Vascular endothelial profilin-1 drives a protumorigenic tumor microenvironment and tumor progression in renal cancer.

Overexpression of actin-binding protein profilin-1 (Pfn1) correlates with advanced disease features and adverse clinical outcome of patients with clear cell renal carcinoma, the most prevalent form of renal cancer. We previously reported that Pfn1 is predominantly overexpressed in tumor-associated vascular endothelial cells in human clear cell renal carcinoma. In this study, we combined in vivo strategies involving endothelial cell-specific depletion and overexpression of Pfn1 to demonstrate a role of vascular endothelial Pfn1 in promoting tumorigenicity and enabling progressive growth and metastasis of renal carcinoma cells in a syngeneic orthotopic mouse model of kidney cancer. We established an important role of endothelial Pfn1 in tumor angiogenesis and further identified endothelial Pfn1-dependent regulation of several pro- (VEGF, SERPINE1, CCL2) and anti-angiogenic factors (platelet factor 4) in vivo. Endothelial Pfn1 overexpression increases tumor infiltration by macrophages and concomitantly diminishes tumor infiltration by T cells including CD8+ T cells in vivo, correlating with the pattern of endothelial Pfn1-dependent changes in tumor abundance of several prominent immunomodulatory cytokines. These data were also corroborated by multiplexed quantitative immunohistochemistry and immune deconvolution analyses of RNA-seq data of clinical samples. Guided by Upstream Regulator Analysis of tumor transcriptome data, we further established endothelial Pfn1-induced Hif1α elevation and suppression of STAT1 activation. In conclusion, this study demonstrates for the first time a direct causal relationship between vascular endothelial Pfn1 dysregulation, immunosuppressive tumor microenvironment, and disease progression with mechanistic insights in kidney cancer. Our study also provides a conceptual basis for targeting Pfn1 for therapeutic benefit in kidney cancer.

The role of profilin-1 in cardiovascular diseases.

Dynamic remodeling of the actin cytoskeleton is an essential feature for virtually all actin-dependent cellular processes, including cell migration, cell cycle progression, chromatin remodeling and gene expression, and even the DNA damage response. An altered actin cytoskeleton is a structural hallmark associated with numerous pathologies ranging from cardiovascular diseases to immune disorders, neurological diseases and cancer. The actin cytoskeleton in cells is regulated through the orchestrated actions of a myriad of actin-binding proteins. In this Review, we provide a brief overview of the structure and functions of the actin-monomer-binding protein profilin-1 (Pfn1) and then discuss how dysregulated expression of Pfn1 contributes to diseases associated with the cardiovascular system.

Using smart phone technology to improve daily living skills for individuals with intellectual disabilities.

BACKGROUND: Individuals with intellectual disabilities need continued supports in completing daily living tasks to increase the likelihood of achieving independence. Fortunately, research has shown that assistive technology, and particularly video prompting helps support independent living for individuals with intellectual disabilities. AIMS: This study investigated the efficacy of a highly customizable task analysis smartphone application in assisting three young adults with intellectual disabilities learn how to cook three different multistep recipes. MATERIALS & METHODS: Three young adults with intellectual disabilities enrolled in a four-year postsecondary education program (PSE) participated in a multiple probe design across participants to examine the effect of a Task Analysis app on the participants' completion of three cooking tasks. RESULTS: In this present study, the use of video prompting to teach a daily living skill resulted in large and meaningful effect size gains of 99%-100% for all three participants, as measured by Tau-U. DISCUSSION: Video prompting is an effective instructional strategy which allows the user to self-prompt and manage their ability to successfully complete daily living skills. In this current study, video prompting made a substantial difference in the safety of participants. CONCLUSION: The use of video prompting can decrease the reliance on others (e.g., teachers and caregivers), improve self-confidence of the user, and improve the user's level of autonomy.

Disruption of profilin1 function suppresses developmental and pathological retinal neovascularization.

Angiogenesis-mediated neovascularization in the eye is usually associated with visual complications. Pathological angiogenesis is particularly prominent in the retina in the settings of proliferative diabetic retinopathy, in which it can lead to permanent loss of vision. In this study, by bioinformatics analyses, we provide evidence for elevated expression of actin-binding protein PFN1 (profilin1) in the retinal vascular endothelial cells (VECs) of individuals with proliferative diabetic retinopathy, findings further supported by gene expression analyses for PFN1 in experimentally induced abnormal retinal neovascularization in an oxygen-induced retinopathy murine model. We observed that in a conditional knockout mouse model, postnatal deletion of the Pfn1 gene in VECs leads to defects in tip cell activity (marked by impaired filopodial protrusions) and reduced vascular sprouting, resulting in hypovascularization during developmental angiogenesis in the retina. Consistent with these findings, an investigative small molecule compound targeting the PFN1-actin interaction reduced random motility, proliferation, and cord morphogenesis of retinal VECs in vitro and experimentally induced abnormal retinal neovascularization in vivo In summary, these findings provide the first direct in vivo evidence that PFN1 is required for formation of actin-based protrusive structures and developmental angiogenesis in the retina. The proof of concept of susceptibility of abnormal angiogenesis to small molecule intervention of PFN1-actin interaction reported here lays a conceptual foundation for targeting PFN1 as a possible strategy in angiogenesis-dependent retinal diseases.

Actin-binding protein profilin1 promotes aggressiveness of clear-cell renal cell carcinoma cells.

Clear-cell renal cell carcinoma (ccRCC), the most common subtype of renal cancer, has a poor clinical outcome. A hallmark of ccRCC is genetic loss-of-function of VHL (von Hippel-Lindau) that leads to a highly vascularized tumor microenvironment. Although many ccRCC patients initially respond to antiangiogenic therapies, virtually all develop progressive, drug-refractory disease. Given the role of dysregulated expressions of cytoskeletal and cytoskeleton-regulatory proteins in tumor progression, we performed analyses of The Cancer Genome Atlas (TCGA) transcriptome data for different classes of actin-binding proteins to demonstrate that increased mRNA expression of profilin1 (Pfn1), Arp3, cofilin1, Ena/VASP, and CapZ, is an indicator of poor prognosis in ccRCC. Focusing further on Pfn1, we performed immunohistochemistry-based classification of Pfn1 staining in tissue microarrays, which indicated Pfn1 positivity in both tumor and stromal cells; however, the vast majority of ccRCC tumors tend to be Pfn1-positive selectively in stromal cells only. This finding is further supported by evidence for dramatic transcriptional up-regulation of Pfn1 in tumor-associated vascular endothelial cells in the clinical specimens of ccRCC. In vitro studies support the importance of Pfn1 in proliferation and migration of RCC cells and in soluble Pfn1's involvement in vascular endothelial cell tumor cell cross-talk. Furthermore, proof-of-concept studies demonstrate that treatment with a novel computationally designed Pfn1-actin interaction inhibitor identified herein reduces proliferation and migration of RCC cells in vitro and RCC tumor growth in vivo Based on these findings, we propose a potentiating role for Pfn1 in promoting tumor cell aggressiveness in the setting of ccRCC.

Perioperative pain protocols following surgery for adolescent idiopathic scoliosis: a snapshot of current treatments utilized by attending orthopedic surgeons.

PURPOSE: Perioperative management after adolescent idiopathic scoliosis (AIS) surgery varies extensively between surgeons and institutions. We devised a questionnaire to assess surgeon baseline characteristics, practice settings, and pain regimens to assess what factors contribute to perioperative pain protocols. METHODS: A multiple-choice questionnaire including 130 independent variables regarding baseline characteristics, practice environments, and pain regimen protocols was distributed to elicit information among surgeons performing AIS fusion surgery. Pairwise bivariate analysis between practice location, length of practice, and practice environment vs. type of post-operative analgesia was completed using two-tailed Fisher's exact test. RESULTS: 85 respondents participated, all identified as practicing orthopedic surgeons. The largest group of respondents reported 20-40% of their total practice was dedicated to AIS (36%). Respondents were predominantly hospital-employed academic physicians (67%). The most common pain medication administered preoperatively was gabapentin (54%). Postoperative regimens were highly varied. Discharge pain regimens most commonly included short-acting opiates (89%), acetaminophen (86%), antispasmodics (59%), and NSAIDs (51%). Bivariate analysis revealed that fentanyl PCA was significantly associated with practice location (p < 0.05). Utilization of NSAIDs was significantly associated with length in training, with older physicians utilizing anti-inflammatories more regularly than younger physicians (p < 0.05). CONCLUSION: This study identifies common perioperative regimens utilized in AIS surgery. Of interest, younger surgeons are less likely to prescribe NSAIDs post-operatively than surgeons who have been in practice for longer periods of time, which may represent a bias against anti-inflammatory medications in younger surgeons.

Epidemiology of pediatric trauma and fractures during and beyond the COVID-19 pandemic.

Purpose: Previous literature has shown decreases in pediatric trauma during the COVID-19 outbreak, but few have analyzed beyond the peak of the pandemic. This study assesses the epidemiology of pediatric trauma cases in a high-volume teaching hospital in New York City before, during, and after the height of the COVID-19 pandemic. Methods: Institutional data on pediatric trauma orthopedic cases from January 1, 2018 to November 30, 2021 were extracted. The following time frames were studied: (1) April 1-June 22 in 2018 and 2019 (pre-pandemic), (2) April 1-June 22, 2020 (peak pandemic), and (3) April 1-June 22, 2021 (post-peak pandemic). Inferential statistics were used to compare patient and trauma characteristics. Results: Compared to the pre-pandemic cohort (n = 6770), the peak pandemic cohort (n = 828) had a greater proportion of fractures (p < 0.01) and had a significantly decreased overall traumas per week rate (p < 0.01) and fractures per week rate (p < 0.01). These decreased trauma (p < 0.01) and fracture rates (p < 0.01) persisted for the post-peak pandemic cohort (n = 2509). Spatial analysis identified zip code clusters throughout New York City with higher rates of emergency department presentation during the peak pandemic compared to pre-pandemic, and these areas aligned with lower-income neighborhoods. Conclusion: During the peak of the pandemic, overall trauma and fracture volumes decreased, the types of prevalent injuries changed, and neighborhoods of different economic resources were variably impacted. These trends have mostly persisted for 12 months post-peak pandemic. This longitudinal analysis helps inform and improve long-term critical care and public health resource allocation for the future. Level of evidence: Level III.

Risk factors for 90-day readmission and prolonged length of stay after hip surgery in children with cerebral palsy.

Objective: Spastic hip dysplasia is a common complication of cerebral palsy in children, and surgical intervention is usually warranted. While current literature has primarily analyzed single institution outcomes, this study utilized a national database to describe readmission rates and factors correlated with readmission for children with cerebral palsy undergoing hip surgery in order to treat this population more effectively. Methods: This study queried the Nationwide Readmissions Database (2014-2018) for pediatric patients with cerebral palsy who underwent hip surgery. Patient demographics, pre-operative comorbidities, length of stay (LOS), treatment complications, and readmission data were collected for each patient and analyzed with inferential statistics. Results: Of the 1225 patients included, the average age was 9.3 ± 3.8 years and 42.8% were female. Approximately 26.3% patients had a prolonged LOS (≥5 days) and 14.2% patients required readmission within 90-days of surgery. Medical complications, cardiac arrhythmias, and iron deficiency anemia were all significantly associated with elongated LOS as well as 90-day readmission. Patients with Medicaid were more frequently associated with an inpatient medical complication and the overall complication rate was 5.5%. Conclusions: While current literature has analyzed common risk factors and complications associated with hip surgery in the pediatric cerebral palsy patient, this study identifies a national readmission rate (14.2%) as well as preoperative comorbidities associated with readmission within 90-days and/or elongated LOS. Notably, complications are more frequently associated with patients using Medicaid. These results further exemplify the importance of equitable access to care and thorough selection of pediatric cerebral palsy patients appropriate for hip surgery.

Building better pediatric surgeons: A sentiment analysis of online physician review websites.

Purpose: Physician review websites are a heavily utilized patient tool for finding, rating, and reviewing surgeons. Natural language processing such as sentiment analysis provides a comprehensive approach to better understand the nuances of patient perception. This study utilizes sentiment analysis to examine how specific patient sentiments correspond to positive and negative experiences in online reviews of pediatric orthopedic surgeons. Methods: The online written reviews and star ratings of pediatric surgeons belonging to the Pediatric Orthopaedic Society of North America were obtained from healthgrades.com. A sentiment analysis package obtained compound scores of each surgeon's reviews. Inferential statistics analyzed relationships between demographic variables and star/sentiment scores. Word frequency analyses and multiple logistic regression analyses were performed on key terms. Results: A total of 749 pediatric surgeons (3830 total online reviews) were included. 80.8% were males and 33.8% were below 50 years of age. Male surgeons and younger surgeons had higher mean star ratings. Surgeon attributes including "confident" (p < 0.01) and "comfortable" (p < 0.01) improved the odds of positive reviews, while "rude" (p < 0.01) and "unprofessional" (p < 0.01) decreased these odds. Comments regarding "pain" lowered the odds of positive reviews (p < 0.01), whereas "pain-free" increased these odds (p < 0.01). Conclusion: Pediatric surgeons who were younger, communicated effectively, eased pain, and curated a welcoming office setting were more likely to receive positively written online reviews. This suggests that a spectrum of interpersonal and ancillary factors impact patient experience and perceptions beyond surgical skill. These outcomes can advise pediatric surgeons on behavioral and office qualities that patients and families prioritize when rating/recommending surgeons online. Level of evidence: IV.

Substance use during COVID-19 pandemic: impact on the underserved communities.

The number of overdose deaths are on the rise all over the world. An estimate of 93,000 drug overdose deaths have been estimated in the United States in 2020. COVID-19 pandemic has exacerbated the drug crisis. Factors, such as existing health disparities among underserved communities, lack of resources for people of color, lack of belief in available resources, social isolation and economic burden, limited access to treatment, regulatory barriers in telehealth, and stress from the on-going COVID-19 pandemic have been identified as some of the key factors behind the acute health effects of people with substance use disorder. These interrelated factors exacerbate the impact of already existing disparities in the underserved communities. Policy and regulatory changes around telehealth and access of treatment for substance use disorder are warranted. Evidence-based strategies and other safer drug practices should be implemented to mitigate the impact on human health. Investment in programs that increase access to treatment, will be useful for potential future pandemics, where increasing mental health services and overall access to healthcare in disadvantaged communities would lessen the disparities in physical and mental ailments. In this review, we are evaluating and summarizing the acute health effects of the ongoing COVID-19 pandemic on individuals with substance use disorder.

The Role of the Orthopaedic Surgeon in the Identification and Management of Nonaccidental Trauma.

Nonaccidental trauma (NAT) has short- and long-term effects on the patient. The importance of identification and management of injuries related to NAT can be vital to the disruption of patterns of abuse and can ultimately save lives. The purpose of this review is to educate the orthopaedic surgeon as the frontline provider in the treatment of these patients. Understanding the risk factors for abuse, sentinel signs on clinical examination, and questionable fracture patterns related to NAT enables providers to better identify and address children at risk of abuse or neglect. Equally as important, understanding pathologic states that can mimic NAT allows the orthopaedic surgeon the tools necessary to address all patients with multiple and/or unexplained fractures. Understanding the legal expectations and protections for practicing orthopaedic surgeons as mandated reporters and the importance of a multidisciplinary approach to intervention in these settings will improve patient experiences and outcomes.

Evidence that knock down of GSK-3ß in Chronic Myelogenous Leukemia cells augments IFN-γ-induced apoptosis.

The role of interferon-gamma (IFN-γ) in Chronic Myelogenous/Myeloid Leukemia (CML) and in the treatment of CML remains unclear; specifically, the effect of IFN-γ on apoptosis. There is reported interplay between IFN-γ and glycogen synthase kinase-3 (GSK-3), a kinase which has been implicated in both cell death and, conversely, cell survival. Thus, we utilized the CML-derived HAP1 cell line and a mutant HAP1 GSK-3ß knocked-down cell line (GSK-3ß 31bp) to investigate whether GSK-3 modulates IFN-γ's action on CML cells. Significantly less GSK-3ß 31bp cells, relative to HAP1 cells, were present after 48 h treatment with IFN-γ. IFN-γ treatment significantly decreased GSK-3ß 31bp substrate adhesiveness (relative to HAP1 cells); an observation often correlated with cell death. Fluorescence microscopy revealed that IFN-γ induces a modest level of apoptosis in the HAP1 cells and that IFN-γ induced apoptosis is significantly enhanced in GSK-3ß 31bp cells. Utilizing a complementary GSK-3ß knocked-down cell line (8bp) we found, via flow cytometric analysis, that IFN-γ induced apoptosis is significantly enhanced in GSK-3ß 8bp cells relative to HAP1 cells. Combined, our findings suggest that IFN-γ induces apoptosis of CML cells and that loss of GSK-3ß significantly augments IFN-γ-induced apoptosis.

Inhibiting Autophagy in Renal Cell Cancer and the Associated Tumor Endothelium.

The clear cell subtype of kidney cancer encompasses most renal cell carcinoma cases and is associated with the loss of von Hippel-Lindau gene function or expression. Subsequent loss or mutation of the other allele influences cellular stress responses involving nutrient and hypoxia sensing. Autophagy is an important regulatory process promoting the disposal of unnecessary or degraded cellular components, tightly linked to almost all cellular processes. Organelles and proteins that become damaged or that are no longer needed in the cell are sequestered and digested in autophagosomes upon fusing with lysosomes, or alternatively, released via vesicular exocytosis. Tumor development tends to disrupt the regulation of the balance between this process and apoptosis, permitting prolonged cell survival and increased replication. Completed trials of autophagic inhibitors using hydroxychloroquine in combination with other anticancer agents including rapalogues and high-dose interleukin 2 have now been reported. The complex nature of autophagy and the unique biology of clear cell renal cell carcinoma warrant further understanding to better develop the next generation of relevant anticancer agents.

Surgical management of Gorham disease involving the upper cervical spine with occipito-cervical-thoracic fusion: a case report.

BACKGROUND CONTEXT: Gorham disease is a rare osteolytic condition, with only 17 reported cases primarily affecting the cervical spine. PURPOSE: We report the case of a 31-year-old woman with severe kyphotic instability and myelopathy of the upper cervical spine secondary to massive osteolysis of the posterior elements. The surgical management, clinical outcome, and review of the relevant literature are discussed. STUDY DESIGN: This is a case report and a literature review. PATIENT SAMPLE: This is a report of one patient with Gorham disease of the cervical spine. METHODS: We report the case of a 31-year-old woman with Gorham disease affecting the upper cervical spine. Combined posterior and anterior stabilization and fusion was performed from the occiput to the thoracic spine. RESULTS: Six-year follow-up with annual computed tomography imaging showed solid fusion from the occiput to T2. No spread or local recurrence has developed to date. CONCLUSIONS: Correction of severe instability with myelopathy due to Gorham disease of the upper cervical spine was achieved through posterior and anterior occipito-cervical-thoracic fusion.

Patient-Reported Outcome Instruments in Pediatric Deformity Surgery: A Bibliometric Analysis.

STUDY DESIGN: Bibliometric analysis. OBJECTIVES: To identify patient-reported outcomes instruments (PROIs) used in pediatric deformity surgery research over the past decade and their frequency and usage trends. SUMMARY OF BACKGROUND DATA: The emphasis on PROIs is increasing along with the demand for evidence-based medicine and cost-effectiveness research. Therefore, investigators and PROI consensus writers should be aware of the PROIs used in pediatric deformity and usage trends. METHODS: Five top orthopedics journals were reviewed from 2004 to 2013 for clinical studies of surgical intervention in pediatric deformity that report PROIs. Publication year, level of evidence (LOE), and PROIs were reported for each article. Mean and range scores for the most frequently used PROIs were analyzed at 2-year follow-up. RESULTS: A total of 79 studies using PROIs were published in the pediatric deformity literature over the period studied. The researchers identified 21 named PROIs and 6 additional custom questionnaires. The Scoliosis Research Society (SRS)-22 was the most frequently used instrument (32.9%), followed by the SRS-24 (29.1%), Oswestry disability index (17.7%), visual analog scale (12.7%), SRS-30 (10.1%), and Short Form-36 (6.3%). Level of evidence III was most common (39.2%) and 1 LOE I study was identified. Mean preoperative and postoperative SRS instrument scores were 4.0 (95% confidence interval, 3.8-4.1) and 4.5 (95% confidence interval, 4.4-4.6), respectively, in SRS-22r equivalents. No studies met the criteria for mean and range calculation for the other top instruments. CONCLUSIONS: Scoliosis Research Society instruments are used in 74.7% of pediatric deformity studies reporting PROIs. Therefore, there is a consensus that SRS instruments should be used in pediatric deformity outcome studies; yet, consistent use of the most up-to-date version, the SRS-22r, is still needed. General health questionnaires are currently underused in pediatric deformity research. Version reporting and use of the latest versions of PROIs need to be improved in future studies.