Loss of Rai1 enhances hippocampal excitability and epileptogenesis in mouse models of Smith-Magenis syndrome.

Hyperexcitability of brain circuits is a common feature of autism spectrum disorders (ASDs). Genetic deletion of a chromatin-binding protein, retinoic acid induced 1 (RAI1), causes Smith-Magenis syndrome (SMS). SMS is a syndromic ASD associated with intellectual disability, autistic features, maladaptive behaviors, overt seizures, and abnormal electroencephalogram (EEG) patterns. The molecular and neural mechanisms underlying abnormal brain activity in SMS remain unclear. Here we show that panneural Rai1 deletions in mice result in increased seizure susceptibility and prolonged hippocampal seizure duration in vivo and increased dentate gyrus population spikes ex vivo. Brain-wide mapping of neuronal activity pinpointed selective cell types within the limbic system, including the hippocampal dentate gyrus granule cells (dGCs) that are hyperactivated by chemoconvulsant administration or sensory experience in Rai1-deficient brains. Deletion of Rai1 from glutamatergic neurons, but not from gamma-aminobutyric acidergic (GABAergic) neurons, was responsible for increased seizure susceptibility. Deleting Rai1 from the Emx1Cre-lineage glutamatergic neurons resulted in abnormal dGC properties, including increased excitatory synaptic transmission and increased intrinsic excitability. Our work uncovers the mechanism of neuronal hyperexcitability in SMS by identifying Rai1 as a negative regulator of dGC intrinsic and synaptic excitability.

Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals.

Sensitive detection of two biological events in vivo has long been a goal in bioluminescence imaging. Antares, a fusion of the luciferase NanoLuc to the orange fluorescent protein CyOFP, has emerged as a bright bioluminescent reporter with orthogonal substrate specificity to firefly luciferase (FLuc) and its derivatives such as AkaLuc. However, the brightness of Antares in mice is limited by the poor solubility and bioavailability of the NanoLuc substrate furimazine. Here, we report a new substrate, hydrofurimazine, whose enhanced aqueous solubility allows delivery of higher doses to mice. In the liver, Antares with hydrofurimazine exhibited similar brightness to AkaLuc with its substrate AkaLumine. Further chemical exploration generated a second substrate, fluorofurimazine, with even higher brightness in vivo. We used Antares with fluorofurimazine to track tumor size and AkaLuc with AkaLumine to visualize CAR-T cells within the same mice, demonstrating the ability to perform two-population imaging with these two luciferase systems.

RFWD3-Dependent Ubiquitination of RPA Regulates Repair at Stalled Replication Forks.

We have used quantitative proteomics to profile ubiquitination in the DNA damage response (DDR). We demonstrate that RPA, which functions as a protein scaffold in the replication stress response, is multiply ubiquitinated upon replication fork stalling. Ubiquitination of RPA occurs on chromatin, involves sites outside its DNA binding channel, does not cause proteasomal degradation, and increases under conditions of fork collapse, suggesting a role in repair at stalled forks. We demonstrate that the E3 ligase RFWD3 mediates RPA ubiquitination. RFWD3 is necessary for replication fork restart, normal repair kinetics during replication stress, and homologous recombination (HR) at stalled replication forks. Mutational analysis suggests that multisite ubiquitination of the entire RPA complex is responsible for repair at stalled forks. Multisite protein group sumoylation is known to promote HR in yeast. Our findings reveal a similar requirement for multisite protein group ubiquitination during HR at stalled forks in mammalian cells.

Quantitative Proteomic Atlas of Ubiquitination and Acetylation in the DNA Damage Response.

Execution of the DNA damage response (DDR) relies upon a dynamic array of protein modifications. Using quantitative proteomics, we have globally profiled ubiquitination, acetylation, and phosphorylation in response to UV and ionizing radiation. To improve acetylation site profiling, we developed the strategy FACET-IP. Our datasets of 33,500 ubiquitination and 16,740 acetylation sites provide valuable insight into DDR remodeling of the proteome. We find that K6- and K33-linked polyubiquitination undergo bulk increases in response to DNA damage, raising the possibility that these linkages are largely dedicated to DDR function. We also show that Cullin-RING ligases mediate 10% of DNA damage-induced ubiquitination events and that EXO1 is an SCF-Cyclin F substrate in the response to UV radiation. Our extensive datasets uncover additional regulated sites on known DDR players such as PCNA and identify previously unknown DDR targets such as CENPs, underscoring the broad impact of the DDR on cellular physiology.

Early adolescent Rai1 reactivation reverses transcriptional and social interaction deficits in a mouse model of Smith-Magenis syndrome.

Haploinsufficiency of Retinoic Acid Induced 1 (RAI1) causes Smith-Magenis syndrome (SMS), a syndromic autism spectrum disorder associated with craniofacial abnormalities, intellectual disability, and behavioral problems. There is currently no cure for SMS. Here, we generated a genetic mouse model to determine the reversibility of SMS-like neurobehavioral phenotypes in Rai1 heterozygous mice. We show that normalizing the Rai1 level 3-4 wk after birth corrected the expression of genes related to neural developmental pathways and fully reversed a social interaction deficit caused by Rai1 haploinsufficiency. In contrast, Rai1 reactivation 7-8 wk after birth was not beneficial. We also demonstrated that the correct Rai1 dose is required in both excitatory and inhibitory neurons for proper social interactions. Finally, we found that Rai1 heterozygous mice exhibited a reduction of dendritic spines in the medial prefrontal cortex (mPFC) and that optogenetic activation of mPFC neurons in adults improved the social interaction deficit of Rai1 heterozygous mice. Together, these results suggest the existence of a postnatal temporal window during which restoring Rai1 can improve the transcriptional and social behavioral deficits in a mouse model of SMS. It is possible that circuit-level interventions would be beneficial beyond this critical window.

Why humility is vital to effective humanitarian aid leadership: a review of the literature.

Organisational scientists are paying increasing attention to humility, following a larger trend in scholarship highlighting the relational and interdependent nature of leadership and business. A growing body of evidence identifies humility as vital to effective organisational leadership, facilitating positive organisational outcomes, such as lower voluntary turnover and greater follower job satisfaction. To date, research on the subject has focused on certain specific organisational contexts, including businesses, hospitals, and schools. This paper reviews the existing literature and explores why humility may be an especially important leader trait in international humanitarian aid organisations and relief work-a context that is not only uniquely challenging, but also one that would seemingly stand to benefit keenly from the quality. It argues that humility is essential for effective leadership because it is normative of good character, it is predictive of positive outcomes, and it corresponds to a genuine representation of the nature of humanitarian aid.

Characterization of Indeterminate Liver Lesions on CT and MRI With Contrast-Enhanced Ultrasound: What Is the Evidence?

OBJECTIVE. CT or MRI is most commonly used for characterizing focal hepatic lesions. However, findings on CT and MRI are occasionally indeterminate. Contrast-enhanced ultrasound (CEUS), with its unique characteristics as a purely intravascular contrast agent and real-time evaluation of enhancement, is a useful next step. The purpose of this article is to review the evidence for performing CEUS in the assessment of indeterminate hepatic lesions seen on CT and MRI. CONCLUSION. CEUS is a useful problem-solving tool in the evaluation of liver lesions that are indeterminate on CT and MRI. Uses include detection of arterial phase hyperenhancement; differentiation between hepatocellular carcinoma and intrahepatic cholangiocarcinoma; determination of benign versus malignant tumor thrombus, benign versus neoplastic cystic hepatic lesions, and hepatocellular adenoma versus focal nodular hyperplasia; and monitoring for recurrence in postablative therapies. CEUS can help establish a confident diagnosis and determine the need for further invasive diagnosis or treatment.

Ruptured Degenerating Fibroid Causing Preterm Labour and Systemic Inflammatory Response Syndrome.

BACKGROUND: Fibroids are present in at least 10% of pregnancies and are recognized to cause a variety of complications. A few case reports have described fibroids as an etiological factor in uterine rupture, sometimes with life-threatening hemorrhage. CASE: A 28-year-old G1, P0 woman at 20 weeks gestation developed systemic inflammatory response syndrome with acute renal failure and massive ascites secondary to a ruptured degenerated fibroid. This resulted in preterm delivery and neonatal death. At 6 weeks postpartum, she successfully underwent an abdominal myomectomy. CONCLUSION: This is a rare case of uterine fibroid rupture causing preterm labour and systemic inflammatory response syndrome. This report discusses the diagnosis of uterine rupture related to the fibroid with imaging and subsequent management, which included fertility-preserving surgery.

Muscle-specific AMPK ß1ß2-null mice display a myopathy due to loss of capillary density in nonpostural muscles.

AMP-activated protein kinase (AMPK) is a master regulator of metabolism. While muscle-specific AMPK ß1ß2 double-knockout (ß1ß2M-KO) mice display alterations in metabolic and mitochondrial capacity, their severe exercise intolerance suggested a secondary contributor to the observed phenotype. We find that tibialis anterior (TA), but not soleus, muscles of sedentary ß1ß2M-KO mice display a significant myopathy (decreased myofiber areas, increased split and necrotic myofibers, and increased centrally nucleated myofibers. A mitochondrial- and fiber-type-specific etiology to the myopathy was ruled out. However, ß1ß2M-KO TA muscles displayed significant (P<0.05) increases in platelet aggregation and apoptosis within myofibers and surrounding interstitium (P<0.05). These changes correlated with a 45% decrease in capillary density (P<0.05). We hypothesized that the ß1ß2M-KO myopathy in resting muscle resulted from impaired AMPK-nNOSµ signaling, causing increased platelet aggregation, impaired vasodilation, and, ultimately, ischemic injury. Consistent with this hypothesis, AMPK-specific phosphorylation (Ser1446) of nNOSµ was decreased in ß1ß2M-KO compared to wild-type (WT) mice. The AMPK-nNOSµ relationship was further demonstrated by administration of 5-aminoimidazole-4-carboxamide 1-ß-D-ribofuranoside (AICAR) to ß1ß2-MKO muscles and C2C12 myotubes. AICAR significantly increased nNOSµ phosphorylation and nitric oxide production (P<0.05) within minutes of administration in WT muscles and C2C12 myotubes but not in ß1ß2M-KO muscles. These findings highlight the importance of the AMPK-nNOSµ pathway in resting skeletal muscle.

Imaging of physeal bars in children.

The growth plate, also known as the physis or epiphyseal plate, is essential for longitudinal growth of bones in the immature skeleton. A variety of insults to the growth plate from trauma to infection to idiopathic causes can lead to physeal bar formation, an interruption in normal growth plate cartilage, where a bony or fibrous bridge develops between the metaphysis and epiphysis. This bridge restricts subsequent bone growth, leading to limb shortening and/or angular deformities. Early recognition of the presence of a physeal bar can help direct appropriate surgical management to restore linear growth of the bone.

Embryonically Active Piriform Cortex Neurons Promote Intracortical Recurrent Connectivity during Development.

Neuronal activity plays a critical role in the maturation of circuits that propagate sensory information into the brain. How widely does early activity regulate circuit maturation across the developing brain? Here, we used Targeted Recombination in Active Populations (TRAP) to perform a brain-wide survey for prenatally active neurons in mice and identified the piriform cortex as an abundantly TRAPed region. Whole-cell recordings in neonatal slices revealed preferential interconnectivity within embryonically TRAPed piriform neurons and their enhanced synaptic connectivity with other piriform neurons. In vivo Neuropixels recordings in neonates demonstrated that embryonically TRAPed piriform neurons exhibit broad functional connectivity within piriform and lead spontaneous synchronized population activity during a transient neonatal period, when recurrent connectivity is strengthening. Selectively activating or silencing of these neurons in neonates enhanced or suppressed recurrent synaptic strength, respectively. Thus, embryonically TRAPed piriform neurons represent an interconnected hub-like population whose activity promotes recurrent connectivity in early development.

Longitudinal processes among humility, social justice activism, transcendence, and well-being.

Introduction: Existing research shows positive associations between humility and well-being, and between civic engagement and well-being. Rarely have humility, civic engagement, and well-being been examined together. We build off of previous cross-sectional findings and a prior longitudinal study that used three waves of data and found significant positive bivariate correlations between humility and the presence of life purpose across time points. Methods: Extending these previous findings, we used six waves of data obtained from graduate students at 18 seminaries across North America (N = 574; Mage = 31.54; 46.7% female; 65.3% White) to explore the dynamic associations among humility and life purpose, along with horizontal transcendence (an indicator of the attitudinal dimension of civic engagement) and social justice activism (an indicator for the behavioral dimension). We explored reciprocal short-run processes and dynamic long-run effects using a general cross-lagged panel model. Results and discussion: We found robust evidence for a reciprocal influence between the presence of life purpose and horizontal transcendence, and long-run effects for initial levels of life purpose to influence later levels of horizontal transcendence. We also found long-run effects for the influence of initial levels of life purpose on later levels of humility, and initial levels of social justice activism on later levels of horizontal transcendence. Implications center on the use of the findings for planning future one-time life purpose and social justice interventions to affect changes in humility and horizontal transcendence.

Catheter ablation of left ventricular tachycardia through internal jugular vein: refining the continuous line.

A retrograde approach to the left ventricle (LV) from the femoral artery is most commonly used for left ventricular tachycardia (VT) ablation. However, as the patient population gets older, the prevalence of peripheral vascular disease and aortic valve disease increases, hampering the retrograde access to the LV, which may result in significant technical difficulties. The transseptal approach from the femoral vein is an alternative access to the LV. This approach has been effective for addressing LV VT originating from the posterior and posteroseptal region. However, the transseptal approach from the femoral vein is impossible if there are any obstacles in the inferior vena cava. There are also significant limitations in approaching the basal-lateral region of the LV from this approach. We demonstrated the safety and feasibility of catheter ablation of LV VT via the transseptal approach from the right internal jugular vein.

The multilevel correlates, contributions, and consequences of leader humility in humanitarian aid work.

Objective: Leader humility has been linked to many positive outcomes but not examined in humanitarian aid work. Three studies examined the multilevel correlates, contributions, and consequences of leader humility in Medair-a large, multinational, faith-based aid organization. Study 1 examined correlates of leader humility in a sample of 308 workers and 167 leaders. Study 2 explored multilevel contributions of leader humility in 96 teams comprised of 189 workers. Study 3 utilized a subsample (50 workers, 34 leaders) to explore consequences of Time 1 leader and team humility on outcomes 6 months later. Method: Participants completed measures of humility (general, relational, team), leader and team attributions (e.g., effectiveness, cohesion, and growth-mindedness), organizational outcomes (e.g., job engagement and satisfaction; worker and team performance), and psychological outcomes (e.g., depression, anxiety, compassion satisfaction, and flourishing). Results: Leader and team humility contributed to multilevel positive attributions about leaders (as effective and impactful), teams (as cohesive, psychologically safe, and growth-minded), and oneself (as humble), and those attributions contributed to organizational and psychological outcomes. Teams' shared attributions of their leader's humility contributed to higher worker job satisfaction and team performance. Longitudinally, for workers and leaders, leader and team humility were associated with some positive organizational and psychological outcomes over time. Conclusion: In humanitarian organizations, leader humility seems to act as an attributional and motivational social contagion that affects aid personnel's positive attributions about their leaders, teams, and themselves. In turn, these multilevel positive attributions contribute to several positive team, organizational, and psychological outcomes among workers and leaders.

AFP peptide (AFPep) as a potential growth factor for prostate cancer.

Prostate cancer is the most common cancer among men in the USA. A peptide derived from the active site of alpha-fetoprotein (AFP), known as AFPep, has been shown to be efficacious in inhibiting breast cancer growth. The role of this derived peptide AFPep in the development of prostate cancer has yet to be studied. To investigate the role of AFPep on prostate cancer, we used the PC-3 and DU-145 cell lines. We found that through key anti-apoptosis and pro-proliferation molecules, AFPep enhances the proliferation of DU-145 prostate cancer cells. The anti-proliferative molecules p18, p21, and p27, along with the pro-apoptotic molecules Fas and Bax, were all down-regulated in DU-145 cell lines treated with AFPep. Conversely, AFPep was not found to have a proliferative effect on the PC-3 prostate cancer cell line. This finding suggests the effects of AFPep to be cell line-specific in prostate cancer. Further investigation into the effects of AFPep could lead to new areas of treating prostate cancer.

Biopsychosocial and Spiritual Implications of Patients With COVID-19 Dying in Isolation.

Critically ill patients with the Coronavirus disease 2019 (COVID-19) are dying in isolation without the comfort of their family or other social support in unprecedented numbers. Recently, healthcare teams at COVID-19 epicenters have been inundated with critically ill patients. Patients isolated for COVID-19 have had no contact with their family or loved ones and may have likely experienced death without closure. This situation highlights concerns about patients' psychological and spiritual well-being with COVID-19 and their families, as they permanently part ways. While palliative care has advanced to adequately address these patients' needs, the COVID-19 pandemic presents several barriers that force healthcare teams to deprioritize these essential aspects of patient care. The severe acute respiratory syndrome (SARS) outbreak in 2003 gave us a glimpse of these challenges as these patients were also isolated in hospitals. Here, we discuss the importance of the biopsychosocial spiritual model in end-of-life care and its implications on patients dying with COVID-19. Furthermore, we outline an integrative approach to address the unique and holistic needs of critically ill patients dying with COVID-19. These include intentional and increased coordination with trained palliative care staff, early and frequent goals of care including discussion of end-of-life plans, broader use of technology to improve connectedness, and shared decision making with patients' families.

Peripherally inserted central catheter placement using the Sonic Flashlight.

The Sonic Flashlight is an ultrasound (US) device that projects real-time US images into patients with use of a semireflective/transparent mirror. The present study evaluated the feasibility of use of the Sonic Flashlight for clinical peripherally inserted central catheter placements, originally with the mirror located inside a sterile cover (n = 15), then with the mirror outside (n = 11). Successful access was obtained in all cases. Results show that this new design improved visibility, as judged subjectively firsthand and in photographs. The study demonstrated the feasibility of the Sonic Flashlight and the new design to help assure sterility without degrading visibility, allowing further clinical trials involving physicians and nurses.

Comprehensive Imaging and Surgical Review of Urinary Diversions: What the Radiologist Needs to Know.

Urinary diversions are surgical procedures that reconstruct the lower urinary tract following cystectomy. The 2 common surgical approaches are based on the continence status of the urinary tract. Incontinent diversions have continuous urine drainage through a cutaneous stoma, whereas continent diversions offer the patient the ability to self-void either via stoma catheterization or with the patient's own urethra. Given the large number of diversion procedures available, postsurgical anatomy may be complex. Multiple imaging modalities can be used to assess the postprocedural anatomy, potential complications, and for on-going oncologic monitoring. The purpose of this review is to describe the common surgical techniques and associated complications.