ObjectiveThe association between aggressive resection and improved survival for adult spinal chordoma patients has not been well characterized in the geriatric population. Thus, the present study aimed to elucidate the relationship between gross total resection (GTR) and survival outcomes for patients across different age groups.MethodsThe authors isolated all adult patients diagnosed with spinal chordoma from the 2000-2019 Surveillance, Epidemiology, and End Results database and divided patients into three surgical subgroups: no surgery, subtotal resection (STR), and GTR. Kaplan-Meier curves with a log-rank test were used to discern differences in overall survival (OS) between surgical subgroups. Univariate and multivariate analyses were used to identify prognostic factors of mortality.ResultsThere were 771 eligible patients: 227 (29.4%) received no surgery, 267 (34.6%) received STR, and 277 (35.9%) received GTR. Patients receiving no surgery had the lowest 5-year OS (45.2%), 10-year OS (17.6%), and mean OS (72.1 months). After stratifying patients by age, our multivariate analysis demonstrated that patients receiving GTR aged 40-59 (HR=0.26, CI=0.12-0.55, p<0.001), 60-79 (HR=0.51, CI=0.32-0.82, p=0.005), and 80-99 (HR=0.14, CI=0.05-0.37, p<0.001) had a lower risk of mortality compared to patients undergoing no surgery. The frequency of receiving GTR also decreased as a function of age (16.4% [80-99 years] vs. 43.2% [20-39 years]; p<0.001), but the frequency of receiving radiotherapy was comparable across all age groups (48.3% [80-99 years] vs. 45.5% [20-39 years]; p=0.762).ConclusionGTR is associated with improved survival for middle-aged and elderly patients with spinal chordoma. Therefore, patients should not be excluded from aggressive resection on the basis of age alone. Rather, the decision to pursue surgery should be decided on an individual basis.

BackgroundThe demand for discreet and low-compliance appliances has driven innovation in orthodontics, particularly with technological advances in artificial intelligence, robotics, and CAD/CAM technology. The evolution of Programmed Non-Sliding Mechanics for precise, automated tooth movement is the latest innovation.AimIn this article, we aimed to demonstrate the application of a novel orthodontic lingual appliance, The Gen 2 InBrace system (InBrace, Irvine, CA), in the orthodontic-restorative treatment of an adult patient with anterior tooth-size discrepancies, anterior openbite, and incisor proclination. The Gen 2 InBrace system uses a new generation of CAD/CAM Smartwires, including pre-programmed customized multiloop nickel-titanium lingual archwires, which enable light, continuous, and frictionless tooth movements.ConclusionThe Gen 2 InBrace system can enhance esthetic and occlusal outcomes in patients requiring combined orthodontic-restorative treatment via automatic space management with maximum esthetic and minimal compliance.Clinical SignificancePrecision dentistry and orthodontics using esthetic custom appliances have become a reality. The Gen 2 InBrace system, a new generation custom lingual appliance, offers effective treatment solutions for adults who have previously avoided preprosthetic orthodontics due to the esthetic concerns of wearing traditional labial braces or the compliance challenges associated with clear aligners.

Background: Cosmetic plastic surgery in the United States is underutilized by African American and Hispanic populations compared with their White and Asian counterparts. This study evaluated whether microeconomic spending traits as a representation of financial stability can inform trends in cosmetic procedure volumes by racial group. Methods: Annual volumes for the top five cosmetic surgical and cosmetic minimally invasive procedures by racial/ethnic group from 2012 to 2020 were collected from the American Society of Plastic Surgeons' annual reports. Factor analysis was used to calculate inflexible and flexible consumer spending by racial/ethnic groupings from the US Bureau of Labor Statistics' consumer expenditure data. All four factors were calculated across US Bureau of Labor Statistics-defined racial/ethnic groupings and standardized so they could be interpreted relative to each other. Results: Compared with the other groupings, the White/Asian/other grouping spent significantly more on average for inflexible consumer spending (P = 0.0097), flexible consumer spending (P < 0.0001), cosmetic surgical procedures (P < 0.0001), and cosmetic minimally invasive procedures (P = 0.0006). In contrast, African American people spent significantly less on average for all four factors (all P < 0.01). For Hispanic people, values were significantly less on average for flexible consumer spending (P = 0.0023), cosmetic surgical procedures (P < 0.0001), and cosmetic minimally invasive procedures (P = 0.0002). Conclusions: This study demonstrates that inflexible and flexible consumer spending follow trends in utilization of cosmetic surgical and minimally invasive procedures by racial/ethnic groups. These microeconomic spending inequities may help further contextualize the racial/ethnic variation in access to cosmetic surgery.

BACKGROUND:Contemporary fixed orthodontic appliances are shifting from non-customized pre-adjusted appliances to custom-designed and printed appliances with novel digital setup systems. We are one step closer to precision dentistry and orthodontics using personalized mechanics and custom appliances. However, despite the evidential enhancement and other improvements to fixed appliances, tooth movement is still limited to five degrees of freedom. Opening or closing spaces still requires manually placing elastomeric chains or coil springs. AIM:In this article, we aimed to demonstrate how advancements in CAD/CAM technology, reverse engineering, and digital customization are helping orthodontics constantly evolve, enabling treatment with enhanced esthetics and minimal compliance. The clinical system (InBrace®, Irvine, CA) described in this article uses a patient-specific, digitally designed multiloop NITI wire that delivers friction-free, light, and continuous forces and activates automatically whenever the malocclusion deviates from the digital setup. CONCLUSION:Through digital customization, InBrace allows for automated tooth movement in all six degrees of freedom, including space opening or closure, via programmed non-sliding mechanics. CLINICAL SIGNIFICANCE:Precision orthodontics and personalized treatment have been significant developments in orthodontics recently. This article focuses on how a technologically advanced lingual appliance system could achieve targeted cosmetic results methodically via automation and personalization.

SARS-CoV-2 variants of concern (VOCs) emerged during the COVID-19 pandemic. Here, we used unbiased systems approaches to study the host-selective forces driving VOC evolution. We discovered that VOCs evolved convergent strategies to remodel the host by modulating viral RNA and protein levels, altering viral and host protein phosphorylation, and rewiring virus-host protein-protein interactions. Integrative computational analyses revealed that although Alpha, Beta, Gamma, and Delta ultimately converged to suppress interferon-stimulated genes (ISGs), Omicron BA.1 did not. ISG suppression correlated with the expression of viral innate immune antagonist proteins, including Orf6, N, and Orf9b, which we mapped to specific mutations. Later Omicron subvariants BA.4 and BA.5 more potently suppressed innate immunity than early subvariant BA.1, which correlated with Orf6 levels, although muted in BA.4 by a mutation that disrupts the Orf6-nuclear pore interaction. Our findings suggest that SARS-CoV-2 convergent evolution overcame human adaptive and innate immune barriers, laying the groundwork to tackle future pandemics.

BACKGROUND:The relationship between marital status and overall survival (OS) in adult patients with craniopharyngioma has not been explored in depth. We aimed to elucidate the impact of marital status on the prognosis of craniopharyngioma patients excluding bias from baseline demographics and treatment. METHODS:We extracted 1539 patients diagnosed with craniopharyngioma between 2000 and 2019 from the Surveillance, Epidemiology, and End Results database and divided patients into 4 marital subgroups: married, single, divorced/separated, and widowed. Kaplan-Meier curves with a log-rank test were used to discern differences in OS between marital subgroups. Univariate and multivariate Cox regression were used to identify independent prognostic factors of mortality. RESULTS:There were 1539 eligible patients: 863 (56.1%) were married, 466 (30.3%) were single, 135 (8.8%) were divorced/separated, and 75 (4.9%) were widowed. Widowed patients had the worst mean OS, 5-year OS and 10-year OS at 84.2 months, 58.0% and 26.9%, respectively. After stratifying patients by age, our multivariate analysis showed that marital status was an independent predictor of mortality in middle-aged craniopharyngioma patients (40-60 years, P < 0.001), but not in young adults (18-39 years, P = 0.646) or elderly patients (>60 years, P = 0.076). Among middle-aged patients, single (hazard ratio 1.72, confidence interval 1.19-2.47, P = 0.004) and divorced/separated patients (hazard ratio = 2.29, confidence interval = 1.49-3.54, P < 0.001) showed a higher risk of mortality compared to married patients (reference). CONCLUSIONS:Marital status is an independent prognostic factor predicting OS for middle-aged patients with craniopharyngioma. Providing additional social and psychological support for single and divorced/separated patients may improve outcomes for this vulnerable population.

Parkinson's disease (PD) is a multi-stage neurodegenerative disorder with largely unknown etiology. Recent findings have identified PD-associated autoimmune features including roles for T cells. To further characterize the role of T cells in PD, we performed RNA sequencing on PBMC and peripheral CD4 and CD8 memory T cell subsets derived from PD patients and age-matched healthy controls. When the groups were stratified by their T cell responsiveness to alpha-synuclein (α-syn) as a proxy for an ongoing inflammatory autoimmune response, the study revealed a broad differential gene expression profile in memory T cell subsets and a specific PD associated gene signature. We identified significant enrichment of transcriptomic signatures previously associated with PD, including for oxidative stress, phosphorylation, autophagy of mitochondria, cholesterol metabolism and inflammation, and the chemokine signaling proteins CX3CR1, CCR5, and CCR1. In addition, we identified genes in these peripheral cells that have previously been shown to be involved in PD pathogenesis and expressed in neurons, such as LRRK2, LAMP3, and aquaporin. Together, these findings suggest that features of circulating T cells with α-syn-specific responses in PD patients provide insights into the interactive processes that occur during PD pathogenesis and suggest potential intervention targets.

ABSTRACTA series of SARS-CoV-2 variants of concern (VOCs) have evolved in humans during the COVID-19 pandemic—Alpha, Beta, Gamma, Delta, and Omicron. Here, we used global proteomic and genomic analyses during infection to understand the molecular responses driving VOC evolution. We discovered VOC-specific differences in viral RNA and protein expression levels, including for N, Orf6, and Orf9b, and pinpointed several viral mutations responsible. An analysis of the host response to VOC infection and comprehensive interrogation of altered virus-host protein-protein interactions revealed conserved and divergent regulation of biological pathways. For example, regulation of host translation was highly conserved, consistent with suppression of VOC replication in mice using the translation inhibitor plitidepsin. Conversely, modulation of the host inflammatory response was most divergent, where we found Alpha and Beta, but not Omicron BA.1, antagonized interferon stimulated genes (ISGs), a phenotype that correlated with differing levels of Orf6. Additionally, Delta more strongly upregulated proinflammatory genes compared to other VOCs. Systematic comparison of Omicron subvariants revealed BA.5 to have evolved enhanced ISG and proinflammatory gene suppression that similarly correlated with Orf6 expression, effects not seen in BA.4 due to a mutation that disrupts the Orf6-nuclear pore interaction. Our findings describe how VOCs have evolved to fine-tune viral protein expression and protein-protein interactions to evade both innate and adaptive immune responses, offering a likely explanation for increased transmission in humans.One sentence summarySystematic proteomic and genomic analyses of SARS-CoV-2 variants of concern reveal how variant-specific mutations alter viral gene expression, virus-host protein complexes, and the host response to infection with applications to therapy and future pandemic preparedness.

Gamma-delta (gamma delta) T cells contribute to the pathology of many immune-related diseases; however, no ex vivo assays to study their activities are currently available. Here, we established a methodology to characterize human allergen-reactive gamma delta T cells in peripheral blood using an activation-induced marker assay targeting upregulated 4-1BB and CD69 expression. Broad and reproducible ex vivo allergen-reactive gamma delta T cell responses were detected in donors sensitized to mouse, cockroach, house dust mite, and timothy grass, but the response did not differ from that in non-allergic participants. The reactivity to 4 different allergen extracts was readily detected in 54.2%-100% of allergic subjects in a donor- and allergen-specific pattern and was abrogated by T cell receptor (TCR) blocking. Analysis of CD40L upregulation and intracellular cytokine staining revealed a T helper type 1 (Th1)-polarized response against mouse and cockroach extract stimulation. These results support the existence of allergen-reactive gamma delta T cells and their potential use in rebalancing dysregulated Th2 responses in allergic diseases.

Orthodontic tooth movement occurs in six degrees of freedom, which includes opening and closing spaces. Traditionally, opening and closing spaces are achieved with auxiliaries such as power chains or springs because all traditional bracket systems cannot achieve this tooth movement by themselves. The InBrace system has the capability to program tooth movement in all six degrees of freedom, including opening and closing spaces, through digital customization and its use of Programmed Non-Sliding Mechanics.

Despite the prevalence and medical significance of human cytomegalovirus (HCMV) infections, a systematic analysis of the targets of T cell recognition in humans that spans the entire genome and includes recently described potential novel open reading frames (ORFs) is not available. Here, we screened a library of epitopes predicted to bind HLA class II that spans over 350 different HCMV ORFs and includes ∼150 previously described and ∼200 recently described potential novel ORFs by using an ex vivo gamma interferon (IFN-γ) FluoroSpot assay. We identified 235 unique HCMV-specific epitopes derived from 100 ORFs, some previously described as immunodominant and others that were not previously described to be immunogenic. Of those, 41 belong to the set of recently reported novel ORFs, thus providing evidence that at least some of these are actually expressed in vivo in humans. These data reveal that the breadth of the human T cell response to HCMV is much greater than previously thought. The ORFs and epitopes identified will help elucidate how T cell immunity relates to HCMV pathogenesis and instruct ongoing HCMV vaccine research. IMPORTANCE To understand the crucial role of adaptive immunity in controlling cytomegalovirus infection and disease, we systematically analyzed the CMV "ORFeome" to identify new CMV epitopes targeted primarily by CD4 T cells in humans. Our study identified >200 new T cell epitopes derived from both canonical and novel ORFs, highlighting the substantial breadth of the anti-CMV T cell response and providing new targets for vaccine design.

The self-antigen α-synuclein (α-syn) was recently shown to be associated with Parkinson’s disease (PD). Here we mapped the T cell receptor (TCR) repertoire of α-syn-specific T cells from six PD patients. The self-antigen α-syn-specific repertoire was compared to the repertoire of T cells specific for pertussis (PT), as a representative foreign antigen that most individuals are exposed to, revealing that the repertoire for α-syn was as diverse as the repertoire for PT. The diversity of PT-specific clonotypes was similar between individuals with PD diagnosis and age-matched healthy controls. We found that the TCR repertoire was specific to each PD patient, and no shared TCRs among patients were defined, likely due to differences in HLA expression that select for different subsets of epitope-specific TCR rearrangements. This study provides the first characterization of α-syn-specific TCR clonotypes in individuals with PD. Antigen-specific TCRs can serve as immunotherapeutics and diagnostics, and means to track longitudinal changes in specific T cells, and disease progression.

AbstractDespite the prevalence and medical significance of human cytomegalovirus (HCMV) infections, a systematic analysis of the targets of T cell recognition in humans that spans the entire genome and includes recently described potential novel ORFs is not available. Here, we screened a library of epitopes predicted to bind HLA class II that spans over 350 different HCMV ORFs and includes ∼150 previously described and ∼200 recently described potential novel ORFs using an ex vivo IFNγ fluorospot assay. We identified 235 unique HCMV specific epitopes derived from 100 ORFs, some previously described as immunodominant and others that were not previously described to be immunogenic. Of those, 41 belong to the set of recently reported novel ORFs, thus providing evidence that at least some of these are actually expressed in vivo in humans. These data reveal that the breadth of the human T cell response to HCMV is much greater than previously thought. The ORFs and epitopes identified will help elucidate how T cell immunity relates to HCMV pathogenesis and instruct ongoing HCMV vaccine research.ImportanceTo understand the crucial role of adaptive immunity in controlling cytomegalovirus infection and disease, we systematically analyzed the CMV ’ORFeome’ to identify new CMV epitopes targeted primarily by CD4 T cells in humans. Our study identified >200 new T cell epitopes derived from both canonical and novel ORFs, highlighting the substantial breadth of anti-CMV T cell response and providing new targets for vaccine design.

Nitric oxide (NO) is an important signaling molecule that is involved in a wide range of physiological and pathological events in biology. Metal coordination chemistry, especially with iron, is at the heart of many biological transformations involving NO. A series of heme proteins, nitric oxide synthases (NOS), soluble guanylate cyclase (sGC), and nitrophorins, are responsible for the biosynthesis, sensing, and transport of NO. Alternatively, NO can be generated from nitrite by heme- and copper-containing nitrite reductases (NIRs). The NO-bearing small molecules such as nitrosothiols and dinitrosyl iron complexes (DNICs) can serve as an alternative vehicle for NO storage and transport. Once NO is formed, the rich reaction chemistry of NO leads to a wide variety of biological activities including reduction of NO by heme or non-heme iron-containing NO reductases and protein post-translational modifications by DNICs. Much of our understanding of the reactivity of metal sites in biology with NO and the mechanisms of these transformations has come from the elucidation of the geometric and electronic structures and chemical reactivity of synthetic model systems, in synergy with biochemical and biophysical studies on the relevant proteins themselves. This review focuses on recent advancements from studies on proteins and model complexes that not only have improved our understanding of the biological roles of NO but also have provided foundations for biomedical research and for bio-inspired catalyst design in energy science.

The mosquito-borne Zika virus (ZIKV) has spread rapidly into regions where dengue virus (DENV) is endemic, and flavivirus cross-reactive T cell responses have been observed repeatedly in animal models and in humans. Preexisting cellular immunity to DENV is thought to contribute to protection in subsequent ZIKV infection, but the epitope targets of cross-reactive T cell responses have not been comprehensively identified. Using human blood samples from the regions of Nicaragua and Sri Lanka where DENV is endemic that were collected before the global spread of ZIKV in 2016, we employed an in vitro expansion strategy to map ZIKV T cell epitopes in ZIKV-unexposed, DENV-seropositive donors. We identified 93 epitopes across the ZIKV proteome, and we observed patterns of immunodominance that were dependent on antigen size and sequence identity to DENV. We confirmed the immunogenicity of these epitopes through a computational HLA binding analysis, and we showed that cross-reactive T cells specifically recognize ZIKV peptides homologous to DENV sequences. We also found that these CD4 responses were derived from the memory T cell compartment. These data have implications for understanding the dynamics of flavivirus-specific T cell immunity in areas of endemicity. IMPORTANCE Multiple flaviviruses, including Zika virus (ZIKV) and the four serotypes of dengue virus (DENV), are prevalent in the same large tropical and equatorial areas, which are inhabited by hundreds of millions of people. The interplay of DENV and ZIKV infection is especially relevant, as these two viruses are endemic in largely overlapping regions, have significant sequence similarity, and share the same arthropod vector. Here, we define the targets of preexisting immunity to ZIKV in unexposed subjects in areas where dengue is endemic. We demonstrate that preexisting immunity to DENV could shape ZIKV-specific responses, and DENV-ZIKV cross-reactive T cell populations can be expanded by stimulation with ZIKV peptides. The issue of potential ZIKV and DENV cross-reactivity is of relevance for understanding patterns of natural immunity, as well as for the development of diagnostic tests and vaccines.

A diagnosis of motor Parkinson’s disease (PD) is preceded by a prolonged premotor phase with accumulating neuronal damage. Here we examined the temporal relation between α-synuclein (α-syn) T cell reactivity and PD. A longitudinal case study revealed that elevated α-syn-specific T cell responses were detected prior to the diagnosis of motor PD, and declined after. The relationship between T cell reactivity and early PD in two independent cohorts showed that α-syn-specific T cell responses were highest shortly after diagnosis of motor PD and then decreased. Additional analysis revealed significant association of α-syn-specific T cell responses with age and lower levodopa equivalent dose. These results confirm the presence of α-syn-reactive T cells in PD and show that they are most abundant immediately after diagnosis of motor PD. These cells may be present years before the diagnosis of motor PD, suggesting avenues of investigation into PD pathogenesis and potential early diagnosis.

Mouse allergies are prevalent in inner city households and laboratory workers. Mouse allergens trigger allergic symptoms from atopic dermatitis/conjunctivitis to rhinitis and asthma, thus their research is of great clinical value. The aim of this study was to characterize the mouse-specific T cell response changes in non-allergic laboratory workers before and after vacation. Peripheral blood mononuclear cells (PBMC) were collected pre-break, pre re-exposure (after break), and after re-exposure to mouse, and were stimulated with peptides identified from high molecular weight fractions of mouse allergen extracts (HiMO). Non-allergics had sizeable reacitivity and majority had decreased T cell responses upon re-exposure. In addition, the decreased magnitudes were positively correlated with time of re-exposure but not length of break. Interestingly, the suppressive activity was dominant and specific for mouse responses and did not affect bystander responses. CD4 T cell responses was down-regulated temporally for a period of time (40–160 days) and eventually went back to baseline. Ex vivo immunophenotyping revealed a trend towards down-regulation of B cells and classical Tregs. Mouse-specific CD4 effector cells were reduced while total CD4 effector cells were not. The results indicate that non-allergic status is an active process which involves allergen-specific modulation of responses upon re-exposure to allergens. Uncovering the molecular programs associated with this modulation by transcriptomic study will be the next step to better understand health and allergic diseases.

Alzheimer's disease (AD), a chronic multifactorial and complex neurodegenerative disorder is a leading cause of dementia. Recently, neuroinflammation has been hypothesized as a contributing factor to AD pathogenesis. The role of adaptive immune responses against neuronal antigens, which can either confer protection or induce damage in AD, has not been fully characterized. Here, we measured T cell responses to several potential antigens of neural origin including amyloid precursor protein (APP), amyloid beta (Aβ), tau, α-synuclein, and transactive response DNA binding protein (TDP-43) in patients with AD and age-matched healthy controls (HC). Antigen-specific T cell reactivity was detected for all tested antigens, and response to tau-derived epitopes was particularly strong, but no significant differences between individuals with AD and age-matched HC were identified. We also did not observe any correlation between the antigen-specific T cell responses and clinical variables including age, gender, years since diagnosis and cognitive score. Additionally, further characterization did not reveal any differences in the relative frequency of major Peripheral Blood Mononuclear Cells (PBMC) subsets, or in the expression of genes between AD patients and HC. These observations have not identified a key role of neuronal antigen-specific T cell responses in AD.