Evaluating implicit gender bias at Canadian otolaryngology meetings through use of professional title.

Objectives: Increasing numbers of women enter medical school annually. The number of female physicians in leadership positions has been much slower to equalize. There are also well-documented differences in the treatment of women as compared to men in professional settings. Female presenters are less likely to be introduced by their professional title ("Doctor") for grand rounds and conferences, especially with a man performing the introduction. This study reviewed the Canadian Society of Otolaryngology-Head and Neck Surgery (CSOHNS) meetings from 2017 to 2020 to determine the proportion of presenters introduced by their professional title and whether this varied by gender. Methods: Recordings from CSOHNS meetings were reviewed and coded for introducer and presenter demographics, including leadership positions and gender. Chi-squared tests of proportion and multivariate logistic regression was used to compare genders and identify factors associated with professional versus unprofessional forms of address. Results: No significant association was found between professional title use and introducer or presenter gender. Female presenters were introduced with professional title 69.6% of the time, while male presenters were introduced with professional title 67.6% of the time (P = 0.69). Residents were introduced with a professional title with the most frequency (75.8%), while attending staff were introduced with a professional title with the least frequency (63.0%) (P = 0.02). Conclusions: The lack of gender bias in speaker introductions at recent CSOHNS meetings demonstrates progress in achieving gender equity in medicine. Research efforts should continue to define additional forms of unconscious bias that may be contributing to gender inequity in leadership positions.

Sortase-Modified Cholera Toxoids Show Specific Golgi Localization.

Cholera toxoid is an established tool for use in cellular tracing in neuroscience and cell biology. We use a sortase labeling approach to generate site-specific N-terminally modified variants of both the A2-B5 heterohexamer and B5 pentamer forms of the toxoid. Both forms of the toxoid are endocytosed by GM1-positive mammalian cells, and while the heterohexameric toxoid was principally localized in the ER, the B5 pentamer showed an unexpectedly specific localization in the medial/trans-Golgi. This study suggests a future role for specifically labeled cholera toxoids in live-cell imaging beyond their current applications in neuronal tracing and labeling of lipid rafts in fixed cells.

An unexpected case of broncholithiasis secondary to pulmonary nocardiosis.

Persistent productive cough despite appropriate treatment warrants consideration of flexible bronchoscopy to obtain bronchial specimens for culture. Endobronchial examination of airways may reveal signs of infection in the form of purulent secretions, sputum plugs or in this case, an unexpected finding of a calcified broncholithiasis secondary to Nocardia infection.

E7-mediated repression of miR-203 promotes LASP1-dependent proliferation in HPV-positive cervical cancer.

Human papillomaviruses (HPV) are a major cause of malignancy, contributing to ∼5% of all human cancers worldwide, including most cervical cancer cases and a growing number of ano-genital and oral cancers. The major HPV viral oncogenes, E6 and E7, manipulate many host cellular pathways that promote cell proliferation and survival, predisposing infected cells to malignant transformation. Despite the availability of highly effective vaccines, there are still no specific anti-viral therapies targeting HPV or treatments for HPV-associated cancers. As such, a better understanding of viral-host interactions may allow the identification of novel therapeutic targets. Here, we demonstrate that the actin-binding protein LASP1 is upregulated in cervical cancer and significantly correlates with a poorer overall survival. In HPV positive cervical cancer, LASP1 depletion significantly inhibited proliferation in vitro , whilst having minimal effects in HPV negative cervical cancer cells. Furthermore, we show that the LASP1 SH3 domain is essential for LASP1-mediated proliferation in these cells. Mechanistically, we show that HPV E7 regulates LASP1 at the post-transcriptional level by repressing the expression of miR-203, which negatively regulated LASP1 mRNA levels by binding to its 3'UTR. Finally, we demonstrated that LASP1 expression is required for the growth of HPV positive cervical cancer cells in an in vivo tumourigenicity model. Together, these data demonstrate that HPV induces LASP1 expression to promote proliferation and survival role in cervical cancer, thus identifying a potential therapeutic target in these cancers.

Chemokines form complex signals during inflammation and disease that can be decoded by extracellular matrix proteoglycans.

Chemokine-driven leukocyte recruitment is a key component of the immune response and of various diseases. Therapeutically targeting the chemokine system in inflammatory disease has been unsuccessful, which has been attributed to redundancy. We investigated why chemokines instead have specific, specialized functions, as demonstrated by multiple studies. We analyzed the expression of genes encoding chemokines and their receptors across species, tissues, and diseases. This analysis revealed complex expression patterns such that genes encoding multiple chemokines that mediated recruitment of the same leukocyte type were expressed in the same context, such as the genes encoding the CXCR3 ligands CXCL9, CXCL10, and CXCL11. Through biophysical approaches, we showed that these chemokines differentially interacted with extracellular matrix glycosaminoglycans (ECM GAGs), which was enhanced by sulfation of specific GAGs. Last, in vivo approaches demonstrated that GAG binding was critical for the CXCL9-dependent recruitment of specific T cell subsets but not of others, irrespective of CXCR3 expression. Our data demonstrate that interactions with ECM GAGs regulated whether chemokines were presented on cell surfaces or remained more soluble, thereby affecting chemokine availability and ensuring specificity of chemokine action. Our findings provide a mechanistic understanding of chemokine-mediated immune cell recruitment and identify strategies to target specific chemokines during inflammatory disease.

Synthesis of the Marine Alkaloid Cylindricine C and Serendipitous Synthesis of Its 2,13-Di-epi Stereoisomer.

A new approach to the marine alkaloid cylindricine C afforded its previously unreported (±)-2,13-di-epi stereoisomer as the major product along with a minor amount of the racemic parent alkaloid. Key steps included a stereoselective dianion alkylation of a monoester of 1,2-cyclohexanedicarboxylic acid and an annulation based on the tandem conjugate addition of a primary amine to an acetylenic sulfone, followed by intramolecular acylation of the resulting sulfone-stabilized carbanion. The cis-azadecalin moiety thus formed, comprising the cyclohexane A-ring and enaminone B-ring of the products, was further elaborated by the selenenyl chloride-induced cyclofunctionalization of a pendant butenyl substituent with the enaminone moiety, followed by a seleno-Pummerer reaction. Desulfonylation and enaminone reduction afforded the final products. Molecular modeling and X-ray crystallography provided further insight into these processes.

Corrigendum: Dynamics of host immune response development during Schistosoma mansoni infection.

[This corrects the article DOI: 10.3389/fimmu.2022.906338.].

CREB1 activation promotes human papillomavirus oncogene expression and cervical cancer cell transformation.

Human papillomaviruses (HPVs) infect the oral and anogenital mucosa and can cause cancer. The high-risk (HR)-HPV oncoproteins, E6 and E7, hijack cellular factors to promote cell proliferation, delay differentiation and induce genomic instability, thus predisposing infected cells to malignant transformation. cAMP response element (CRE)-binding protein 1 (CREB1) is a master transcription factor that can function as a proto-oncogene, the abnormal activity of which is associated with multiple cancers. However, little is known about the interplay between HPV and CREB1 activity in cervical cancer or the productive HPV lifecycle. We show that CREB is activated in productively infected primary keratinocytes and that CREB1 expression and phosphorylation is associated with the progression of HPV+ cervical disease. The depletion of CREB1 or inhibition of CREB1 activity results in decreased cell proliferation and reduced expression of markers of epithelial to mesenchymal transition, coupled with reduced migration in HPV+ cervical cancer cell lines. CREB1 expression is negatively regulated by the tumor suppressor microRNA, miR-203a, and CREB1 phosphorylation is controlled through the MAPK/MSK pathway. Crucially, CREB1 directly binds the viral promoter to upregulate transcription of the E6/E7 oncogenes, establishing a positive feedback loop between the HPV oncoproteins and CREB1. Our findings demonstrate the oncogenic function of CREB1 in HPV+ cervical cancer and its relationship with the HPV oncogenes.

Comparative phenotype of circulating versus tissue immune cells in human lung and blood compartments during health and disease.

The lung is a dynamic mucosal surface constantly exposed to a variety of immunological challenges including harmless environmental antigens, pollutants, and potentially invasive microorganisms. Dysregulation of the immune system at this crucial site is associated with a range of chronic inflammatory conditions including asthma and Chronic Pulmonary Obstructive Disease (COPD). However, due to its relative inaccessibility, our fundamental understanding of the human lung immune compartment is limited. To address this, we performed flow cytometric immune phenotyping of human lung tissue and matched blood samples that were isolated from 115 donors undergoing lung tissue resection. We provide detailed characterization of the lung mononuclear phagocyte and T cell compartments, demonstrating clear phenotypic differences between lung tissue cells and those in peripheral circulation. Additionally, we show that CD103 expression demarcates pulmonary T cells that have undergone recent TCR and IL-7R signalling. Unexpectedly, we discovered that the immune landscape from asthmatic or COPD donors was broadly comparable to controls. Our data provide a much-needed expansion of our understanding of the pulmonary immune compartment in both health and disease.

Human footprint is associated with shifts in the assemblages of major vector-borne diseases.

Predicting how increasing intensity of human-environment interactions affects pathogen transmission is essential to anticipate changing disease risks and identify appropriate mitigation strategies. Vector-borne diseases (VBDs) are highly responsive to environmental changes, but such responses are notoriously difficult to isolate because pathogen transmission depends on a suite of ecological and social responses in vectors and hosts that may differ across species. Here we use the emerging tools of cumulative pressure mapping and machine learning to better understand how the occurrence of six medically important VBDs, differing in ecology from sylvatic to urban, respond to multidimensional effects of human pressure. We find that not only is human footprint-an index of human pressure, incorporating built environments, energy and transportation infrastructure, agricultural lands and human population density-an important predictor of VBD occurrence, but there are clear thresholds governing the occurrence of different VBDs. Across a spectrum of human pressure, diseases associated with lower human pressure, including malaria, cutaneous leishmaniasis and visceral leishmaniasis, give way to diseases associated with high human pressure, such as dengue, chikungunya and Zika. These heterogeneous responses of VBDs to human pressure highlight thresholds of land-use transitions that may lead to abrupt shifts in infectious disease burdens and public health needs.

Exploitation of ATP-sensitive potassium ion (KATP) channels by HPV promotes cervical cancer cell proliferation by contributing to MAPK/AP-1 signalling.

Persistent infection with high-risk human papillomaviruses (HPVs) is the causal factor in multiple human malignancies, including >99% of cervical cancers and a growing proportion of oropharyngeal cancers. Prolonged expression of the viral oncoproteins E6 and E7 is necessary for transformation to occur. Although some of the mechanisms by which these oncoproteins contribute to carcinogenesis are well-characterised, a comprehensive understanding of the signalling pathways manipulated by HPV is lacking. Here, we present the first evidence to our knowledge that the targeting of a host ion channel by HPV can contribute to cervical carcinogenesis. Through the use of pharmacological activators and inhibitors of ATP-sensitive potassium ion (KATP) channels, we demonstrate that these channels are active in HPV-positive cells and that this activity is required for HPV oncoprotein expression. Further, expression of SUR1, which forms the regulatory subunit of the multimeric channel complex, was found to be upregulated in both HPV+ cervical cancer cells and in samples from patients with cervical disease, in a manner dependent on the E7 oncoprotein. Importantly, knockdown of SUR1 expression or KATP channel inhibition significantly impeded cell proliferation via induction of a G1 cell cycle phase arrest. This was confirmed both in vitro and in in vivo tumourigenicity assays. Mechanistically, we propose that the pro-proliferative effect of KATP channels is mediated via the activation of a MAPK/AP-1 signalling axis. A complete characterisation of the role of KATP channels in HPV-associated cancer is now warranted in order to determine whether the licensed and clinically available inhibitors of these channels could constitute a potential novel therapy in the treatment of HPV-driven cervical cancer.

m6A Regulates the Stability of Cellular Transcripts Required for Efficient KSHV Lytic Replication.

The epitranscriptomic modification N6-methyladenosine (m6A) is a ubiquitous feature of the mammalian transcriptome. It modulates mRNA fate and dynamics to exert regulatory control over numerous cellular processes and disease pathways, including viral infection. Kaposi's sarcoma-associated herpesvirus (KSHV) reactivation from the latent phase leads to the redistribution of m6A topology upon both viral and cellular mRNAs within infected cells. Here we investigate the role of m6A in cellular transcripts upregulated during KSHV lytic replication. Our results show that m6A is crucial for the stability of the GPRC5A mRNA, whose expression is induced by the KSHV latent-lytic switch master regulator, the replication and transcription activator (RTA) protein. Moreover, we demonstrate that GPRC5A is essential for efficient KSHV lytic replication by directly regulating NFκB signalling. Overall, this work highlights the central importance of m6A in modulating cellular gene expression to influence viral infection.

Long-term trends in yield variance of temperate managed grassland.

The management of climate-resilient grassland systems is important for stable livestock fodder production. In the face of climate change, maintaining productivity while minimizing yield variance of grassland systems is increasingly challenging. To achieve climate-resilient and stable productivity of grasslands, a better understanding of the climatic drivers of long-term trends in yield variance and its dependence on agronomic inputs is required. Based on the Park Grass Experiment at Rothamsted (UK), we report for the first time the long-term trends in yield variance of grassland (1965-2018) in plots given different fertilizer and lime applications, with contrasting productivity and plant species diversity. We implemented a statistical model that allowed yield variance to be determined independently of yield level. Environmental abiotic covariates were included in a novel criss-cross regression approach to determine climatic drivers of yield variance and its dependence on agronomic management. Our findings highlight that sufficient liming and moderate fertilization can reduce yield variance while maintaining productivity and limiting loss of plant species diversity. Plots receiving the highest rate of nitrogen fertilizer or farmyard manure had the highest yield but were also more responsive to environmental variability and had less plant species diversity. We identified the days of water stress from March to October and temperature from July to August as the two main climatic drivers, explaining approximately one-third of the observed yield variance. These drivers helped explain consistent unimodal trends in yield variance-with a peak in approximately 1995, after which variance declined. Here, for the first time, we provide a novel statistical framework and a unique long-term dataset for understanding the trends in yield variance of managed grassland. The application of the criss-cross regression approach in other long-term agro-ecological trials could help identify climatic drivers of production risk and to derive agronomic strategies for improving the climate resilience of cropping systems. Supplementary Information: The online version contains supplementary material available at 10.1007/s13593-023-00885-w.

Spontaneous pulmonary herniation in COVID-19.

Spontaneous pulmonary hernia (SPH) is a rare condition. We report a case secondary to extreme coughing and COVID-19 infection. The patient displayed several clinical features typical of this diagnosis; difficult to manage pain on coughing, flank haematoma and bulging of the chest wall on coughing. Clinicians should be aware of the risk factors and clinical features of SPH to aid diagnosis of this rare condition.

Mycobacterium vaccae NCTC 11659, a Soil-Derived Bacterium with Stress Resilience Properties, Modulates the Proinflammatory Effects of LPS in Macrophages.

Inflammatory conditions, including allergic asthma and conditions in which chronic low-grade inflammation is a risk factor, such as stress-related psychiatric disorders, are prevalent and are a significant cause of disability worldwide. Novel approaches for the prevention and treatment of these disorders are needed. One approach is the use of immunoregulatory microorganisms, such as Mycobacterium vaccae NCTC 11659, which have anti-inflammatory, immunoregulatory, and stress-resilience properties. However, little is known about how M. vaccae NCTC 11659 affects specific immune cell targets, including monocytes, which can traffic to peripheral organs and the central nervous system and differentiate into monocyte-derived macrophages that, in turn, can drive inflammation and neuroinflammation. In this study, we investigated the effects of M. vaccae NCTC 11659 and subsequent lipopolysaccharide (LPS) challenge on gene expression in human monocyte-derived macrophages. THP-1 monocytes were differentiated into macrophages, exposed to M. vaccae NCTC 11659 (0, 10, 30, 100, 300 µg/mL), then, 24 h later, challenged with LPS (0, 0.5, 2.5, 250 ng/mL), and assessed for gene expression 24 h following challenge with LPS. Exposure to M. vaccae NCTC 11659 prior to challenge with higher concentrations of LPS (250 ng/mL) polarized human monocyte-derived macrophages with decreased IL12A, IL12B, and IL23A expression relative to IL10 and TGFB1 mRNA expression. These data identify human monocyte-derived macrophages as a direct target of M. vaccae NCTC 11659 and support the development of M. vaccae NCTC 11659 as a potential intervention to prevent stress-induced inflammation and neuroinflammation implicated in the etiology and pathophysiology of inflammatory conditions and stress-related psychiatric disorders.

T helper 2 cells control monocyte to tissue-resident macrophage differentiation during nematode infection of the pleural cavity.

The recent revolution in tissue-resident macrophage biology has resulted largely from murine studies performed in C57BL/6 mice. Here, using both C57BL/6 and BALB/c mice, we analyze immune cells in the pleural cavity. Unlike C57BL/6 mice, naive tissue-resident large-cavity macrophages (LCMs) of BALB/c mice failed to fully implement the tissue-residency program. Following infection with a pleural-dwelling nematode, these pre-existing differences were accentuated with LCM expansion occurring in C57BL/6, but not in BALB/c mice. While infection drove monocyte recruitment in both strains, only in C57BL/6 mice were monocytes able to efficiently integrate into the resident pool. Monocyte-to-macrophage conversion required both T cells and interleukin-4 receptor alpha (IL-4Rα) signaling. The transition to tissue residency altered macrophage function, and GATA6+ tissue-resident macrophages were required for host resistance to nematode infection. Therefore, during tissue nematode infection, T helper 2 (Th2) cells control the differentiation pathway of resident macrophages, which determines infection outcome.

The role of the host gut microbiome in the pathophysiology of schistosomiasis.

The pathophysiology of schistosomiasis is linked to the formation of fibrous granulomas around eggs that become trapped in host tissues, particularly the intestines and liver, during their migration to reach the lumen of the vertebrate gut. While the development of Schistosoma egg-induced granulomas is the result of finely regulated crosstalk between egg-secreted antigens and host immunity, evidence has started to emerge of the likely contribution of an additional player-the host gut microbiota-to pathological processes that culminate with the formation of these tissue lesions. Uncovering the role(s) of schistosome-mediated changes in gut microbiome composition and function in granuloma formation and, more broadly, in the pathophysiology of schistosomiasis, will shed light on the mechanisms underlying this three-way parasite-host-microbiome interplay. Such knowledge may, in turn, pave the way towards the discovery of novel therapeutic targets and control strategies.

Head and Neck Radiation Therapy Patterns of Practice Variability Identified as a Challenge to Real-World Big Data: Results From the Learning from Analysis of Multicentre Big Data Aggregation (LAMBDA) Consortium.

Purpose: Outside of randomized clinical trials, it is difficult to develop clinically relevant evidence-based recommendations for radiation therapy (RT) practice guidelines owing to lack of comprehensive real-world data. To address this knowledge gap, we formed the Learning from Analysis of Multicenter Big Data Aggregation consortium to cooperatively implement RT data standardization, develop software solutions for data analysis, and recommend clinical practice change based on real-world data analyzed. The first phase of this "Big Data" study aimed at characterizing variability in clinical practice patterns of dosimetric data for organs at risk (OARs) that would undermine subsequent use of large-scale, electronically aggregated data to characterize associations with outcomes. Evidence from this study was used as the basis for practical recommendations to improve data quality. Methods and Materials: Dosimetric details of patients with head and neck cancer treated with radiation therapy between 2014 and 2019 were analyzed. Institutional patterns of practice were characterized, including structure nomenclature, volumes, and frequency of contouring. Dose volume histogram (DVH) distributions were characterized and compared with institutional constraints and literature values. Results: Plans for 4664 patients treated to a mean plan dose of 64.4 ± 13.2 Gy in 32 ± 4 fractions were aggregated. Before implementation of TG-263 guidelines in each institution, there was variability in OAR nomenclature across institutions and structures. With evidence from this study, we identified a targeted and practical set of recommendations aimed at improving the quality of real-world data. Conclusions: Quantifying similarities and differences among institutions for OAR structures and DVH metrics is the launching point for next steps to investigate potential relationships between DVH parameters and patient outcomes.

The posterior crescent sign on MRI and MR arthrography: is it a marker of hip dysplasia and instability?

OBJECTIVE: To evaluate the prevalence of the 'posterior crescent sign' in symptomatic patients referred for MRI/MR arthrogram of the hip and identify any correlation with imaging features of joint pathology. MATERIALS AND METHODS: Retrospective imaging assessment of a cohort of 1462 hips, from 1380 included MR examinations (82 bilateral) retrieved from a search of all examinations in patients 16-50 years old from June 2018 to June 2021, with median age 45.8 years (range 17.8-50.0) and 936 hips (64%) in women. Radiographic and MR findings related to hip dysplasia, femoroacetabular impingement and osteoarthritis were assessed. RESULTS: Fifty-one hips (3.5%) were positive for the posterior crescent sign, median age of 45.8 years (range 17.8-50.0) and 29 (58%) in women. Radiographic findings included the following: mean lateral centre edge angle (LCEA) 22.2° (± 7.8°) with LCEA < 20° in 15 (31%) and LCEA 20-25° in 17 (35%) and mean acetabular index (AI) of 13.1° (± 5.8°) with AI > 13° in 22 (45%). MR findings included the following: mean anterior acetabular sector angle (AASA) 54.3° (± 9.8°), mean posterior acetabular sector angle (PASA) 92.7° (± 7.0°), labral tear at 3-4 o'clock in 20 (39%), high-grade acetabular chondral loss in 42 (83%) and ligamentum teres abnormality in 20 (39%). CONCLUSION: The posterior crescent sign occurs in 3.5% of symptomatic young and middle-aged adults on MR. It is associated with overt and borderline hip dysplasia and other findings of hip instability. It is also associated with osteoarthritis in some cases and should be interpreted with caution in these patients.