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Phosphorylated signal transducer and activator of transcription 3 (pSTAT3), which is related to anti‐apoptosis, cellular proliferation, invasion and migration of tumours, has prognostic significance in malignant tumours in humans as well as in canine melanoma. However, the significance of pSTAT3 in canine liver tissues has not yet been evaluated. This study's objective was to compare its expression in canine normal, non‐neoplastic hepatic disease and hepatocellular carcinoma (HCC) tissues by immunohistochemical analysis. Furthermore, the association between pSTAT3 immunostaining and clinicopathological factors was investigated. Overall, 68 canine liver tissues, including 10 normal liver tissues, 30 non‐neoplastic hepatic disease tissues and 28 HCC tissues were examined, revealing distinct differences in pSTAT3 immunostaining among the groups. (p < 0.001). Additionally, high pSTAT3 immunostaining was significantly associated with increased tumour size (5 > cm) (p = 0.041), and metastasis (p = 0.046). Furthermore, Kaplan–Meier survival curve analysis revealed a correlation between high pSTAT3 immunostaining and poor disease‐free survival (p = 0.013) and overall survival (p = 0.011). These findings suggest that overactivation of STAT3 is associated with poor prognosis in canine HCC. Therefore, pSTAT3 is considered a potential prognostic marker and therapeutic target for canine HCC. [ABSTRACT FROM AUTHOR]

Purpose: The objective of the present work was to test the feasibility of in vivo imaging of hyperpolarized 50‐nm silicon‐29 (29Si) nanoparticles. Methods: Commercially available, crystalline 50‐nm nanoparticles were hyperpolarized using dynamic polarization transfer via the endogenous silicon oxide–silicon defects without the addition of exogenous radicals. Phantom experiments were used to quantify the effect of sample dissolution and various surface coating on T1 and T2 relaxation. The in vivo feasibility of detecting hyperpolarized silicon‐29 was tested following intraperitoneal, intragastric, or intratumoral injection in mice and compared with the results obtained with previously reported, large, micrometer‐size particles. The tissue clearance of SiNPs was quantified in various organs using inductively coupled plasma optical emission spectroscopy. Results: In vivo images obtained after intragastric, intraperitoneal, and intratumoral injection compare favorably between small and large SiNPs. Improved distribution of small SiNPs was observed after intraperitoneal and intragastric injection as compared with micrometer‐size SiNPs. Sufficient clearance of nanometer‐size SiNPs using ex vivo tissue sample analysis was observed after 14 days following injection, indicating their safe use. Conclusion: In vivo MRI of hyperpolarized small 50‐nm SiNPs is feasible with polarization levels and room‐temperature relaxation times comparable to large micrometer‐size particles. [ABSTRACT FROM AUTHOR]

Virtual histopathology is an emerging technology in medical imaging that utilizes advanced computational methods to analyze tissue images for more precise disease diagnosis. Traditionally, histopathology relies on manual techniques and expertise, often resulting in time-consuming processes and variability in diagnoses. Virtual histopathology offers a more consistent, and automated approach, employing techniques like machine learning, deep learning, and image processing to simulate staining and enhance tissue analysis. This review explores the strengths, limitations, and clinical applications of these methods, highlighting recent advancements in virtual histopathological approaches. In addition, important areas are identified for future research to improve diagnostic accuracy and efficiency in clinical settings. [ABSTRACT FROM AUTHOR]

Feline hyperthyroidism (FHT) is a debilitating disease affecting > 10% of elderly cats. It is generally characterised by chronic elevation of thyroid hormone in the absence of circulating TSH. Understanding of the molecular pathogenesis of FHT is currently limited. However, FHT shares clinical and histopathological similarities with human toxic multinodular goitre, which has been associated with activating mutations in TSH receptor (TSHR) and Gsα encoding genes. Using RNA-seq transcriptomic analysis of thyroid tissue from hyperthyroid and euthyroid cats, we identified differentially expressed genes and dysregulated pathways in FHT, many of which are downstream of TSHR. In addition, we detected missense variants in thyroid RNA-seq reads that alter the structure of both TSHR and Gsα. All FHT-associated mutations were absent in germline sequence from paired blood samples. Only a small number of hyperthyroid cats demonstrated TSHR variation, however all thyroids from advanced cases of FHT carried at least one missense variant affecting Gsα. The activating nature of the acquired Gsα mutations was demonstrated by increased cAMP production in vitro. These data indicate that constitutive activation of signalling downstream of TSHR is central to the TSH-independent production of thyroid hormone in FHT, offering a novel therapeutic target pathway in this common disease. [ABSTRACT FROM AUTHOR]

Zinc finger protein 384 (ZNF384) is a highly conserved transcribed gene associated with the development of multiple tumors, however, its role and mechanism in serous ovarian cancer (SOC) are unknown. We first confirmed that ZNF384 was abnormally highly expressed in SOC tissues by bioinformatics analysis and immunohistochemistry. We further used lentivirus packaging and transfection techniques to construct ZNF384 overexpression or knockdown cell lines, and through a series of cell function experiments, gradually verified that ZNF384 promoted a series of malignant behaviors of SOC cell proliferation, migration, and invasion. By establishing a xenotransplantation model in nude mice, it was confirmed that ZNF384 promoted the progress of SOC in vivo. Mechanistically, Overexpression of ZNF384 enhanced the transcriptional activity of Lin-28 homolog B (LIN28B), which promoted the malignant behavior of SOC cells. In addition, LIN28B could regulate the expression of the downstream factor ubiquitin D (UBD) in SOC cells, further promoting the development of SOC. This study shows that ZNF384 aggravates the malignant behavior of SOC cells through the LIN28B/UBD axis, which may be used as a diagnostic biomarker for patients with SOC. [ABSTRACT FROM AUTHOR]

Background: Drug discovery strongly relies on the thorough evaluation of preclinical experimental studies. In the context of pulmonary fibrosis, micro-computed tomography (µCT) and histology are well-established and complementary tools for assessing, in animal models, disease progression and response to treatment. µCT offers dynamic, real-time insights into disease evolution and the effects of therapies, while histology provides a detailed microscopic examination of lung tissue. Here, we present a semi-automatic pipeline that integrates these readouts by matching individual µCT volume slices with the corresponding histological sections, effectively linking densitometric data with Ashcroft score measurements. Methods: The tool first geometrically aligns the vertical axis of the µCT volume with the cutting plane used to prepare the histological sample. Then, focusing on the left lung, it computes the affine registration that identifies the µCT coronal slice that best matches the histological section. Finally, quantitative µCT imaging parameters are extracted from the selected slice. In a proof-of-concept test, the tool was applied to a bleomycin-induced mouse model of lung fibrosis. Results: The proposed approach demonstrated high accuracy and time effectiveness in matching µCT and histological sections minimizing manual intervention, with an overall success rate of 95%, and reduced time required to align µCT and histological data from 40 to 5 min. Significant correlations were found between quantitative data derived from µCT and histology data. Conclusions: The precise combination of microscopic ex-vivo information with 3D in-vivo data enhances the accuracy and representativeness of tissue analysis and provides a structural context for omic studies, serving as the foundation for a multi-layer platform. By facilitating a detailed and objective view of disease progression and treatment response, this approach has the potential to accelerate the development of effective therapies for lung fibrosis. [ABSTRACT FROM AUTHOR]

Simple Summary: Antibody Drug Conjugates (ADCs) are a newer class of therapeutic agents which present a promising therapeutic approach for non-small cell lung cancer (NSCLC). Several ADCs are in clinical trial, targeting various antigens including TROP2, HER2, MET and others. Despite the promise, ADCs present unique challenges including toxicity like interstitial lung disease (ILD). A more thorough analysis of target expression could lead to a better understanding of adverse events, such as ILD. This improved understanding may help reduce the occurrence of these adverse effects. Our study aims to analyze the ADC target expression in lung tissue to help us understand the relation between treatment with ADCs and onset of interstitial lung disease. Introduction: Non-small-cell lung cancer (NSCLC) remains a leading cause of cancer-related mortality worldwide, despite advances in immune checkpoint inhibitors and targeted therapies. Antibody–drug conjugates (ADCs) represent a promising therapeutic approach by delivering cytotoxic agents specifically to cancer cells, potentially reducing harm to healthy tissues. This study aims to explore the effectiveness and challenges associated with ADCs in NSCLC, with a focus on drug-induced interstitial lung disease (D-ILD). Methods: A comprehensive literature review was conducted across MEDLINE (Ovid), Embase (Elsevier), CENTRAL (Cochrane Library), and other sources up to March 2023, to identify ADCs used in NSCLC treatment and their associated risk of D-ILD. The incidence of ILD was analyzed from clinical trial data, while ADC target expression was examined through RNA and protein levels in normal and tumor lung tissues. Discussion: Our findings highlight the therapeutic potential of ADCs in NSCLC, as evidenced by significant clinical outcomes. However, the occurrence of D-ILD presents a notable challenge, as its incidence was not directly correlated with the expression levels of the target antigens. This suggests that D-ILD may result from factors beyond antigen expression, including the cytotoxic payload and linker characteristics of ADCs. Conclusion: ADCs offer a promising avenue for NSCLC treatment. Nonetheless, the risk of D-ILD necessitates a balanced approach in ADC development, focusing on optimizing linker and payload properties to mitigate this adverse effect. Further research is essential to better understand and manage D-ILD, ensuring the safe and effective use of ADCs in clinical practice. [ABSTRACT FROM AUTHOR]

The presence of microplastics in the environment is now becoming a challenge for many scientific disciplines. Molecular diversity and spatial migration make it difficult to find plastic-free areas. Their negative, often toxic, effects affect plants and animals to varying degrees, causing many biochemical disorders, species degradation, and population changes. This study aimed to determine the possibility of accumulation of spherical low-density polyethylene particles of 38–63 µm (38–45 µm 1.00 g/cm3, and 53–63 µm 1.00 g/cm3) with fluorescent properties in muscle tissues of the cosmopolitan earthworm species Lumbricus terrestris, exposed to plastic contained in the soil at a concentration of 0.1% dry weight for 3 months. Analysis of the tissues by Raman microscopy included the estimation of mapping area size, sampling density, accumulation time, spectra, laser line, and laser power to detect plastic in the samples effectively. Our results demonstrate the ability of low-density polyethylene microparticles to accumulate in earthworm tissues and are presented graphically for the mapping area and images with plastic detection sites marked. In addition, this article highlights the potential of using Raman microscopy for research in the field of tissue analysis. [ABSTRACT FROM AUTHOR]

Transplantable ready-made microvessels have therapeutic potential for tissue regeneration and cell replacement therapy. Inspired by the natural rapid angiogenic sprouting of microvessels in vivo, engineered injectable 3D microvessel networks are created using thermoplastic elastomer (TPE) microfluidic devices. The TPE material used here is flexible, optically transparent, and can be robustly yet reversibly bonded to a variety of plastic substrates, making it a versatile choice for microfluidic device fabrication because it overcomes the weak self-adhesion properties and limited manufacturing options of poly(dimethylsiloxane) (PDMS). By leveraging the reversible bonding characteristics of TPE material templates, we present their utility as an organ-on-a-chip platform for forming and handling microvessel networks, and demonstrate their potential for animal-free tissue generation and transplantation in clinical applications. We first show that TPE-based devices have nearly 6-fold higher bonding strength during the cell culture step compared to PDMS-based devices while simultaneously maintaining a full reversible bond to (PS) culture plates, which are widely used for biological cell studies. We also demonstrate the successful generation of perfusable and interconnected 3D microvessel networks using TPE–PS microfluidic devices on both single and multi-vessel loading platforms. Importantly, after removing the TPE slab, microvessel networks remain intact on the PS substrate without any structural damage and can be effectively harvested following gel digestion. The TPE-based organ-on-a-chip platform offers substantial advantages by facilitating the harvesting procedure and maintaining the integrity of microfluidic-engineered microvessels for transplant. To the best of our knowledge, our TPE-based reversible bonding approach marks the first confirmation of successful retrieval of organ-specific vessel segments from the reversibly-bonded TPE microfluidic platform. We anticipate that the method will find applications in organ-on-a-chip and microphysiological system research, particularly in tissue analysis and vessel engraftment, where flexible and reversible bonding can be utilized. [ABSTRACT FROM AUTHOR]

Introduction: Distinct androgen formulations have been used as gender-affirming hormone treatment, but little is known about the specific changes that may occur in the ovary depending on the testosterone preparation used. The study aims to evaluate the histological modifications of the ovarian tissue and the hormonal changes after gender-affirming surgery based on the testosterone preparation employed, such as testosterone cypionate or undecanoate. Design: Unicenter transversal cohort study. Materials and methods: Sixty transmasculine persons before and after gender-affirming surgery. A histological examination of the ovaries was conducted, including the follicular population and the characterization of the ovarian stroma. Hormonal status (testosterone, estradiol, FSH, and LH) were also assessed before and after the procedure. Results: The median age of participants was similar between the two groups (27.9 vs. 26.7 years, p = 0.27). There were no differences in all hormonal determinations before gender-affirming surgery between the groups. After surgery, FSH levels increased significantly, especially in the testosterone undecanoate group compared to the cypionate group (72.3 vs. 38.3 U/L, p = 0.02), consistent with LH determinations (43.0 vs. 23.4 U/L, p = 0.02). However, no regimen modification was required for any individual. No statistical differences were observed in any parameter concerning the follicular population, nor were there any variances in the thickness of the tunica albuginea (p = 0.85) or the proportion of luteinized stromal cells. Nevertheless, there was a tendency toward increased luteinization in the testosterone cypionate group (88.2% vs. 76.9%, p > 0.05). Conclusions: In a cohort of transmasculine individuals using different androgen preparations, histological analysis of ovarian tissue revealed comparable findings. Both groups exhibited similar follicular populations and comparable modifications in stromal tissue. However, significant differences were observed in hormonal profiles, although no modification in testosterone dosage was needed. [ABSTRACT FROM AUTHOR]

Background Purpose Methods Results Conclusion Total‐body positron emission tomography (PET), already well‐established in the pre‐clinical setting, makes it possible to study multi‐parameters in biological systems as a whole, rather than focusing on single tissues analysis. Simultaneous kinetic analysis of multiple organs poses some daunting new challenges.To explore quantifying the pharmacokinetics of Na[18F]F in multiple dissimilar murine organs simultaneously in vivo with total‐body PET imaging using different compartmental models for each organ and a shared cardiovascular system.Six mice underwent a 60‐min total‐body PET scan following intravenous bolus injection of Na[18F]F. Compartmental models were constructed for each organ (heart, lungs, liver, kidneys, and bone) using an image derived input function. Non‐linear least squares fitting of a model that connects the five organs to a shared cardiovascular system was used to analyze both the first 3 min of data and the full hour. Analysis was repeated 5000 times using different initial parameter values for each duration, permitting analysis of correlations between parameters.The models give a good qualitative account of the activity curves irrespective of the duration of the data; however, the quality of the fits to 3 min of data (average χν2$\chi _\nu ^2$ is 2.72) was generally better. Comparison of perfusion values to literature values was possible for the liver and lungs with the former (liver, 0.540 ± 0.177 mL/ml/min) being well‐above expectations and the latter (lungs, 0.184 ± 0.413 mL/ml/min) in rough agreement. Correlations between microparameter values (especially affecting k2) caused very noticeable problems for data modeling from both the kidneys and the femur.The present study demonstrates an approach to performing kinetic modeling for multiple organs simultaneously with Na[18F]F. The observed correlations between microparameter values remain a challenge. Nonetheless, many microparameters can be estimated reliably with a quantitative analysis of perfusion being possible for some organs. [ABSTRACT FROM AUTHOR]

Background: To understand the connection between Muc5AC expression and the likelihood of rhinosinusitis, with the goal of providing insights into its prospective use as a biomarker. Methods: We searched PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, and Wanfang databases for studies up to November 2023 to conduct a literature review. After screening and quality assessment, eligible studies meeting the criteria were included. Muc5AC expression and rhinosinusitis association was analyzed by STATA 14.0. Results: Including weighted mean difference and 95% confidence interval, were reported. The meta‐analysis included 16 studies with 1448 rhinosinusitis patients. MUC5AC expression was significantly up‐regulated in both chronic rhinosinusitis with nasal polyps (CRSwNP; WMD: 0.52; 95% CI: 0.41–0.63) and chronic rhinosinusitis without nasal polyps (CRSsNP; WMD: 0.42; 95% CI: 0.28–0.56) patients compared to controls. IHC positive area analysis corroborated these findings, with elevated MUC5AC levels in CRSwNP (WMD: 25.61; 95% CI: 22.41–28.81) and CRSsNP (WMD: 39.74; 95% CI: 25.6–53.88) patients. Subgroup analysis based on tissue type (nasal tissue fluid and sinus mucosa) consistently supported the overall results. Conclusion: Our meta‐analysis robustly demonstrates a significant association between elevated MUC5AC expression and rhinosinusitis risk. This finding underscores the potential of MUC5AC as a molecular marker, providing valuable insights for future research and potential therapeutic interventions in rhinosinusitis management. Systematic review registration: CRD42024518932. [ABSTRACT FROM AUTHOR]

Diffuse midline gliomas present a particularly intricate and challenging clinical scenario. This rare case involves a patient with comutant H3F3A and FGFR diffuse midline glioma with a clinical presentation of fulminant leptomeningitis. A 22‐year‐old male presented with fatal and fulminant diffuse leptomeningitis. Next‐generation sequencing of plasma and cerebrospinal circulating tumour DNA revealed diffuse midline gliomas with H3F3A and FGFR mutations. Methylome analysis of meningeal tissue collected during autopsy confirmed the diagnosis. Liquid biopsy plays a crucial role in the diagnosis of diffuse midline gliomas, mainly those with exclusively leptomeningeal presentations. [ABSTRACT FROM AUTHOR]

The development of new drug therapies for Alzheimer's disease (AD) is an important research topic today, but the pathogenesis of AD has not been thoroughly studied, and there are still several shortcomings in existing drug therapies. Therefore, this study aimed to explore the molecular mechanism of lactoferrin (LF) in the treatments of AD and ulcerative colitis (UC) that is susceptible to AD, starting from the principle of one drug, two diseases, and the same treatment. This study used pathological staining and specific indicator staining to preliminarily evaluate the interventions of LF on UC injury and AD progression. We also used 16s RNA full-length sequencing to investigate the effect of LF on the abundance of intestinal microbiota in AD mice. Intestinal tissue and brain tissue metabolomics analysis were then used to screen specific metabolic pathways and preliminarily verify the metabolic mechanism of LF in alleviating the 2 diseases by regulating certain specific metabolites. Moreover, LF significantly changed the types and abundance of gut microbiota in AD mice complicated by UC. To conclude, this study proved the clinical phenomenon of AD susceptibility to UC, and verified the therapeutic effect of LF on 2 diseases. More importantly, we revealed the possible molecular mechanism of LF: Not only does it enrich the cognitive level of LF in alleviating AD by regulating the gut microbiota through the brain gut axis from the perspective of the theory of food nutrition promoting human health, but it also provides a practical basis for the subsequent research and development of LF and drug validation from the perspective of drug food homology. The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. [ABSTRACT FROM AUTHOR]

Purpose: We hypothesized that contrast-enhanced ultrasound (CEUS) using a microbubble technique to quantify microvascular changes and Nakagami imaging for tissue characterization would provide a new approach for diagnosing and differentiating benign and malignant choroidal lesions. Methods: Five patients with choroidal melanoma (CM) and five patients with choroidal hemangioma (CH) were selected. Definity®, which contains perflutren microbubbles, was administered as a slow IV bolus (1 ml). CEUS was performed for 1 min postinjection of the contrast agent with ultrasound radiofrequency data acquired from 10 s to 60 s. The contrast value was calculated for the whole tumor region. A gradient magnitude method was used for each postcontrast frames with 1-second interval, and the time-averaged value in pixel intensity gradient of postinjection frames was estimated and reported. Based on the Nakagami statistical distribution model, two Nakagami parameters, m and Ω, where m (shape parameter), representing tissue heterogeneity, and Ω (scale parameter), representing the average energy of backscattered signals, were studied. Results: CEUS analysis showed that the time-averaged estimated contrast was significantly higher (p = 0.008) for CH compared to CM. Furthermore, the time-averaged contrast within the normal choroidal region was significantly higher than the choroidal tumor region for both CH and CM (p = 0.001 for CH cases and p < 0.0001 for CM cases). Nakagami analysis showed that the m estimates were significantly higher (p = 0.032) for CH (m = 0.61) than for CM (m = 0.28), indicating that CH is a more heterogeneous tumor than CM. The Ω estimates were significantly higher (p = 0.0019) for CH (Ω = 0.15) compared to CM (Ω = 0.03). These results may be due to the more vascular structures in CH compared to CM. Conclusions: Quantitative intensity-based perfusion analysis using CEUS and backscattering tissue analysis using Nakagami imaging can provide valuable insights to differentiate benign and malignant choroidal lesions. [ABSTRACT FROM AUTHOR]

To review the efficacy and safety of oral vismodegib (Erivedge; Genentech) in the management of locally advanced orbital and periorbital basal cell carcinoma (BCC). A literature search was conducted last in September 2023 in the PubMed database for English language original research that evaluated the effect of oral vismodegib on orbital and periorbital BCC. Sixty articles were identified and 16 met the inclusion criteria. Most studies demonstrated high response rates, with up to 100% of patients responding to the medication in individual studies and initial complete regression occurring in up to 88% of patients. Vismodegib treatment resulted in significant reductions in tumor volume, resulting in globe preservation for most patients. However, in 12% of patients, the response was partial. Recurrences also occurred with substantial frequency, even after an initial complete response. As such, up to 79.4% of patients required surgical intervention, and up to 23% of patients still required exenteration. Use of these agents resulted in reductions in tumor volume that may delay or prevent the need for exenteration in some, but not all, patients. Importantly, molecular analysis of tissue excised after vismodegib therapy revealed persistent tumor in all patients, with frequent accumulation of mutations that may confer resistance to further hedgehog inhibitor therapy. Although most adverse events were rated as level I or II, side effects were common, with up to 100% of patients in studies experiencing at least 1 event. Muscle cramps, alopecia, weight loss, fatigue, and dysgeusia were the most common adverse events, and several patients discontinued therapy because of them. Furthermore, 1 patient died of sepsis that may have resulted from the therapy. Although level I and II evidence are lacking, most studies indicate a benefit from the use of oral vismodegib to treat orbital and periorbital BCC tumor volume. However, patients should be cautioned about the adverse side effects of treatment and the persistence of tumor cells with mutations that may cause long-term resistance. Use of vismodegib as short-term neoadjuvant therapy may be effective in shrinking tumor volume to reduce surgical morbidity while reducing the frequency and severity of side effects. The author(s) have no proprietary or commercial interest in any materials discussed in this article. [ABSTRACT FROM AUTHOR]

Spinal cord injury results in significant sensorimotor deficits, currently, there is no curative treatment for the symptoms induced by spinal cord injury. Basic and pre-clinical research on spinal cord injury relies on the development and characterization of appropriate animal models. These models should replicate the symptoms observed in human, allowing for the exploration of functional deficits and investigation into various aspects of physiopathology of spinal cord injury. Non-human primates, due to their close phylogenetic association with humans, share more neuroanatomical, genetic, and physiological similarities with humans than rodents. Therefore, the responses to spinal cord injury in nonhuman primates most likely resemble the responses to traumatism in humans. In this review, we will discuss nonhuman primate models of spinal cord injury, focusing on in vivo assessments, including behavioral tests, magnetic resonance imaging, and electrical activity recordings, as well as ex vivo histological analyses. Additionally, we will present therapeutic strategies developed in non-human primates and discuss the unique specificities of non-human primate models of spinal cord injury. [ABSTRACT FROM AUTHOR]

Image-guided biopsy is an integral step in the diagnosis and management of suspicious image-detected breast or axillary lesions, allowing for accurate diagnosis and, if indicated, treatment planning. Tissue sampling can be performed under guidance of a full spectrum of breast imaging modalities, including stereotactic, tomosynthesis, sonographic, and MRI, each with its own set of advantages and limitations. Procedural planning, which includes consideration of technical, patient, and lesion factors, is vital for diagnostic accuracy and limitation of complications. The purpose of this paper is to review and provide guidance for breast imaging radiologists in selecting the best procedural approach for the individual patient to ensure accurate diagnosis and optimal patient outcomes. Common patient and lesion factors that may affect successful sampling and contribute to postbiopsy complications are reviewed and include obesity, limited patient mobility, patient motion, patients prone to vasovagal reactions, history of anticoagulation, and lesion location, such as proximity to vital structures or breast implant. [ABSTRACT FROM AUTHOR]

Confocal Raman spectroscopy (CRS) has been widely used for noninvasive biochemical analysis of biological tissue in vivo. Currently, reflectance confocal microscopy (RCM) has been integrated with CRS as visual guidance for a square-/rectangular-shaped region of interest or a point of interest micro-Raman measurement. However, biological structures often have various morphologies; it is highly desirable to acquire a representative Raman spectrum from and only from the area that covers the whole target biological micro-structure. To achieve this goal, we developed a method to acquire Raman spectrum from a region of interest with any (arbitrary) shape under simultaneous RCM imaging guidance. [ABSTRACT FROM AUTHOR]

Horse bone is rich in collagen, with a composition similar to that of human collagen. Collagen peptides supply nutrients needed for human growth that act as antioxidants, lower blood pressure. This study explored the extraction of collagen and the preparation of collagen short peptides from Mongolian horse bones. Bones were collected from horses of varying ages, and the collagen content along with calcium salt distribution were observed through staining and imaging analyses. Next, the bones were processed into a powder and then subjected to ultra-high-pressure processing for degreasing. The degreasing conditions were optimised by single-factor and orthogonal tests. Following this, collagen was extracted using an acid-enzymatic method, and its structural characteristics and thermal stability were assessed. The collagen short peptides were extracted from the collagen samples, and the effects of the enzymatic hydrolysis time, temperature, pH, and enzyme amount on the extraction rate were evaluated. Finally, the resulting collagen peptides were analysed for antioxidant activity. In summary, this experiment optimised the extraction conditions for horse bone collagen, demonstrating that the ultra-high-pressure method minimally affects collagen structure, and the extraction rate was high. Hence our method has significant development potential. [ABSTRACT FROM AUTHOR]

Spatial transcriptomics is an emerging technology that enables the profiling of gene expression in tissues while preserving spatial location information. This innovative approach is anticipated to provide a comprehensive understanding of the spatial distribution of different cells within tissues and facilitate in-depth analysis of tissue structure. To accurately recognize spatial domains from spatial transcriptomics, we have introduced a generalized deep learning method called GAAEST (Graph Attention-based Autoencoder for Spatial Transcriptomics). Our proposed approach effectively integrates both spatial location information and gene expression data from spatial transcriptomics. Specifically, it leverages spatial location details to construct a neighborhood graph and employs a graph attention network-based encoder to embed gene expression information into a spatially informed space. At the same time, to further optimize the learned potential embedding, self-supervised contrastive learning is introduced to capture spatial information at three levels: local, global and contextual feature of spots. Finally, the decoder reconstructs gene expressions, which are then clustered to identify spatial domains with similar expression patterns and spatial proximity. Based on our experiments conducted on multiple datasets, GAAEST consistently outperforms existing state-of-the-art methods. The proposed GAAEST demonstrates excellent capabilities in spatial domain recognition, positioning it as an ideal tool for advancing spatial transcriptomics research. The graph attention based automatic encoder utilizing contrastive learning effectively combines spatial location information and gene expression data from spatial transcriptomics for enhanced domain recognition capacity. [ABSTRACT FROM AUTHOR]

Introduction: RECONCILE (ClinicalTrials.gov:NCT04340245) will identify molecular and radiomic markers associated with clinical progression and radiological progression events in a cohort of localised, newly diagnosed Gleason 3 + 4 tumours. Molecular markers will be correlated against standard of care MRI-targeted histology and oncological outcomes. Methods: RECONCILE is an ethics approved (20/LO/0366) single centre, prospective, longitudinal, observational cohort study of recently diagnosed (within 12 months), organ-confined Gleason 3 + 4 cancers (MCCL ≤10mm) currently under active surveillance. 60 treatment-naïve participants with a concordant MRI lesion (Likert score 4 or 5) and PSA ≤ 15 ng/ml will be recruited. Blood, urine and targeted prostate tissue cores will be subject to next generation sequencing at baseline and one year in all participants. Semen will be collected from a specified sub-population. Baseline and interval MR images will be extracted from standard of care prostate MRI ahead of radiomic analysis. Data extracted from radiological and biological samples will be used to derive the association of molecular change and radiological progression, the primary outcome of the study. To compensate for spatial intratumoral heterogeneity and inherent sampling bias, a molecular index will be derived for each participant using the molecular profile of tumour tissue at both baseline (MolBL) and one year (MolFU). We will extract a ΔMolBL:MolFU score for each participant. Molecular progression will be defined as a MolBL:MolFU score >95% CI of the combined ΔMolBL scores. Radiological progression is defined as a PRECISE score of 4 or 5. The study is powered to detect an association with a statistical power of 80%. Results: Recruitment began in July 2020 (n = 62). To date, 37 participants have donated tissue for analysis. Conclusion: We have designed and implemented a prospective, longitudinal study to evaluate the underlying molecular landscape of intermediate risk, MR-visible prostate tumours. Recruitment is ongoing. [ABSTRACT FROM AUTHOR]

The Borrelia burgdorferi sensu lato (s.l.) complex includes a group of spirochete bacteria that are involved in transmission cycles with vertebrates and the ticks associated with them. Rodents play an essential role in the ecoepidemiology of Borrelia, acting as reservoirs for the bacteria and hosts for ticks. To identify potential reservoir vertebrate hosts of Borrelia spirochetes in Argentina, we conducted molecular analyses on tissues obtained from a population of Sigmodontinae rodents inhabiting the Delta and Parana Islands ecoregion. We report the presence of B. burgdorferi s.l. complex genotypes infecting Akodon azarae and Oxymycterus rufus in this ecoregion. Phylogenetic analysis revealed at least three flaB haplotypes related to ‘Candidatus Borrelia paulista’ and Borrelia sp. strain Pampa from Brazil and to several Borrelia haplotypes from Uruguay. Additionally, we identified different alleles of the plasmid‐borne gene ospC in B. burgdorferi s.l. infecting A. azarae and Ox. rufus. The ospC allele present in Borrelia from Ox. rufus was also detected in Borrelia from Oligoryzomys mattogrossae, indicating the ability of Borrelia harbouring this ospC allele to infect multiple Sigmodontinae species, whereas the ospC allele from Borrelia infecting A. azarae represents a novel variant. Further research is needed to determine host specificity of ospC alleles present in South America. [ABSTRACT FROM AUTHOR]

This work aimed to evaluate the lactoferrin (LF) effect on arsenic (As) and imidacloprid (IMI) toxicity in broiler chicks. One-week old broiler chicks (n=105) were divided into seven groups (x15 each). The animals were orally supplemented with As, IMI, and/or LF for 4 weeks as follows: Control (G1) no supplements, G2 supplemented with As, G3 supplemented with IMI, G4 supplemented with As+IMI, G5 supplemented with As+LF, G6 supplemented with IMI+LF, G7 supplemented with As+IMI+LF. Body weight and weight gain were recorded on weekly interval. Blood, serum, liver, kidney, and muscle samples were collected at the end of the experimental period for biochemical and histopathological examination. Body weight performance, hematological, serum, and liver tissue biochemical analysis revealed adverse changes in G2, G3, and G4 compared to control, G5, G6, and G7. There was higher tissue residue of As and IMI in G4 and G5 compared to G5, G6, and G7. Liver histopathological changes in the groups supplemented with As and/or IMI were observed with necrosis, congestion, and inflammatory cell aggregates. The use of LF in broiler chicks improves weight gain performance and modulates the adverse effects of As and/or IMI toxicity. [ABSTRACT FROM AUTHOR]

A nanoscale-based 2D-photonic crystal (PhC) biosensor with silicon rods arranged in a triangular lattice structure is proposed in this work. The unique characteristic of the proposed structure is the design of dual nanocavity where two different rod radii are used within the ring resonator. The plane-wave expansion (PWE) method is used to analyse photonic band gaps (PBGs) and the sensing parameters are analysed using finite difference time domain (FDTD) techniques. The proposed biosensor is aimed at detecting the brain tissues, human immune deficiency virus (HIV)–infected blood samples, and sickle cell anaemia. The sensor proved its efficiency in detecting and distinguishing accurately between the brain tissues that are normal and aberrant (tumorourous, malignant, and damaged tissues). The proposed sensor achieves a high-quality factor (QF) of 9867, a high sensitivity of 1105 nm/RIU with an extremely low detection limit (DL) of 0.70 × 10−5, towards brain tissue analysis. This sensor can distinguish between a normal blood sample and HIV-infected samples. The QF, sensitivity, and DL of the biosensor for the HIV-infected sample are 2720, 1034 nm/RIU, and 2.61 × 10−5 RIU respectively. The sensor also proved its efficiency in analysing sickle cell anaemia in the blood sample. The QF and sensitivity of the biosensor are 6428 and 1071 nm/RIU respectively towards anaemia prediction. The proposed nanoscale photonic sensor could be a promising platform for other biomedical applications, such as the detection of various diseases at the cellular level. [ABSTRACT FROM AUTHOR]

Determination of infarct and scar size following myocardial infarction (MI) is commonly used to evaluate the efficacy of potential cardioprotective treatments in animal models. However, histological methods to determine morphological features in the infarcted heart have barely improved since implementation while still consuming large parts of the tissue and offering little options for parallel analyses. We aim to develop a new fluorescence technology for determining infarct area and area at risk that is comparable to 2,3,5-triphenyltetrazolium chloride (TTC) staining but allows for multiple analyses on the same heart tissue. For early and late time points following MI, we compared classical histochemical approaches with fluorescence staining methods. Reperfused MI was induced in male mice, the hearts were extracted 24 h, 7-, 21-, or 28-days later and fluorescently stained by combining Hoechst and phalloidin. This approach allowed for clear visualization of the infarct area, the area at ischemic risk and the remote area not affected by MI. The combined fluorescence staining correlated with the classic TTC/Evans Blue staining 24 h after MI (r = 0.8334). In later phases (>7 d) post-MI, wheat germ agglutinin (WGA) is equally accurate as classical Sirius Red (r = 0.9752), Masson's (r = 0.9920) and Gomori's Trichrome (r = 0.8082) staining for determination of scar size. Additionally, feasibility to co-localize fluorescence-stained immune cells in specific regions of the infarcted myocardium was demonstrated with this protocol. In conclusion, this new procedure for determination of post-MI infarct size is not inferior to classical TTC staining, yet provides substantial benefits, including the option for unbiased software-assisted analysis while sparing ample residual tissue for additional analyses. Overall, this enhances the data quality and reduces the required animal numbers consistent with the 3R concept of animal experimentation. [ABSTRACT FROM AUTHOR]

Simple Summary: A liquid biopsy (LB) is now considered to be a valuable diagnostic tool for advanced and metastatic nonsquamous nonsmall cell lung cancer (NS-NSCLC) patients. The analysis of circulating tumor DNA (ct-DNA) for NS-NSCLC has been shown to increase the chance of identifying the presence of targetable mutations and has been adopted by many clinicians around the world. However, a tissue biopsy (TB) is still the "golden standard" for lung cancer diagnosis and molecular profiling, which is essential for decision-making regarding treatment of NS-NSCLC. Both approaches should be considered as complementary and their sequential or simultaneous use for next-generation sequencing (NGS) can be questionable at tumor diagnosis and/or at tumor progression. The targeted therapies and immunotherapies in thoracic oncology, particularly for NS-NSCLC, are associated with an increase in the number of predictive biomarkers to be assessed in routine clinical practice. These treatments are administered thanks to marketing authorization for use in daily practice or are evaluated during clinical trials. Since the molecular targets to be identified are more and more complex and numerous, it is now mandatory to use NGS. NGS can be developed from both tissue and fluid (mainly blood). The blood tests in oncology, so-called "liquid biopsies" (LB), are performed with plasmatic circulating free DNA (cf-DNA) and are complementary to the molecular testing performed with a TB. LB use in lung cancer is associated with international guidelines, but additional algorithms could be set up. However, even if useful for better care of patients, notably with advanced and metastatic NS-NSCLC, until now LB are not often integrated into daily practice, at least in Europe and notably in France. The purpose of this review is to describe the different opportunities and algorithms leading to the identification of the molecular signature of NS-NSCLC, using both tissue and liquid biopsies, and to introduce the principle limitations but also some perspectives in this field. [ABSTRACT FROM AUTHOR]

The distinctive aroma of onions, consisting primarily of sulfur‐containing compounds, is one of the reasons for the popularity of the vegetable. The rapid enzymatic and chemical reactions that occur after the destruction of onion bulb tissue render the analysis of the volatile profile challenging. Therefore, sample preparation is a crucial step in the analysis of the onion volatilome, but it often does not receive the necessary attention in the literature. In this work, we focused on two aspects: Firstly, we compared different sample preparation approaches to maximize the volatile yield and to enable the description of the onion volatile profile as it would emerge after a solid‐phase microextraction (SPME) Arrow sampling procedure. For headspace (HS)‐gas chromatography‐mass spectrometry analysis, onion juice with the addition of an ammonium sulfate solution proved to be the best option. Secondly, we optimized the HS sampling and desorption parameters (enrichment time, enrichment temperature, agitator speed, desorption time) for onion volatiles using the efficient design of experiments (DoE) approach. The optimal conditions for the analysis with HS‐SPME Arrow were an enrichment time of 75 min at 60°C with an agitator speed of 713 rpm and a desorption time of 120 s. [ABSTRACT FROM AUTHOR]

Objectives: Cell models have shown great promise as tools for research, potentially providing intriguing alternatives to animal models. However, the original tissue characteristics must be maintained in culture, a fact that is often assumed, but seldom assessed. We aimed to follow the retention of the original tissue identities of cleft lip‐derived skin and mucosa keratinocytes in vitro. Methods: Cleft lip‐derived keratinocytes were isolated from discarded tissue along the cleft margins during cheiloplasty. Cell identities were assessed by immunohistochemistry and quantitative real‐time PCR for tissue‐specific markers and compared with native lip tissue. Moreover, keratinocytes were regularly analyzed for the retention of the original tissue characteristics by the aforementioned methods as well as by differentiation assays. Results: The various anatomical zones of the human lip could be distinguished using a panel of differentiation and functional‐based markers. Using these markers, retention of the original tissue identities could be followed and confirmed in the corresponding primary keratinocytes in culture. Conclusions: Our findings promote patient‐derived cells retaining their original identities as astonishing and clinically relevant in vitro tools. Such cells allow a better molecular understanding of various lip‐associated pathologies as well as their modeling in vitro, including but not restricted to orofacial clefts. [ABSTRACT FROM AUTHOR]

Patients with brain tumours are motivated to participate in clinical trials involving repeat tissue sampling. Normalising the use of neoadjuvant and staged surgical trials necessitates collaboration among patients, regulatory agencies, and researchers. Initial and repetitive tissue sampling plays a crucial role in enhancing our understanding of resistance mechanisms and vulnerabilities in brain tumour therapy. Standardising biopsy techniques and ensuring technical uniformity across institutions are vital for effective interinstitutional collaboration. Although liquid biopsy technologies hold promise, they are not yet ready to replace tissue analysis. Clear communication about the risks and benefits of biopsies is essential, particularly regarding potential postoperative deficits. Changes in mindset and neurosurgical culture are imperative to achieve much needed breakthroughs in the development of new, effective therapies for brain tumours. [ABSTRACT FROM AUTHOR]

The circadian dynamics of the expression of key genes of carotenoid metabolism (PSY2, LCYE, CrtRB1, and NCED1) in the photosynthetic tissue of tomato Solanum lycopersicum L. (cultivar Korneevsky) plants was characterized. An in silico analysis of the gene expression pattern was carried out and a high level of their transcripts was detected in the leaf tissue. qRT-PCR analysis of gene expression was performed at six time points during the day and showed the highest levels of PSY2, LCYE, and NCED1 transcripts in the second half of the light phase and CrtRB1 at the end of the dark phase. The content and composition of carotenoids in leaf tissue in the middle of the day was determined; it was shown that the leaf accumulates 1.5 times more compounds of the ɛ/β-branch of carotenoid biosynthesis pathway than compounds of the β/β-branch. [ABSTRACT FROM AUTHOR]

Background: Many infectious diseases are diagnosed in emergency departments (ED) and patients are prescribed antimicrobial therapy. Results from cultures typically take a few days to become finalized. Following up on these results is necessary when medication changes are indicated due to results that show bacteria are resistant to the prescribed antibiotics. Involving pharmacists in assessing the culture and sensitivity results, and making interventions when needed, is an innovative way to ensure that patients receive appropriate antimicrobial therapy based on the culture and sensitivity data. This study analyzed the impact of pharmacist involvement in the ED's post-discharge positive culture review process on ED re-visits and hospitalizations. Methods : This single-center, pre- and post-implementation study examined the impact of pharmacist involvement in the post-ED visit culture review process on ED re-visits and hospitalizations. Positive microbiological results included documented growth from urine, skin and soft tissue, throat, blood, or stool cultures. Patients included in the study were of 18 years of age or older and had a positive culture result post ED-discharge. Patients were excluded from the study if they were admitted to the hospital or transferred to another facility. The primary outcomes included ED re-visits within 7 days and hospital readmissions within 30 days for the same condition. The secondary outcomes were percentage of pharmacist interventions accepted and types of pharmacist interventions implemented. Results : A total of 141 patients were included in the study, with 65 in the pre-implementation group and 76 in the post-implementation group. The primary outcome of ED re-visits within 7 days for the same condition occurred in 11 (17%) patients in the pre-implementation group and 5 (7%) patients in the post-implementation group (P =.0454). The primary outcome of hospitalizations within 30 days for the same condition occurred in 5 (8%) patients in the pre-implementation group and 1 (1%) patient in the post-implementation group (P =.0137). Seventeen (94%) out of the 18 pharmacist interventions were accepted and implemented. The intervention types implemented were to recommend to: change antibiotic (35%), not initiate antibiotic (24%), initiate antibiotic (24%), and continue antibiotic (18%). Conclusion : Pharmacist involvement in the ED post-discharge positive culture review process showed a decrease in ED re-visits and hospitalizations for the same condition. [ABSTRACT FROM AUTHOR]

Background: Formaldehyde is a reactive chemical that bonds randomly with various cellular elements. This substance, which is also found in the natural structure of the organism, is used in many areas, from industry to household materials, from the production of coatings in dentistry to the fixation of cadavers in laboratories. Formaldehyde is commonly used in laboratories to fix tissues. Objectives: In this study, the staining properties of tissues fixed with date molasses, a natural sugar, using the May-Grunwald-Giemsa (MGG) staining method were compared with the staining properties of tissues fixed with buffered formol-saline. Methods: Liver, spleen, kidney, skin, testicle, small intestine, large intestine, brain, cerebellum, and lung tissue samples of 4 rats of both genders were used as materials in the study. Each tissue underwent routine histological tissue tracing and was stained with MGG. Results: When all tissues were evaluated anatomically, the general morphology of the tissues showed that the tissue integrity was preserved. There was no color change in the tissues. In addition, tissue hardness was at the required level, supporting previous studies. Histologically, regarding chromatin distribution, nucleus separation and cytoplasm staining, tissues fixed with date molasses and tissues fixed with buffered formal saline showed similar properties, except for the tissues subjected to intense maceration (skin and small intestines). Pale staining was observed in some tissues fixed with buffered formal saline (liver, skin, heart), but the nucleus and cytoplasm structures were better distinguished at higher magnifications. Considering the chromatin distribution, nuclear separation and cytoplasm staining in other tissues (spleen, kidney, testicle, large intestine, brain, cerebellum and lung), tissues fixed with date molasses showed similar properties to tissues fixed with buffered formal-saline. Conclusion: We recommend that researchers interested in the subject prolong the fixation period (6 months) and compare these results with those of tissues fixed for 24 hours. [ABSTRACT FROM AUTHOR]

Heavy metals are an important group of toxic substances harmful for many organisms. Of these, mercury is one of the most monitored in the environment. Several matrices are used for the monitoring of environmental load, including a range of organisms; bats, however, have only been examined rarely. Insectivorous bats are apex predators threatened by several human interventions in their natural environment, including heavy metal pollution. The aim of this study was to analyze the content of total mercury in the fur, flight membrane, and pectoral muscle of greater mouse-eared bats (Myotis myotis). Total mercury concentrations were also measured in carabid beetles from the catch locality Zastávka u Brna. Samples were obtained from 43 bat carcasses at two different localities in the Czech Republic (Zastávka u Brna, Malá Morávka). Total mercury content varied between 1.76–72.20 µg/g in fur, 0.04–0.14 µg/g in skin, and 0.05–0.20 µg/g in muscle. Total mercury values in the fur of some individuals from Malá Morávka exceeded the recognized toxicity limit. Furthermore, there was a significant difference (p < 0.001) in content of total mercury in fur between localities, and there was a clear effect of age on concentrations in fur, skin, and muscle, the concentrations being significantly correlated (fur and skin rs = 0.783; fur and muscle rs = 0.716; skin and muscle rs = 0.884). These findings confirm the usefulness of fur samples from living bats for biomonitoring mercury burden in the environment. [ABSTRACT FROM AUTHOR]

Automatic assessment of brain regions in an MR image has emerged as a pivotal tool in advancing diagnosis and continual monitoring of neurological disorders through different phases of life. Nevertheless, current solutions often exhibit specificity to particular age groups, thereby constraining their utility in observing brain development from infancy to late adulthood. In our research, we introduce a novel approach for segmenting and classifying neonatal brain images. Our methodology capitalizes on minimum spanning tree (MST) segmentation employing the Manhattan distance, complemented by a shrunken centroid classifier empowered by the Brier score. This fusion enhances the accuracy of tissue classification, effectively addressing the complexities inherent in age-specific segmentation. Moreover, we propose a novel threshold estimation method utilizing the Brier score, further refining the classification process. The proposed approach yields a competitive Dice similarity index of 0.88 and a Jaccard index of 0.95. This approach marks a significant step toward neonatal brain tissue segmentation, showcasing the efficacy of our proposed methodology in comparison to the latest cutting-edge methods. [ABSTRACT FROM AUTHOR]

Background: Neutrophilic cells are among the most abundant immune populations within the head and neck tumor microenvironment (TME) and harbor multiple mechanisms of immunosuppression. Despite these important features, neutrophilic cells may be underrepresented in contemporary studies that aim to comprehensively characterize the immune landscape of the TME due to discrepancies in tissue processing and analysis techniques. Here, we review the role of pathologically activated neutrophilic cells within the TME and pitfalls of various approaches used to study their frequency and function in clinical samples. Methods: The literature was identified by searching PubMed for "immune landscape" and "tumor immune microenvironment" in combination with keywords describing solid tumor malignancies. Key publications that assessed the immune composition of solid tumors derived from human specimens were included. The tumor and blood processing methodologies in each study were reviewed in depth and correlated with the reported abundance of neutrophilic cells. Results: Neutrophilic cells do not survive cryopreservation, and many studies fail to identify and study neutrophilic cell populations due to cryopreservation of clinical samples for practical reasons. Additional single‐cell transcriptomic studies filter out neutrophilic cells due to low transcriptional counts. Conclusions: This report can help readers critically interpret studies aiming to comprehensively study the immune TME that fail to identify and characterize neutrophilic cells. [ABSTRACT FROM AUTHOR]

Background: A challenge in addressing neonatal opioid withdrawal syndrome (NOWS) is knowing who has been exposed and needs monitoring for withdrawal. Women do not always get asked about or disclose opioid use, and biologic testing is neither universal nor infallible. We investigate the prevalence and effectiveness of methods for identifying prenatal opioid exposure. Methods: A review of medical charts at five delivery hospitals identified newborns with known exposure (i.e., NOWS diagnosis) for study inclusion. Results: Over 95% of the mothers had 1+ urine drug screening (UDS) during pregnancy, 38% had UDS at delivery, 94% had documentation of self-report inquiry, and 81% of the newborns had biologic testing of meconium and/or cord tissue. Pregnancy UDS detected opioid use for 17% of the sample, UDS at delivery detected for 32%. A self-report of the use prenatally/at delivery identified 85%. Cord tissue testing was positive for 78%, meconium for 61%, and infant UDS 15%. 96% of the newborns were positive on 1+ exposure variable. Conclusions: With drug testing only when indicated, almost all exposed pregnancies included inquiry about and biologic testing for opioid use/exposure. Reliance on any one assessment method may miss exposed newborns, but consideration of all information may identify most if not all newborns needing monitoring for NOWS. [ABSTRACT FROM AUTHOR]

Objective: To investigate potential mechanisms of anti-atherosclerosis by berberine (BBR) using ApoE−/− mice. Methods: Eight 8-week-old C57BL/6J mice were used as a blank control group (normal), and 56 8-week-old AopE−/− mice were fed a high-fat diet for 12 weeks, according to a completely random method, and were divided into the model group, BBR low-dose group (50 mg/kg, BBRL), BBR medium-dose group (100 mg/kg, BBRM), BBR high-dose group (150 mg/kg, BBRH), BBR+nuclear factor erythroid 2-related factor 2 (NRF2) inhibitor group (100 mg/kg BBR+30 mg/kg ML385, BBRM+ML385), NRF2 inhibitor group (30 mg/kg, ML385), and positive control group (2.5 mg/kg, atorvastatin), 8 in each group. After 4 weeks of intragastric administration, samples were collected and serum, aorta, heart and liver tissues were isolated. Biochemical kits were used to detect serum lipid content and the expression levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in all experimental groups. The pathological changes of atherosclerosis (AS) were observed by aorta gross Oil Red O, aortic sinus hematoxylin-eosin (HE) and Masson staining. Liver lipopathy was observed in mice by HE staining. The morphology of mitochondria in aorta cells was observed under transmission electron microscope. Flow cytometry was used to detect reactive oxygen species (ROS) expression in aorta of mice in each group. The content of ferrous ion Fe2+ in serum of mice was detected by biochemical kit. The mRNA and protein relative expression levels of NRF2, glutathione peroxidase 4 (GPX4) and recombinant solute carrier family 7 member 11 (SLC7A11) were detected by quantitative real time polymerase chain reaction (RT-qPCR) and Western blot, respectively. Results: BBRM and BBRH groups delayed the progression of AS and reduced the plaque area (P<0.01). The characteristic morphological changes of ferroptosis were rarely observed in BBR-treated AS mice, and the content of Fe2+ in BBR group was significantly lower than that in the model group (P<0.01). BBR decreased ROS and MDA levels in mouse aorta, increased SOD activity (P<0.01), significantly up-regulated NRF2/SLC7A11/GPX4 protein and mRNA expression levels (P<0.01), and inhibited lipid peroxidation. Compared with the model group, the body weight, blood lipid level and aortic plaque area of ML385 group increased (P<0.01); the morphology of mitochondria showed significant ferroptosis characteristics; the serum Fe2+, MDA and ROS levels increased (P<0.05 or P<0.01), and the activity of SOD decreased (P<0.01). Compared with BBRM group, the iron inhibition effect of BBRM+ML385 group was significantly weakened, and the plaque area significantly increased (P<0.01). Conclusion: Through NRF2/SLC7A11/GPX4 pathway, BBR can resist oxidative stress, inhibit ferroptosis, reduce plaque area, stabilize plaque, and exert anti-AS effects. [ABSTRACT FROM AUTHOR]

N6-methyladenosine (m6A) is a prevalent mRNA modification known for its implications in various cancer types, yet its role in chromophobe renal cell carcinoma (chRCC) remains largely unexplored. In this study, we performed m6A-SEAL-seq and RNA-seq analyses on tissues from three chRCC subjects, aiming to uncover m6A alterations in chRCC. Our findings revealed reduced expression levels of four m6A regulators in chRCC tissues and highlighted differences in m6A levels compared to normal tissues. Furthermore, we identified specific genes and cancer-related pathways affected by these differences, including notable candidates like NOTCH1 and FGFR1, implicated in chRCC development. Additionally, we developed a predictive model based on the expression level of m6A associated genes, demonstrating promising prognostic capabilities for patient survival prediction. Overall, our study provides valuable insights into the role of m6A in chRCC and its potential as a prognostic indicator. [ABSTRACT FROM AUTHOR]

Traumatic brain injury (TBI) is an alteration of brain function caused by a sudden transmission of an external force to the head. The biomechanical impact induces acute and chronic metabolic changes that highly contribute to injury evolution and outcome. TBI heterogeneity calls for approaches allowing the mapping of regional molecular and metabolic changes underpinning disease progression, with mass spectrometry imaging (MSI) as an efficient tool to study the spatial distribution of small metabolites. In this study, we applied an innovative targeted atmospheric pressure-MALDI mass spectrometry imaging (AP-MALDI MSI) approach, starting from an extensive list of metabolites, representative of different metabolic pathways, individually validated on the tissue under analysis with original standards using 2,5-dihydroxybenzoic acid (DHB), to characterize the impact of TBI on regional changes to small metabolites in the brain. Brains from sham and TBI mice obtained 21 days post-injury were analyzed to examine the spatial metabolic profile of small metabolites belonging to different metabolic pathways. By a whole brain analysis, we identified four metabolites (alanine, lysine, histidine, and inosine) with higher abundance in TBI than sham mice. Within the TBI group, lysine, histidine, and inosine were higher in the hemisphere ipsilateral to the biomechanical impact vs. the contralateral one. Images showed a major involvement of the ipsilateral thalamus characterized by the increase of arginine, lysine, histidine, and inosine and a significant reduction of glutamic acid, and N-acetylaspartic acid compared to the contralateral thalamus. These findings indicate high-resolution imaging mass spectrometry as a powerful tool to identify region-specific changes after a TBI to understand the metabolic changes underlying brain injury evolution. [ABSTRACT FROM AUTHOR]

AbstractFlorida sugarcane production is mostly on organic soils, but the cultivation area on mineral soils is also expanding. Silicon (Si) is beneficial for sugarcane growth and development on weathered soil. However, the low solubility and availability of soluble Si is an issue in Florida’s mineral soils. This study aimed to assess soil Si extraction methods based on sugarcane yield response to calcium silicate application. It also examined the relationship between sugarcane yield with extracted plant-available soil Si, alongside plant Si concentrations. The experiment followed a randomized complete block design involving 2 fields, with 24 plots each, and 6 replications. Four rates of calcium silicate (0 control, 2.24, 4.48, and 6.67 Mg ha−1) were applied before planting sugarcane. Soil samples were collected prior to planting and after each harvest. Soil Si was extracted using 0.5 M acetic acid, 0.7 N NH4OAc, Mehlich 3, and 0.01 M CaCl2 extractants. Leaf samples were collected over a three-year sugarcane life cycle (from 2019 to 2022) to evaluate the relationship between soil Si and leaf Si concentrations. The relationships between parameters were analyzed using linear and nonlinear regression models. Results indicated that higher Si application rates increased Si concentrations in sugarcane leaves in both fields. However, only one field showed a yield response to Si, likely due to high pre-plant soil Si in the other field. The best extractant should correlate strongly with either yield or leaf Si content, but no single extractant showed a strong correlation with either, making it infeasible to identify the best extractant. [ABSTRACT FROM AUTHOR]

Background: Idiopathic pulmonary fibrosis (IPF) is an age-related disease severely affecting life quality with its prevalence rising as the population ages, yet there is still no effective treatment available. Cell therapy has emerged as a promising option for IPF, however, the absence of mature and stable animal models for IPF immunodeficiency hampers preclinical evaluations of human cell therapies, primarily due to rapid immune clearance of administered cells. This study aims to establish a reliable pulmonary fibrosis (PF) model in immunodeficient mice that supports autologous cell therapy and to investigate underlying mechanism. Methods: We utilized thirty 5-week-old male NOD/SCID mice, categorizing them into three age groups: 12weeks, 32 weeks and 43 weeks, with 6 mice euthanized randomly from each cohort for lung tissue analysis. We assessed fibrosis using HE staining, Masson's trichrome staining, α-SMA immunohistochemistry and hydroxyproline content measurement. Further, β-galactosidase staining and gene expression analysis of MMP9, TGF-β1, TNF-α, IL-1β, IL-6, IL-8, SOD1, SOD2, NRF2, SIRT1, and SIRT3 were performed. ELISA was employed to quantify protein levels of TNF-α, TGF-β1, and IL-8. Results: When comparing lung tissues from 32-week-old and 43-week-old mice to those from 12-week-old mice, we noted a marked increase in inflammatory infiltration, fibrosis severity, and hydroxyproline content, alongside elevated expression levels of α-SMA and MMP9. Notably, the degree of fibrosis intensified with age. Additionally, β-galactosidase staining became more pronounced in older mice. Quantitative PCR analyses revealed age-related, increases in the expression of senescence markers (GLB1, P16, P21), and proinflammatory genes (TGF-β1, TNF-α, IL-1β, IL-6, and IL-8). Conversely, the expression of anti-oxidative stress-related genes (SOD1, SOD2, NRF2, SIRT1, and SIRT3) declined, showing statistically significant differences (*P < 0.05, **P < 0.01, ***P < 0.001). ELISA results corroborated these findings, indicating a progressive rise in the protein levels of TGF-β1, TNF-α, and IL-8 as the mice aged. Conclusions: The findings suggest that NOD/SCID mice aged 32 weeks and 43 weeks effectively model pulmonary fibrosis in an elderly context, with the disease pathogenesis likely driven by age-associated inflammation and oxidative stress. [ABSTRACT FROM AUTHOR]

The objective of this work is to showcase the ortho-positronium lifetime as a probe for soft-tissue characterization. We employed positron annihilation lifetime spectroscopy to experimentally measure the three components of the positron annihilation lifetime—para-positronium (p-Ps), positron, and ortho-positronium (o-Ps)—for three types of porcine, non-fixated soft tissues ex vivo: adipose, hepatic, and muscle. Then, we benchmarked our measurements with X-ray phase-contrast imaging, which is the current state-of-the-art for soft-tissue analysis. We found that the o-Ps lifetime in adipose tissues (2.54 ± 0.12 ns) was approximately 20% longer than in hepatic (2.04 ± 0.09 ns) and muscle (2.03 ± 0.12 ns) tissues. In addition, the separation between the measurements for adipose tissue and the other tissues was better from o-Ps lifetime measurement than from X-ray phase-contrast imaging. This experimental study proved that the o-Ps lifetime is a viable non-invasive probe for characterizing and classifying the different soft tissues. Specifically, o-Ps lifetime as a soft-tissue characterization probe had a strong sensitivity to the lipid content that can be potentially implemented in commercial positron emission tomography scanners that feature list-mode data acquisition. [ABSTRACT FROM AUTHOR]

Background: The roles of the transcriptional factor SIX2 have been identified in several tumors. However, its roles in gastric cancer (GC) progression have not yet been revealed. Our objective is to explore the impact and underlying mechanisms of SIX2 on the stemness of GC cells. Methods: Lentivirus infection was employed to establish stable expression SIX2 or PFN2 in GC cells. Gain- and loss-of-function experiments were conducted to detect changes of stemness markers, flow cytometry profiles, tumor spheroid formation, and tumor-initiating ability. ChIP, RNA-sequencing, tissue microarray, and bioinformatics analysis were performed to reveal the correlation between SIX2 and PFN2. The mechanisms underlying the SIX2/PFN2 loop-mediated effects were elucidated through tissue microarray analysis, RNA stability assay, IP-MS, Co-Immunoprecipitation, and inhibition of the JNK signaling pathway. Results: The stemness of GC cells was enhanced by SIX2. Mechanistically, SIX2 directly bound to PFN2's promoter and promoted PFN2 activity. PFN2, in turn, promoted the mRNA stability of SIX2 by recruiting RNA binding protein YBX-1, subsequently activating the downstream MAPK/JNK pathway. Conclusion: This study unveils the roles of SIX2 in governing GC cell stemness, defining a novel SIX2/PFN2 regulatory loop responsible for this regulation. This suggests the potential of targeting the SIX2/PFN2 loop for GC treatment (Graphical Abstracts). [ABSTRACT FROM AUTHOR]

The discovery of biomarkers, essential for successful drug development, is often hindered by the limited availability of tissue samples, typically obtained through core needle biopsies. Standard ‘omics platforms can consume significant amounts of tissue, forcing scientist to trade off spatial context for high-plex assays, such as genome-wide assays. While bulk gene expression approaches and standard single-cell transcriptomics have been valuable in defining various molecular and cellular mechanisms, they do not retain spatial context. As such, they have limited power in resolving tissue heterogeneity and cell–cell interactions. Current spatial transcriptomics platforms offer limited transcriptome coverage and have low throughput, restricting the number of samples that can be analyzed daily or even weekly. While the Digital Spatial Profiling (DSP) method does not provide single-cell resolution, it presents a significant advancement by enabling scalable whole transcriptome and ultrahigh-plex protein analysis from distinct tissue compartments and structures using a single tissue slide. These capabilities overcome significant constraints in biomarker analysis in solid tissue specimens. These advancements in tissue profiling play a crucial role in deepening our understanding of disease biology and in identifying potential therapeutic targets and biomarkers. To enhance the use of spatial biology tools in drug discovery and development, the DSP Scientific Consortium has created best practices guidelines. These guidelines, built on digital spatial profiling data and expertise, offer a practical framework for designing spatial studies and using current and future spatial biology platforms. The aim is to improve tissue analysis in all research areas supporting drug discovery and development. [ABSTRACT FROM AUTHOR]

This paper reports results obtained using white light diffraction phase microscopy (wDPM) on captured images of breast and colon tissue samples, marking a contribution to the advancement in biomedical imaging. Unlike conventional brightfield microscopy, wDPM offers the capability to capture intricate details of biological specimens with enhanced clarity and precision. It combines high resolution, enhanced contrast, and quantitative capabilities with non-invasive, label-free imaging. These features make it a useful tool for tissue imaging, providing detailed and accurate insights into tissue structure and dynamics without compromising the integrity of the samples. Our findings underscore the potential of quantitative phase imaging in histopathology, in the context of automating the process of tissue analysis and diagnosis. Of particular note are the insights gained from the reconstructed phase images, which provide physical data regarding peripheral glandular cell membranes. These observations serve to focus attention on pathologies involving the basal membrane, such as early invasive carcinoma. Through our analysis, we aim to contribute to catalyzing further advancements in tissue (breast and colon) imaging. [ABSTRACT FROM AUTHOR]

Deep learning-based virtual staining has the potential to replace chemical staining in histology and to provide more sustainable, rapid, and cost-effective pipelines for histopathology. Virtual staining will allow multiple outputs of stained images from a single input image of a tissue section, hence enabling virtual multiplexing and single-cell resolution between histological assessments that currently require separate tissue sections. Deep learning-based virtual staining models require large amounts of training data, and quantitative computational and rigorous histological validation is required for each intended use. In pathology and biomedical research, histology is the cornerstone method for tissue analysis. Currently, the histological workflow consumes plenty of chemicals, water, and time for staining procedures. Deep learning is now enabling digital replacement of parts of the histological staining procedure. In virtual staining, histological stains are created by training neural networks to produce stained images from an unstained tissue image, or through transferring information from one stain to another. These technical innovations provide more sustainable, rapid, and cost-effective alternatives to traditional histological pipelines, but their development is in an early phase and requires rigorous validation. In this review we cover the basic concepts of virtual staining for histology and provide future insights into the utilization of artificial intelligence (AI)-enabled virtual histology. [ABSTRACT FROM AUTHOR]

Background and aims: Evaluation of the programmed cell death ligand‐1 (PD‐L1) combined positive score (CPS) is vital to predict the efficacy of the immunotherapy in triple‐negative breast cancer (TNBC), but pathologists show substantial variability in the consistency and accuracy of the interpretation. It is of great importance to establish an objective and effective method which is highly repeatable. Methods: We proposed a model in a deep learning‐based framework, which at the patch level incorporated cell analysis and tissue region analysis, followed by the whole‐slide level fusion of patch results. Three rounds of ring studies (RSs) were conducted. Twenty‐one pathologists of different levels from four institutions evaluated the PD‐L1 CPS in TNBC specimens as continuous scores by visual assessment and our artificial intelligence (AI)‐assisted method. Results: In the visual assessment, the interpretation results of PD‐L1 (Dako 22C3) CPS by different levels of pathologists have significant differences and showed weak consistency. Using AI‐assisted interpretation, there were no significant differences between all pathologists (P = 0.43), and the intraclass correlation coefficient (ICC) value was increased from 0.618 [95% confidence interval (CI) = 0.524–0.719] to 0.931 (95% CI = 0.902–0.955). The accuracy of interpretation result is further improved to 0.919 (95% CI = 0.886–0.947). Acceptance of AI results by junior pathologists was the highest among all levels, and 80% of the AI results were accepted overall. Conclusion: With the help of the AI‐assisted diagnostic method, different levels of pathologists achieved excellent consistency and repeatability in the interpretation of PD‐L1 (Dako 22C3) CPS. Our AI‐assisted diagnostic approach was proved to strengthen the consistency and repeatability in clinical practice. [ABSTRACT FROM AUTHOR]

The science of dental tissue grafting is evolving, with an increased understanding of factors influencing graft behavior. Despite the widespread clinical use of soft tissue grafts, the histological characteristics of different gingival harvesting sites are still underexplored. This study aimed to fill this gap by analyzing 50 tissue samples harvested from 25 patients across three sites: the hard palate, maxillary tuberosity, and palatal rugae. Each sample underwent thorough histological and histomorphometric analysis. Conventional statistical analysis was performed using SPSS, while predictive modeling was conducted with RapidMiner Studio. The study identified significant histological differences among the graft sites, with notable variations in total graft height, epithelial height, and interdigitation perimeter. These findings underscore the importance of donor site selection in influencing graft success. Pair plots and principal component analysis (PCA) further highlighted the distinct histological features of each tissue type. The random forest classifier identified total graft height, epithelial height, and perimeter as the most influential factors in predicting graft site behavior. This study offers valuable insights into the histological characteristics of soft tissue grafts, potentially leading to more predictable clinical outcomes. [ABSTRACT FROM AUTHOR]

Ultrasound‐based elastography techniques are increasingly available on modern ultrasound systems. The application of elastography extends the utility of ultrasound, yielding new information about the impact of disease on tissue biomechanics. By modifying the B‐mode imaging pulse, an Acoustic Radiation Force Impulse produces shear waves in selected tissue. Quantifying the speed of the resultant shear wave provides an indirect assessment of tissue stiffness, with stiffer tissue producing faster shear wave movement than softer tissue. Two main types of ultrasound shear wave elastography are currently in use. Point shear wave elastography uses one or two pulses to measure shear wave speeds over a small fixed area. Alternatively, two‐dimensional shear‐wave elastography provides a colour elastogram map over a larger area, allowing the operator to select specific regions of interest to measure shear wave speed. This article will discuss the essential physics principles, and clinical uses of two‐dimensional shear wave elastography, aiming to provide sonographers with a basic understanding of the technology and current applications. [ABSTRACT FROM AUTHOR]