IGF2BP3 as a Prognostic Biomarker in Well-Differentiated/Dedifferentiated Liposarcoma.

BACKGROUND: Although IGF2BP3 has been implicated in tumorigenesis and poor outcomes in multiple cancers, its role in soft-tissue sarcoma (STS) remains unknown. Preliminary data have suggested an association with IGF2BP3 expression among patients with well-differentiated/dedifferentiated liposarcoma (WD/DD LPS), a disease where molecular risk stratification is lacking. METHODS: We examined the survival associations of IGF2BP3 via univariate and multivariate Cox regression in three unique datasets: (1) the Cancer Genome Atlas (TCGA), (2) an in-house gene microarray, and (3) an in-house tissue microarray (TMA). A fourth dataset, representing an independent in-house TMA, was used for validation. RESULTS: Within the TCGA dataset, IGF2BP3 expression was a poor prognostic factor uniquely in DD LPS (OS 1.6 vs. 5.0 years, p = 0.009). Within the microarray dataset, IGF2BP3 expression in WD/DD LPS was associated with worse survival (OS 7.7 vs. 21.5 years, p = 0.02). IGF2BP3 protein expression also portended worse survival in WD/DD LPS (OS 3.7 vs. 13.8 years, p < 0.001), which was confirmed in our validation cohort (OS 2.7 vs. 14.9 years, p < 0.001). In the multivariate model, IGF2BP3 was an independent risk factor for OS, (HR 2.55, p = 0.034). CONCLUSION: IGF2BP3 is highly expressed in a subset of WD/DD LPS. Across independent datasets, IGF2BP3 is also a biomarker of disease progression and worse survival.

Proteomes of Micro- and Nanosized Carriers Engineered from Red Blood Cells.

Red blood cell (RBC)-derived systems offer a potential platform for delivery of biomedical cargos. Although the importance of specific proteins associated with the biodistribution and pharmacokinetics of these particles has been recognized, it remains to be explored whether some of the key transmembrane and cytoskeletal proteins responsible for immune-modulatory effects and mechanical integrity of the particles are retained. Herein, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and quantitative tandem mass tag mass spectrometry in conjunction with bioinformatics analysis, we have examined the proteomes of micro- and nanosized erythrocyte ghosts doped with indocyanine green and compared them with those of RBCs. We identified a total of 884 proteins in each set of RBCs, micro-, and nanosized particles, of which 8 and 45 proteins were expressed at significantly different relative abundances when comparing micro-sized particles vs RBCs and nanosized particles vs RBCs, respectively. We found greater differences in relative abundances of some mechano-modulatory proteins, such as band 3 and protein 4.2, and immunomodulatory proteins like CD44, CD47, and CD55 in nanosized particles as compared to RBCs. Our findings highlight that the methods utilized in fabricating RBC-based systems can induce substantial effects on their proteomes. Mass spectrometry data are available at ProteomeXchange with the identifier PXD038780.

Demographic and HIV Status Diversities as Mechanisms of Social Integration and Segregation Among Black Sexual and Gender Minorities Enrolled in a Community-Based Social Network Intervention.

Drawing on a social integration and intersectionality framework, this study advances a dynamic network understanding of the mechanisms that enable differential patterns of within-group social integration and segregation among Black sexual and gender minorities (BSGM). Specifically, in a cohort of BSGM (18-35 years of age, n = 340) participating in a community-based network intervention for HIV prevention, we examine how sexual, gender, age, and HIV status diversities contribute to friendship formation and maintenance patterns over the 12-month study enrollment period. We found attenuated social integration (or social activity) among non-gay-identified and older BSGM and evidence of social segregation (or homophily) on the basis sexual identity and age similarities. Accounting for the moderating effects of the intervention revealed that the attenuated integration of non-gay-identified and older BSGM were stronger for participants who received the peer leadership training and integration challenges were also found for transgender BSGM who received the peer leadership training. Meanwhile, BSGM living with HIV who received the peer leadership training were significantly more integrated than their counterparts in the control arm. These findings help us understand the complicated social fabric among BSGM and the dynamics that interventions for this community may have to contend with or alter.

Multi-window approach enables two-fold improvement in OCT axial resolution with strong side-lobe suppression and improved phase sensitivity.

A common processing approach for optical coherence tomography (OCT) uses a window function (e.g., Hann or rectangular window) for spectral shaping prior to calculating the Fourier transform. Here we build on a multi-window approach [Opt. Express8, 5267 (2017)10.1364/BOE.8.005267] that enables improved resolution while still suppressing side-lobe intensity. The shape of the window function defines the trade-off between main-lobe width (resolution) and side-lobe intensity. We have extended the approach to include the interferometric phase for phase-sensitive applications like vibrometry and Doppler OCT. Using the Hann window as a reference, we show that 11 Taylor windows are sufficient to achieve 50% improvement in axial resolution, -31 dB side-lobe intensity, and 20% improvement in phase sensitivity with low computational cost.

Social Media Communication and Network Correlates of HIV Infection and Transmission Risks Among Black Sexual Minority Men: Cross-sectional Digital Epidemiology Study.

BACKGROUND: In the United States, HIV disproportionately affects Black cisgender sexual minority men (BSMM). Although epidemiological and behavioral surveillance are integral to identifying BSMM at risk of HIV infection and transmission, overreliance on self-reported data, inability to observe social contexts, and neglect of populations with limited engagement in health care systems limits their effectiveness. Digital epidemiological approaches drawing on social media data offer an opportunity to overcome these limitations by passively observing in organic settings activities, beliefs, behaviors, and moods that indicate health risks but are otherwise challenging to capture. OBJECTIVE: The primary aim of this study was to determine whether features of Facebook communication and networks were associated with biological, behavioral, and psychological indicators of HIV infection and transmission risk. METHODS: Facebook and survey data were collected from BSMM aged 18 to 35 years living in Chicago (N=310). Participants' Facebook posts were characterized using 4 culturally tailored topic dictionaries related to aspects of HIV protection and risk among BSMM (sexual health; substance use; sex behavior; and ballroom culture, a salient subculture in lesbian, gay, bisexual, transgender, and queer communities of color). Social network methods were used to capture structural features of BSMM's Facebook friendships (centrality, brokerage, and local clustering) and Facebook group affiliations. Multivariable regressions revealed relationships between these Facebook features and 5 ground truth indicators of HIV infection and transmission risk (sexually transmitted infection incidence, condomless sex, sex drug use, biomedical prevention, and depression). RESULTS: Although analysis of participants' Facebook posts revealed that HIV-related topics occupied a small portion of the total messages posted by each participant, significant associations were found between the following HIV risk indicators and Facebook features: Condomless sex, including communication about sexual health (odds ratio [OR] 1.58, 95% CI 1.09-2.29), ballroom culture (OR 0.76, 95% CI 0.63-0.93), and friendship centrality (OR 0.69, 95% CI 0.52-0.92); Sex drug use, including communication about substance use (OR 1.81, 95% CI 1.17-2.79) and friendship centrality (OR 0.73, 95% CI 0.55-0.96) and brokerage (OR 0.71, 95% CI 0.51-0.99); Biomedical prevention, including communication about ballroom culture (OR 0.06, 95% CI 0.01-0.71); and Depression, including communication about sexual health (ß=-0.72, 95% CI -1.42 to -0.02), ballroom culture (ß=.80, 95% CI 0.27-1.34), friendship centrality (ß=-0.90, 95% CI -1.60 to -0.21), and Facebook group affiliations (ß=.84, 95% CI 0.25-1.43). Facebook features provided no significant explanatory value for sexually transmitted infection incidence. CONCLUSIONS: Finding innovative strategies to detect BSMM at risk of contracting or transmitting HIV is critical to eliminating HIV disparities in this community. The findings suggest that social media data enable passive observance of social and communicative contexts that would otherwise go undetected using traditional HIV surveillance methods. As such, social media data are promising complements to more traditional data sources.

Hypophosphatemia Due to Increased Effector Cell Metabolic Activity Is Associated with Neurotoxicity Symptoms in CD19-Targeted CAR T-cell Therapy.

A major complication of chimeric antigen receptor (CAR) T-cell therapy is immune effector cell-associated neurotoxicity syndrome (ICANS), which presents as aphasia, confusion, weakness, somnolence, seizures, and coma. This is similar to the neurologic manifestations of hypophosphatemia, which can result from sudden increases in metabolic demand for phosphorylated intermediates (e.g., refeeding syndrome and sepsis). Given these similarities, we investigated whether CAR T-cell effector metabolic activity is associated with increased extracellular phosphate consumption and a possible association between hypophosphatemia and ICANS. In vitro 4-1BB and CD28 CD19-targeted CAR T-cell effector activity was found to be associated with increased consumption of media phosphorus, which was temporally associated with increased single-cell effector secretomic activity and increased phosphorus-dependent metabolic demand of the CAR T cells. A clinical cohort of 77 patients treated with CD19-targeted CAR T-cell therapy demonstrated a significant anticorrelation between serum phosphorus and ICANS incidence and severity, with earlier onset of hypophosphatemia after CAR T-cell infusion more likely to result in neurotoxicity. These results imply phosphorous level monitoring could alert to the development of ICANS in clinical scenarios. See related Spotlight by Tobin et al., p. 1422.

Membrane Cholesterol Enrichment of Red Blood Cell-Derived Microparticles Results in Prolonged Circulation.

Particles fabricated from red blood cells (RBCs) can serve as vehicles for delivery of various biomedical cargos. Flipping of phosphatidylserine (PS) from the inner to the outer membrane leaflet normally occurs during the fabrication of such particles. PS externalization is a signal for phagocytic removal of the particles from circulation. Herein, we demonstrate that membrane cholesterol enrichment can mitigate the outward display of PS on microparticles engineered from RBCs. Our in-vitro results show that the phagocytic uptake of cholesterol-enriched particles by murine macrophages takes place at a lowered rate, resulting in reduced uptake as compared to RBC-derived particles without cholesterol enrichment. When administered via tail-vein injection into healthy mice, the percent of injected dose (ID) per gram of extracted blood for cholesterol-enriched particles was ∼1.5 and 1.8 times higher than the particles without cholesterol enrichment at 4 and 24 h, respectively. At 24 h, ∼43% ID/g of the particles without cholesterol enrichment was eliminated or metabolized while ∼94% ID/g of the cholesterol-enriched particles were still retained in the body. These results indicate that membrane cholesterol enrichment is an effective method to reduce PS externalization on the surface of RBC-derived particles and increase their longevity in circulation.

Phototheranostics Using Erythrocyte-Based Particles.

There has been a recent increase in the development of delivery systems based on red blood cells (RBCs) for light-mediated imaging and therapeutic applications. These constructs are able to take advantage of the immune evasion properties of the RBC, while the addition of an optical cargo allows the particles to be activated by light for a number of promising applications. Here, we review some of the common fabrication methods to engineer these constructs. We also present some of the current light-based applications with potential for clinical translation, and offer some insight into future directions in this exciting field.

CARD8 is an inflammasome sensor for HIV-1 protease activity.

HIV-1 has high mutation rates and exists as mutant swarms within the host. Rapid evolution of HIV-1 allows the virus to outpace the host immune system, leading to viral persistence. Approaches to targeting immutable components are needed to clear HIV-1 infection. Here, we report that the caspase recruitment domain-containing protein 8 (CARD8) inflammasome senses HIV-1 protease activity. HIV-1 can evade CARD8 sensing because its protease remains inactive in infected cells before viral budding. Premature intracellular activation of the viral protease triggered CARD8 inflammasome-mediated pyroptosis of HIV-1-infected cells. This strategy led to the clearance of latent HIV-1 in patient CD4+ T cells after viral reactivation. Thus, our study identifies CARD8 as an inflammasome sensor of HIV-1, which holds promise as a strategy for the clearance of persistent HIV-1 infection.

RBC-Derived Optical Nanoparticles Remain Stable After a Freeze-Thaw Cycle.

Nanosized carriers engineered from red blood cells (RBCs) provide a means for delivering various cargos, including drugs, biologics, and imaging agents. We have engineered nanosized particles from RBCs, doped with the near-infrared (NIR) fluorochrome, indocyanine green (ICG). An important issue related to clinical translation of RBC-derived nanocarriers, including these NIR nanoparticles, is their stability postfabrication. Freezing may provide a method for long-term storage of these and other RBC-derived nanoparticles. Herein, we have investigated the physical and optical stability of these particles in response to a single freeze-thaw cycle. Nanoparticles were frozen to -20 °C, stored frozen for up to 8 weeks, and then thawed at room temperature. Our results show that the hydrodynamic diameter, zeta potential, optical density, and NIR fluorescence emission of these nanoparticles are retained following the freeze-thaw cycle. The ability of these nanoparticles in NIR fluorescence imaging of ovarian cancer cells, as well as their biodistribution in reticuloendothelial organs of healthy Swiss Webster mice after the freeze-thaw cycle is similar to that for freshly prepared nanoparticles. These results indicate that a single cycle of freezing the RBC-derived nanoparticles to -20 °C followed by thawing at room temperature is an effective method to retain the physical and optical characteristics of the nanoparticles, and their interactions with biological systems without the need for use of cryoprotectants.

Dendritic Cell Paucity Leads to Dysfunctional Immune Surveillance in Pancreatic Cancer.

Here, we utilized spontaneous models of pancreatic and lung cancer to examine how neoantigenicity shapes tumor immunity and progression. As expected, neoantigen expression during lung adenocarcinoma development leads to T cell-mediated immunity and disease restraint. By contrast, neoantigen expression in pancreatic ductal adenocarcinoma (PDAC) results in exacerbation of a fibro-inflammatory microenvironment that drives disease progression and metastasis. Pathogenic TH17 responses are responsible for this neoantigen-induced tumor progression in PDAC. Underlying these divergent T cell responses in pancreas and lung cancer are differences in infiltrating conventional dendritic cells (cDCs). Overcoming cDC deficiency in early-stage PDAC leads to disease restraint, while restoration of cDC function in advanced PDAC restores tumor-restraining immunity and enhances responsiveness to radiation therapy.

Intravital Vascular Phototheranostics and Real-Time Circulation Dynamics of Micro- and Nanosized Erythrocyte-Derived Carriers.

Erythrocyte-based carriers can serve as theranostic platforms for delivery of imaging and therapeutic payloads. Engineering these carriers at micro- or nanoscales makes them potentially useful for broad clinical applications ranging from vascular diseases to tumor theranostics. Longevity of these carriers in circulation is important in delivering a sufficient amount of their payloads to the target. We have investigated the circulation dynamics of micro (∼4.95 µm diameter) and nano (∼91 nm diameter) erythrocyte-derived carriers in real time using near-infrared fluorescence imaging, and evaluated the effectiveness of such carrier systems in mediating photothermolysis of cutaneous vasculature in mice. Fluorescence emission half-lives of micro- and nanosized carriers in response to a single intravenous injection were ∼49 and ∼15 min, respectively. A single injection of microsized carriers resulted in a 3-fold increase in signal-to-noise ratio that remained nearly persistent over 1 h of imaging time. Our results also suggest that a second injection of the carriers 7 days later can induce a transient inflammatory response, as manifested by the apparent leakage of the carriers into the perivascular tissue. The administration of the carriers into the mice vasculature reduced the threshold laser fluence to induce photothermolysis of blood vessels from >65 to 20 J/cm2. We discuss the importance of membrane physicochemical and mechanical characteristics in engineering erythrocyte-derived carriers and considerations for their clinical translation.

Factors Associated with Publication of Research Projects from a Canadian Master of Science Degree Programme in Physical Therapy.

Purpose: The purpose of this study was to describe the nature and extent of publications and to evaluate whether lead advisor role and experience, data collection tool, sample size, and research topic predict publication for research projects completed as part of a Canadian Master of Science in Physical Therapy (MScPT) programme. Method: We conducted a quantitative, cross-sectional, retrospective review of projects completed between 2003 and 2015 and confirmed publication status through citations of published work, a literature search, and a survey of advisors. We used descriptive statistics to describe the nature and extent of publications and logistic regression to analyze potential predictor variables. Results: Between 2003 and 2015, 44.5% of the 218 projects completed were associated with at least one peer-reviewed journal publication, and there was a seven-fold increase in annual publication rate. Projects led by a scientist or researcher ([OR] = 3.09; 95% CI: 1.15, 8.35), qualitative projects with 10 or more participants ([OR] 6.22; 95% CI: 1.96, 19.78), and quantitative projects with more than 50 participants ([OR] = 2.29; 95% CI: 1.14, 4.63) were associated with an increased likelihood of publication. Conclusions: MScPT research is published at a moderate rate, and annual publication rates increased between 2003 and 2015. Encouragement to obtain adequate sample sizes and additional support for clinician-led projects may enhance publication rates and, ultimately, bridge gaps in research-to-practice integration.

Objectif : décrire la nature et la portée des publications et évaluer si le rôle et l'exercice de conseiller principal, l'outil de collecte de données utilisé la taille de l'échantillon et le sujet retenu prédisent la publication des projets de recherche réalisés dans le cadre d'une maîtrise ès sciences en physiothérapie au Canada. Méthodologie : analyse rétrospective quantitative et transversale de projets terminés entre 2003 et 2015, et confirmation de la publication par des citations déjà publiées, recherche documentaire et enquête auprès des conseillers. Les auteurs ont utilisé des statistiques descriptives pour décrire la durée et l'étendue des publications, de même que la régression logistique pour analyser les variables prédictives potentielles. Résultats : le taux annuel de publications a septuplé entre 2003 et 2015, et 44,5 % des 218 projets terminés s'associaient à au moins une publication dans une revue dotée d'un comité de lecture. Les projets dirigés par un scientifique ou un chercheur (rapport de cotes [RC] 3,09; IC à 95 % : 1,15, 8,35), les projets qualitatifs comptant au moins dix participants (RC 6,22; IC à 95 % : 1,96, 19,78) et les projets quantitatifs comptant plus de 50 participants (RC 2,29; IC à 95 % : 1,14, 4,63) étaient plus susceptibles d'être publiés. Conclusion : la publication des recherches de maîtrise est modérée, et le taux annuel de publications a augmenté entre 2003 et 2015. L'encouragement à utiliser des tailles d'échantillon suffisantes et le soutien accru des projets dirigés par un clinicien pourraient accroître le taux de publications et, au bout du compte, corriger les lacunes relatives à l'intégration de la recherche à la pratique.

Parkinson's disease protein DJ-1 regulates ATP synthase protein components to increase neuronal process outgrowth.

Familial Parkinson's disease (PD) protein DJ-1 mutations are linked to early onset PD. We have found that DJ-1 binds directly to the F1FO ATP synthase ß subunit. DJ-1's interaction with the ß subunit decreased mitochondrial uncoupling and enhanced ATP production efficiency while in contrast mutations in DJ-1 or DJ-1 knockout increased mitochondrial uncoupling, and depolarized neuronal mitochondria. In mesencephalic DJ-1 KO cultures, there was a progressive loss of neuronal process extension. This was ameliorated by a pharmacological reagent, dexpramipexole, that binds to ATP synthase, closing a mitochondrial inner membrane leak and enhancing ATP synthase efficiency. ATP synthase c-subunit can form an uncoupling channel; we measured, therefore, ATP synthase F1 (ß subunit) and c-subunit protein levels. We found that ATP synthase ß subunit protein level in the DJ-1 KO neurons was approximately half that found in their wild-type counterparts, comprising a severe defect in ATP synthase stoichiometry and unmasking c-subunit. We suggest that DJ-1 enhances dopaminergic cell metabolism and growth by its regulation of ATP synthase protein components.

Near-Infrared-Fluorescent Erythrocyte-Mimicking Particles: Physical and Optical Characteristics.

Exogenous fluorescent materials activated by near-infrared (NIR) light can offer deep optical imaging with subcellular resolution, and enhanced image contrast. We have engineered NIR particles by doping hemoglobin-depleted erythrocyte ghosts (EGs) with indocyanine green (ICG). We refer to these optical particles as NIR erythrocyte-mimicking transducers (NETs). A particular feature of NETs is that their diameters can be tuned from micrometer to nanometer scale, thereby, providing a capability for broad NIR biomedical imaging applications. Herein, we investigate the effects of ICG concentration on key material properties of micrometer-sized NETs, and nanometer-sized NETs fabricated by either sonication or mechanical extrusion of EGs. The zeta potentials of NETs do not vary significantly with ICG concentration, suggesting that ICG is encapsulated within NETs regardless of particle size or ICG concentration. Loading efficiency of ICG into the NETs monotonically decreases with increasing values of ICG concentration. Based on quantitative analyses of the fluorescence emission spectra of the NETs, we determine that 20 µM ICG utilized during fabrication of NETs presents an optimal concentration that maximizes the integrated fluorescence emission for micrometer- and nanometer-sized NETs. Encapsulation of the ICG in these constructs also enhances the fluorescence stability and quantum yield of ICG. These results guide the engineering of NETs with maximal NIR emission for imaging applications such as fluorescence-guided tumor resection and real-time angiography.

An alternatively spliced form affecting the Marked Box domain of Drosophila E2F1 is required for proper cell cycle regulation.

Across metazoans, cell cycle progression is regulated by E2F family transcription factors that can function as either transcriptional activators or repressors. For decades, the Drosophila E2F family has been viewed as a streamlined RB/E2F network, consisting of one activator (dE2F1) and one repressor (dE2F2). Here, we report that an uncharacterized isoform of dE2F1, hereon called dE2F1b, plays an important function during development and is functionally distinct from the widely-studied dE2F1 isoform, dE2F1a. dE2F1b contains an additional exon that inserts 16 amino acids to the evolutionarily conserved Marked Box domain. Analysis of de2f1b-specific mutants generated via CRISPR/Cas9 indicates that dE2F1b is a critical regulator of the cell cycle during development. This is particularly evident in endocycling salivary glands in which a tight control of dE2F1 activity is required. Interestingly, close examination of mitotic tissues such as eye and wing imaginal discs suggests that dE2F1b plays a repressive function as cells exit from the cell cycle. We also provide evidence demonstrating that dE2F1b differentially interacts with RBF1 and alters the recruitment of RBF1 and dE2F1 to promoters. Collectively, our data suggest that dE2F1b is a novel member of the E2F family, revealing a previously unappreciated complexity in the Drosophila RB/E2F network.

Evaluation of Invisalign treatment effectiveness and efficiency compared with conventional fixed appliances using the Peer Assessment Rating index.

INTRODUCTION: The purpose of this retrospective case-control study was to compare the treatment effectiveness and efficiency of the Invisalign system with conventional fixed appliances in treating orthodontic patients with mild to moderate malocclusion in a graduate orthodontic clinic. METHODS: Using the peer assessment rating (PAR) index, we evaluated pretreatment and posttreatment records of 48 Invisalign patients and 48 fixed appliances patients. The 2 groups of patients were controlled for general characteristics and initial severity of malocclusion. We analyzed treatment outcome, duration, and improvement between the Invisalign and fixed appliances groups. RESULTS: The average pretreatment PAR scores (United Kingdom weighting) were 20.81 for Invisalign and 22.79 for fixed appliances (P = 1.0000). Posttreatment weighted PAR scores between Invisalign and fixed appliances were not statistically different (P = 0.7420). On average, the Invisalign patients finished 5.7 months faster than did those with fixed appliances (P = 0.0040). The weighted PAR score reduction with treatment was not statistically different between the Invisalign and fixed appliances groups (P = 0.4573). All patients in both groups had more than a 30% reduction in the PAR scores. Logistic regression analysis indicated that the odds of achieving "great improvement" in the Invisalign group were 0.329 times the odds of achieving "great improvement" in the fixed appliances group after controlling for age (P = 0.0150). CONCLUSIONS: Our data showed that both Invisalign and fixed appliances were able to improve the malocclusion. Invisalign patients finished treatment faster than did those with fixed appliances. However, it appears that Invisalign may not be as effective as fixed appliances in achieving "great improvement" in a malocclusion. This study might help clinicians to determine appropriate patients for Invisalign treatment.