International guidelines for intratumoral and intranodal injection of NBTXR3 nanoparticles in head and neck cancers.

BACKGROUND: An international multidisciplinary panel of experts aimed to provide consensus guidelines describing the optimal intratumoral and intranodal injection of NBTXR3 hafnium oxide nanoparticles in head and neck squamous cell carcinoma (HNSCC) of the oral cavity, oropharynx, and cervical lymph nodes and to review data concerning safety, feasibility, and procedural aspects of administration. METHODS: The Delphi method was used to determine consensus. A 4-member steering committee and a 10-member monitoring committee wrote and revised the guidelines, divided into eight sections. An independent 3-member reading committee reviewed the recommendations. RESULTS: After two rounds of voting, strong consensus was obtained on all recommendations. Intratumoral and intranodal injection was deemed feasible. NBTXR3 volume calculation, choice of patients, preparation and injection procedure, potential side effects, post injection, and post treatment follow-up were described in detail. CONCLUSIONS: Best practices for the injection of NBTXR3 were defined, thus enabling international standardization of intratumoral nanoparticle injection.

Safety and reliability of the internal jugular vein for venous anastomoses in head and neck oncological reconstruction: A retrospective study.

This study aimed to assess the efficacy of utilizing the internal jugular vein (IJV) as the primary recipient site for venous anastomoses in head and neck oncological reconstruction. Patients who underwent a free flap reconstruction of the head and neck were retrospectively included. Venous anastomoses were preferentially performed less than 1 cm from the IJV, either end-to-side (EtS) on the IJV, or end-to-end (EtE) on the origin of the thyrolingofacial venous (TLF) trunk. When the pedicle length was insufficient to reach the IJV, anastomoses were performed EtE to a size-matched cervical vein. Of the 246 venous anastomoses, 216 (87.8%) were performed less than 1 cm from the IJV, including 150 EtS on the IJV (61.0%), and 66 EtE on the TLF trunk (26.8%). Thirty veins (12.1%) were anastomosed EtE on other cervical veins more than 1 cm from the IJV. Two venous thromboses occurred (0.9%) and were successfully managed after revision surgery. There was no evidence of an increased thrombosis rate in high-risk or pre-irradiated patients. These findings suggest that the internal jugular vein is safe and reliable as a first-choice recipient vessel for free flap transfers in head and neck oncological reconstruction.

Randomized phase II study of preoperative afatinib in untreated head and neck cancers: predictive and pharmacodynamic biomarkers of activity.

There is no strong and reliable predictive biomarker in head and neck squamous cell carcinoma (HNSCC) for EGFR inhibitors. We aimed to identify predictive and pharmacodynamic biomarkers of efficacy of afatinib, a pan-HER tyrosine kinase inhibitor, in a window-of-opportunity trial (NCT01415674). Multi-omics analyses were carried out on pre-treatment biopsy and surgical specimen for biological assessment of afatinib activity. Sixty-one treatment-naïve and operable HNSCC patients were randomised to afatinib 40 mg/day for 21-28 days versus no treatment. Afatinib produced a high rate of metabolic response. Responders had a higher expression of pERK1/2 (P = 0.02) and lower expressions of pHER4 (P = 0.03) and pRB1 (P = 0.002) in pre-treatment biopsy compared to non-responders. At the cellular level, responders displayed an enrichment of tumor-infiltrating B cells under afatinib (P = 0.02). At the molecular level, NF-kappa B signaling was over-represented among upregulated genes in non-responders (P < 0.001; FDR = 0.01). Although exploratory, phosphoproteomics-based biomarkers deserve further investigations as predictors of afatinib efficacy.

Large-scale analysis of cell-cell communication reveals angiogenin-dependent tumor progression in clear cell renal cell carcinoma.

Cellular crosstalk in the tumor microenvironment (TME) is still largely uncharacterized, while it plays an essential role in shaping immunosuppression or anti-tumor response. Large-scale analyses are needed to better decipher cell-cell communication in cancer. In this work, we used original and publicly available single-cell RNA sequencing (scRNAseq) data to characterize in-depth the communication networks in human clear cell renal cell carcinoma (ccRCC). We identified 50 putative communication channels specifically used by cancer cells to interact with other cells, including two novel angiogenin-mediated interactions. Expression of angiogenin and its receptors was validated at the protein level in primary ccRCC. Mechanistically, angiogenin enhanced ccRCC cell line proliferation and down-regulated secretion of IL-6, IL-8, and MCP-1 proinflammatory molecules. This study provides novel biological insights into molecular mechanisms of ccRCC, and suggests angiogenin and its receptors as potential therapeutic targets in clear cell renal cancer.

Breastfeeding of preterm newborns in a neonate hospitalization unit.

OBJECTIVE: To analyze the factors associated with breastfeeding of preterm infants at discharge. METHOD: Cross-sectional study with newborns with gestational age <37 weeks, admitted to a university hospital. Data obtained from the medical records of 180 participants, from August/2019 to August/2020. To assess an association between categorical variables, Pearson's chi-square and Fisher's exact tests were used. The significance level adopted was 5% (p ≤ 0.05). RESULTS: Mean gestational age was 32.8 ± 2.7 weeks and mean birth weight was 1,890 grams ± 682 grams. During hospitalization, (n=166) 28.3% received predominantly breast milk. At discharge, (n=164) 84.1% received breast milk and, of these, 2.4% were exclusively breastfed. Breastfeeding at discharge was associated with gestational age ≥ 33.5 weeks, higher weight at birth, and shorter hospitalization. CONCLUSION: The study showed that during hospitalization, about a third of the participants were breastfed. However, at the time of discharge, there was a predominance of breastfeeding in most cases, and the associated factors were higher weight at birth and shorter hospital stay.

Selective STING stimulation in dendritic cells primes antitumor T cell responses.

T cells that recognize tumor antigens are crucial for mounting antitumor immune responses. Induction of antitumor T cells in immunogenic tumors depends on STING, the intracellular innate immune receptor for cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) and related cyclic dinucleotides (CDNs). However, the optimal way to leverage STING activation in nonimmunogenic tumors is still unclear. Here, we show that cGAMP delivery by intratumoral injection of virus-like particles (cGAMP-VLP) led to differentiation of circulating tumor-specific T cells, decreased tumor regulatory T cells (Tregs), and antitumoral responses that synergized with PD1 blockade. By contrast, intratumoral injection of the synthetic CDN ADU-S100 led to tumor necrosis and systemic T cell activation but simultaneously depleted immune cells from injected tumors and induced minimal priming of circulating tumor-specific T cells. The antitumor effects of cGAMP-VLP required type 1 conventional dendritic cells (cDC1), whereas ADU-S100 eliminated cDC1 from injected tumors. cGAMP-VLP preferentially targeted STING in dendritic cells at a 1000-fold smaller dose than ADU-S100. Subcutaneous administration of cGAMP-VLP showed synergy when combined with PD1 blockade or a tumor Treg-depleting antibody to elicit systemic tumor-specific T cells and antitumor activity, leading to complete and durable tumor eradication in the case of tumor Treg depletion. These findings show that cell targeting of STING stimulation shapes the antitumor T cell response and identify a therapeutic strategy to enhance T cell-targeted immunotherapy.

The predictive power of insomnia symptoms on other aspects of mental health during the COVID-19 pandemic: a longitudinal study.

Symptoms of insomnia are an important risk factor for the development of mental disorders, especially during stressful life periods such as the coronavirus disease 2019 (COVID-19) pandemic. However, up to now, most studies have used cross-sectional data, and the prolonged impact of insomnia symptoms during the pandemic on later mental health remains unclear. Therefore, we investigated insomnia symptoms as a predictor of other aspects of mental health across 6 months, with altogether seven assessments (every 30 days, t0-t6), in a community sample (N = 166-267). Results showed no mean-level increase of insomnia symptoms and/or deterioration of mental health between baseline assessment (t0) and the 6- month follow-up (t6). As preregistered, higher insomnia symptoms (between persons) across all time points predicted reduced mental health at the 6-month follow-up. Interestingly, contrary to our hypothesis, higher insomnia symptoms at 1 month, within each person (i.e., compared to that person's symptoms at other time points), predicted improved rather than reduced aspects of mental health 1 month later. Hence, we replicated the predictive effect of averagely increased insomnia symptoms on impaired later mental health during the COVID-19 pandemic. However, we were surprised that increased insomnia symptoms at 1 month predicted aspects of improved mental health 1 month later. This unexpected effect might be specific for our study population and a consequence of our study design. Overall, increased insomnia symptoms may have served as a signal to engage in, and successfully implement, targeted countermeasures, which led to better short-term mental health in this healthy sample.

Meta-Analysis of Human Cancer Single-Cell RNA-Seq Datasets Using the IMMUcan Database.

The development of single-cell RNA sequencing (scRNA-seq) technologies has greatly contributed to deciphering the tumor microenvironment (TME). An enormous amount of independent scRNA-seq studies have been published representing a valuable resource that provides opportunities for meta-analysis studies. However, the massive amount of biological information, the marked heterogeneity and variability between studies, and the technical challenges in processing heterogeneous datasets create major bottlenecks for the full exploitation of scRNA-seq data. We have developed IMMUcan scDB (https://immucanscdb.vital-it.ch), a fully integrated scRNA-seq database exclusively dedicated to human cancer and accessible to nonspecialists. IMMUcan scDB encompasses 144 datasets on 56 different cancer types, annotated in 50 fields containing precise clinical, technological, and biological information. A data processing pipeline was developed and organized in four steps: (i) data collection; (ii) data processing (quality control and sample integration); (iii) supervised cell annotation with a cell ontology classifier of the TME; and (iv) interface to analyze TME in a cancer type-specific or global manner. This framework was used to explore datasets across tumor locations in a gene-centric (CXCL13) and cell-centric (B cells) manner as well as to conduct meta-analysis studies such as ranking immune cell types and genes correlated to malignant transformation. This integrated, freely accessible, and user-friendly resource represents an unprecedented level of detailed annotation, offering vast possibilities for downstream exploitation of human cancer scRNA-seq data for discovery and validation studies. SIGNIFICANCE: The IMMUcan scDB database is an accessible supportive tool to analyze and decipher tumor-associated single-cell RNA sequencing data, allowing researchers to maximally use this data to provide new insights into cancer biology.

Breastfeeding of preterm newborns in a neonate hospitalization unit / Lactancia del recién nacidoprematuro en la unidad de admisión neonatal / Aleitamento materno de recém-nascido prematuro em unidade de internação neonatal

ABSTRACT Objective: To analyze the factors associated with breastfeeding of preterm infants at discharge. Method: Cross-sectional study with newborns with gestational age <37 weeks, admitted to a university hospital. Data obtained from the medical records of 180 participants, from August/2019 to August/2020. To assess an association between categorical variables, Pearson's chi-square and Fisher's exact tests were used. The significance level adopted was 5% (p ≤ 0.05). Results: Mean gestational age was 32.8 ± 2.7 weeks and mean birth weight was 1,890 grams ± 682 grams. During hospitalization, (n=166) 28.3% received predominantly breast milk. At discharge, (n=164) 84.1% received breast milk and, of these, 2.4% were exclusively breastfed. Breastfeeding at discharge was associated with gestational age ≥ 33.5 weeks, higher weight at birth, and shorter hospitalization. Conclusion: The study showed that during hospitalization, about a third of the participants were breastfed. However, at the time of discharge, there was a predominance of breastfeeding in most cases, and the associated factors were higher weight at birth and shorter hospital stay.

RESUMEN Objetivo: Analizar los factores asociados a la lactancia materna de prematuros al alta. Método: Estudio transversal compuesto por recién nacidos con edad gestacional <37 semanas, internados en un hospital universitario. Datos obtenidos de las historias clínicas de 180 participantes, entre agosto/2019 y agosto/2020. Para evaluar una asociación entre las variables categóricas, se utilizaron las pruebas de chi-cuadrado de Pearson o exacta de Fisher. El nivel de significación adoptado fue del 5% (p ≤ 0,05). Resultados: La edad gestacional media fue de 32,8 ± 2,7 semanas y el peso medio al nacer fue de 1.890 gramos ± 682 gramos. Durante la hospitalización, (n=166) 28,3% los niños recibieron predominantemente leche materna. Al alta, (n=164) 84,1% recibieron leche materna y, de estos, 2,4% estaban en lactancia materna exclusiva. La lactancia materna al alta se asoció con una edad gestacional ≥ 33,5 semanas; mayor peso al nacer y hospitalización más corta. Conclusión: El estudio mostró que, durante la hospitalización, alrededor de un tercio de los participantes recibieron leche materna. Sin embargo, al momento del alta prevaleció la lactancia materna en la mayoría de los casos, y los factores asociados fueron mayor peso al nacer y menor estancia hospitalaria.

RESUMO Objetivo: Analisar os fatores associados ao aleitamento materno do pré-termo na alta. Método: Estudo transversal composto por recém-nascidos de idade gestacional menor que 37 semanas, internados em hospital universitário. Dados obtidos dos prontuários dos180 participantes, incluídos de agosto/2019 a agosto/2020. Para avaliar a associação entre as variáveis categóricas, os testes qui-quadrado de Pearson ou exato de Fisher foram utilizados. O nível de significância adotado foi de 5% (p ≤ 0,05). Resultados: A idade gestacional média foi de 32,8 ± 2,7 semanas e peso ao nascer médio de 1.890 gramas ± 682 gramas. Na internação, (n=166) 28,3% receberam leite materno predominantemente. Na alta, (n=164) 84,1% recebiam leite materno e, desses, 2,4% estavam em aleitamento materno exclusivo. O aleitamento materno na alta foi associado à idade gestacional maior/igual a 33,5 semanas, maior peso ao nascer e menor tempo de internação. Conclusão: O estudo evidenciou que durante a internação, cerca de um terço dos participantes foram alimentados com leite materno. Entretanto, no momento da alta, houve prevalência de alimentação com leite materno na maioria dos casos, sendo que os fatores associados foram maior peso ao nascer e menor tempo de hospitalização.

GM-CSF-activated human dendritic cells promote type 1 T follicular helper cell polarization in a CD40-dependent manner.

T follicular helper (Tfh) cells regulate humoral responses and present a marked phenotypic and functional diversity. Type 1 Tfh (Tfh1) cells were recently identified and associated with disease severity in infection and autoimmune diseases. The cellular and molecular requirements to induce human Tfh1 differentiation are not known. Here, using single-cell RNA sequencing (scRNAseq) and protein validation, we report that human blood CD1c+ dendritic cells (DCs) activated by GM-CSF (also known as CSF2) drive the differentiation of naive CD4+ T cells into Tfh1 cells. These Tfh1 cells displayed typical Tfh molecular features, including high levels of PD-1 (encoded by PDCD1), CXCR5 and ICOS. They co-expressed BCL6 and TBET (encoded by TBX21), and secreted large amounts of IL-21 and IFN-γ (encoded by IFNG). Mechanistically, GM-CSF triggered the emergence of two DC sub-populations defined by their expression of CD40 and ICOS ligand (ICOS-L), presenting distinct phenotypes, morphologies, transcriptomic signatures and functions. CD40High ICOS-LLow DCs efficiently induced Tfh1 differentiation in a CD40-dependent manner. In patients with mild COVID-19 or latent Mycobacterium tuberculosis infection, Tfh1 cells were positively correlated with a CD40High ICOS-LLow DC signature in scRNAseq of peripheral blood mononuclear cells or blood transcriptomics, respectively. Our study uncovered a novel CD40-dependent Tfh1 axis with potential physiopathological relevance to infection. This article has an associated First Person interview with the first author of the paper.

Motivational Interdependence in Couple Relationships.

This article presents an integrative conceptual model of motivational interdependence in couples, the MIC model. Based on theoretical tenets in motivation psychology, personality psychology, and research on interpersonal perception, the MIC model postulates that two partners' motive dispositions fundamentally interact in shaping their individual motivation and behavior. On a functional level, a partner's motivated behavior is conceptualized as an environmental cue that can contribute to an actor's motive expression and satisfaction. However, the partner's motivated behavior is considered to gain this motivational relevance only via the actor's subjective perception. Multilevel analyses of an extensive experience sampling study on partner-related communal motivation (N = up to 60,803 surveys from 508 individuals nested in 258 couples) supported the MIC model. Participants, particularly those with strong communal motive dispositions, behaved more communally at moments when they perceived their partners to behave more communally. In addition, participants experienced momentary boosts in satisfaction when they behaved more communally and, at the same time, perceived their partners' behavior as similarly communal. Broader implications of the MIC model for research on romantic relationships are discussed.

Pseudocowpox virus, a novel vector to enhance the therapeutic efficacy of antitumor vaccination.

Objective: Antitumor viral vaccines, and more particularly poxviral vaccines, represent an active field for clinical development and translational research. To improve the efficacy and treatment outcome, new viral vectors are sought, with emphasis on their abilities to stimulate innate immunity, to display tumor antigens and to induce a specific T-cell response. Methods: We screened for a new poxviral backbone with improved innate and adaptive immune stimulation using IFN-α secretion levels in infected PBMC cultures as selection criteria. Assessment of virus effectiveness was made in vitro and in vivo. Results: The bovine pseudocowpox virus (PCPV) stood out among several poxviruses for its ability to induce significant secretion of IFN-α. PCPV produced efficient activation of human monocytes and dendritic cells, degranulation of NK cells and reversed MDSC-induced T-cell suppression, without being offensive to activated T cells. A PCPV-based vaccine, encoding the HPV16 E7 protein (PCPV-E7), stimulated strong antigen-specific T-cell responses in TC1 tumor-bearing mice. Complete regression of tumors was obtained in a CD8+ T-cell-dependent manner after intratumoral injection of PCPV-E7, followed by intravenous injection of the cancer vaccine MVA-E7. PCPV also proved active when injected repeatedly intratumorally in MC38 tumor-bearing mice, generating tumor-specific T-cell responses without encoding a specific MC38 antigen. From a translational perspective, we demonstrated that PCPV-E7 effectively stimulated IFN-γ production by T cells from tumor-draining lymph nodes of HPV+-infected cancer patients. Conclusion: We propose PCPV as a viral vector suitable for vaccination in the field of personalised cancer vaccines, in particular for heterologous prime-boost regimens.

ß-Boronic Acid-Substituted Bodipy Dyes for Fluorescence Anisotropy Analysis of Carbohydrate Binding.

Boronic acids are widely used for labeling catechols and carbohydrates in analytical (bio)chemistry due to their high binding affinities for diols. Here, we present two asymmetrically substituted Bodipy dyes with a boronic acid at the ß-position (BBB). We present a green-emitting BBB, gBBB, and, by expanding the conjugated system of the Bodipy core at α-position, a red-emitting rBBB. Especially, gBBB shows a bathochromic shift of the electronic spectra upon binding to saccharides and polyols, whereas the fluorescence lifetime of rBBB is more sensitive to hydroxy-ligand binding. Moreover, gBBB constantly shows higher binding affinities than rBBB, reaching Kb ≈ 103 M-1 at pH 8.5 for fructose. Finally, time-resolved fluorescence anisotropy allows to distinguish the number of saccharide units of oligosaccharides as the bond along the transition dipole moment ensures that the fluorescence anisotropy only decays due to the rotational diffusion of labeled carbohydrates. ß-substituted BODIPY dyes are, thus, foreseen as fluorescence anisotropy labels for macromolecule sizing, where conventional fluorophores fail to discriminate due to the chemical similarity of recognition sites.

PD-L1 and ICOSL discriminate human Secretory and Helper dendritic cells in cancer, allergy and autoimmunity.

Dendritic cells (DC) are traditionally classified according to their ontogeny and their ability to induce T cell response to antigens, however, the phenotypic and functional state of these cells in cancer does not necessarily align to the conventional categories. Here we show, by using 16 different stimuli in vitro that activated DCs in human blood are phenotypically and functionally dichotomous, and pure cultures of type 2 conventional dendritic cells acquire these states (termed Secretory and Helper) upon appropriate stimuli. PD-L1highICOSLlow Secretory DCs produce large amounts of inflammatory cytokines and chemokines but induce very low levels of T helper (Th) cytokines following co-culturing with T cells. Conversely, PD-L1lowICOSLhigh Helper DCs produce low levels of secreted factors but induce high levels and a broad range of Th cytokines. Secretory DCs bear a single-cell transcriptomic signature indicative of mature migratory LAMP3+ DCs associated with cancer and inflammation. Secretory DCs are linked to good prognosis in head and neck squamous cell carcinoma, and to response to checkpoint blockade in Melanoma. Hence, the functional dichotomy of DCs we describe has both fundamental and translational implications in inflammation and immunotherapy.

Unmet Needs and Perspectives in Oral Cancer Prevention.

Oral potentially malignant disorders (OPMD) may precede oral squamous cell carcinoma (OSCC). Reported rates of malignant transformation of OPMD range from 3 to 50%. While some clinical, histological, and molecular factors have been associated with a high-risk OPMD, they are, to date, insufficiently accurate for treatment decision-making. Moreover, this range highlights differences in the clinical definition of OPMD, variation in follow-up periods, and molecular and biological heterogeneity of OPMD. Finally, while treatment of OPMD may improve outcome, standard therapy has been shown to be ineffective to prevent OSCC development in patients with OPMD. In this perspective paper, several experts discuss the main challenges in oral cancer prevention, in particular the need to (i) to define an OPMD classification system by integrating new pathological and molecular characteristics, aiming (ii) to better identify OPMD at high risk of malignant transformation, and (iii) to develop treatment strategies to eradicate OPMD or prevent malignant transformation.

Nanoparticle therapy for head and neck cancers.

PURPOSE OF REVIEW: The current review focuses on the therapeutic use of nanoparticles in head and neck cancer (HNC), highlighting nanoparticles at the most advanced clinical development stages. RECENT FINDINGS: Literature review covers the three main approaches for therapeutic use of nanoparticles in HNC: first, enhancing radiotherapy effect; second, performing targeted delivery of chemotherapy, immunotherapy, or genome editing molecules; third, photothermal therapy. SUMMARY: Nanoparticles are spherical nanoscale objects that have application in cancer therapies. Nanoparticles have diverse and often composite structure composition to ensure their function, increase their bioavailability in tumor tissues, and decrease off-target effects, sometimes by means of activating internal or external stimuli. Hafnium oxide nanoparticles are being tested in phase I to III trials for radiotherapy enhancement. Nanoparticle-based delivery of paclitaxel, cisplatin, and of the immune activator CpG-A DNA is being evaluated in phase II trials. No nanoparticle is currently approved for HNC treatment.

Exploratory window-of-opportunity trial to investigate the tumor pharmacokinetics/pharmacodynamics of the IAP antagonist Debio 1143 in patients with head and neck cancer.

Inhibitor of apoptosis proteins (IAPs) regulate apoptosis and modulate NF-κB signaling thereby driving expression of genes involved in immune/inflammatory responses. The orally available IAP antagonist Debio 1143 has potential to enhance tumor response to chemoradiotherapy and/or immunotherapy. Patients with pre-operative squamous cell carcinomas of the head and neck (SCCHN) received: Debio 1143 monotherapy (200 mg/day [D]1-15 +/- 2); Debio 1143 (200 mg/day D1-15 +/- 2) plus cisplatin (40 mg/m2 D 1 and 8); cisplatin alone (40 mg/m2 D 1 and 8; EudraCT: 2014-004655-31). Pharmacokinetic/pharmacodynamic effects were assessed in plasma and resected tumors. Primary end point; effect of Debio 1143 on cellular IAP-1 (cIAP-1). Levels of cIAP-1/-2, X-linked inhibitor of apoptosis protein (XIAP), tumor infiltrating lymphocytes (TILs), including CD8+ T cells, programmed cell death protein 1 (PD-1), PD-ligand 1 (PD-L1), and gene expression were also analyzed. Twenty-three of 26 patients completed treatment. In the Debio 1143 monotherapy cohort (n = 13), mean tumor concentrations of Debio 1143 were 18-fold (maximum 55.2-fold) greater than in plasma, exceeding the half-maximal inhibitory concentration for cIAPs and XIAP by 100 to 1000-fold, with significant engagement/degradation of cIAP-1 (p < 0.05). Overall, levels of CD8+ TILs, PD-1, and PD-L1 positive immune cells increased significantly (p < 0.05) following Debio 1143 treatment. Changes were observed in the expression of genes related to NF-κB signaling. Treatments were well-tolerated. Debio 1143 penetrated SCCHN tumors, engaged cIAP-1, and induced immune inflammatory changes in the tumor microenvironment. Based on the mode of action demonstrated here and in previous studies, these data support future combinations of Debio 1143 with immune-checkpoint agents.

Measuring motivational relationship processes in experience sampling: A reliability model for moments, days, and persons nested in couples.

The investigation of within-person process models, often done in experience sampling designs, requires a reliable assessment of within-person change. In this paper, we focus on dyadic intensive longitudinal designs where both partners of a couple are assessed multiple times each day across several days. We introduce a statistical model for variance decomposition based on generalizability theory (extending P. E. Shrout & S. P. Lane, 2012), which can estimate the relative proportion of variability on four hierarchical levels: moments within a day, days, persons, and couples. Based on these variance estimates, four reliability coefficients are derived: between-couples, between-persons, within-persons/between-days, and within-persons/between-moments. We apply the model to two dyadic intensive experience sampling studies (n1 = 130 persons, 5 surveys each day for 14 days, ≥ 7508 unique surveys; n2 = 508 persons, 5 surveys each day for 28 days, ≥ 47764 unique surveys). Five different scales in the domain of motivational processes and relationship quality were assessed with 2 to 5 items: State relationship satisfaction, communal motivation, and agentic motivation; the latter consists of two subscales, namely power and independence motivation. Largest variance components were on the level of persons, moments, couples, and days, where within-day variance was generally larger than between-day variance. Reliabilities ranged from .32 to .76 (couple level), .93 to .98 (person level), .61 to .88 (day level), and .28 to .72 (moment level). Scale intercorrelations reveal differential structures between and within persons, which has consequences for theory building and statistical modeling.

Prognostic value of intratumoral Fusobacterium nucleatum and association with immune-related gene expression in oral squamous cell carcinoma patients.

Changes in the oral microbiome, particularly Fusobacterium nucleatum, are associated with oral squamous cell carcinoma (OSCC). F. nucleatum has been reported to modulate local immunity in cancers. We aimed to assess the association between intratumoral F. nucleatum and clinico-pathological features, relapse, and overall survival (OS) in two independent cohorts of patients with OSCC, and to explore the interplay with immune-related genes. We retrospectively analyzed tissue samples from a first cohort of 122 patients with head and neck squamous cell carcinoma, including 61 OSCC (cohort #1), and a second cohort of 90 additional OSCC (cohort #2). We then performed a sensitivity analysis on the merged cohort of OSCC patients (N = 151). F. nucleatum 16S rRNA gene sequences were quantified using real-time quantitative PCR. The presence of gram-negative bacteria and macrophages was confirmed by LPS and CD163 immunostainings, respectively. F. nucleatum positivity was associated with older age, less alcohol and combined alcohol plus tobacco consumption, and less frequent lymph node invasion. There was a trend for a lower recurrence rate in F. nucleatum-positive cases, with less metastatic relapses compared to F. nucleatum-negative tumors, and significantly longer OS, relapse-free and metastasis-free survival. F. nucleatum status was independently associated with OS in multivariate analysis. Immune-related gene and immunohistochemistry analyses showed that gram-negative bacteria load inversely correlated with M2 macrophages. F. nucleatum-associated OSCC has a specific immune microenvironment, is more frequent in older, non-drinking patients, and associated with a favorable prognosis.