rosettes are transient epithelial structures that serve as important cellular intermediates in the formation of diverse organs. Using the zebrafish posterior lateral line primordium (pLLP) as a model system, we investigated the role of the RhoA GEF Mcf2lb in rosette morphogenesis. The pLLP is a group of ∼150 cells that migrates along the zebrafish trunk and is organized into epithelial rosettes; these are deposited along the trunk and will differentiate into sensory organs called neuromasts (NMs). Using single-cell RNA-sequencing and whole-mount in situ hybridization, we showed that mcf2lb is expressed in the pLLP during migration. Live imaging and subsequent 3D analysis of mcf2lb mutant pLLP cells showed disrupted apical constriction and subsequent rosette organization. This resulted in an excess number of deposited NMs along the trunk of the zebrafish. Cell polarity markers ZO-1 and Par-3 were apically localized, indicating that pLLP cells are properly polarized. In contrast, RhoA activity, as well as signaling components downstream of RhoA, Rock2a and non-muscle Myosin II, were diminished apically. Thus, Mcf2lb-dependent RhoA activation maintains the integrity of epithelial rosettes.

Lamellipodia-like protrusions and focal adhesions contribute to collective cell migration in zebrafish.

Collective cell migration is a process where cohorts of cells exhibit coordinated migratory behavior. During individual and collective cellular migration, cells must extend protrusions to interact with the extracellular environment, sense chemotactic cues, and act as points of attachment. The mechanisms and regulators of protrusive behavior have been widely studied in individually migrating cells; however, how this behavior is regulated throughout collectives is not well understood. To address this, we used the zebrafish posterior lateral line primordium (pLLP) as a model. The pLLP is a cluster of ~150 â€‹cells that migrates along the zebrafish trunk, depositing groups of cells that will become sensory organs. To define protrusive behavior, we performed mosaic analysis to sparsely label pLLP cells with a transgene marking filamentous actin. This approach revealed an abundance of brush-like protrusions throughout the pLLP that orient in the direction of migration. Formation of these protrusions depends on the Arp2/3 complex, a regulator of dendritic actin. This argues that these brush-like protrusions are an in vivo example of lamellipodia. Mosaic analysis demonstrated that these lamellipodia-like protrusions are located in a close proximity to the overlying skin. Immunostaining revealed an abundance of focal adhesion complexes surrounding the pLLP. Disruption of these complexes specifically in pLLP cells led to impaired pLLP migration. Finally, we show that Erk signaling, a known regulator of focal adhesions, is required for proper formation of lamellipodia-like protrusions and pLLP migration. Altogether, our results suggest a model where the coordinated dynamics of lamellipodia-like protrusions, making contact with either the overlying skin or the extracellular matrix through focal adhesions, promotes migration of pLLP cells.

Early life patterns of criminal legal system involvement: Inequalities by race/ethnicity, gender, and parental education.

BACKGROUND: Contacts with the criminal legal system have consequences for a host of outcomes. Still, early life age patterns of system involvement remain to be better understood. OBJECTIVE: We estimate cumulative risks of arrest, probation, and incarceration from childhood through early adulthood and assess disparities by race/ethnicity, gender, and parental education. METHODS: Data come from the Transition to Adulthood Supplement of the Panel Study of Income Dynamics (n = 2,736). We use Kaplan-Meier curves and Cox regression models to estimate cumulative risks of arrest, probation, and incarceration across the early life course and document disparities by race/ethnicity, gender, and parental education, as well as at their intersections. RESULTS: Criminal legal system involvement is common among recent cohorts, but Black and Latinx boys and young men face especially high risks. Among Black men whose highest-educated parent completed high school or less, an estimated six in ten had been arrested, four in ten had experienced probation, and four in ten had been incarcerated by age 26. Among Latinx men whose highest-educated parent completed high school or less, an estimated four in ten had been arrested and one in four had been incarcerated by age 26. Black women also experienced high risks, with an estimated one in four arrested by age 26. CONTRIBUTION: We document early life patterns of criminal legal system involvement among young people who came of age during the expansion of proactive policing and mass incarceration in the United States, providing important context for understanding the role of the system in generating and exacerbating life course inequalities.

RhoA GEF Mcf2lb regulates rosette integrity during collective cell migration.

During development, multicellular rosettes serve as important cellular intermediates in the formation of diverse organ systems. Multicellular rosettes are transient epithelial structures that are defined by the apical constriction of cells towards the rosette center. Due to the important role these structures play during development, understanding the molecular mechanisms by which rosettes are formed and maintained is of high interest. Utilizing the zebrafish posterior lateral line primordium (pLLP) as a model system, we identify the RhoA GEF Mcf2lb as a regulator of rosette integrity. The pLLP is a group of ~150 cells that migrates along the zebrafish trunk and is organized into epithelial rosettes; these are deposited along the trunk and will differentiate into sensory organs called neuromasts (NMs). Using single-cell RNA sequencing and whole-mount in situ hybridization, we showed that mcf2lb is expressed in the pLLP during migration. Given the known role of RhoA in rosette formation, we asked whether Mcf2lb plays a role in regulating apical constriction of cells within rosettes. Live imaging and subsequent 3D analysis of mcf2lb mutant pLLP cells showed disrupted apical constriction and subsequent rosette organization. This in turn resulted in a unique posterior Lateral Line phenotype: an excess number of deposited NMs along the trunk of the zebrafish. Cell polarity markers ZO-1 and Par-3 were apically localized, indicating that pLLP cells are normally polarized. In contrast, signaling components that mediate apical constriction downstream of RhoA, Rock-2a and non-muscle Myosin II were diminished apically. Altogether our results suggest a model whereby Mcf2lb activates RhoA, which in turn activates downstream signaling machinery to induce and maintain apical constriction in cells incorporated into rosettes.

Increase in the level of ß-HCG after uterine artery embolization in the context of partial molar pregnancy: A case report.

This report documents the case of a patient with a previously diagnosed partial molar pregnancy evacuated by dilation and suction curettage with appropriately declining post-operative levels of beta-human chorionic gonadotropin (beta-hCG), who, one month later, underwent uterine artery embolization in the setting of acute bleeding and imaging concerning for arteriovenous malformation. After embolization, beta-hCG levels increased, prompting concern for gestational trophoblastic neoplasia and referral to gynecologic oncology. With further workup, the elevation was found to be transient and benign - a phenomenon not previously described.

Minimally invasive adnexa-sparing surgery for benign ovarian and paratubal masses in children.

BACKGROUND: The precision of minimally invasive surgery (MIS) to resect benign ovarian and paratubal masses while preserving adnexa in children is unclear. This study evaluated the integrity of laparoscopy to spare adnexa while resecting benign pathologies in children. METHODS: The institutional pathology database was queried to identify patients aged 18 years and younger having any ovarian or tubal lesion resected at a comprehensive children's hospital between 2006 and 2021. Adnexa-sparing surgery was defined as preserving both the ovary and tube from which the lesion was resected. Postoperative ultrasounds were reviewed to document ovarian follicles, blood flow, volumes, and lesion recurrence. RESULTS: Adnexal preservation was implemented in 168 of 328 pathological resections. MIS approach was used in 149 cases. Median age was 13.4 [11.0-15.3]. Among sparing surgeries, MIS associated with benign pathology, shorter operative time (median minutes: 78 MIS [59-111.5]; 130 open [92.8-149.8]; 174 MIS-to-open [132.8-199.5]; p = 0.010), and reduced hospital stay (median days: 1 MIS (Lindfors, 1971; Lovvorn III et al., 1998) [1-2]; 2 open [2-2.9], 2 MIS-to-open [1-3.3]; p = 0.001). Postoperative ovarian volume ipsilateral to the MIS procedure (median, 7.6 ml [4.3-12.1]) was relatively smaller than the contralateral ovary (median, 9.1 ml [5.5-15.0]). Blood flow was documented to the ovary in 70/94 (74.4%) of patients after MIS adnexal-sparing surgery. Distinct follicles were detected in 48/74 (64.8%) of post-menarchal patients after MIS adnexal-sparing surgery. Five ovarian cysts recurred. CONCLUSIONS: MIS preserves adnexa reliably, with postoperative ovarian follicles and blood flow detected in most patients, and maintains ∼80% of contralateral volume, while achieving definitive tumor resection. LEVEL OF EVIDENCE: III.

Vicarious Exposure to the Criminal Legal System Among Parents and Siblings.

Objective: This study documents life course patterns of vicarious exposure to the criminal legal system among parents and siblings in the United States. Background: The criminal legal system shapes family outcomes in important ways. Still, life course patterns of vicarious exposure to the system-especially to lower-level contacts-among parents and siblings are not well documented. Method: Using longitudinal data from the Panel Study of Income Dynamics, Kaplan-Meier survival curves, and Cox regression models, we estimate cumulative risks of vicarious exposure to arrest, probation, and incarceration among parents (n=3,885 parents; 185,444 person-years) and siblings (n=1,875; 44,766 person-years) and examine disparities by race-ethnicity, gender, and education, and at their intersections. Results: Vicarious exposure to the system is common-but highly unequal-among parents and siblings. Racially minoritized parents and siblings had greater levels and earlier risks of exposure. For example, by age 50, an estimated one in five Black parents experienced having a child incarcerated, a risk about twice as high as White and 50% higher than Latinx parents. By age 26, an estimated six in 10 Black young people with brothers experienced having a brother arrested; more than four in 10 experienced a brother on probation; and more than three in 10 experienced brother incarceration. For many estimates, racialized inequities in risks of vicarious system exposure widened at higher levels of education. Conclusion: These findings provide essential context for understanding the role of the criminal legal system in maintaining and exacerbating family inequality.

Genotype classification of Moraxella bovis using MALDI-TOF MS profiles.

Moraxella bovis (M. bovis) is regarded as a causative agent of infectious bovine keratoconjunctivitis (IBK), the most common ocular disease of cattle. Recently, whole genome sequencing identified the presence of two distinct genotypes within M. bovis that differ in chromosome content, potential virulence factors, as well as prophage and plasmid profiles. It is unclear if the genotypes equally associate with IBK or if one is more likely to be isolated from IBK lesions. We utilized 39 strains of M. bovis that had previously undergone whole genome sequencing and genotype classification to determine the utility of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) to accurately genotype M. bovis strains. We successfully developed two biomarker models that accurately classified strains according to genotype with an overall accuracy of 85.8-100% depending upon the model and sample preparation method used. These models provide a practical tool to enable studies of genotype associations with disease, allow for epidemiological studies at the sub-species level, and can be used to enhance disease prevention strategies.

Using Zebrafish to Study Collective Cell Migration in Development and Disease.

Cellular migration is necessary for proper embryonic development as well as maintenance of adult health. Cells can migrate individually or in groups in a process known as collective cell migration. Collectively migrating cohorts maintain cell-cell contacts, group polarization, and exhibit coordinated behavior. This mode of migration is important during numerous developmental processes including tracheal branching, blood vessel sprouting, neural crest cell migration and others. In the adult, collective cell migration is important for proper wound healing and is often misappropriated during cancer cell invasion. A variety of genetic model systems are used to examine and define the cellular and molecular mechanisms behind collective cell migration including border cell migration and tracheal branching in Drosophila melanogaster, neural crest cell migration in chick and Xenopus embryos, and posterior lateral line primordium (pLLP) migration in zebrafish. The pLLP is a group of about 100 cells that begins migrating around 22 hours post-fertilization along the lateral aspect of the trunk of the developing embryo. During migration, clusters of cells are deposited from the trailing end of the pLLP; these ultimately differentiate into mechanosensory organs of the lateral line system. As zebrafish embryos are transparent during early development and the pLLP migrates close to the surface of the skin, this system can be easily visualized and manipulated in vivo. These advantages together with the amenity to advance genetic methods make the zebrafish pLLP one of the premier model systems for studying collective cell migration. This review will describe the cellular behaviors and signaling mechanisms of the pLLP and compare the pLLP to collective cell migration in other popular model systems. In addition, we will examine how this type of migration is hijacked by collectively invading cancer cells.

Nothing to Be Sniffed At: Anosmin1 Tunes Fgf Diffusivity.

The extracellular matrix plays both positive and negative roles in growth factor diffusion, a process critical for organ formation. In this issue of Developmental Cell, Wang et al. (2018) identify the extracellular matrix protein Anosmin1 as a key regulator of Fgf diffusion during sensory organ formation in zebrafish.

Lariat sign: An MRI finding associated with common peroneal nerve rupture.

Traumatic knee dislocation represents a catastrophic orthopedic injury with potentially devastating vascular and neurologic injuries. We report a case of common peroneal nerve rupture sustained during a knee dislocation with novel radiographic findings that we describe as a lariat sign. At the site of rupture, the distal nerve loops back on itself forming a lasso shape or lariat. This thickened nerve's abnormal course should not be misinterpreted as a vessel.