Despite the astonishing diversity of life, evolutionary biologists have documented numerous instances where species converge in how they behave, look and function. Given the importance of happenstance in evolution, it is often assumed that mechanisms driving independent evolution of similar phenotypes (convergence) are distinct; yet recent discoveries suggest that conserved genomic mechanisms can underlie convergent traits. Here, we generate forebrain transcriptomes from six sympatric Labridae wrasses that vary in mutualistic cleaning behaviour and apply differential gene expression, co-expression and phylogenetic comparative analyses to address three questions. First, do neurotranscriptomes vary among species with distinct cleaning behaviour? Second, do species that have independently evolved facultative cleaning share neurotranscriptomic patterns? Third, are transcriptomic signatures linked to facultative cleaning also expressed in obligate cleaners? We identify shared neurotranscriptomic patterns in the repeated evolution of facultative cleaning, including 25 novel candidate genes. We then find that transcriptomic patterns associated with facultative cleaning are shared in our focal obligate cleaner species, with more genes than expected by chance with concordant expression patterns in obligate and facultative cleaners. Our results illuminate the neuromolecular basis of cooperative behaviour and demonstrate the predictive potential of comparative transcriptomics to unravel mechanistic underpinnings of the repeated evolution of complex phenotypes.

Long-term memory formation transiently activates Ca2+-calmodulin kinase IIα and atypical protein kinase C isoform iota/lambda, whereas persistent activation of the other atypical PKC, protein kinase M zeta (PKMζ), together with its interacting partner, the scaffolding-protein KIBRA (Wwc1), are necessary for maintaining potentiated synapses and memory. Here, we use immediate-early gene (IEG) Arc activation during active place avoidance memory expression to tag memory-activated neurons with EYFP-ChR2. PKMζ immunohistochemistry identified which hippocampal synaptic pathways are persistently altered. EYFP-PKMζ colocalization persistently increases in the hippocampal trisynaptic pathway (dentate gyrus [DG]→CA3→CA1) tracing a one-month-old PKMζ engram. DG, CA3, and CA1 transcriptional profiling identifies that memory persistence correlates with upregulated immediate-early-genes Arc, Fos, and NPas4 in DG, but not with PKMζ or most genes known to be crucial for LTP and memory. This rules out strong memory-related transcriptional regulation, but not regulation of mRNA translation or altered stability of "shadow proteins" like PKMζ that, despite being crucial for memory maintenance, evade detection by unbiased transcriptome profiling. In contrast, our method Correlation Signal Co-cluster Reduction (C-SCoRe) incorporates weak linear and non-linear gene correlations and highlights network interaction changes predicting memory, and related IEG and Prkcz/Wwc1 expression. Manifold transcriptional relationships can reveal shadow molecular components of long-term memory.

Mothers, fathers, and offspring clash over how much care offspring receive. Offspring beg to solicit more food — but how much begging is rewarded can depend on who is listening. While both parents benefit from provisioning offspring, each benefits from their partner shouldering more of the burden of care, leading to sexual conflict. Additionally, if the costs and/or benefits of provisioning differ by sex, parent–offspring conflict should vary by sex. Whether there are sex differences in males’ and females’ responses to begging in birds, and to which extent evolutionary conflicts affect sex differences in parental response to begging, is not known. We therefore conducted a meta-analysis on 31 bird species, comparing sex differences in parental responses to begging to social and physiological traits that may influence conflict. Within species with stable bonds and/or low extra-pair paternity, and thus lower sexual and paternal-offspring conflict, males’ provisioning effort is more strongly correlated with offspring begging than females’. The opposite holds within species that change partners between breeding bouts and/or have high extra-pair paternity: females respond more to begging, perhaps compensating for males’ lower responsiveness. These results suggest that sex differences in parent–offspring communication can emerge from both sexual and parent–offspring conflict.

For small lung carcinomas, sublobar resections allow the preservation of a greater pulmonary reserve than after lobectomy. It was unclear for a long time to what extent this would jeopardize the goal of curative, radical tumor removal. Current studies show under what conditions a sublobar resection should be carried out and under which circumstances lobectomy continues to be the required standard.

Zusammenfassung Primäre Trachealmalignome sind selten, die beiden häufigsten histologischen Subtypen sind das Plattenepithelkarzinom und das adenoidzystische Karzinom. Beide Tumorentitäten unterscheiden sich hinsichtlich des lokalen Wachstumsmusters und der Prognose. Den diagnostischen Standard bilden Bronchoskopie und radiologische Schnittbildgebung. Bei fehlender Fernmetastasierung und technischer Resektabilität ist die Operation die Therapie der Wahl. Auch im Falle einer inkompletten Resektion kann, insbesondere beim adenoidzystischen Karzinom, durch eine postoperative Radiotherapie ein zufriedenstellendes Langzeitergebnis erzielt werden. Im Kontext der palliativen Symptomlinderung hat neben der Strahlentherapie die interventionelle Bronchoskopie (Tumorabtragung, Stentimplantation) einen relevanten Stellenwert.

Complex behavioral phenotypes, such as social status, emerge from the genome across biological levels, with many of the fundamental neural mechanisms shared across vertebrates. While various aspects of the brain have been implicated in modulating social behavior, critical regulators include cells of the preoptic area (POA) and hypothalamus, which by applying cellular- resolution transcriptomic approaches allows for greater exploration of cellular dynamics in these cells. Yet, how complex gene networks function between and within cell types to regulate complex social behavior is still poorly understood. Importantly, when considering functionally relevant neuronal classes of genes such as neuropeptides, understanding the inherent complexity that emerges from the interaction of these genes in the transcriptomic neuropeptidome can provide unique insight into how social behavior is regulated. Here, we used single-nucleus RNA-sequencing in the hypothalamus and POA of a highly social cichlid fish, Astatotilapia burtoni , to understand the effect that social status has on cellular-level transcriptomic profiles. Males of this species are well known for their highly plastic phenotypes related to social status, which allows for a hypothesis-driven approach. We demonstrate how social status manifests in changes of gene co-expression networks across neuronal populations and highlight transcriptomic signatures of social dominance when targeting known functional differences among AVP neuronal cell types. We implement a novel approach to relate how differences in social state translate to the integration of the transcriptomic neuropeptidome. Taken together, this research provides insights into how gene expression networks that modulate social behavior, including neuropeptide networks, function at the cellular level. Significance Statement:Here, we used single-nucleus RNA-seq in the hypothalamus and POA of Astatotilapia burtoni to understand the effect that social status has on cellular gene expression. We demonstrate how social status manifests, from changes in broader neuronal gene networks to targeted changes between known socially-relevant neurons. For the first time, we assess the entirety of the transcriptomic neuropeptidome to understand the interaction of neuropeptide gene networks with social dominance. These findings provide a valuable resource for future functional work and an analytical framework for comparative studies on the evolution of the neural mechanisms of social behavior. Insights into the transcriptomic networks that modulate social status, specifically with neuropeptides, aid in our understanding of the complexity inherent in social behavior.

Social hierarchies are a common form of social organization across species. Although hierarchies are largely stable across time, animals may socially ascend or descend within hierarchies depending on environmental and social challenges. Here, we develop a novel paradigm to study social ascent and descent within male CD-1 mouse social hierarchies. We show that mice of all social ranks rapidly establish new stable social hierarchies when placed in novel social groups with animals of equivalent social status. Seventy minutes following social hierarchy formation, males that were socially dominant prior to being placed into new social hierarchies exhibit higher increases in plasma corticosterone and vastly greater transcriptional changes in the medial amygdala (MeA), which is central to the regulation of social behavior, compared to males who were socially subordinate prior to being placed into a new hierarchy. Specifically, the loss of social status in a new hierarchy (social descent) is associated with reductions in MeA expression of myelination and oligodendrocyte differentiation genes. Maintaining high social status is associated with high expression of genes related to cholinergic signaling in the MeA. Conversely, gaining social status in a new hierarchy (social ascent) is related to relatively few unique rapid changes in the MeA. We also identify novel genes associated with social transition that show common changes in expression when animals undergo either social descent or social ascent compared to maintaining their status. Two genes, Myosin binding protein C1 (Mybpc1) and μ-Crystallin (Crym), associated with vasoactive intestinal polypeptide (VIP) and thyroid hormone pathways respectively, are highly upregulated in socially transitioning individuals. Further, increases in genes associated with synaptic plasticity, excitatory glutamatergic signaling and learning and memory pathways were observed in transitioning animals suggesting that these processes may support rapid social status changes.

Mate choice plays a fundamental role in speciation, yet we know little about the molecular mechanisms that underpin this crucial decision-making process. Stickleback fish differentially adapted to limnetic and benthic habitats are reproductively isolated and females of each species use different male traits to evaluate prospective partners and reject heterospecific males. Here, we integrate behavioural data from a mate choice experiment with gene expression profiles from the brains of females actively deciding whether to mate. We find substantial gene expression variation between limnetic and benthic females, regardless of behavioural context, suggesting general divergence in constitutive gene expression patterns, corresponding to their genetic differentiation. Intriguingly, female gene co-expression modules covary with male display traits but in opposing directions for sympatric populations of the two species, suggesting male displays elicit a dynamic neurogenomic response that reflects known differences in female preferences. Furthermore, we confirm the role of numerous candidate genes previously implicated in female mate choice in other species, suggesting evolutionary tinkering with these conserved molecular processes to generate divergent mate preferences. Taken together, our study adds important new insights to our understanding of the molecular processes underlying female decision-making critical for generating sexual isolation and speciation.

Background/Objectives: Lung cancer remains a global health concern, with substantial variation in patient survival. Despite advances in detection and treatment, the genetic basis for the divergent outcomes is not understood. We investigated germline polymorphisms that modulate overall survival in 1464 surgically resected lung adenocarcinoma patients. Methods: A multivariable Cox proportional hazard model was used to assess the association of more than seven million polymorphisms with overall survival at the 60-month follow-up, considering age, sex, pathological stage, decade of surgery and principal components as covariates. Genes in which variants were identified were studied in silico to investigate functional roles. Results: Six germline variants passed the genome-wide significance threshold. These single nucleotide polymorphisms were mapped to non-coding (intronic) regions on chromosomes 2, 3, and 5. The minor alleles of rs13000315, rs151212827, and rs190923216 (chr. 2, 3 and 5, respectively) were found to be independent negative prognostic factors. All six variants have been reported to regulate the expression of nine genes, seven of which are protein-coding, in different tissues. Survival-associated variants on chromosomes 2 and 3 were already reported to regulate the expression of NT5DC2 and NAGK, with high expression associated with the minor alleles. High NT5DC2 and NAGK expression in lung adenocarcinoma tissue was already shown to correlate with poor overall survival. Conclusions: This study highlights a potential regulatory role of the identified polymorphisms in influencing outcome and suggests a mechanistic link between these variants, gene expression regulation, and lung adenocarcinoma prognosis. Validation and functional studies are warranted to elucidate the mechanisms underlying these associations.

OBJECTIVES:The study aimed to assess the opinion of pathologists and thoracic surgeons of the International Association for the Study of Lung Cancer regarding the application and interpretation of the residual tumor (R) classification for lung cancer. METHODS:On the basis of their membership profiles, a total of 623 pathologists and thoracic surgeons were identified and contacted by email with a cover letter and a link to an online survey. The questionnaire consisted of 12 questions about various aspects of the application and interpretation of the R classification for lung cancer. The response rate (to at least one question) was 72% (144 pathologists and 303 surgeons). RESULTS:The frequency of use of the R classification varies by geographic region. Although R status is regularly reported in Europe and Asia, 70% of pathologists in the United States and Canada never include R status in reports. Similar variations exist about who assigns the R category for the resection-in Europe and the United Kingdom, it is mainly the pathologist, whereas in China, Japan and the United States, it is the surgeon. There are some good agreements about margins examined and how to manage staple lines. The category "uncertain resection" has not been practically implemented in most of the world, except at some centers in Japan and the United Kingdom. CONCLUSION:This survey shows that surgical resection margins are part of routine reporting in most institutions; but the assignment of an R category is not always part of the pathology report, with considerable variation between countries. Despite the International Association for the Study of Lung Cancer proposals, the application of the uncertain resection category has not been taken up by most institutions, and further evidence is needed.

Among vertebrates, ray-finned fishes (Actinopterygii) display the highest diversity in parental care, and their diversification has been hypothesized to be related to phylogenetic changes in fertilization modes. Using the most comprehensive, sex-specific data from 7,600 species of 62 extant orders of ray-finned fishes, we inferred ancestral states and transitions among care types and caring episodes (i.e., the stage of offspring development). Our work has uncovered 3 novel findings. First, transitions among different care types (i.e., male-only care, female-only care, biparental care, and no care) are common, and the frequencies of these transitions show unusually diverse patterns concerning fertilization modes (external, or internal via oviduct, mouth, or brood pouch). Second, both oviduct and mouth fertilization are selected for female-biased care, whereas fertilization in a brood pouch is selected for male-biased care. Importantly, internal fertilization without parental care is extremely unstable phylogenetically. Third, we show that egg care in both sexes is associated with nest building (which is male-biased) and fry care (which is female-biased). Taken together, the aquatic environment, which supports considerable flexibility in care, facilitated the diversification of parenting behavior, creating the evolutionary bases for more comprehensive parenting to protect offspring in semiterrestrial or terrestrial environments.

The goal of surgical resection is to completely remove a cancer; it is useful to have a system to describe how well this was accomplished. This is captured by the residual tumor (R) classification, which is separate from the TNM classification that describes the anatomic extent of a cancer independent of treatment. The traditional R-classification designates as R0 a complete resection, as R1 a macroscopically complete resection but with microscopic tumor at the surgical margin, and as R2 a resection that leaves gross tumor behind. For lung cancer, an additional category encompasses situations in which the presence of residual tumor is uncertain. This paper represents a comprehensive review of evidence regarding these R categories and the descriptors thereof, focusing on studies published after the year 2000 and with adjustment for potential confounders. Consistent discrimination between complete, uncertain, and incomplete resection is demonstrated with respect to overall survival. Evidence regarding specific descriptors is generally somewhat limited and only partially consistent; nevertheless, the data suggests retaining all descriptors but with clarifications to address ambiguities. Based on this review, the R-classification for the 9th edition of stage classification of lung cancer is proposed to retain the same overall framework and descriptors, with more precise definitions of descriptors. These refinements should facilitate application as well as further research.

Abstract Background Reports from case series suggest that operative outcomes are comparable amongst different age groups following surgery with curative intent for non-small cell lung cancer (NSCLC). The purpose of this study was to compare morbidity and mortality after NSCLC surgery in older patients (≥ 75 years) versus younger patients (< 75 years) and identify independent predictive risk factors. Methods We identified 2015 patients with postoperative stages IA to IIIA according to AJCC/UICC 7th edition who had undergone NSCLC surgery with curative intent at a single specialized lung cancer center from January 2010 to December 2015. A matched-pair analysis was performed on 227 older patients and corresponding 227 younger patients. Short-term surgical outcomes were postoperative morbidity, length of hospital stay, 30-day and 90-day mortality. Long-term operative outcomes were disease-free and overall survival. Results 454 patients were included in the matched-pair analysis. 36% of younger patients developed postoperative complications versus 42% in older patients (p = 0.163). Age was not significantly associated with the occurrence of postoperative complications. Median length of hospital stay was 14 days in older patients and 13 days in younger patients (p = 0.185). 90-day mortality was 2.2% in younger patients compared to 4% in older patients (p = 0.424). In patients aged 75 and older impaired performance status (ECOG ≥ 1) was associated with decreased overall survival (HR = 2.15, CI 1.34–3.46), as were preoperative serum C-reactive protein / albumin ratio ≥ 0.3 (HR = 1.95, CI 1.23–3.11) and elevated preoperative serum creatinine levels ≥ 1.1 mg/dl (HR = 1.84, CI 1.15–2.95). In the younger cohort male sex (HR = 2.26, CI 1.17–4.36), postoperative stage III disease (HR 4.61, CI 2.23–9.54) and preoperative anemia (hemoglobin < 12 g/dl) (HR 2.09, CI 1.10–3.96) were associated with decreased overall survival. Conclusions Lung resection for NSCLC in older patients is associated with postoperative morbidity and mortality comparable to those of younger patients. In older patients, physical activity, comorbidities and nutritional status are related to survival and should influence the indication for surgery rather than age alone.

The process of implementing early detection of lung cancer with low-dose CT (LDCT) in Germany has gained significant momentum in recent years. It is expected that the ordinance of the Federal Ministry for the Environment, Nature Conservation, Nuclear Safety and Consumer Protection (BMUV) on early detection of lung cancer, which has been commented on by the professional societies, will come into effect by the end of 2023. Based on this regulation, the Federal Joint Committee (G-BA) will set up a program for early lung cancer detection with LDCT in the near future. In this position paper, the specialist societies involved in lung cancer screening present concrete cornerstones for a uniform, structured and quality-assured early detection program for lung cancer in Germany to make a constructive contribution to this process.

Abstract Background In 2022, an update of the German lung cancer guideline, first published in 2010 and revised in 2018, was released. This article aims to show the process of updating, developing, and implementing guideline-based quality indicators (QI) into the certification system for lung cancer centers (LCC). Methods A multidisciplinary and interprofessional working group revised the guideline QIs from 2018 using the strong recommendations of the guideline update, a systematic review for QIs, and the results of the implemented QIs from LCC. Results For 4 out of 8 indicators from the 2018 guideline, the LCC showed an improved implementation of the requirements in the last 3 years (2018–2020). For 3 indicators, the median of the results was constant at a very high level (≥96% or 100%). Only the “adjuvant cisplatin-based chemotherapy” indicator showed declining values between 2018 and 2020. The target values and plausibility limits were well achieved by LCC. After updating the guideline, one QI from 2018 was not included in the new QI set due to the small denominator population. Based on the new strong recommendations, 8 new QIs were defined. From the QI set of the guideline update, 13 of 15 indicators (7 since 2018 and 6 from 2022 on) were adopted into the certification program. Conclusions The guideline recommendations are implemented by LCC at a high level. The process presented confirms the successful implementation of the so-called quality cycle in oncology. The QIs developed by the German Guideline Program in Oncology (GGPO) are adopted by the certification program. The implementation of the QI is measured in LCC, evaluated by the German Cancer Society (DKG), and reflected back to the GGPO. The “real world” data have led to the deletion of one QI and show a high implementation of most QIs in LCC.

How does complex behavior arise from the genome across biological levels? Specific cell types have evolutionarily conserved functional roles in regulating and maintaining various aspects of social behavior. There is a long history of research demonstrating how specific cells in the hypothalamus and POA are critical in regulating social status and reproductive behavior. This cell function is dependent not just on one gene, but the entire gene expression network working within each cell. The brain is a heterogenous tissue made up of a broad diversity of cells. Targeting the cellular-level transcriptomes in socially relevant brain regions enables the identification of social status mediators. Single-nuclei RNA-sequencing (snRNA-seq) provides the resolution to identify cell-specific transcriptomes, with more practical benefits compared to single-cell approaches. Single-nucleus sequencing of neuronal tissue has remained fairly limited to model mammalian organisms. Here we performed snRNA-seq in the hypothalamus and POA of Astatotilapia burtoni, a fish system ideal for social neuroscience. Male A. burtoni are characterized by plastic phenotypes associated with social status, dominant and subordinate, with well-established differences in neural gene expression patterns and specific neuron morphology. This provides a novel opportunity to apply snRNA-seq in a complex tissue of a non-model system using hypothesis driven approaches. We show how changes in social status are linked to distinct transcriptomic profiles at the cellular level. ### Competing Interest Statement The authors have declared no competing interest.

Malignant primary tracheal tumours are rare. The most common histological subtypes are squamous cell carcinoma and adenoid cystic carcinoma. These two entities have different prognoses and growth patterns. Tracheobronchoscopy and thoracic sectional imaging are standard diagnostic tools for tumour staging and local evaluation. Complete surgical resection of the affected tracheal segment is the treatment of choice for limited disease without distant metastases. Incomplete gross tumour resection with additional irradiation is an acceptable therapeutic option for adenoid cystic carcinoma. Interventional endoscopy with tumour debulking or tracheal stenting and/or definitive mediastinal radiotherapy are treatment alternatives in either a locally advanced or palliative setting.

Contextual fear conditioning has been shown to activate a set of "fear ensemble" cells in the hippocampal dentate gyrus (DG) whose reactivation is necessary and sufficient for expression of contextual fear. We previously demonstrated that extinction learning suppresses reactivation of these fear ensemble cells and activates a competing set of DG cells - the "extinction ensemble." Here, we tested whether extinction was sufficient to suppress reactivation in other regions and used single nucleus RNA sequencing (snRNA-seq) of cells in the dorsal dentate gyrus to examine how extinction affects the transcriptomic activity of fear ensemble and fear recall-activated cells. Our results confirm the suppressive effects of extinction in the dorsal and ventral dentate gyrus and demonstrate that this same effect extends to fear ensemble cells located in the dorsal CA1. Interestingly, the extinction-induced suppression of fear ensemble activity was not detected in ventral CA1. Our snRNA-seq analysis demonstrates that extinction training markedly changes transcription patterns in fear ensemble cells and that cells activated during recall of fear and recall of extinction have distinct transcriptomic profiles. Together, our results indicate that extinction training suppresses a broad portion of the fear ensemble in the hippocampus, and this suppression is accompanied by changes in the transcriptomes of fear ensemble cells and the emergence of a transcriptionally unique extinction ensemble.

Zusammenfassung Der Prozess zur Implementierung der Lungenkrebsfrüherkennung mit Niedrigdosis-CT (LDCT) in Deutschland hat in den letzten Jahren deutlich an Dynamik gewonnen. Es wird erwartet, dass der von den Fachgesellschaften kommentierte Referentenentwurf des Bundesministeriums für Umwelt, Naturschutz, nukleare Sicherheit und Verbraucherschutz (BMUV) zur Lungenkrebsfrüherkennung noch bis Ende 2023 in Kraft tritt. Basierend auf dieser Verordnung wird der Gemeinsame Bundesausschuss (G-BA) zeitnah ein Programm zur Lungenkrebsfrüherkennung mit LDCT aufsetzten. In diesem Positionspapier stellen die im Lungenkrebsscreening beteiligten Fachgesellschaften erstmals konkrete Eckpunkte für ein einheitliches, strukturiertes und qualitätsgesichertes Früherkennungsprogramm für Lungenkrebs in Deutschland vor, um zu diesem Prozess konstruktiv beizutragen.