The scent gland composition of the Mangshan pit viper, Protobothrops mangshanensis.

The Mangshan pit viper, Protobothrops mangshanensis, is a rare, highly endangered snake native to the mountainous regions of Hunan Province in China. Snakes possess abdominal scent glands, which have been chemically studied in several species. These glands can contain various lipids and peptides, but very often also complex mixtures of carboxylic acids. We report here the occurrence of novel methyl-branched unsaturated acids found in the secretions of six captive individuals living in a zoo. The structures of these compounds, 4.6-dimethylalk-5-enoates in a homologous series from C11-C16, were characterized by GC-MS and GC-IR analysis and various microderivatization reactions including hydrogenation and esterification leading to methyl and pyridylmethyl esters. In addition, dimethyloxazoline formation helped to localize the double bond. The synthesis of methyl 4,6-dimethyldodec-5-enoate allowed the correct assignment of structures and showed the (E)-configuration of the double bond for the major naturally occurring diastereomers. These acids occur in small amounts compared to the major glandular components, cholesterol, and 1-O-hexadecylglycerol, as well as other common long-chain alcohols and amides. Although a general defensive function has been proposed for snake abdominal scent glands, the specific chemistry and moderate amounts of acids reported here may suggest a function in chemical signaling for the Mangshan pit viper. In addition, proline-containing diketopiperazines were identified for the first time in snake scent glands, although an artificial formation from amino acids likely present in the secretion cannot be excluded.

Identification of unique highly hetero-substituted benzenes as chemical weapons of springtails by a combination of trace analytical methods with DFT calculations and synthesis.

Springtails (Collembola) are important members of the soil mesofauna. They are small, often less than 1-2 mm in length. A typical escape response of most surface-living species is to jump, using their furca. However, some species also use chemical defence against predators. While the defence chemistry of higher insects has been well studied, reports from the basal Collembola are rare, linked to the difficulties in obtaining enough biomass. We herein report on the identification and repellent activity of compounds detected in Ceratophysella denticulata. Extracts with various solvents obtained from only 50 individuals were sufficient for analysis by GC/MS, GC/HR-MS, and GC/IR. The large number of candidate structures of the major components were then prioritised by DFT calculations of IR spectra. Finally, the total synthesis of the top candidates confirmed the structures of the three major compounds to be 4-methoxy-5-(methylthio)benzo-1,3-dioxolane, 5,6,7-trimethoxybenzo-1,3-oxathiolane, and 8-amino-5,6,7-trimethoxybenzo-1,3-oxathiolane, the latter being the first naturally occurring fully hetero-substituted benzene. These highly substituted benzenes have no precedence in nature and carry structural motifs rare in nature, such as the benzo-1,3-oxathiolane ring system or the occurrence of O-, N-, and S-substituents at the same benzene core. Another novel natural compound, 2-methyl-1H-imidazo[4,5-b]pyridine, is used by Hypogastrura viatica. 4-Methoxy-5-(methylthio)benzo-1,3-dioxolane showed significant activity in deterrence assays with the ant Lasius niger. The data indicate that the title compounds are used in the chemical defence of these springtails, thus adding a new compound class to the known antipredator defences of arthropods. The results underline the difference in defence chemistry between Collembola and insects.

Processing of Short-Form Content in Clinical Narratives: Systematic Scoping Review.

BACKGROUND: Clinical narratives are essential components of electronic health records. The adoption of electronic health records has increased documentation time for hospital staff, leading to the use of abbreviations and acronyms more frequently. This brevity can potentially hinder comprehension for both professionals and patients. OBJECTIVE: This review aims to provide an overview of the types of short forms found in clinical narratives, as well as the natural language processing (NLP) techniques used for their identification, expansion, and disambiguation. METHODS: In the databases Web of Science, Embase, MEDLINE, EBMR (Evidence-Based Medicine Reviews), and ACL Anthology, publications that met the inclusion criteria were searched according to PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines for a systematic scoping review. Original, peer-reviewed publications focusing on short-form processing in human clinical narratives were included, covering the period from January 2018 to February 2023. Short-form types were extracted, and multidimensional research methodologies were assigned to each target objective (identification, expansion, and disambiguation). NLP study recommendations and study characteristics were systematically assigned occurrence rates for evaluation. RESULTS: Out of a total of 6639 records, only 19 articles were included in the final analysis. Rule-based approaches were predominantly used for identifying short forms, while string similarity and vector representations were applied for expansion. Embeddings and deep learning approaches were used for disambiguation. CONCLUSIONS: The scope and types of what constitutes a clinical short form were often not explicitly defined by the authors. This lack of definition poses challenges for reproducibility and for determining whether specific methodologies are suitable for different types of short forms. Analysis of a subset of NLP recommendations for assessing quality and reproducibility revealed only partial adherence to these recommendations. Single-character abbreviations were underrepresented in studies on clinical narrative processing, as were investigations in languages other than English. Future research should focus on these 2 areas, and each paper should include descriptions of the types of content analyzed.

The unique epicuticular chemistry of Collembola - A cross-species analysis.

Springtails (Collembola), tiny hexapod arthropods, are abundant in the soil of most ecosystems, but our knowledge of their secondary metabolites is limited, in contrast to that of insects. In insects, the outer cuticle is usually covered by mixtures of long-chain hydrocarbons serving different functions, such as water regulation or chemical communication. In contrast, the knowledge of the epicuticular chemistry of springtails is scarce. We analyzed the cuticular lipids of 23 species covering different lineages. The often complicated structures were elucidated using gas chromatography/mass spectrometry, microderivatization, and synthesis. In contrast to insects, the terpene biosynthetic pathway is used for many of these lipids, producing unprecedented higher terpenes. In addition, evidence for de novo cholesterol biosynthesis in springtails was found, which is absent in insects. Finally, diverse non-insect linear compounds originating from the fatty acid biosynthetic pathway were identified. Our comparative analysis showed clear differences compared to insects and shed light on phylogenetic relationships.

GRK2 kinases in the primary cilium initiate SMOOTHENED-PKA signaling in the Hedgehog cascade.

During Hedgehog (Hh) signal transduction in development and disease, the atypical G protein-coupled receptor (GPCR) SMOOTHENED (SMO) communicates with GLI transcription factors by binding the protein kinase A catalytic subunit (PKA-C) and physically blocking its enzymatic activity. Here, we show that GPCR kinase 2 (GRK2) orchestrates this process during endogenous mouse and zebrafish Hh pathway activation in the primary cilium. Upon SMO activation, GRK2 rapidly relocalizes from the ciliary base to the shaft, triggering SMO phosphorylation and PKA-C interaction. Reconstitution studies reveal that GRK2 phosphorylation enables active SMO to bind PKA-C directly. Lastly, the SMO-GRK2-PKA pathway underlies Hh signal transduction in a range of cellular and in vivo models. Thus, GRK2 phosphorylation of ciliary SMO and the ensuing PKA-C binding and inactivation are critical initiating events for the intracellular steps in Hh signaling. More broadly, our study suggests an expanded role for GRKs in enabling direct GPCR interactions with diverse intracellular effectors.

From Clinical Information Systems to Personalized Health Knowledge Graphs.

This paper presents a versatile solution to formally represent the contents of electronic health records. It is based on the knowledge graph paradigm, and semantic web standards RDF and OWL. It employs the established semantic standards SNOMED CT and FHIR, which warrant international interoperability. A graph-based form is not only useful to feed different target visualizations, but it can also be subject to AI-powered services such as (fuzzy) retrieval and summarization.

Key phosphorylation sites for robust ß-arrestin2 binding at the MOR revisited.

Desensitisation of the mu-opioid receptor (MOR) is proposed to underlie the initiation of opioid analgesic tolerance and previous work has shown that agonist-induced phosphorylation of the MOR C-tail contributes to this desensitisation. Moreover, phosphorylation is important for ß-arrestin recruitment to the receptor, and ligands of different efficacies induce distinct phosphorylation barcodes. The C-tail 370TREHPSTANT379 motif harbours Ser/Thr residues important for these regulatory functions. 375Ser is the primary phosphorylation site of a ligand-dependent, hierarchical, and sequential process, whereby flanking 370Thr, 376Thr and 379Thr get subsequently and rapidly phosphorylated. Here we used GRK KO cells, phosphosite specific antibodies and site-directed mutagenesis to evaluate the contribution of the different GRK subfamilies to ligand-induced phosphorylation barcodes and ß-arrestin2 recruitment. We show that both GRK2/3 and GRK5/6 subfamilies promote phosphorylation of 370Thr and 375Ser. Importantly, only GRK2/3 induce phosphorylation of 376Thr and 379Thr, and we identify these residues as key sites to promote robust ß-arrestin recruitment to the MOR. These data provide insight into the mechanisms of MOR regulation and suggest that the cellular complement of GRK subfamilies plays an important role in determining the tissue responses of opioid agonists.

Tumor Board Visualization: Integrating Clinical and Laboratory Insights.

We present a data visualization tool for tumor boards, merging clinical with molecular and multi-omics data to refine precision oncology decisions. The tool offers a holistic patient perspective, facilitating personalized treatment strategies. By integrating clinical and laboratory datasets, it enables intuitive navigation through complex information. Clinicians are supported in their decision-making by user-friendly visualizations. Future studies are needed to evaluate its real-world impact and usability in precision oncology settings.

'SNOMEDizing' Questionnaires for Standardizing Stroke Registry Data.

International interoperability of healthcare and research data requires a commitment to standards. To this end, SNOMED CT was evaluated for representing questionnaire items of the European Registry of Stroke Care Quality using a complex annotation protocol. The agreement between validators and annotators was 72.4%. At least 64% of the information could be represented by using SNOMED CT only, including complex post-coordinations. 9% of the information would require an information model, and 14% the addition of new content to SNOMED CT. Next steps will be the creation of an annotation guideline for questionnaires, a specific reference set, and the combination of both with an information model such as HL7 FHIR.

Term Candidate Generation to Enrich Clinical Terminologies with Large Language Models.

Annotated language resources derived from clinical routine documentation form an intriguing asset for secondary use case scenarios. In this investigation, we report on how such a resource can be leveraged to identify additional term candidates for a chosen set of ICD-10 codes. We conducted a log-likelihood analysis, considering the co-occurrence of approximately 1.9 million de-identified ICD-10 codes alongside corresponding brief textual entries from problem lists in German. This analysis aimed to identify potential candidates with statistical significance set at p < 0.01, which were used as seed terms to harvest additional candidates by interfacing to a large language model in a second step. The proposed approach can identify additional term candidates at suitable performance values: hypernyms MAP@5=0.801, synonyms MAP@5 = 0.723 and hyponyms MAP@5 = 0.507. The re-use of existing annotated clinical datasets, in combination with large language models, presents an interesting strategy to bridge the lexical gap in standardized clinical terminologies and real-world jargon.

Chronic morphine treatment induces sex- and synapse-specific cellular tolerance on thalamo-cortical mu opioid receptor signaling.

How cellular adaptations give rise to opioid analgesic tolerance to opioids like morphine is not well understood. For one, pain is a complex phenomenon comprising both sensory and affective components, largely mediated through separate circuits. Glutamatergic projections from the medial thalamus (MThal) to the anterior cingulate cortex (ACC) are implicated in processing of affective pain, a relatively understudied component of the pain experience. The goal of this study was to determine the effects of chronic morphine exposure on mu-opioid receptor (MOR) signaling on MThal-ACC synaptic transmission within the excitatory and feedforward inhibitory pathways. Using whole cell patch-clamp electrophysiology and optogenetics to selectively target these projections, we measured morphine-mediated inhibition of optically evoked postsynaptic currents in ACC layer V pyramidal neurons in drug-naïve and chronically morphine-treated mice. We found that morphine perfusion inhibited the excitatory and feedforward inhibitory pathways similarly in females but caused greater inhibition of the inhibitory pathway in males. Chronic morphine treatment robustly attenuated morphine presynaptic inhibition within the inhibitory pathway in males, but not females, and mildly attenuated presynaptic inhibition within the excitatory pathway in both sexes. These effects were not observed in MOR phosphorylation-deficient mice. This study indicates that chronic morphine treatment induces cellular tolerance to morphine within a thalamo-cortical circuit relevant to pain and opioid analgesia. Furthermore, it suggests this tolerance may be driven by MOR phosphorylation. Overall, these findings improve our understanding of how chronic opioid exposure alters cellular signaling in ways that may contribute to opioid analgesic tolerance.NEW & NOTEWORTHY Opioid signaling within the anterior cingulate cortex (ACC) is important for opioid modulation of affective pain. Glutamatergic medial thalamus (MThal) neurons synapse in the ACC and opioids, acting through mu opioid receptors (MORs), acutely inhibit synaptic transmission from MThal synapses. However, the effect of chronic opioid exposure on MThal-ACC synaptic transmission is not known. Here, we demonstrate that chronic morphine treatment induces cellular tolerance at these synapses in a sex-specific and phosphorylation-dependent manner.

Identification by Synthesis: Imidacins, Urocanate-Derived Alkaloids from the Myxobacterium Stigmatella aurantiaca.

Innovative discovery approaches such as genome-mining and metabolomics-inspired methods have reshaped the natural product research field, complementing traditional bioactivity-based screens and allowing hitherto unseen compounds to be uncovered from previously investigated producers. In line with these trends, we report here imidacins, a novel class of secondary metabolites specific to the myxobacterial genus Stigmatella. A combination of secondary metabolome analysis, genome-mining techniques, spectroscopic analysis, and finally total synthesis was used to allow structure elucidation. Imidacins are urocanate-derived aliphatic acids with an adjacent cyclopropane moiety, structural features unprecedented in natural products to date.

Enantioselective synthesis of all stereoisomers of geosmin and of biosynthetically related natural products.

Synthetic routes to geosmin and its enantiomer are well established, but the enantioselective synthesis of stereoisomers of geosmin is unknown. Here a stereoselective synthesis of all stereoisomers of geosmin is reported, yielding all compounds in high enantiomeric purity. Furthermore, the stereoselective synthesis of a geosmin derivative isolated from a mangrove associated streptomycete was performed, establishing the absolute configuration of the natural product. Finally, a new side product of the geosmin synthase from Streptomyces ambofaciens was isolated and its structure was elucidated by NMR spectroscopy. The absolute configuration of this new compound was determined through a stereoselective synthesis.

Process Development of Aluminum Electroplating from an Ionic Liquid on 150 mm Wafer Level.

This paper focuses on the development of electroplating on 150 mm wafer level for microsystem technology applications from 1-Ethyl-3-methylimidazolium chloride (EMImCl) with Aluminumtrichloride (AlCl3). The deposition was carried out on 150 mm wafers with Au or Al seed layers deposited by physical vapor deposition (PVD). The electrodeposition was carried out using pattern plating. On the Au seed layer, bipolar pulse plating was applied. Compared to the Au seed layer, the electrodeposition on the Al seed layer was favorable, with lower current densities and pulsing frequencies. Utilizing the recurrent galvanic pulses and avoiding ionic liquid convection, inhomogeneities lower than 15% were achieved with a laboratory plating cell. One major aspect of this study was the removal of the native Al oxide prior to deposition. It was investigated on the chip and wafer levels using either current- or potential-controlled removal pulses. This process step was affected by the plasma treatment of the wafer, thus the surface free energy, prior to plating. It turned out that a higher surface free energy hindered proper oxide removal at a potential of 3 V. The theory of oxide breakdown based on electrostriction force via the electrical field was applied to discuss the findings and to derive conclusions for future plating experiments.

Disambiguation of acronyms in clinical narratives with large language models.

OBJECTIVE: To assess the performance of large language models (LLMs) for zero-shot disambiguation of acronyms in clinical narratives. MATERIALS AND METHODS: Clinical narratives in English, German, and Portuguese were applied for testing the performance of four LLMs: GPT-3.5, GPT-4, Llama-2-7b-chat, and Llama-2-70b-chat. For English, the anonymized Clinical Abbreviation Sense Inventory (CASI, University of Minnesota) was used. For German and Portuguese, at least 500 text spans were processed. The output of LLM models, prompted with contextual information, was analyzed to compare their acronym disambiguation capability, grouped by document-level metadata, the source language, and the LLM. RESULTS: On CASI, GPT-3.5 achieved 0.91 in accuracy. GPT-4 outperformed GPT-3.5 across all datasets, reaching 0.98 in accuracy for CASI, 0.86 and 0.65 for two German datasets, and 0.88 for Portuguese. Llama models only reached 0.73 for CASI and failed severely for German and Portuguese. Across LLMs, performance decreased from English to German and Portuguese processing languages. There was no evidence that additional document-level metadata had a significant effect. CONCLUSION: For English clinical narratives, acronym resolution by GPT-4 can be recommended to improve readability of clinical text by patients and professionals. For German and Portuguese, better models are needed. Llama models, which are particularly interesting for processing sensitive content on premise, cannot yet be recommended for acronym resolution.

Expression of free fatty acid receptor 2 in normal and neoplastic tissues.

OBJECTIVE: Little information is available concerning protein expression of the free fatty acid receptor 2 (FFAR2), especially in tumours. Therefore, the aim of the present study was to comprehensively characterise the expression profile of FFAR2 in a large series of human normal and neoplastic tissues using immunohistochemistry thus providing a basis for further in-depth investigations into its potential diagnostic or therapeutic importance. METHODS: We developed a novel rabbit polyclonal anti-FFAR2 antibody, 0524, directed against the C-terminal region of human FFAR2. Antibody specificity was confirmed via Western blot analyses and immunocytochemistry using the FFAR2-expressing cell line BON-1 and FFAR2-specific small interfering RNA as well as native and FFAR2-transfected HEK-293 cells. The antibody was then used for immunohistochemical analyses of various formalin-fixed, paraffin-embedded specimens of normal and neoplastic human tissues. RESULTS: In normal tissues, FFAR2 was mainly present in distinct cell populations of the cerebral cortex, follicular cells and C cells of the thyroid, cardiomyocytes of the heart, bronchial epithelia and glands, hepatocytes and bile duct epithelia of the liver, gall bladder epithelium, exocrine and ß-cells of the endocrine pancreas, glomerular mesangial cells and podocytes as well as collecting ducts of the kidney, intestinal mucosa (particularly enteroendocrine cells), prostate epithelium, seminiferous tubules of the testicles, and placental syncytiotrophoblasts. In neoplastic tissues, FFAR2 was particularly prevalent in papillary thyroid carcinomas, parathyroid adenomas, and gastric, colon, pancreatic, hepatocellular, cholangiocellular, urinary bladder, breast, cervical, and ovarian carcinomas. CONCLUSIONS: We generated and characterised a novel rabbit polyclonal anti-human FFAR2 antibody that is well-suited for visualising FFAR2 expression in human routine pathology tissues. This antibody is also suitable for Western blot and immunocytochemistry experiments. To our knowledge, this antibody enabled the first broad FFAR2 protein expression profile in various normal and neoplastic human tissues.

Patient informed consent, ethical and legal considerations in the context of digital vulnerability with smart, cardiac implantable electronic devices.

Advancements in digitalisation with cardiac implantable electronic devices (CIEDs) allow patients opportunities for improved autonomy, quality of life, and a potential increase in life expectancy. However, with the digital and functional practicalities of CIEDs, there exists also cyber safety issues with transferring wireless information. If a digital network were to be hacked, a CIED patient could experience both the loss of sensitive data and the loss of functional control of the CIED due to an unwelcome party. Moreover, if a CIED patient were to become victim of a cyber attack, which resulted in a serious or lethal event, and if this information were to become public, the trust in healthcare would be impacted and legal consequences could result. A cyber attack therefore poses not only a direct threat to the patient's health but also the confidentiality, integrity, and availability of the CIED, and these cyber threats could be considered "patient-targeted threats." Informed consent is a key component of ethical care, legally concordant practice, and promoting patient-as-partner therapeutic relationships [1]. To date, there are no standardised guidelines for listing cybersecurity risks within the informed consent or for discussing them during the consent process. Providers are responsible for adhering to the ethical principles of autonomy, beneficence, non-maleficence, and justice, both in medical practice generally and the informed consent process specifically. At present, the decision to include cybersecurity risks is mainly left to the provider's discretion, who may also have limited cyber risk information. Without effective and in-depth communication about all possible cybersecurity risks during the consent process, CIED patients can be left unaware of the privacy and physical risks they possess by carrying such a device. Therefore, cyber risk factors should be covered within the patients' informed consent and reviewed on an ongoing basis as new risk information becomes available. By including cyber risk information in the informed consent process, patients are given the autonomy to make the best-informed decision.

Rapid elucidation of agonist-driven regulation of the neurokinin 1 receptor using a GPCR phosphorylation immunoassay.

Agonist-induced phosphorylation is a crucial step in the activation/deactivation cycle of G protein-coupled receptors (GPCRs), but direct determination of individual phosphorylation events has remained a major challenge. We have recently developed a bead-based immunoassay for the quantitative assessment of agonist-induced GPCR phosphorylation that can be performed entirely in 96-well plates, thus eliminating the need for western blot analysis. In the present study, we adapted this assay to three novel phosphosite-specific antibodies directed against the neurokinin 1 (NK1) receptor, namely pS338/pT339-NK1, pT344/pS347-NK1, and pT356/pT357-NK1. We found that substance P (SP) stimulated concentration-dependent phosphorylation of all three sites, which could be completely blocked in the presence of the NK1 receptor antagonist aprepitant. The other two endogenous ligands of the tachykinin family, neurokinin A (NKA) and neurokinin B (NKB), were also able to induce NK1 receptor phosphorylation, but to a much lesser extent than substance P. Interestingly, substance P promoted phosphorylation of the two distal sites more efficiently than that of the proximal site. The proximal site was identified as a substrate for phosphorylation by protein kinase C. Analysis of GPCR kinase (GRK)-knockout cells revealed that phosphorylation was mediated by all four GRK isoforms to similar extents at the T344/S347 and the T356/T357 cluster. Knockout of all GRKs resulted in abolition of all phosphorylation signals highlighting the importance of these kinases in agonist-mediated receptor phosphorylation. Thus, the 7TM phosphorylation assay technology allows for rapid and detailed analyses of GPCR phosphorylation.

Chemical species recognition in an adaptive radiation of Hawaiian Tetragnatha spiders (Araneae: Tetragnathidae).

Studies of adaptive radiations have played a central role in our understanding of reproductive isolation. Yet the focus has been on human-biased visual and auditory signals, leaving gaps in our knowledge of other modalities. To date, studies on chemical signals in adaptive radiations have focused on systems with multimodal signalling, making it difficult to isolate the role chemicals play in reproductive isolation. In this study we examine the use of chemical signals in the species recognition and adaptive radiation of Hawaiian Tetragnatha spiders by focusing on entire communities of co-occurring species, and conducting behavioural assays in conjunction with chemical analysis of their silks using gas chromatography-mass spectrometry. Male spiders significantly preferred the silk extracts of conspecific mates over those of sympatric heterospecifics. The compounds found in the silk extracts, long chain alkyl methyl ethers, were remarkably species-specific in the combination and quantity. The differences in the profile were greatest between co-occurring species and between closely related sibling species. Lastly, there were significant differences in the chemical profile between two populations of a particular species. These findings provide key insights into the role chemical signals play in the attainment and maintenance of reproductive barriers between closely related co-occurring species.