Telomere Length Measurement in Human Tissue Sections by Quantitative Fluorescence In Situ Hybridization (Q-FISH).

Telomeres in most somatic cells shorten with each cell division, and critically short telomeres lead to cellular dysfunction, cell cycle arrest, and senescence. Thus, telomere shortening is an important hallmark of human cellular senescence. Quantitative fluorescence in situ hybridization (Q-FISH) using formalin-fixed paraffin-embedded (FFPE) tissue sections allows the estimation of telomere lengths in individual cells in histological sections. In our Q-FISH method, fluorescently labelled peptide nucleic acid (PNA) probes are hybridized to telomeric and centromeric sequences in FFPE human tissue sections, and relative telomere lengths (telomere signal intensities relative to centromere signal intensities) are measured. This chapter describes our Q-FISH protocols for assessing relative telomere lengths in FFPE human tissue sections.

The links between experiential learning and 4E cognition.

In this article, we explore the connections between two distinct approaches: experiential learning (EL) and 4E cognition. EL emphasizes the role of concrete experiences as the building blocks of learning, whereas 4E cognition views cognition as arising from the interactions that an individual has with their physical and social environment. Despite their divergent theoretical frameworks, methodologies, and interests, we argue that both frameworks share a common vision of cognition and that their integration could mutually enhance their respective fields. This article outlines the historical origins and underlying assumptions of both frameworks, highlighting the potential links that can be established between them. Specifically, we explore the significance of embodiment, embeddedness, extended cognition, and enactive processes in learning and cognition. To bridge these frameworks, we propose employing the concept of "concrete experience" as an active engagement of individuals with their physical and social surroundings. By encompassing the essential aspects assigned to concrete experiences in EL, as well as the embodiment, situatedness, extended cognition, and enactive features of 4E cognition, this notion serves as a unifying element. Ultimately, the article suggests that combining the insights from EL and 4E cognition can offer a richer, more holistic understanding of representation.

Enhancing vaccine efficacy: Evaluating the superiority of cationic liposome-embedded squalene adjuvant against PCV2 infection.

Cationic liposome-embedded squalene (CLS) is a promising adjuvant that enhances antigen stability and mobility and improves immune response. This study compares the efficacy of a CLS-adjuvant porcine circovirus type 2 (PCV2) vaccine (CSV) with a conventional vaccine against PCV2. The CSV vaccine showed superior stability and was effective against PCV2-induced growth decline. It significantly increased serum immunoglobulin and cytokine levels, reduced serum PCV2 DNA, shortened the duration of viremia, and provided robust protection. CSV outperformed conventional vaccines, highlighting its potential for innovative vaccine development.

Unveiling STRs instability in a colorectal cancer FFPE sample: a case report.

In forensic genetics, sometimes formalin-fixed paraffin-embedded (FFPE) biopsy material taken during life is the only biological sample available for individual identification or paternity testing. In most cases, this biological tissue is characterized by the presence of tumor cells characterized by instability and loss of heterozygosity of microsatellites (MSI/LOH) compared to the DNA present in cells of normal tissue.In this case report, two FFPE samples from the same male subject were available for genetic investigation: one sample with colorectal cancer tissue and the other with normal tissue (no cancerous histopathological features). The comparison of the genetic profiles obtained from DNA extracted from the two tissues showed in the tumor tissue the presence of three genomic instability phenomena affecting FGA, CSF1P0, D21S2055 loci, located on three distinct autosomal chromosomes, and one duplication phenomenon affecting the DYS438. Therefore, due to the MSI/LOH phenomena, the genetic profile acquired from the tumor tissue was distorted and thus generated a fictitious genetic profile, not corresponding to the subject's real one (normal tissue free of tumor cells).

Dual-Mode Embedded Impulse-Radio Ultra-Wideband Radar System for Biomedical Applications.

This paper presents a real-time and non-contact dual-mode embedded impulse-radio (IR) ultra-wideband (UWB) radar system designed for microwave imaging and vital sign applications. The system is fully customized and composed of three main components, an RF front-end transmission block, an analog signal processing (ASP) block, and a digital processing block, which are integrated in an embedded system. The ASP block enables dual-path receiving for image construction and vital sign detection, while the digital part deals with the inverse scattering and direct current (DC) offset issues. The self-calibration technique is also incorporated into the algorithm to adjust the DC level of each antenna for DC offset compensation. The experimental results demonstrate that the IR-UWB radar, based on the proposed algorithm, successfully detected the 2D image profile of the object as confirmed by numerical derivation. In addition, the radar can wirelessly monitor vital sign behavior such as respiration and heartbeat information.

Streamlining YOLOv7 for Rapid and Accurate Detection of Rapeseed Varieties on Embedded Device.

Real-time seed detection on resource-constrained embedded devices is essential for the agriculture industry and crop yield. However, traditional seed variety detection methods either suffer from low accuracy or cannot directly run on embedded devices with desirable real-time performance. In this paper, we focus on the detection of rapeseed varieties and design a dual-dimensional (spatial and channel) pruning method to lighten the YOLOv7 (a popular object detection model based on deep learning). We design experiments to prove the effectiveness of the spatial dimension pruning strategy. And after evaluating three different channel pruning methods, we select the custom ratio layer-by-layer pruning, which offers the best performance for the model. The results show that using custom ratio layer-by-layer pruning can achieve the best model performance. Compared to the YOLOv7 model, this approach results in mAP increasing from 96.68% to 96.89%, the number of parameters reducing from 36.5 M to 9.19 M, and the inference time per image on the Raspberry Pi 4B reducing from 4.48 s to 1.18 s. Overall, our model is suitable for deployment on embedded devices and can perform real-time detection tasks accurately and efficiently in various application scenarios.

Quantification of the Uncertainty in Ultrasonic Wave Speed in Concrete: Application to Temperature Monitoring with Embedded Transducers.

This article presents an overall examination of how small temperature fluctuations affect P-wave velocity (Vp) measurements and their uncertainties in concrete using embedded piezoelectric transducers. This study highlights the fabrication of custom transducers tailored for long-term concrete monitoring. Accurate and reliable estimation of ultrasonic wave velocities is challenging, since they can be impacted by multiple experimental and environmental factors. In this work, a reliable methodology incorporating correction models is introduced for the quantification of uncertainties in ultrasonic absolute and relative velocity measurements. The study identifies significant influence quantities and suggests uncertainty estimation laws, enhancing measurement accuracy. Determining the onset time of the signal is very time-consuming if the onset is picked manually. After testing various methods to pinpoint the onset time, we selected the Akaike Information Criterion (AIC) due to its ability to produce sufficiently reliable results. Then, signal correlation was used to determine the influence of temperature (20 °C to 40 °C) on Vp in different concrete samples. This technique proved effective in evaluating velocity changes, revealing a persistent velocity decrease with temperature increases for various concrete compositions. The study demonstrated the capability of ultrasonic measurements to detect small variations in the state of concrete under the influence of environmental variables like temperature, underlining the importance of incorporating all influencing factors.

Overview of Embedded Rust Operating Systems and Frameworks.

Embedded Operating Systems (OSs) are often developed in the C programming language. Developers justify this choice by the performance that can be achieved, the low memory footprint, and the ease of mapping hardware to software, as well as the strong adoption by industry of this programming language. The downside is that C is prone to security vulnerabilities unknowingly introduced by the software developer. Examples of such vulnerabilities are use-after-free, and buffer overflows. Like C, Rust is a compiled programming language that guarantees memory safety at compile time by adhering to a set of rules. There already exist a few OSs and frameworks that are entirely written in Rust, targeting sensor nodes. In this work, we give an overview of these OSs and frameworks and compare them on the basis of the features they provide, such as application isolation, scheduling, inter-process communication, and networking. Furthermore, we compare the OSs on the basis of the performance they provide, such as cycles and memory usage.

Aqueous extraction of formalin-fixed paraffin-embedded tissue and detection of prion disease using real-time quaking-induced conversion.

OBJECTIVE: The goal of the research presented here is to determine if methods previously developed for the aqueous extraction of PrPSc from formalin-fixed paraffin-embedded tissue (FFPET) are applicable to the detection PrPSc by real-time quaking induced conversion (RT-QuIC). Previous work has utilized aqueous extraction of FFPET for detection of transmissible spongiform encephalopathies (TSEs) utilizing western blot and ELISA. This research extends the range of suitable methods for detection of TSEs in FFPET to RT-QuIC, which is arguably the most sensitive method to detect TSEs. RESULTS: We found complete agreement between the TSE status and the results from RT-QuIC seeded with the aqueous extract of FFPET samples. The method affords the diagnostic assessment TSE status by RT-QuIC of FFPET without the use of organic solvents that would otherwise create a mixed chemical-biological waste for disposal.

Microbiological investigation of pregnancies following vaginal radical trachelectomy using 16S rRNA sequencing of FFPE placental specimens.

This study examined the risk of intrauterine infection associated with radical trachelectomy (RT) in early-stage cervical cancer patients. This procedure preserves fertility but is linked to increased risk of intrauterine infection due to cervical defects during pregnancy. DNA was extracted from the formalin-fixed paraffin-embedded (FFPE) placental specimens of 23 pregnant post-RT patients and 16S rRNA gene sequencing was used for bacterial identification. The prevalence of Lactobacillus crispatus and Burkholderia stabilis was significantly higher in the non-chorioamnionitis group. In contrast, alpha diversity analysis using the PD index showed significantly higher diversity in the chorioamnionitis group (P = 0.04). The demonstrated relationship between chorioamnionitis and microbial diversity affirms the importance of controlling the genital bacterial flora in pregnancies following RT.

A set of pretreatment reagents including improved formula fixation and decalcification facilitating immunohistochemistry and DNA analyses of formalin-fixed paraffin-embedded bone marrow trephine biopsy.

Bone marrow biopsy depends on tissue morphology, immunohistochemical staining, and moleculardetection. Tissue pretreatment is required for bone marrow samples, from clinical specimen acquisition to pathological reporting, but during the process, proteins and nucleic acids are often altered because of the acid in fixation and decalcification solutions. In our study, we present an easy and effective pretreatment protocol and compared this novel pretreatment protocol (Set 2) with an existing traditional pretreatment process (Set 1) using tissue morphology, IHC staining, and molecular pathological analyses. Granulocytic IHC markers showed more intensive staining in samples of Set 2 than in those of Set 1. The Set 2 protocol provided a higher DNA yield and less fragmentation; moreover, samples processed with the Set 2 protocol could be subsequently used in FISH and DNA sequencing assays. Our optimized novel pretreatment protocol could better protect proteins and DNA molecules while maintaining good cell morphology compared to traditional pretreatment The novel pretreatment reagents could role as a reference by more laboratories for pretreating bone marrow biopsy samples and scientific research.

Concurrent and prolonged embedded earring at the earlobe as a possible factor of erosive lichen planus.

Key Clinical Message: Embedded earrings in adults have been reported to be a possible cause of erosive oral lichen planus. Abstract: Erosive lichen planus is rare and its cause cannot be determined. Embedded earring in the earlobes is rare clinical presentation in adult. The concurrent nature of these two conditions has not been previously, described in the published literature. Unexpectedly, we removed the earring from the earlobe and successfully treated erosive oral lichen planus. We hypothesized that prolonged embedded earring is a possible factor that induced erosive lichen planus in this case.

Retrospective clinical and microbiologic analysis of metagenomic next-generation sequencing in the microbiological diagnosis of cutaneous infectious granulomas.

BACKGROUND: Cutaneous infectious granulomas (CIG) are localized and chronic skin infection caused by a variety of pathogens such as protozoans, bacteria, worms, viruses and fungi. The diagnosis of CIG is difficult because microbiological examination shows low sensitivity and the histomorphological findings of CIG caused by different pathogens are commonly difficult to be distinguished. OBJECTIVE: The objective of this study is to explore the application of mNGS in tissue sample testing for CIG cases, and to compare mNGS with traditional microbiological methods by evaluating sensitivity and specificity. METHODS: We conducted a retrospective study at the Department of Dermatology of Sun Yat-sen Memorial Hospital, Sun Yat-sen University from January 1st, 2020, to May 31st, 2024. Specimens from CIG patients with a clinical presentation of cutaneous infection that was supported by histological examination were retrospectively enrolled. Specimens were delivered to be tested for microbiological examinations and mNGS. RESULTS: Our data show that mNGS detected Non-tuberculosis mycobacteria, Mycobacterium tuberculosis, fungi and bacteria in CIG. Compared to culture, mNGS showed a higher positive rate (80.77% vs. 57.7%) with high sensitivity rate (100%) and negative predictive value (100%). In addition, mNGS can detect more pathogens in one sample and can be used to detect variable samples including the samples of paraffin-embedded tissue with shorter detective time. Of the 21 patients who showed clinical improvement within a 30-day follow-up, eighteen had their treatments adjusted, including fifteen who continued treatment based on the results of mNGS. CONCLUSIONS: mNGS could provide a potentially rapid and effective alternative detection method for diagnosis of cutaneous infectious granulomas and mNGS results may affect the clinical prognosis resulting from enabling the patients to initiate timely treatment.

How to build a better 4E cognition.

Mougenot and Matheson outline a theoretical approach to cognitive neuroscience that combines the commitments of embodied cognition with a mechanistic approach to scientific explanation. They argue that this theoretical approach provides several general benefits, including enabling researchers to develop more robust theories and ontologies that do not require either neuroscientific reductionism or the complete autonomy of psychology from neuroscience. In this commentary, I argue that the sort of embodied cognitive neuroscience that they envision has a more specific benefit: it has the potential to help resolve internal tensions within 4E cognition.

Cognitive Function of Climbers: An Exploratory Study of Working Memory and Climbing Performance.

Sport climbing requires a combination of physical and cognitive skills, with working memory (WM) playing a crucial role in performance. This study aimed to investigate the association between WM capacity and climbing ability, while considering potential confounding factors including sex, age, education level, and climbing experience. Additionally, the study compared prefrontal cortex (PFC) hemodynamic responses among different climbing ability groups and sex during WM performance. Twenty-eight climbers participated, with WM assessed using the eCorsi task and PFC hemodynamic responses measured with near infrared spectroscopy (NIRS). Initial linear regression analyses revealed no association between WM and climbing ability. However, significant associations were found after adjustment for covariates. Specifically, sex (p = .014), sex in conjunction with age (p = .026), sex combined with climbing experience (p = .022), and sex along with education level (p = .038) were identified as significant predictors of differences in WM between Expert and Elite climbers. Additionally, notable differences in PFC hemodynamic responses were observed between Expert and Elite climbers, as well as between sexes during the WM task, providing support for differences in WM capacity. This study contributes to understanding the complex relationship between WM capacity and climbing performance, emphasizing the need to account for influencing factors in assessments.

Effect of Nitrite-Embedded Packaging Film on Growth of Listeria monocytogenes in Nitrite-free and Conventionally-cured Bologna Sausage.

Listeria monocytogenes is a pathogen frequently associated with ready-to-eat (RTE) meat and poultry products. Nitrite is a key antimicrobial additive that can offer some degree of protection against L. monocytogenes when included in meat product formulations. The objectives of this study were to determine the potential of nitrite-embedded film to affect the growth of L. monocytogenes following postthermal processing of conventionally-cured and nitrite-free bologna. Two bologna treatment formulations, a conventionally-cured control formulation (CON) and a nitrite-free formulation (UCC), were manufactured, packaged in conventional (CF) or nitrite-embedded (NEF) film, inoculated with 3.5 log CFU/cm2 of a cocktail of L. monocytogenes strains, and stored at 10 ± 1 °C. CON-NEF and UCC-NEF treatments had significantly slower (P < 0.05) growth of L. monocytogenes than CON-CF and UCC-CF, with populations in UCC-CF (which contained no nitrite) increasing by 3.4 logs after 10 d of storage in UCC-CF and 3.6 logs after 50 d in CON-CF (which had formulated nitrite only), while in the NEF-packaged samples, with or without formulated nitrite, they did not exceed the inoculum level until after day 40. Initial (day 0) residual nitrite was significantly greater (P < 0.05) in the control formulation. Packaging in NEF, however, resulted in an increase of 27-28 ppm by day 3, regardless of formulation, after which it decreased rapidly. Results suggest NEF can be used as a post-lethality antimicrobial intervention in food safety intervention strategies, in both cured and uncured processed meat products.

Engaging athletes as research participants. A document analysis of published sport science literature.

Sport science practitioners utilise findings from peer reviewed research to inform practice. Fewer studies are conducted with high performance athletes, however, than those involving recreationally active participants. Noting that research findings from recreational athletes may not be generalisable to the elite, there is a need to engage the latter cohort in research with better potential to influence health and performance. This study identified methods used to engage and recruit highly trained, elite and world class athletes as research participants. A document analysis was conducted using a purposive sample of peer-reviewed sport science literature. All articles published in 2022 from 18 highly ranked sport science journals were screened for inclusion. Studies investigating athletes ranked as highly trained/national level or above were included. All details related to participant recruitment were extracted from included articles, with the content being coded and thematically analysed using an interpretivist approach. A total of 439 studies from the 2356 screened were included in the analysis. Five primary themes of recruitment strategies were identified, beneath an overarching strategy of purposeful, convenience sampling. Recruitment themes related to the use of a gatekeeper, the research environment providing convenient access to athletes, promoting the study electronically, utilising professional networks and recruiting at training or competition. Engaging athletes through a gatekeeper is a prominent strategy to involve elite athletes in research. It is suggested that researchers work collaboratively with team or organisation personnel to promote recruitment, creating co-designed approaches that address issues most relevant to athletes and staff.

A Novel Acoustic Modulation of Oscillating Thin Elastic Membrane for Enhanced Streaming in Microfluidics and Nanoscale Liposome Production.

Liposomes are widely utilized in therapeutic nanosystems as promising drug carriers for cancer treatment, which requires a meticulous synthesis approach to control the nanoprecipitation process. Acoustofluidic platforms offer a favorable synthesis environment by providing robust agitation and rapid mixing. Here, a novel high-throughput acoustofluidic micromixer is presented for a solvent and solvent-free synthesis of ultra-small and size-tunable liposomes. The size-tunability is achieved by incorporating glycerol as a new technique into the synthesis reagents, serving as a size regulator. The proposed device utilizes the synergistic effects of vibrating trapped microbubbles and an oscillating thin elastic membrane to generate vigorous acoustic microstreaming. The working principle and mixing mechanism of the device are explored numerically and experimentally. The platform exhibits remarkable mixing efficacy for aqueous and viscous solutions at flow rates up to 8000 µL/h, which makes it unique for high-throughput liposome formation and preventing aggregation. As a proof of concept, this study investigates the impact of phospholipid type and concentration, flow rate, and glycerol on the size and size distribution of liposomes. The results reveal a significant size reduction, from ≈900 nm to 40 nm, achieved by merely introducing 75% glycerol into the synthesis reagents, highlighting an innovative approach toward size-tunable liposomes.

Challenges and solutions in FISH for formalin-fixed paraffin-embedded tissue: A scoping review.

Fluorescence in situ hybridization (FISH) has revolutionized molecular cytogenetic analysis since the 1980s, enabling precise localization of DNA sequences in cells and tissues. Despite its relevance, applying FISH to formalin-fixed paraffin-embedded (FFPE) tissue samples encounters significant technical challenges. This review addresses the main issues encountered in this context, such as inadequate fixation, contamination, block and slide age, inadequate pretreatment, and FISH technique. Proposed solutions include optimized pretreatment protocols, monitoring of blockage, careful selection of probes, and thorough analysis of results. Implementing good laboratory practices and quality control strategies are essential to ensure reliable results. Additionally, the use of emerging technologies such as artificial intelligence and digital pathology offers new perspectives for improving the efficiency and accuracy of FISH in FFPE samples. This review highlights the importance of a careful and personalized approach to overcome the technical challenges associated with FISH in FFPE samples, strengthening its role in research and clinical diagnosis. RESEARCH HIGHLIGHTS: Few FISH studies on FFPE: The scarcity of studies specifically addressing FISH applications in FFPE tissues highlights a critical gap in the literature. Troubleshooting FISH in FFPE tissues: Identifying and addressing common challenges in FISH techniques when applied to FFPE samples, such as signal quality and hybridization efficiency. Critical aspects of FISH technique: Discuss the main technical considerations crucial for successful FISH in FFPE tissues, including sample preparation, probe selection, and protocol optimization.