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Abstract Mapping neuronal networks is a central focus in neuroscience. While volume electron microscopy (vEM) can reveal the fine structure of neuronal networks (connectomics), it does not provide molecular information to identify cell types or functions. We developed an approach that uses fluorescent single-chain variable fragments (scFvs) to perform multiplexed detergent-free immunolabeling and volumetric-correlated-light-and-electron-microscopy on the same sample. We generated eight fluorescent scFvs targeting brain markers. Six fluorescent probes were imaged in the cerebellum of a female mouse, using confocal microscopy with spectral unmixing, followed by vEM of the same sample. The results provide excellent ultrastructure superimposed with multiple fluorescence channels. Using this approach, we documented a poorly described cell type, two types of mossy fiber terminals, and the subcellular localization of one type of ion channel. Because scFvs can be derived from existing monoclonal antibodies, hundreds of such probes can be generated to enable molecular overlays for connectomic studies.

Here, we present the first analysis of the connectome of a small volume of the Octopus vulgaris vertical lobe (VL), a brain structure mediating the acquisition of long-term memory in this behaviorally advanced mollusk. Serial section electron microscopy revealed new types of interneurons, cellular components of extensive modulatory systems, and multiple synaptic motifs. The sensory input to the VL is conveyed via~1.8 × 106 axons that sparsely innervate two parallel and interconnected feedforward networks formed by the two types of amacrine interneurons (AM), simple AMs (SAMs) and complex AMs (CAMs). SAMs make up 89.3% of the~25 × 106VL cells, each receiving a synaptic input from only a single input neuron on its non-bifurcating primary neurite, suggesting that each input neuron is represented in only~12 ± 3.4SAMs. This synaptic site is likely a ‘memory site’ as it is endowed with LTP. The CAMs, a newly described AM type, comprise 1.6% of the VL cells. Their bifurcating neurites integrate multiple inputs from the input axons and SAMs. While the SAM network appears to feedforward sparse ‘memorizable’ sensory representations to the VL output layer, the CAMs appear to monitor global activity and feedforward a balancing inhibition for ‘sharpening’ the stimulus-specific VL output. While sharing morphological and wiring features with circuits supporting associative learning in other animals, the VL has evolved a unique circuit that enables associative learning based on feedforward information flow.

Connectomics is fundamental in propelling our understanding of the nervous system's organization, unearthing cells and wiring diagrams reconstructed from volume electron microscopy (EM) datasets. Such reconstructions, on the one hand, have benefited from ever more precise automatic segmentation methods, which leverage sophisticated deep learning architectures and advanced machine learning algorithms. On the other hand, the field of neuroscience at large, and of image processing in particular, has manifested a need for user-friendly and open source tools which enable the community to carry out advanced analyses. In line with this second vein, here we propose mEMbrain, an interactive MATLAB-based software which wraps algorithms and functions that enable labeling and segmentation of electron microscopy datasets in a user-friendly user interface compatible with Linux and Windows. Through its integration as an API to the volume annotation and segmentation tool VAST, mEMbrain encompasses functions for ground truth generation, image preprocessing, training of deep neural networks, and on-the-fly predictions for proofreading and evaluation. The final goals of our tool are to expedite manual labeling efforts and to harness MATLAB users with an array of semi-automatic approaches for instance segmentation. We tested our tool on a variety of datasets that span different species at various scales, regions of the nervous system and developmental stages. To further expedite research in connectomics, we provide an EM resource of ground truth annotation from four different animals and five datasets, amounting to around 180 h of expert annotations, yielding more than 1.2 GB of annotated EM images. In addition, we provide a set of four pre-trained networks for said datasets. All tools are available from https://lichtman.rc.fas.harvard.edu/mEMbrain/. With our software, our hope is to provide a solution for lab-based neural reconstructions which does not require coding by the user, thus paving the way to affordable connectomics.

Abstract Elevated osteoclast (OC) activity is a major contributor to inflammatory bone loss (IBL) during chronic inflammatory diseases. However, the specific OC precursors (OCPs) responding to inflammatory cues and the underlying mechanisms leading to IBL are poorly understood. We identified two distinct OCP subsets: Ly6ChiCD11bhi inflammatory OCPs (iOCPs) induced during chronic inflammation, and homeostatic Ly6ChiCD11blo OCPs (hOCPs) which remained unchanged. Functional and proteomic characterization revealed that while iOCPs were rare and displayed low osteoclastogenic potential under normal conditions, they expanded during chronic inflammation and generated OCs with enhanced activity. In contrast, hOCPs were abundant and manifested high osteoclastogenic potential under normal conditions but generated OCs with low activity and were unresponsive to the inflammatory environment. Osteoclasts derived from iOCPs expressed higher levels of resorptive and metabolic proteins than those generated from hOCPs, highlighting that different osteoclast populations are formed by distinct precursors. We further identified the TNF-α and S100A8/A9 proteins as key regulators that control the iOCP response during chronic inflammation. Furthermore, we demonstrated that the response of iOCPs but not that of hOCPs was abrogated in tnf-α −/− mice, in correlation with attenuated IBL. Our findings suggest a central role for iOCPs in IBL induction. iOCPs can serve as potential biomarkers for IBL detection and possibly as new therapeutic targets to combat IBL in a wide range of inflammatory conditions.

This article explores the ability of ChatGPT to function as a virtual colleague in helping to design materials for higher education design students. Using a self-study methodology, two university educators attempted to collaborate with ChatGPT to create course materials targeted at higher education design students, before reflecting on its strengths and weaknesses during the process. Contextualising ChatGPT as the latest acute example of digital disruptors that design practices and processes have faced, the authors evaluated its current and potential threats and opportunities for the creation of design-focused learning content. The authors found that ChatGPT was a competent partner with regard to saving time, structuring textual content and documentation, and as a brainstorming tool. However, ChatGPT’s weaknesses included content generation that was often generic, usually requiring much human prompting, cajoling, and manual editing to produce desirable outcomes. Overall, ChatGPT was found to excel at its stated functionality as a language model, with some potentially useful functionality for the creation of higher education design course materials and outlines, as well as limitations. The reflections discussed can be used to inform design educators who may want to work with ChatGPT when designing course materials. However, acknowledging limitations and potential ethical challenges, the authors’ caution that educators may have to evaluate for themselves whether ChatGPT’s potential advantages outweigh its disadvantages.

The study aimed to investigate the role of RvD1 in acute and prolonged sterile inflammation and bone remodeling. A mouse model of sterile inflammation that involves bone resorption was used to examine endogenous RvD1 kinetics during inflammation. Application of exogenous RvD1 significantly inhibited bone remodeling via osteoclast reduction, alongside an anti-inflammatory secretome shift, increased macrophages recruitment and reduction of T-cytotoxic cells. In vitro and in vivo, RvD1 led to significant reduction in RANK expression which reduce osteoclastogenesis in a dose-dependent manner. Taken together, the data shows a dual role for RvD1, as a potent immunoresolvent agent alongside an osteoresolvent role, showing a potential therapeutic agent in bone resorption associated inflammatory conditions.

Synthetic surgical meshes are commonly used in abdominal wall reconstruction surgeries to strengthen a weak abdominal wall. Common mesh-related complications include local infection and inflammatory processes. Because cannabigerol (CBG) has both antibacterial and anti-inflammatory properties, we proposed that coating VICRYL (polyglactin 910) mesh with a sustained-release varnish (SRV) containing CBG would prevent these complications. We used an in vitro infection model with Staphylococcus aureus and an in vitro inflammation model of lipopolysaccharide (LPS)-stimulated macrophages. Meshes coated with either SRV-placebo or SRV-CBG were exposed daily to S. aureus in tryptic soy medium (TSB) or macrophage Dulbecco’s modified eagle medium (DMEM). Bacterial growth and biofilm formation in the environment and on the meshes were assessed by changes in optical density, bacterial ATP content, metabolic activity, crystal violet staining, spinning disk confocal microscopy (SDCM), and high-resolution scanning electron microscopy (HR-SEM). The anti-inflammatory effect of the culture medium that was exposed daily to the coated meshes was analyzed by measuring the release of the cytokines IL-6 and IL-10 from LPS-stimulated RAW 264.7 macrophages with appropriate ELISA kits. Additionally, a cytotoxicity assay was performed on Vero epithelial cell lines. We observed that compared with SRV-placebo, the segments coated with SRV-CBG inhibited the bacterial growth of S. aureus in the mesh environment for 9 days by 86 ± 4% and prevented biofilm formation and metabolic activity in the surroundings for 9 days, with respective 70 ± 2% and 95 ± 0.2% reductions. The culture medium that was incubated with the SRV-CBG-coated mesh inhibited LPS-induced secretion of IL-6 and IL-10 from the RAW 264.7 macrophages for up to 6 days without affecting macrophage viability. A partial anti-inflammatory effect was also observed with SRV-placebo. The conditioned culture medium was not toxic to Vero epithelial cells, which had an IC50 of 25 µg/mL for CBG. In conclusion, our data indicate a potential role of coating VICRYL mesh with SRV-CBG in preventing infection and inflammation in the initial period after surgery.

Summary:. Deep sternal wound infection (DSWI) is one of the most complex and devastating complications post cardiac surgery. We present here the modified sternoplasty, a novel surgical technique for treating DSWI post cardiac surgery. The modified sternoplasty includes debridement and sternal refixation via bilateral longitudinal stainless-steel wires that are placed parasternally along the ribs at the midclavicular or anterior axillary line, followed by six to eight horizontal stainless-steel wires that are anchored laterally and directly into the ribs. On top of that solid structure, wound reconstruction is performed by the use of bilateral pectoralis muscle flaps followed by subcutaneous tissue and skin closure. We reported mortality rates and length of hospitalization of patients who underwent the modified sternoplasty. In total, 68 patients underwent the modified sternoplasty. Two of these critically ill patients died (2.9%). The average length of hospitalization from the diagnosis of DSWI was 24.63 ± 22.09 days. The modified sternoplasty for treating DSWI is a more complex surgery compared with other conventional sternoplasty techniques. However, this technique was demonstrated to be more effective, having a lower rate of mortality, and having a length of hospitalization lower than or comparable to other techniques previously reported in the literature.

Vertical green living walls (VGWs)—growing plants on vertical walls inside or outside buildings—have been suggested as a nature-based solution to improve air quality and comfort in modern cities. However, as with other greenery systems (e.g., agriculture), managing VGW systems requires adequate temporal and spatial monitoring of the plants as well as the surrounding environment. Remote sensing cameras and small, low-cost sensors have become increasingly valuable for conventional vegetation monitoring; nevertheless, they have rarely been used in VGWs. In this descriptive paper, we present a first-of-its-kind remote sensing high-throughput monitoring system in a VGW workplace. The system includes low- and high-cost sensors, thermal and hyperspectral remote sensing cameras, and in situ gas-exchange measurements. In addition, air temperature, relative humidity, and carbon dioxide concentrations are constantly monitored in the operating workplace room (scientific computer lab) where the VGW is established, while data are continuously streamed online to an analytical and visualization web application. Artificial Intelligence is used to automatically monitor changes across the living wall. Preliminary results of our unique monitoring system are presented under actual working room conditions while discussing future directions and potential applications of such a high-throughput remote sensing VGW system.

Wildfire simulations depend on fuel representation. Present fuel models are mainly based on the density and properties of different vegetation types. This study aims to improve the accuracy of WRF-Fire wildfire simulations, by using synthetic-aperture radar (SAR) data to estimate the fuel load and the trend of vegetation index to estimate the dryness of woody vegetation. We updated the chaparral and timber standard woody fuel classes in the WRF-Fire fuel settings. We used the ESA global above-ground biomass (AGB) based on SAR data to estimate the fuel load, and the Landsat normalized difference vegetation index (NDVI) trends of woody vegetation to estimate the fuel moisture content. These fuel sub-parameters represent the dynamic changes and spatial variability of woody fuel. We simulated two wildfires in Israel while using three different fuel models: the original 13 Anderson Fire Behavior fuel model, and two modified fuel models introducing AGB alone, and AGB and dryness. The updated fuel model (the basic fuel model plus the AGB and dryness) improved the simulation results significantly, i.e., the Jaccard similarity coefficient increased by 283% on average. Our results demonstrate the potential of combining satellite SAR data and Landsat NDVI trends to improve WRF-Fire wildfire simulations.

Objectives: To compare patient-reported pain, bleeding, and device safety between intrauterine contraceptive device (IUD) insertion procedures employing a suction cervical stabilizer or single-tooth tenaculum., Study Design: This was a randomized, prospective, single-blinded study conducted at two centers, enrolling women aged 18 years or older, eligible for IUD insertion. The primary end point measure was patient-reported pain, measured on a 100-mm Visual Analogue Scale. Safety was assessed on the amount of bleeding, adverse events, and serious adverse events., Results: One hundred women were randomized, 48 to the investigational device and 52 to control. There were no statistically significant differences between the groups in factors potentially associated with pain on IUD insertion. IUD insertion was successful in 94% of all subjects. Subjects in the investigational device group reported pain scores ≥14 points lower than in the control group at cervix grasping (14.9 vs 31.3; p < 0.001) and traction (17.0 vs 35.9; p < 0.001), and smaller differences in pain scores at the IUD insertion (31.5 vs 44.9; p = 0.021) and cervix-release (20.6 vs 30.9; p = 0.049) steps. Nulliparous women experienced the greatest pain differences to control. Mean blood loss was 0.336 (range 0.022-2.189) grams in the investigational device group and 1.336 (range 0.201-11.936) grams in the control group, respectively (p = 0.03 for the comparison). One adverse event (bruising and minor bleeding) in the investigational device group was considered causally related to the study device., Conclusions: The suction cervical stabilizer had a reassuring safety profile and its use was associated with significant reductions in pain during the IUD insertion procedure compared with standard single-tooth tenaculum use, particularly among nulliparous women., Implications: Pain can be an important barrier to greater use of IUD devices among prescribers and users, particularly nulliparous women. The suction cervical stabilizer may provide an appealing alternative to currently available tenacula, filling an important unmet need., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

This article translates and synthesizes a reference guide containing iconographic material of the pre- and post-pubertal female external genitals with and without genital mutilation/cutting (FGM/C). The literature focuses on adults, whereas FGM/C are usually performed before the age of 15. Signs of FGM/C can be subtle and depend on the type of mutilation as well as the experience of the examiner. Published in 2022 and produced with the collaboration of 23 professionals, this illustrated guide "Female Genital Mutilation/Cutting in Children and Adolescents: Illustrated Guide to Diagnose, Assess, Inform and Report, https://link.springer.com/book/10.1007/978-3-030-81736-7 " is currently available in open access. It is aimed at facilitating the training of health professionals in making diagnosis, clinical management, and reporting to child safeguarding/law enforcement agencies, where required., Competing Interests: Les auteur-es n’ont déclaré aucun conflit d’intérêts en relation avec cet article.

Heavy menstrual bleeding is one of the most common causes of consultation in haematology. We present the clinical case of a 20-year-old woman referred by her gynaecologist due to heavy menstrual bleeding since menarche, complicated by iron deficiency anaemia. Haemostasis work-up was initially suggestive of a von Willebrand disease type 1. Genetic analyses by whole exome sequencing lead to a fortuitous discovery of haemophilia by identifying a heterozygous missense mutation in F8 , exon 8 c.1127T>G:p.Val376Gly, previously reported in a patient with mild haemophilia A. The bleeding phenotype worsened by concomitant low von Willebrand factor (VWF) due to VWF variants influencing VWF levels. Our case highlights how whole exome sequencing can help to correct an erroneous diagnosis and identify polymorphisms that eventually contribute to the overall haemostatic balance., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Objective: To evaluate if daily oral 75 µg of Desogestrel (DSG) for 3 months prior to the insertion of etonogestrel-releasing contraceptive implant (ENG-IMPLANT) might help reduce its premature discontinuation., Results: A total of 66 women were randomized in the ENG-IMPLANT group (26) and in the DSG + ENG-IMPLANT group (40), respectively, in the Geneva University Hospitals and Basel University Hospital, from August 15th, 2016 through September 30th, 2019. In the DSG + ENG-IMPLANT group, patients were given a 3 months' supply of 75 µg of DSG before the insertion of the ENG-IMPLANT. All women were seen after 3 months for bleeding and satisfaction evaluation, and at 12 months post ENG-IMPLANT insertion. Higher levels of satisfaction at 12-months were found in the ENG-IMPLANT group compared to the DSG + ENG-IMPLANT group (8.5 ± 1.7 vs. 6.6 ± 2.9, p = 0.012). There were no statistically significant differences regarding tolerance (7.8 ± 2.5 vs 6.8 ± 2.6, p = 0.191) and contraceptive continuation (80% vs 72.4%, p = 0.544) between groups., Conclusion: DSG prior to insertion of the ENG-IMPLANT did not improve its continuation rate neither its satisfaction at 1 year. Trial registration NCT05174195. Retrospectively registered, the 30th December 2021., (© 2023. The Author(s).)

Paediatricians may face the notion of 'virginity' in various situations while caring for children and adolescents, but are often poorly prepared to address this sensitive topic. Virginity is a social construct. Despite medical evidence that there is no scientifically reliable way to determine virginity, misconceptions about the hymen and its supposed association with sexual history persist and lead to unethical practices like virginity testing, certificate of virginity or hymenoplasty, which can be detrimental to the health and well-being of females of all ages. The paediatrician has a crucial role in providing evidence-based information and promoting positive sexual education to children, adolescents and parents. Improving knowledge can help counter misconceptions and reduce harms to girls and women., (© 2022 The Authors. Journal of Paediatrics and Child Health published by John Wiley & Sons Australia, Ltd on behalf of Paediatrics and Child Health Division (The Royal Australasian College of Physicians).)

Study Objective: The FertiSTAT (fertility status awareness) tool provides personalized advice on reducing risk factors for infertility and seeking medical advice on the basis of lifestyle and reproductive profile. The aim of our research was to test the FertiSTAT tool in younger patients (14-24 years). A secondary objective was to screen for and evaluate knowledge of risk factors that affect fertility. DESIGN, SETTING, PARTICIPANTS, INTERVENTIONS, AND MAIN OUTCOME MEASURES: Patients aged 14-24 years attending consultations at Geneva University Hospitals received a quantitative questionnaire before consultations. Questions covered lifestyle, gynecological history, perception of fertility, and pregnancy intent. We investigated respondents' beliefs with regard to risk factors for infertility through "true/false" questions. We selected questions relevant to our population from the original FertiSTAT questionnaire to calculate each respondent's FertiSTAT score. Scores ranged from "blue" (low risk, score 1) to "red" (risk of infertility, score 4)., Results: A total of 279 women aged 14-24 years were included. Nonpregnant patients had overall higher FertiSTAT scores (2.7 ± 0.8). Upon logistic regression analysis, with every additional FertiSTAT point, the odds of being pregnant at the time of survey decreased by 0.48. Risk factors for infertility and knowledge of these risk factors were equally distributed between pregnant and nonpregnant women., Conclusion: Our findings suggest FertiSTAT might be a useful tool in the younger population to whom we extended it, and highlight gaps in knowledge on risk factors for infertility. These findings are of interest when considering FertiSTAT as a starting point to discuss contraception and risk factors for infertility at an age at which risk mitigation would prove most effective in preserving future fertility., (Copyright © 2021 North American Society for Pediatric and Adolescent Gynecology. Published by Elsevier Inc. All rights reserved.)

Background and Purpose: We aimed to examine epidemiological data and identify risk factors for sexual assault. This study included women seeking care at the Geneva University Hospitals emergency obstetrics and gynaecological unit., Method: Retrospective, sociodemographic, and clinical data were collected from the medical reports., Results: We reviewed 836 medical charts and registered 92.8 women and girls per year who consulted the emergency department after sexual assault. The average age was 26 (±12) years, with a median of 23 years. Body lesions were reported in 525 patients (63%) and genital traumatic lesions were reported in 230 (28%) patients. Sexual assault by a stranger was reported in 436 cases (52%). Thirty percent of patients knew their aggressor. Aggressions were predominantly committed on the weekend, accounting for 367 cases (46%), with nearly two-thirds (65%, n = 474) occurring between 10 pm and 6 am. In total, 399 (48%) patients who were sexually assaulted reported having consumed alcohol and 102 (12%) reported having taken drugs prior to the aggression; 80 (10%) patients had consumed both and 286 (34%) had amnesia. Half of the sample sought and received medical care within 24 h from the time the aggression took place., Conclusion: Sexual assault risk factors in our study population in Geneva included use of drugs and alcohol, with amnesia. Future prevention and education interventions should target these areas., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)