Quantification Of Signals Research Articles

P0944 Fluorescently labelled adalimumab to visualise drug targeting in Inflammatory Bowel Disease: a safety, feasibility and dose-finding study

Abstract Background Treatment of inflammatory bowel disease (IBD) with biologicals, such as adalimumab, is hampered by high non-response rates and lack of reliable response prediction tools. Therefore, patients are potentially exposed to ineffective treatment and side effects, while clinical deterioration continues. Moreover, it is unclear whether the drug reaches its target site in adequate concentrations to achieve treatment response. We aim to visualise adalimumab distribution and detect adalimumab target cells using quantified fluorescence molecular endoscopy (qFME). Methods Adalimumab was labelled with IRDye 680LT under cGMP conditions, resulting in a fluorescent tracer suitable for human use. This tracer is used in an ongoing, non-randomized, non-blinded, prospective feasibility study. A total of 21 IBD patients scheduled for an endoscopy will be included. Up until now, eight patients received adalimumab-680LT (4.5 mg: n=3, 15 mg: n=3, 25 mg: n=2), and three patients were included as a control. Two to four days after tracer administration, qFME was performed to gather in vivo fluorescence data of inflamed and non-inflamed tissue. Fluorescent signals were quantified by multi-diameter single fibre reflectance/single fibre fluorescence (MDSFR/SFF) spectroscopy. Furthermore, biopsies were taken from inflamed and non-inflamed mucosa for ex vivo analysis. Results To date, 11 patients were included. Tracer administration was well tolerated and no adverse events occurred in any dose group. Real-time in vivo macroscopic imaging showed clear uptake of adalimumab-680LT in inflamed tissue compared to non-inflamed tissue (figure 1A). Spectroscopy revealed a dose-dependent increase in fluorescent signal for adalimumab-680LT in inflamed tissue, with a significant difference between 4.5 and 15 mg (0.016 [0.011-0.021] vs 0.033 [0.021-0.043] (p=0.036)) (figure 1B). The difference between inflamed and non-inflamed tissue is most noticeable in the 25 mg group. Ex vivo Mean Fluorescent Intensity (MFI) quantification of all biopsies showed a significant increase of fluorescent signal in the 15 and 25 mg dose groups compared to control in inflamed tissue (65.0 [54.0-75.7] vs 32.4 [27.1-38.2] (p=0.0040), and 62.1 [44.1-82.8] vs 32.4 [27.1-38.2] (p=0.0286), respectively) (figure 1C). Conclusion Preliminary results show adalimumab-680LT is safe for visualising adalimumab distribution. Doses of at least 15 mg are sufficient for visualisation and quantification of fluorescent signal in vivo. Furthermore, higher MFI’s and spectroscopy results were measured in inflamed tissue compared to healthy tissue, indicating targeting of the tracer to inflamed tissue. Future ex vivo experiments are ongoing to visualise adalimumab target cells.

The ratio of Wnt signaling activity to Sox2 transcription factor levels predicts neuromesodermal fate potential.

Neuromesodermal progenitors (NMPs) are a vertebrate cell type that contribute descendants to both the spinal cord and the mesoderm. The undifferentiated bipotential NMP state is maintained when both Wnt signaling is active and Sox2 is present. We used transgenic reporter lines to live-image both Wnt activity and Sox2 levels in NMPs and observed a unique cellular ratio in NMPs compared to NMP-derived mesoderm or neural tissue. We used this unique signature to identify the previously unknown anatomical position of a progenitor population that gives rise to the midline tissues of the floor plate of the spinal cord and the mesodermal notochord. Thus, quantification of the active Wnt signaling to Sox2 ratio can be used to predict and identify cells with neuromesodermal potential. We also developed the auxin inducible degron 2 system for use in zebrafish to test the temporal role that Sox2 plays during midline formation. We found ectopic Sox2 in the presence of Wnt activity holds cells in the undifferentiated floor plate/notochord progenitor state, and that degradation of the ectopic Sox2 is required for cells to adopt a notochord fate.

A real-world drug safety surveillance study from the FAERS database of hepatocellular carcinoma patients receiving pembrolizumab alone and plus lenvatinib

Pembrolizumab plus Lenvatinib is regarded as a significant treatment option for advanced unresectable hepatocellular carcinoma (HCC). This study aims to meticulously monitor and identify adverse events (AEs) related to this combined therapy, enhance patient safety, and offer evidence-based recommendations for the appropriate use of these drugs. We gathered adverse drug reactions (ADRs)-related data from the FAERS database for HCC patients who received Pembrolizumab, both alone and in combination with Lenvatinib from the first quarter of 2014 to the fourth quarter of 2023. ADRs signal detection was performed using the ROR, PRR, BCPNN, MHRA, and MGPS methods. We gathered data on 459 and 358 AEs from patients with HCC treated with pembrolizumab alone and in combination with lenvatinib, respectively. Using four signal quantification techniques, we identified 50 and 38 distinct AEs, which were classified into 15 different System organ class (SOC) categories. Notably, the most common AEs associated with pembrolizumab were gastrointestinal disorders and hepatobiliary disorders. In both patient groups, the most frequently reported AEs were hepatic encephalopathy, blood bilirubin increased and diarrhea. We also observed some unexpected significant AEs, such as dehydration, skin ulcers, and intestinal perforation. The countries reporting the highest number of AEs were the United States, followed by China, France, and Japan. The median onset time for AEs related to pembrolizumab alone and its combination with lenvatinib was 80.5 days (interquartile range 20.0–217.3 days) and 77.5 days (interquartile range 19.7–212.3 days), respectively. This study offers new insights into the monitoring and management of ADRs in HCC patients receiving pembrolizumab alone or in combination with lenvatinib. It is crucial to closely monitor the safety of this treatment regimen in HCC patients to avoid serious AEs.

Links Between Variation in Movement-Based Visual Signals and Social Communication Complexity in an Asian Agamid Lizard Phrynocephalus vlangalii.

The social complexity hypothesis suggests that complex social interactions drive the evolution of sophisticated communicative signals. While the relationship between social communication and the complexity of sound and color signals has been extensively studied, the correlation between social communication and movement-based visual signal complexity remains underexplored. In this study, we selected the Asian agamid lizard, Phrynocephalus vlangalii, as our model system. Through a combination of controlled experiments, behavioral observations, and signal quantification, we examined the relationship between social communications and variation in movement-based visual signals and tested our social complexity hypothesis. Our experiments revealed that males significantly decreased the tail coil duration during the mating season to deal with high social interaction. Conversely, females significantly increased the tail display duration and variation in mean tail coil amplitude in line with the intensity of parental care, and tail display duration showed a significant and positive correlation with female reproductive investment. These findings suggest that social communication plays an important role in shaping the changes in movement-based visual signals, providing new evidence for the social complexity hypothesis.

Plasmid Stability Analysis with Open-Source Droplet Microfluidics.

Plasmids play a vital role in synthetic biology by enabling the introduction and expression of foreign genes in various organisms, thereby facilitating the construction of biological circuits and pathways within and between cell populations. For many applications, maintaining functional plasmids without antibiotic selection is critical. This study introduces an open-hardware-based microfluidic workflow for analyzing plasmid retention by culturing single cells in gel microdroplets and quantifying microcolonies using fluorescence microscopy. This approach allows for the parallel analysis of numerous droplets and microcolonies, providing greater statistical power compared to traditional plate counting and enabling the integration of the assay into other droplet microfluidic workflows. By using plasmids expressing fluorescent proteins alongside a non-specific fluorescent DNA stain, single colonies can be identified and differentiated based on plasmid loss or fluorescent marker expression. Notably, this advanced workflow, implemented with open-source hardware, offers precise flow control and temperature management of both the sample and the microfluidic chip. These features enhance the workflow's ease of use, robustness, and accessibility. While the study focuses on Escherichia coli as the experimental model, the method's true potential lies in its versatility. It can be adapted for various studies requiring fluorescence signal quantification from plasmids or stains, as well as for other applications. The adoption of open-source hardware broadens the potential for conducting high-throughput bioanalyses using accessible technology in diverse research settings.

Safety assessment of ezetimibe: real-world adverse event analysis from the FAERS database

ABSTRACT Background Ezetimibe is known for its lipid-lowering safety and tolerability, but its real-world adverse effects have not been fully evaluated. In this study, adverse events associated with ezetimibe were investigated using the FAERS database for the period 2004 to 2023. Research design and methods Adverse events data for ezetimibe, spanning from the first quarter of 2004 to the fourth quarter of 2023, were standardized and analyzed using signal quantification methods like Reporting Odds Ratio (ROR), Proportional Reporting Ratio (PRR), Bayesian Confidence Propagation Neural Network (BCPNN), and Multi-item Gamma Poisson Shrinker (EBGM). Results Among 11,550 adverse drug events reports with ezetimibe as the primary suspect drug, 211 preferred terms (PTs) were identified across 24 different system organ classes (SOCs).Notably, in addition to the adverse reactions already specified in the instruction manual, unstable angina (n = 120, ROR 30.53(25.47-36.58), PRR 30.41(25.49-36.28), IC 4.9(4.64), EBGM 29.85(25.66)), crush syndrome(n = 19,ROR 298.83(182.95-488.09), PRR 298.65(182.96-487.49), IC 7.97(7.29),EBGM 251.12(166.57)), and autoscopy (n = 7, ROR 28.81(13.64-60.85), PRR 28.80(13.68-60.65), IC 4.82(3.81), EBGM 28.30(15.14))were new adverse reactions that emerged as strong signals. Conclusions Ezetimibe is effective in lowering blood lipids, but there is a risk of adverse reactions such as unstable angina, rhabdomyolysis and autoscopy, which require careful monitoring by physicians.

Safety assessment of tafamidis: a real-world adverse event analysis from the FAERS database

ABSTRACT Background Tafamidis emerged as the first FDA-approved drug for treating termed amyloid fibrils. This study aims to analyze adverse events (AEs) related to tafamidis from the second quarter (Q2) of 2019 to the fourth quarter (Q4) of 2023 from the FDA adverse event reporting system (FAERS) database. Research design and methods The AE data related to tafamidis from 2019 Q2 to 2023 Q4 were collected and standardized. Various signal quantification techniques, including reporting odds ratio (ROR), proportional reporting ratio (PRR), Bayesian confidence propagation neural network, and multi-item gamma Poisson shrinker, were employed for analysis. Results Among the 8742 AE reports with tafamidis as the primary suspected drug, 180 preferred terms of AEs spanning 27 different system organ classes were identified. Unique adverse events to tafamidis included renal and urinary disorders and ear and labyrinth disorders, with not mentioned in the official drug label. Additionally, uncommon but significantly strong AE signals, such as blood culture positive and acquired hemophilia, were observed. Common AEs with relatively high occurrence rates included death, off-label use, fall, hypoacusis, and dysphagia. Conclusion These findings offer valuable insights for optimizing the use of tafamidis and reducing potential side effects, thereby facilitating its safe use in clinical settings.

Nanopaper Integrated Smart Device: An Opto-Electrochemical Biosensor for Real-Time Dual On-Field Detection of Organophosphorus Pesticides.

The frequent and excessive use of organophosphorus pesticides in the agriculture industry raises persistent concerns regarding their environmental protection and public health implications. Addressing these issues requires the development of affordable and reliable sensing platforms for on-field monitoring to mitigate their adverse impacts promptly. This study utilizes nanocellulose papers (bacterial and TEMPO-oxidized) combined with butyrylcholinesterase to create a novel reagent-free and orthogonal nanobioplatform featuring smart opto-electrochemical dual outputs. An integrated nano-PAD, preloaded with enzymes and enzymatic substrates, is fabricated using wax-printing and screen-printing technologies. The nano-PAD measures opto-electroactive products, specifically indoxyl and thiocholine, whose concentrations correlate directly with the enzymatic inhibition caused by paraoxon, used as the organophosphate model. To enhance user convenience and meet the requirements for smart real-time point-of-need detection, integration of the nano-PAD with a smartphone-operated miniaturized potentiostat and a self-developed portable smart optical reader is achieved. The developed bioanalytical platform, further supported by a self-developed Android application, enables accurate and efficient quantification of dual signals in real time. The system covers a wide detection range of paraoxon (20-100 ppb) and demonstrates reliable recovery levels (ranging from 98 to 107%) in a real matrix, specifically wastewater. Given these demonstrated capabilities, this innovative biosensing strategy holds substantial potential for practical application in environmental surveillance, facilitating timely and informed environmental management decisions, particularly in resource-limited settings where traditional analytical tools are inaccessible.

An Advanced Combinatorial System from Vitis vinifera Leaves and Propolis Enhances Antioxidants' Skin Delivery and Fibroblasts Functionality.

Background/Objectives: Vine leaves are a bulky by-product that are disposed of and treated as waste in the wine production process. In the present study polyphenols from vine leaves were extracted and simultaneously encapsulated in a new delivery system consisting of liposomes and cyclodextrins. This system was further combined with propolis polyphenols encapsulated in cyclodextrins, resulting in a colloidal suspension for the release of antioxidants in a time-controlled way, the rate of which depends on the ratio of the materials. The result is a raw material that exhibits antioxidant and ECM protective effects when administered in skin fibroblasts (NHDFs). Methods: The antioxidant and ECM promoting efficacy of the produced raw material was assessed by the Folin-Ciocalteu method, DPPH assay, and in cellulo assays in fibroblasts, such as the cell viability assay, scratch assay, cell migration assay, gene expression analysis, and immunofluorescence analysis, for the detection, visualization, and quantification of collagen-I, collagen-IIIa, and elastin signals and collagenase assay. Results: Treatment of NHDFs with the combinatorial delivery system promoted collagen and elastin synthesis and deposition in normal conditions and, upon induced external stress, as assessed by in vitro transcriptomic and proteomic analysis. A significant inhibition of collagenase was also observed, suggesting a multitargeted efficacy of the active ingredients also by preventing collagen degradation. Conclusions: Therefore, this liposome-cyclodextrin encapsulated polyphenol complex represents a novel bioactive ingredient with promising skin applications.

Splenic T2 signal intensity loss on MRI is associated with disease burden in multiple myeloma.

This study aims to evaluate correlations between spleen signal changes in different MRI sequences and bone marrow plasma cell infiltration as potential indicator of disease burden in multiple myeloma (MM) patients. We retrospectively analyzed 45 patients with newly diagnosed MM that underwent whole-body MRI with axial DWI at b-values 50 (b50) and 800 (b800), and coronal T1 and T2 fast spin-echo (T2-TSE) imaging. A subcohort of 39 patients had concomitant [18F]FDG PET/CT. The spleen was segmented in all MRI sequences and signal intensities were normalized. MR signal intensities and ADC values were correlated with bone marrow plasma cell infiltration from biopsy, laboratory markers (Beta 2-microglobulin, M-Protein, Red blood count (RBC), Hemoglobin, Hematocrit, Total protein, Creatinine), clinical data (ISS stages, high-risk chromosomal aberrations), and standardized uptake value (SUV) in the spleen as well as spleen-to-liver and spleen-to-blood pool SUV ratios on [18F]FDG PET-CT. Bone marrow plasma cell infiltration was negatively correlated with (normalized) mean splenic signal intensity on DWI-b50, DWI-b800, and T2-TSE images (r = -0.64, p < 0.001, r = -0.58, p < 0.001, and r = -0.66, p < 0.001, respectively) while there was no correlation with the apparent diffusion coefficient or spleen size (p = 0.52). In the subgroup analysis of 39 patients with concomitant [18F]FDG PET-CT, there was no correlation of normalized splenic [18F]FDG uptake either with MR spleen signal (for T2 p = 0.64) or with bone marrow plasma cell infiltration (p = 0.37). Our findings reveal a significant association between spleen signal intensity especially on normalized T2-weighted images and tumor burden. Question What changes occur in spleen signal on MRI as tumor load marker changes in multiple myeloma (MM)? Findings Spleen signal intensity, particularly on T2-weighted MRI, negatively correlates with bone marrow plasma cell infiltration and laboratory markers of tumor burden. Clinical relevance Standardized quantification of splenic T2 signal is proposed as a new marker for MM disease burden.

A real-world disproportionality analysis of FDA adverse event reporting system (FAERS) events for avatrombopag

Avatrombopag is a next-generation thrombopoietin receptor agonist approved for the treatment of thrombocytopenia in patients with chronic liver disease scheduled to undergo surgery and thrombocytopenia in patients with chronic immune thrombocytopenia. However, realistic data on its post-marketing long-term safety and tolerability in large sample populations are incomplete. The adverse event (AE) reports of avatrombopag were analyzed based on the FAERS database. By extracting large-scale data from the FAERS database, this study used various signal quantification techniques such as reporting odds ratios, proportional reporting ratios, Bayesian confidence propagation neural network, and multi-item gamma Poisson shrinker to calculate and evaluate the ratio and association between avatrombopag and specific AEs. In the FAERS database, data from the second quarter of 2018 to the fourth quarter of 2023 were extracted for this study, a total of 1217 avatrombopag-related AEs reports were included for analysis. Based on four calculation methods, this study identified and 32 preferred terms associated with avatrombopag. Common AEs in the product label such as thrombosis, headache, contusion, petechiae, and gingival bleeding were detected. AEs not mentioned in the product label, such as photopsia and ear discomfort were also detected. The first 30 days after initiation of medication were the most common period for both serious and non-serious AEs. This study demonstrates the common AEs and the potential AEs associated with avatrombopag in real-world practice, which could provide valuable cautionary evidence for clinicians and pharmacists to manage safety issues with avatrombopag.

Exploring Plasmonic Standalone Surface-Enhanced Raman Scattering Nanoprobes for Multifaceted Applications in Biomedical, Food, and Environmental Fields.

Surface-enhanced Raman scattering (SERS) is an innovative spectroscopic technique that amplifies the Raman signals of molecules adsorbed on rough metal surfaces, making it pivotal for single-molecule detection in complex biological and environmental matrices. This review aims to elucidate the design strategies and recent advancements in the application of standalone SERS nanoprobes, with a special focus on quantifiable SERS tags. We conducted a comprehensive analysis of the recent literature, focusing on the development of SERS nanoprobes that employ novel nanostructuring techniques to enhance signal reliability and quantification. Standalone SERS nanoprobes exhibit significant enhancements in sensitivity and specificity due to optimized hot spot generation and improved reporter molecule interactions. Recent innovations include the development of nanogap and core-satellite structures that enhance electromagnetic fields, which are crucial for SERS applications. Standalone SERS nanoprobes, particularly those utilizing indirect detection mechanisms, represent a significant advancement in the field. They hold potential for wide-ranging applications, from disease diagnostics to environmental monitoring, owing to their enhanced sensitivity and ability to operate under complex sample conditions.

Evaluation of an integrated variable flip angle protocol to estimate coil B1 for hyperpolarized MRI

AbstractPurposeThe purpose of this work is to validate a simple and versatile integrated variable flip angle (VFA) method for mapping B1 in hyperpolarized MRI, which can be used to correct signal variations due to coil inhomogeneity.Theory and MethodsSimulations were run to assess performance of the VFA B1 mapping method compared to the currently used constant flip angle (CFA) approach. Simulation results were used to inform the design of VFA sequences, validated in four volunteers for hyperpolarized xenon‐129 imaging of the lungs and another four volunteers for hyperpolarized carbon‐13 imaging of the human brain. B1 maps obtained were used to correct transmit and receive inhomogeneity in the images.ResultsSimulations showed improved performance of the VFA approach over the CFA approach with reduced sensitivity to T1. For xenon‐129, the B1 maps accurately reflected the variation of signal depolarization, but in some cases could not be used to correct for coil receive inhomogeneity due to a lack of transmit‐receive reciprocity resulting from suboptimal coil positioning. For carbon‐13, the B1 maps showed good agreement with a separately acquired B1 map of a phantom and were effectively used to correct coil‐induced signal inhomogeneity.ConclusionA simple, versatile, and effective VFA B1 mapping method was implemented and evaluated. Inclusion of the B1 mapping method in hyperpolarized imaging studies can enable more robust signal quantification.

A real-world pharmacovigilance study of FDA Adverse Event Reporting System events for pralsetinib.

Pralsetinib, a selective oral inhibitor of rearranged during transfection (RET) fusion proteins and oncogenic RET mutants, has shown significant efficacy in treating RET fusion-positive non-small cell lung cancer and thyroid cancer. However, since pralsetinib was approved in the United States in September 2020, there have been limited reports of post-marketing adverse events (AEs). In this study, we aimed to analyze the AE signals with pralsetinib on the basis of the United States Food and Drug Administration (FDA) Adverse Event Reporting System (FAERS) to provide instructions in clinical practice. All AE reports were obtained from the FAERS database from the first quarter (Q3) of 2020 to the second quarter (Q2) of 2024. Various signal quantification techniques were used for analysis, including reporting odds ratios, proportional reporting ratios, Bayesian confidence propagation neural network, and multi-item gamma Poisson shrinker (MGPS)-based empirical Bayesian geometric mean. Out of 8,341,673 case reports in the FAERS database, 1,064 reports of pralsetinib as the "primary suspected (PS)" AEs were recorded, covering 26 system organ classes and 256 preferred terms. Of the reports, 62.5% were from consumers rather than healthcare professionals. The most common systems were general disorders and administration site conditions (n = 704), investigations (n = 516), and gastrointestinal disorders (n = 405). A total of 95 significant disproportionality preferred terms (PTs) conformed to the four algorithms simultaneously. AEs that ranked the top three at the PT level were hypertension (n = 80), asthenia (n = 79), and anemia (n = 65). Of the 95 PTs with significant disproportionation, unexpected significant AEs such as increased blood calcitonin, increased myocardial necrosis marker, and bacterial cystitis were observed, which were not mentioned in the drug's instructions. The median onset time of pralsetinib-associated AEs was 41 days [interquartile range (IQR) 14-86 days]. The majority of the AEs occurred in 30 days (42.86%). Our pharmacovigilance analysis of real-world data from the FEARS database revealed the safety signals and potential risks of pralsetinib usage. These results can provide valuable evidence for further clinical application of pralsetinib and are important in enhancing clinical medication safety.

Drug-induced noninfectious myocarditis: a disproportionality analysis of the FAERS database

ABSTRACT Background This investigation aims to identify and evaluate the most common and critical drugs associated with the risk of noninfectious myocarditis utilizing the US Food and Drug Administration Adverse Event Reporting System (FAERS) database. Methods Data pertaining to noninfectious myocarditis from 2004 Q1 to 2024 Q2 were extracted. Following data standardization, multiple signal quantification techniques, including Reporting Odds Ratio (ROR) and Proportional Reporting Ratio, were employed for analysis. Results The study identified a total of 10,763 adverse event reports associated with noninfectious myocarditis. Disproportionality analysis revealed that the top 5 drugs by ROR were phendimetrazine tartrate (ROR 104.64), trimethoprim + sulfamethoxazole (ROR 67.65), aldesleukin (ROR 52.67), mesalazine (ROR 49.73), and balsalazide disodium (ROR 45.26). Notably, among the 30 drugs with the strongest ROR signals, 8 drugs lacked myocarditis risk information in their package inserts. Conclusion Through comprehensive analysis of the FAERS database, our study identified drugs with a strong signal for myocarditis that are not currently indicated on their labels. The findings suggest that the potential risk of myocarditis associated with these medications is significant and warrants close monitoring in clinical practice.

Quantitative magnetooptical analysis using indicator films for the detection of magnetic field distributions, temperature, and electrical currents

The accurate characterization of local magnetic fields and temperature is vital for the design of electronic systems. To meet this imperative, we present a novel non-contact approach for simultaneous quantitative magnetic field imaging and temperature sensing using magnetooptics and a bismuth-doped yttrium iron garnet film with out-of-plane anisotropy. For the direct signal quantification, a Stokes polarization camera is employed in a conventional magnetooptical microscope. The magnetization in the garnet is modulated with an external magnetic field to continuously image the Faraday rotation at four distinct points along the saturating magnetization loop. The method enables sensing of the magnetooptical signal in saturation, the magnetooptical susceptibility, the temperature, and self-calibrated driftfree imaging of the out-of-plane magnetic field component. A spatial resolution of magnetic field in the micrometer range with millisecond exposure time is demonstrated. The method is verified by analyzing the stray magnetic field distribution of electrical current in a wire simultaneously to the Joule heating induced by the applied current.

Mining and analysis of adverse event signals for alendronate based on the real-world data of FDA adverse event reporting system database

ABSTRACT Objective Our study aims to assess alendronate-related adverse events (AEs) from the US FDA adverse event reporting system database. Methods The AE data associated with alendronate between the first quarter of 2004 and the first quarter of 2024 were selected. Various signal quantification methods, including the ROR, PRR, BCPNN, and EBGM, were applied for analysis. Results In 34,943 reports where alendronate was the primary suspected drug for the AE, 24 affected system organ classes and 1046 significant preferred terms were identified in this study. Several significant AEs beyond drug instructions with strong signals were determined, including low turnover osteopathy, fracture delayed union, fracture nonunion, loss of anatomical alignment after fracture reduction, fracture malunion, periprosthetic fracture, carotid bruit, oral fibroma, traumatic occlusion, and phlebolith. The median time to onset of alendronate-related AEs was 306 days (interquartile range [IQR] 12–1,461 days), and the majority of cases occurred 2 years later (18.80%) and within 30 days (14.49%). Conclusions The current study detected multiple potential new AE signals for alendronate, and more clinical researches are required to further validate our results and clarify their associations.

Inputs of pharmacoepidemiology in addictovigilance: How do they fit together?

The French Addictovigilance Network has been using data from the French Heath insurance since the late 1990s to assess prescription drug abuse. In this narrative review, we illustrate the inputs of pharmacoepidemiology in addictovigilance based on the experience of the French Addictovigilance Network. The review focuses on pharmacoepidemiology using the French National Health Data System. We propose three examples: the MEGADOSE study, which aimed to conduct the first nation-wide, systematic, repeated assessment of doctor shopping; the DANTE study, which aimed to assess trends in analgesic use, focusing on the prevalence of use and the demographic profiles of analgesic users by age and sex; and the ZORRO study, which aimed to assess the impact of secure prescription forms on the use of zolpidem and other sedatives. These examples show how pharmacoepidemiology fits in the multifaceted monitoring conducted by the French Addictovigilance Network and complements the other data sources of this framework. This approach improves signal detection, confirmation, and quantification. It also makes it possible to overcome the limitations of each data source taken individually.

BrightMice: a low-cost do-it-yourself instrument, designed for in vivo fluorescence mouse imaging.

In vivo fluorescent imaging represents a potent means for real-time probe quantification, facilitating insights into disease pathophysiology and therapeutic responses. Nonetheless, accurate signal quantification remains challenging due to inherent factors like light scattering and tissue absorption. Existing imaging systems, though sophisticated, often entail high costs and are typically restricted to well-funded laboratory settings. This study introduces BrightMice, an innovative in vivo fluorescent imaging system that harnesses 3D printing and consumer-grade digital cameras. Tailored for various fluorophores such as EYFP and E2-crimson, the system showcases both adaptability and effectiveness in detecting in vivo fluorescent signals in several reporter mouse strains. Comparative analyses against commercial instruments confirm BrightMice's sensitivity and underscore its potential to democratize in vivo fluorescence imaging. By providing a cost-effective and accessible solution, BrightMice stands to benefit diverse research environments.

Miniaturized protein profiling permits targeted signaling pathway analysis in individual circulating tumor cells to improve personalized treatment

BackgroundTraditional genomic profiling and mutation analysis of single cells like Circulating Tumor Cells (CTCs) fails to capture post-translational and functional alterations of proteins, often leading to limited treatment efficacy. To overcome this gap, we developed a miniaturized ‘protein analysis on the single cell level’ workflow—baptized ZeptoCTC. It integrates established technologies for single-cell isolation with sensitive Reverse Phase Protein Array (RPPA) analysis, thus enabling the comprehensive assessment of multiple protein expression and activation in individual CTCs.MethodsThe ZeptoCTC workflow involves several critical steps. Firstly, individual cells are labeled and isolated. This is followed by cell lysis and the printing of true single cell lysate preparations onto a ZeptoChip using a modified micromanipulator, CellCelector™. The printed lysates then undergo fluorescence immunoassay RPPA protein detection using a ZeptoReader. Finally, signal quantification is carried out with Image J software, ensuring precise measurement of multiple protein levels.ResultsThe efficacy of ZeptoCTC was demonstrated through various applications. Initially, it was used for measuring EpCAM protein expression, a standard marker for CTC detection, revealing higher levels in single MCF-7 over MDA-MB-231 tumor cells. Furthermore, in Capivasertib (Akt-inhibitor)-treated MCF-7 single cells, ZeptoCTC detected a 2-fold increase in the pAkt/Akt ratio compared to control cells, and confirmed co-performed bulk-cell western blot analysis results. Notably, when applied to individual CTCs from metastasized breast cancer patients, ZeptoCTC revealed significant differences in protein activation levels, particularly in measured pAkt and pErk levels, compared to patient-matched WBCs. Moreover, it successfully differentiated between CTCs from patients with different Akt1 genotypes, highlighting its potential to determine the activation status of druggable cancer driving proteins for individual and targeted treatment decision making.ConclusionsThe ZeptoCTC workflow represents a valuable tool in single cell cancer research, crucial for personalized medicine. It permits detailed analysis of key proteins and their activation status of targeted, cancer-driven signaling pathways in single cell samples, aiding in understanding tumor response, progression, and treatment efficacy beyond bulk analysis. The method significantly advances clinical investigations in cancer, improving treatment precision and effectiveness. The workflow will be applicable to protein analysis on other types of single cells like relevant in stem cell, neuropathology and hemopoietic cell research.