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Many solid tumors present with perineural invasion (PNI), and innervation correlates with worsened prognosis. The effects that commonly administered therapies such as radiation therapy (RT) have on PNI status remain unknown. We investigated the contribution of RT on the nervous system and elucidated the implications that increased nerve signaling can have on tumor burden using our previously developed orthotopic murine model of rectal cancer (RC) and our targeted and clinically relevant short-course RT (SCRT) regimen.Medical charts for patients with RC treated at the Wilmot Cancer Institute were obtained and PNI status was analyzed. Human data were accompanied by an orthotopic murine model of RC. Briefly, luciferase-expressing murine colon-38 (MC38-luc) tumor cells were injected orthotopically into the rectal wall of C57BL6 mice. Targeted SCRT (5 gray (Gy) per fraction for 5 consecutive fractions) was administered to the tumor. Intratumoral innervation was determined by immunohistochemistry (IHC), local norepinephrine (NE) concentration was quantified by enzyme-linked immunosorbent assay (ELISA), and β2-adrenergic receptor (B2AR) expression was assessed by flow cytometry. Chronic NE signaling was mirrored by daily isoproterenol treatment, and the effect on tumor burden was determined by overall survival, presence of metastatic lesions, and tumor size. Isoproterenol signaling was inhibited by administration of propranolol.Human RC patients with PNI have decreased overall survival compared with patients without PNI. In our mouse model, SCRT induced the expression of genes involved in neurogenesis, increased local NE secretion, and upregulated B2AR expression. Treating mice with isoproterenol resulted in decreased overall survival, increased rate of metastasis, and reduced SCRT efficacy. Interestingly, the isoproterenol-induced decrease in SCRT efficacy could be abrogated by blocking the BAR through the use of propranolol, suggesting a direct role of BAR stimulation on impairing SCRT responses.Our results indicate that while SCRT is a valuable treatment, it is accompanied by adverse effects on the nervous system that may impede the efficacy of therapy and promote tumor burden. Therefore, we could speculate that therapies aimed at targeting this signaling cascade or impairing nerve growth in combination with SCRT may prove beneficial in future cancer treatment.

Background In predynamic or dynamic scapholunate (SL) instability, standard diagnostic imaging may not identify SL interosseous ligament (SLIL) injury, leading to delayed detection and intervention. This study describes the use of four-dimensional computed tomography (4DCT) in identifying early SLIL injury and following injured wrists to 1-year postoperatively. Description of Technique 4DCT acquires a series of three-dimensional volume data with high temporal resolution (66 ms). 4DCT-derived arthrokinematic data can be used as biomarkers of ligament integrity. Patients and Methods This study presents the use of 4DCT in a two-participant case series to assess changes in arthrokinematics following unilateral SLIL injury preoperatively and 1-year postoperatively. Patients were treated with volar ligament repair with volar capsulodesis and arthroscopic dorsal capsulodesis. Arthrokinematics were compared between uninjured, preoperative injured, and postoperative injured (repaired) wrists. Results 4DCT detected changes in interosseous distances during flexion-extension and radioulnar deviation. Generally, radioscaphoid joint distances were greatest in the uninjured wrist during flexion-extension and radioulnar deviation, and SL interval distances were smallest in the uninjured wrist during flexion-extension and radioulnar deviation. Conclusion 4DCT provides insight into carpal arthrokinematics during motion. Distances between the radioscaphoid joint and SL interval can be displayed as proximity maps or as simplified descriptive statistics to facilitate comparisons between wrists and time points. These data offer insight into areas of concern for decreased interosseous distance and increased intercarpal diastasis. This method may allow surgeons to assess whether (1) injury can be visualized during motion, (2) surgery repaired the injury, and (3) surgery restored normal carpal motion. Level of Evidence Level IV, Case series.

To assess the association of common sleep disorders with female sexual dysfunction.We conducted a cross-sectional analysis using the TriNetX Diamond Network. Adult women diagnosed with insomnia, obstructive sleep apnea, or circadian rhythm sleep disorder were assessed to determine the association with female sexual dysfunction, hypoactive sexual desire disorder, female orgasmic disorder, and female sexual arousal disorder. A propensity-score matched control cohort for age, race, ethnicity, tobacco use, outpatient service utilization, obesity, hyperlipidemia, diabetes mellitus, hypertensive disease, ischemic heart disease, and surgical procedures on the female genital system was generated, excluding those with any sleep disorders, sleep deprivation, or morbid obesity with alveolar hypoventilation. All cohorts excluded those with any antidepressant or antipsychotic prescriptions.Our query yielded 1,317,491 women diagnosed with sleep apnea, 1,538,567 with insomnia, and 58,902 with circadian rhythm sleep disorder. Women with sleep apnea and insomnia had higher odds of hypoactive sexual desire disorder, female sexual arousal disorder, and female orgasmic disorder compared to matched controls. Women with sleep apnea, insomnia, or circadian rhythm sleep disorder all had a significantly higher odds of female sexual dysfunction compared to matched controls.In this large cross-sectional analysis, sleep disorders were strongly associated with female sexual dysfunction. It is thus crucial for providers to screen for poor sleep when conducting a thorough evaluation for sexual dysfunction in women to diagnose highly prevalent sleep disorders and improve overall health.

OBJECTIVE. The accessory sacroiliac joint (ASIJ) is the most common sacroiliac joint anatomical variant; however, its literature-reported prevalence is inconsistent. Previous CT-based studies of the ASIJ have used thick axial slices, which may not adequately detail ASIJ anatomy. The aims of this study are to (1) evaluate ASIJ prevalence and radiographic features in a large age- and sex-balanced cohort using thin-section CT and (2) determine associations between ASIJ anatomy, patient features, and treatment strategies. MATERIALS AND METHODS. Thin-section CTs (0.75 to 2.00 mm) of the pelvis from 800 patients were reviewed by two musculoskeletal radiologists. Degree of degenerative change and ankylosis at ASIJs were detailed. The EMR was used to capture demographics, lower back or sacroiliac joint symptoms, and treatments. RESULTS. The ASIJ was present in 25.8% of patients and bilateral in 53.3% of those with any ASIJ. ASIJs were more common at the S2 than S1 neural foramen level (75.7% and 27.2%). There was a statistically significant difference between age and presence of any ASIJ anatomy (mean (SD) 69.0 (19.8) with ASIJ versus 55.9 (22.1) years without ASIJ). Degenerative changes and ankylosis were found in 93.5% and 20.3% of ASIJs, respectively. There was a higher odds ratio of having received a sacroiliac joint corticosteroid injection in those with ASIJ anatomy. CONCLUSION. Radiologists should be familiar with the ASIJ and consider its age-related association, propensity to show ASIJ degenerative change, and ability to serve as a potential pain generator. Steroid injections may be considered for diagnostic and therapeutic purposes.

Recent research has suggested that mental health is not only the absence of mental illness but includes aspects of well-being. One common psychological assessment used to assess dimensions of well-being is the Mental Health Continuum-Short Form (MHC-SF). The MHC-SF is a 14-item measure that assesses emotional, psychological, and social well-being. The purpose of this study was to assess measurement invariance of the MHC-SF across US military veterans and civilians.First, we examined the factor structure of the MHC-SF separately for veterans (n = 418) and civilians (n = 411). We then conducted multiple group confirmatory factor analysis (MG-CFA) to assess measurement invariance for the two groups.Findings suggested there were three latent factors representing emotional, social, and psychological well-being. Results from MG-CFA suggested that the MHC-SF is invariant across veterans and civilians.Researchers and practitioners can administer the MHC-SF with both groups.

Cognitive impairment is common among individuals with Parkinson's disease (PD). Intraindividual variability (IIV) is a measure of variability across multiple tasks of cognitive functioning. Due to the limited amount of research, particularly among individuals with PD, IIV has been an underutilized metric of cognitive functioning both in research and clinical practice. Previous research demonstrated that individuals with PD have greater variability in cognitive performance relative to controls, and that IIV is predictive of future cognitive impairments. The aim of this study is to investigate the association between baseline IIV and change in cortical and subcortical volumes among individuals with PD.The present study used data from 80 newly diagnosed PD patients who were part of a longitudinal cohort study (Parkinson progression marker initiative [PPMI]). Participants completed neuropsychological measures and underwent T1 structural magnetic resonance imaging (MRI) at baseline and the first annual follow-up. Neuropsychological tests assessed attention, processing speed, visuospatial functioning, verbal fluency learning, and memory. T1 scans were processed using standard Freesurfer protocols for extraction of regional volumes.Greater IIV at baseline was predictive of change in cortical volume in posterior temporal/parietal regions over the 1-year period. Baseline IIV predicted cortical volume changes above and beyond the main effect of motor severity and the baseline statistical mean/global cognition score.Our results provide initial evidence that IIV is a marker of longitudinal cortical volume loss. Evidence is building that IIV is a sensitive marker of cognitive impairment and the underlying neurodegeneration among individuals with PD. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

This qualitative study investigated creative adolescent perceptions of their educational and mental health experiences during the coronavirus disease 2019 (COVID-19) pandemic. Participants were 25 English-speaking adolescents from the Midwest in the United States. They were identified as creative by their teachers according to known creative profiles. Participants attended an all-day creative career workshop in the Spring 2021 semester. The five focus groups guided by semi-structured interviews conducted for this study occurred during the workshop. This study was phenomenological in nature with constructivist and transformative paradigms, and transcripts were analyzed using reflexive thematic analysis by the first, second, and third authors. Creative adolescents' education during the pandemic was marked by classroom changes based on COVID-19 policies, experiences of disconnection, disengagement, and disappointment, as well as feelings of empathy and appreciation for their educators. Creative adolescents experienced mental health challenges related to adjustment issues, powerlessness and hopelessness, and isolation. Implications of results are discussed.

INTRODUCTION: To broadly disseminate 5 user-centered educational videos for patients with inflammatory bowel disease and their family and friends on social media. METHODS: Relevant social media users were iteratively identified based on their online behavior. For each video, 2 different accompanying texts were tested. RESULTS: We reached 4.2 million social media users of whom 320,302 watched at least 50% of the video. A short description resulted in higher view rates than posing an open-ended question. DISCUSSION: We showed the feasibility of large-scale dissemination of health-related educational videos through social media. Our findings can inform future online dissemination approaches of educational content.

The Proterodiplostomidae Dubois, 1936 is a family of digeneans within the superfamily Diplostomoidea Poirier, 1886. Members of the family are distributed mostly in the tropics and subtropics, primarily in crocodilians, although some species are known from other reptiles. Despite their broad geographical distribution, the knowledge of proterodiplostomid diversity remains limited, mostly because a number of potential host species and regions of the world have not been sufficiently explored for these parasites. In this study, we use morphological and molecular data to describe four new genera (Afroproterodiplostomum gen. nov., Dungalabatrema gen. nov., Australiadiplostomum gen. nov. and Nattererodiplostomum gen. nov.) and five new species (Afroproterodiplostomum ingwenyae sp. nov., Australiadiplostomum blairi sp. nov., Dungalabatrema kostadinovae sp. nov., Dungalabatrema snyderi sp. nov. and Pseudoneodiplostomum angustus sp. nov.) of proterodiplostomids from crocodilians in Australia, South Africa and South America. Nattererodiplostomum gen. nov. has been established upon re-evaluation of the status of Proterodiplostomum medusae (Dubois, 1936) from caimans in Brazil using combined morphological and molecular evidence. Only a few previous studies provided DNA sequence data of proterodiplostomids. We generated partial 28S rDNA and cytochrome c subunit (cox1) mtDNA for three previously undescribed proterodiplostomids collected from Crocodylus spp. in Australia and South Africa. The newly generated 28S sequences were used to examine phylogenetic affinities of these taxa. All three newly sequenced proterodiplostomid species appeared in the phylogenetic tree in a strongly supported monophyletic clade comprising exclusively parasites of Crocodylus.

Context Penile prosthesis is a durable and effective treatment for erectile dysfunction (ED). Even as other treatment options for ED have been brought to market, penile prosthetic surgery remains a mainstay for urologists treating ED. No systematic study has yet summarized the global trends in penile prosthetic surgery. Objective To systematically review studies of trends in penile prosthetic surgery to determine global movements in implantation rates, malleable versus inflatable prosthetic surgery, inpatient versus outpatient implantation surgery, proportion of men with ED undergoing penile prosthetic surgery, and prosthetic cost. Evidence acquisition A systematic review of MEDLINE, EMBASE, Cochrane Library, and ClinicalTrials.gov was performed for studies assessing trends in penile prosthetic surgeries and costs associated with penile prosthetic device and inclusive surgical costs. Evidence synthesis Twenty-seven studies were identified during the systematic review, comprising 447,204 penile prosthetic surgeries reported from 1988 to 2019. A trend analysis demonstrates that rates of penile prosthetic surgery declined dramatically in the late 1980s and early 1990s, but have demonstrated modest growth since the mid-2000s. Outpatient inflatable penile prosthetic surgery has strongly trended upward. Costs of penile prosthetic device have matched the rate of inflation, but inclusive surgical cost has radically outpaced inflation. Growth has mainly been seen in the USA, with a more modest global growth. Conclusions Penile prosthesis remains a viable option for the treatment of ED. Trends such as outpatient surgery and inflatable penile prosthesis placement may be driving the recent steady growth of penile prosthetic surgeries, but surging inclusive surgical cost may present a barrier for some patients without insurance coverage. Patient summary Penile prostheses continue to be an important treatment for erectile dysfunction. While the volume of penile prosthetic surgeries dropped when phosphidiesterase-5 inhibitors became available, prosthetic surgery is becoming more patient centric, as seen by increases in inflatable prosthetic placement and outpatient surgery.

Early-onset Alzheimer’s disease (EOAD) is a rare but particularly devastating form of AD. Though notable for its high degree of clinical heterogeneity, EOAD is defined by the same neuropathological hallmarks underlying the more common, late-onset form of AD. In this review, we describe the various clinical syndromes associated with EOAD, including the typical amnestic phenotype as well as atypical variants affecting visuospatial, language, executive, behavioral, and motor functions. We go on to highlight advances in fluid biomarker research and describe how molecular, structural, and functional neuroimaging can be used not only to improve EOAD diagnostic acumen but also enhance our understanding of fundamental pathobiological changes occurring years (and even decades) before the onset of symptoms. In addition, we discuss genetic variation underlying EOAD, including pathogenic variants responsible for the well-known mendelian forms of EOAD as well as variants that may increase risk for the much more common forms of EOAD that are either considered to be sporadic or lack a clear autosomal-dominant inheritance pattern. Intriguingly, specific pathogenic variants in PRNP and MAPT—genes which are more commonly associated with other neurodegenerative diseases—may provide unexpectedly important insights into the formation of AD tau pathology. Genetic analysis of the atypical clinical syndromes associated with EOAD will continue to be challenging given their rarity, but integration of fluid biomarker data, multimodal imaging, and various ‘omics techniques and their application to the study of large, multicenter cohorts will enable future discoveries of fundamental mechanisms underlying the development of EOAD and its varied clinical presentations.

Supplementary Figures S2 and S3 - PDF file 2332K, Figure S2:SG3 tumors are positive for beta-catenin and Myf5 Figure S3:Normal development of Myf5 myoblasts in beta-catenin knockout mice

Supplementary Figure S9 - PDF file 867K,Figure S9: The SYT-SSX2 domain SSXRD executes �-catenin activation and myogenic differentiation

Supplementary Figure S1 - PDF file 554K, Breeding strategy used to generate beta-catenin-null SYT-SSX2 transgenic mice

Supplementary Figure S8 - PDF file 1084K, Figure S8: Effects of SSTC-104 on Wnt-regulated tissues

Supplementary Figures S4, S5, S6, S7 - PDF file 1832K, Figure S4:Pyrvinium effect on beta-catenin in SYT-SSX2 expressing C2C12 and synovial sarcoma cells Figure S5:Phosphorylated �-catenin and full-length APC in HS-SY-II and SYO-1 SS cells Figure S6:SSTC-104 effects on growth and beta-catenin in HS-SY-II synovial sarcoma cells Figure S7: LRP6 depletion in HS-SY-II synovial sarcoma cells induces �-catenin nuclear exit

Supplementary Figure S11 and Tables S1-S3 - PDF file 460K, Figure S11: Regulation of an embryonic Wnt-interactive network by SYT-SSX2 and in human synovial sarcomas Table S1: Differentially regulated Wnt components in mesenchymal precursor cells expressing SYT-SSX2 and in synovial sarcoma tumors Table S2: Differentially regulated Wnt targets in mesenchymal precursor cells expressing SYT-SSX2 and in synovial sarcoma tumors Table S3: The embryonic Wnt interactive network in mesenchymal precursor cells expressing SYTSSX2 and in synovial sarcoma tumors

Supplementary Figure S10 4882K, Figure S10a and S10b: Nuclear beta-catenin accumulation in human synovial sarcoma tumors

Supplementary Methods and Legends - PDF file 130K, Methods:Additional information on experimental techniques Legends: describing supplementary figures

Background: Radioscapholunate (RSL) fusion is performed for painful radiocarpal arthritis that is typically posttraumatic or inflammatory in nature. The purpose of this study was to determine whether patients with inflammatory conditions undergoing RSL fusion had comparable outcomes as those with posttraumatic arthritis and to determine factors impacting union. Methods: This was a retrospective review of all RSL fusions over a 25-year period. Demographic information, wrist range of motion, postoperative complications, and surgical technique, including presence or absence of resection of distal scaphoid pole, data were collected. Radiographs were examined for evidence of healing of the RSL fusion site as well as development of midcarpal arthritis. Results: Fifty-six patients underwent RSL fusion. Eight patients required revision of radiocarpal arthrodesis for painful nonunion. The fusion rate was 85.7%, and median 25th to 75th percentile (time to healing was 4.0 3.2-5.2) months. There were no differences in time to or rate of union by inflammatory arthropathy status. Grip strength increased significantly and pain decreased significantly with surgery. Union rate was 90% with the use of K-wires, screws, or staples, which was significantly greater than plates, fusion cups, or multiple implant types. The use of autologous bone graft significantly decreased the rate of nonunion and significantly decreased time to union by 3 months compared to allograft alone. Extension was significantly improved with scaphoid distal pole resection versus without distal scaphoid pole resection. Conclusions: Radioscapholunate (RSL) fusion is an effective treatment for radiocarpal arthritis but has high nonunion rates. Fixation with K-wires, screws, or staples and use of autograft confers improved union rate.

Alaria, Didelphodiplostomum and Pharyngostomoides are among genera of diplostomid digeneans known to parasitize mammalian definitive hosts. Despite numerous recent molecular phylogenetic studies of diplostomids, limited DNA sequence data is available from diplostomids parasitic in mammals. Herein, we provide the first 28S rDNA and cox1 mtDNA sequences from morphologically identified, adult specimens of Didelphodiplostomum and Pharyngostomoides. Newly generated 28S sequences were used to infer the phylogenetic interrelationships of these two genera among other major lineages of diplostomoideans. The phylogeny based on 28S and a review of morphology clearly suggests that Pharyngostomoides should be considered a junior synonym of Alaria, while Didelphodiplostomum should be considered a junior synonym of Tylodelphys. Pharyngostomoides procyonis (type species), Pharyngostomoides adenocephala and Pharyngostomoides dasyuri were transferred into Alaria as Alaria procyonis comb. nov., Alaria adenocephala comb. nov. and Alaria dasyuri comb. nov.; Didelphodiplostomum variabile (type species) and Didelphodiplostomum nunezae were transferred into Tylodelphys as Tylodelphys variabilis comb. nov. and Tylodelphys nunezae comb. nov. In addition, Alaria ovalis comb. nov. (formerly included in Pharyngostomoides) was restored and transferred into Alaria based on a morphological study of well-fixed, adult specimens and the comparison of cox1 DNA sequences among Alaria spp. The diplostomid genus Parallelorchis was restored based on review of morphology.

COVID orchitis (testicular pain) is reported in 10-15% of men with long COVID. We identified two siblings with COVID orchitis and hypothesized that genetic mutations are associated with susceptibility. Blood samples from five COVID-19 (+) men, three of whom had orchitis were evaluated by whole-exome-sequencing. A rare deletion on chromosome 7 was found in NACAD among the three men with orchitis. Interestingly, circulating ACE2 levels was decreased in men with COVID orchitis. This pilot study generated the hypothesis that men who develop COVID orchitis could have underlying genetic variants and altered levels in circulating ACE2 that may increase their risk.

We sought to compare testosterone formulations and determine the degree that hematocrit increases vary by testosterone therapy formulation. As head-to-head trials are rare, network meta-analysis of the contemporary studies is the only way to compare hematocrit changes by testosterone type, including topical gels and patches, injectables (both short-acting and long-acting) and oral tablets.We conducted a thorough search of listed publications in Scopus®, PubMed®, Embase®, Cochrane CENTRAL, and ClinicalTrials.gov. A total of 29 placebo-controlled randomized trials (3,393 men) met inclusion criteria for analysis of mean hematocrit change after testosterone therapy. Randomized controlled trial data for the following formulations of testosterone were pooled via network meta-analysis: gel, patch, oral testosterone undecanoate, intramuscular testosterone undecanoate, and intramuscular testosterone enanthate/cypionate.All types of testosterone therapies result in statistically significant increases in mean hematocrit when compared with placebo. Meta-analysis revealed all formulations, including gel (3.0%, 95% CI 1.8-4.3), oral testosterone undecanoate (4.3%, 0.7-8.0), patch (1.4%, 0.2-2.6), intramuscular testosterone enanthate/cypionate (4.0%, 2.9-5.1), and intramuscular testosterone undecanoate (1.6%, 0.3-3.0) result in statistically significant increases in mean hematocrit when compared with placebo. When comparing all formulations against one another, intramuscular testosterone cypionate/enanthate were associated with a significantly higher increase in mean hematocrit compared to patch, but no differences in hematocrit between other formulations were detected.All types of testosterone are associated with increased hematocrit; however, the clinical concern of this increase remains questionable, warranting future studies. This is the first network meta-analysis to quantify mean hematocrit change and compare formulations, given the absence of head-to-head trials.

Purpose: Stereotactic body radiotherapy (SBRT) is an emerging treatment modality for pancreatic ductal adenocarcinoma (PDAC), which can effectively prime cytotoxic T cells by inducing immunogenic tumor cell death in preclinical models. SBRT effects on human PDAC have yet to be thoroughly investigated; therefore, this study aimed to characterize immunomodulation in the human PDAC tumor microenvironment following therapy. Experimental Design: Tumor samples were obtained from patients with resectable PDAC. Radiotherapy was delivered a median of 7 days prior to surgical resection, and sections were analyzed by multiplex IHC (mIHC), RNA sequencing, and T-cell receptor sequencing (TCR-seq). Results: Analysis of SBRT-treated tumor tissue indicated reduced tumor cell density and increased immunogenic cell death relative to untreated controls. Radiotherapy promoted collagen deposition; however, vasculature was unaffected and spatial analyses lacked evidence of T-cell sequestration. Conversely, SBRT resulted in fewer tertiary lymphoid structures and failed to lessen or reprogram abundant immune suppressor populations. Higher percentages of PD-1+ T cells were observed following SBRT, and a subset of tumors displayed more clonal T-cell repertoires. Conclusions: These findings suggest that SBRT augmentation of antitumor immunogenicity may be dampened by an overabundance of refractory immunosuppressive populations, and support the continued development of SBRT/immunotherapy combination for human PDAC.

Supplementary Data from Modulation of the Human Pancreatic Ductal Adenocarcinoma Immune Microenvironment by Stereotactic Body Radiotherapy

Purpose:Stereotactic body radiotherapy (SBRT) is an emerging treatment modality for pancreatic ductal adenocarcinoma (PDAC), which can effectively prime cytotoxic T cells by inducing immunogenic tumor cell death in preclinical models. SBRT effects on human PDAC have yet to be thoroughly investigated; therefore, this study aimed to characterize immunomodulation in the human PDAC tumor microenvironment following therapy.Experimental Design:Tumor samples were obtained from patients with resectable PDAC. Radiotherapy was delivered a median of 7 days prior to surgical resection, and sections were analyzed by multiplex IHC (mIHC), RNA sequencing, and T-cell receptor sequencing (TCR-seq).Results:Analysis of SBRT-treated tumor tissue indicated reduced tumor cell density and increased immunogenic cell death relative to untreated controls. Radiotherapy promoted collagen deposition; however, vasculature was unaffected and spatial analyses lacked evidence of T-cell sequestration. Conversely, SBRT resulted in fewer tertiary lymphoid structures and failed to lessen or reprogram abundant immune suppressor populations. Higher percentages of PD-1+ T cells were observed following SBRT, and a subset of tumors displayed more clonal T-cell repertoires.Conclusions:These findings suggest that SBRT augmentation of antitumor immunogenicity may be dampened by an overabundance of refractory immunosuppressive populations, and support the continued development of SBRT/immunotherapy combination for human PDAC.

Tumor necrosis factor (TNF)-α is present in the microenvironment of human tumors, including malignant pleural effusion (MPE). Although the cytokine is produced in the pleural cavity by both tumor and host cells, its effects on MPE formation are unknown. In these studies, we sought to determine the role of TNF-α in the pathogenesis of MPE and to assess the therapeutic effects of its neutralization in a preclinical model. For this, MPEs were generated in immunocompetent mice using intrapleural injection of mouse lung adenocarcinoma cells. The roles of tumor- and host-derived TNF-α were assessed using combined experimentation with TNF-α gene–deficient mice and in vivo TNF-α neutralization. To expand the scope of preclinical data, TNF-α and vascular endothelial growth factor (VEGF) expression were determined in human cancer cell lines and human MPE. In the MPE model, TNF-α of host and tumor origin was present. TNF-α neutralization significantly limited tumor dissemination, effusion formation, vascular hyperpermeability, TNF-α and VEGF expression, and angiogenesis, thereby improving survival. In contrast, these variables were not different between TNF-α gene–sufficient and TNF-α gene–deficient mice. In mouse cancer cells, TNF-α functioned via nuclear factor-κB– and neutral sphingomyelinase–dependent pathways to induce TNF-α and VEGF, respectively. These results were recapitulated in human cancer cells, and a correlation was detected between TNF-α and VEGF content of human MPE. We conclude that tumor-derived TNF-α is important in the development of MPE in mice, and provide preclinical evidence supporting the efficacy of TNF-α blockade against malignant pleural disease. [Cancer Res 2007;67(20):9825–34]

Supplementary Figure Legend from Tumor Necrosis Factor-α Promotes Malignant Pleural Effusion

Figure S2. Deficiency in IL-5 does not affect infiltration of lungs with T and B lymphocytes and NK cells during initial stages of metastatic load of LLC cells.

Figure S1. Lung eosinophils (Eos), identified as CD45+/CD11c-/ F4/80+/siglec-F+ cells using flow cytometry are A) negative for CD68, B) express high levels of CCR3, and C) express low levels of Gr1 compared to the CD45+/Gr1hi neutrophils (PMN).

Supplementary Figure 1 from Tumor Necrosis Factor-α Promotes Malignant Pleural Effusion