Sample Collection Protocols Research Articles

From confiscation to conservation: Wildlife DNA forensic for species identification of confiscated Felidae in Indonesia.

Illegal wildlife trade poses a significant threat to Indonesia's biodiversity, especially among its diverse Felidae species (cats). While molecular methods have proven effective for identifying some Felidae species, there remains a gap in comparing these techniques across different endemic Felidae species in Indonesia, particularly in cases involving multiple species in confiscated wildlife products. This study applies DNA forensic techniques to analyze 38 confiscated Felidae samples, identifying four species: Sumatran tiger (Panthera tigris sumatrae), leopard (Panthera pardus), leopard cat (Prionailurus bengalensis), and clouded leopard (Neofelis nebulosa). Notably, 21 samples identified as Sumatran tigers showed 100 % similarity to the reference sequence, demonstrating the effectiveness of custom-designed primers. The analysis also revealed a diverse range of biological materials, supporting the robustness of DNA forensics in wildlife conservation. A Bayesian phylogenetic tree further confirmed species differentiation with strong bootstrap values. These findings underscore the importance of accurate species identification for prosecuting wildlife crimes and formulating targeted conservation strategies. Despite limitations, including sample size and reliance on existing reference data, the study emphasizes the crucial role of DNA forensics in combating illegal wildlife trade and highlights the need for improved genetic databases. Practical implications include establishing protocols for sample collection, DNA forensic training for law enforcement, and fostering collaboration between forensic laboratories and conservation organizations. Integrating these forensic techniques supports broader conservation goals by enabling targeted enforcement, effective prosecution, and informed conservation planning through accurate subspecies identification.

Factores de riesgo asociados a la contaminación de hemocultivos en pacientes adultos y pediátricos: una revisión bibliográfica

Introduction: bloodstream infections are one of the leading causes of mortality in children and adults worldwide. Blood cultures are essential diagnostic tests for identifying microorganisms and guiding antimicrobial treatment. However, sample contamination reduces diagnostic accuracy due to the introduction of contaminating organisms during sample collection or the patient's clinical condition.Methods: a systematic review was conducted following the PRISMA guidelines on studies found in PubMed, Scielo, Scopus, Web of Science, and BVS, including a total of 19 studies.Results: after reviewing the 19 studies, detailed information was gathered on a standardized protocol for the proper collection of blood samples for blood cultures in adult and pediatric patients. Additionally, internal and external risk factors associated with specimen contamination were analyzed. Some of these factors included the severity of the patient's condition, the immune system, the use of invasive devices, skin antisepsis, technical difficulties, or high workload.Conclusions: this research highlights the importance of rigorous implementation of standardized procedures to minimize risk factors associated with blood culture contamination and, consequently, the prevention of unnecessary treatments.

Sustainable Protocols For Leaf Sample Collection in In-Vitro Culture: Evaluating The Impact of Sample Bags and Hydrogen Peroxide Pre-Sterilization

This study investigates sustainable leaf sample collection protocols for in-vitro culture (IVC) of chili (Capsicum frutescens) and tomato (Solanum lycopersicum) leaves. The research aimed to enhance viability and reduce contamination of leaf explants by evaluating various types of sample bags and different concentrations of hydrogen peroxide (H2O2) pre-sterilization. Specifically, the study compared the effectiveness of normal sealed (NS) bags and vacuum-sealed (VS) bags, including high-action (VSH) and low-action (VSL) vacuum-sealed bags, alongside H2O2 pre-sterilization at various concentrations. Leaf samples were treated with ten different concentrations of H2O2 (5% to 50%) to assess their impact on necrosis and surface contamination over 72 hr. Results showed that higher H2O2 concentrations (above 30%) caused significant necrosis, while concentrations between 10% and 15% provided optimal pre-sterilization for both leaf types, effectively reducing contamination without excessive tissue damage. In the second phase, the research examined the influence of different sample bags on leaf explant sustainability. Vacuum-sealed bags, particularly those with low-action vacuum (VSL), significantly improved leaf longevity and minimized contaminant emergence compared to normal sealed bags. Combined with 15% H2O2 pre-sterilization, VSL bags performed best, maintaining leaf morphology and viability for extended periods. Statistical analyses confirmed the significant impact of sampling bag type and pre-sterilization on contamination levels, necrosis emergence, and leaf longevity. The findings suggest that using low-action vacuum-sealed bags (VSL) with 15% H2O2 pre-sterilization is a promising approach for sustainable leaf sample collection, enhancing the success rate of IVC by minimizing microbial contamination and preserving leaf integrity during transport. This optimized protocol offers valuable insights for researchers and practitioners in plant tissue culture and agriculture, aiming to improve the sustainability and efficiency of leaf sample collection for in-vitro applications.

Logistics for Rapid Isolation of Viruses From Humans.

An important aspect of microbiological surveillance is the ability to access live viruses for microneutralization assays, which enables the study of viral characteristics and mechanisms in vitro and production of positive controls for diagnostic methods. During the COVID-19 pandemic, the Public Health Agency of Sweden established a protocol for the rapid collection of clinical samples and subsequent isolation of novel virus variants.

Chinese guideline for clinical application of fluid biomarkers for Alzheimer's disease(2024 edition)

Alzheimer's disease is the most prevalent neurodegenerative disorder leading to cognitive impairment, but its progression is subtle and the early recognition is difficult. With advancements in disease-modifying therapies, the need for precise early diagnosis of Alzheimer's disease is increasingly pressing. Fluid biomarkers of Alzheimer's disease, detectable in bodily fluid samples, are intricately associated with the disease.It can be used for screening, diagnosis, staging, prediction of disease progression, and clinical trials, playing an increasingly critical role in clinical practice.. This guideline systematically reviews and evaluates the spectrum of fluid biomarkers for Alzheimer's disease, propose standardized protocols for sample collection and processing, and delineates the application standards of fluid biomarkers in disease screening, diagnosis, staging, prognosis of disease progression, and clinical trials. A total of 24 recommendations have been formulated. The publication of this guideline aims to standardize the application of fluid biomarkers in clinical practice, thereby advancing research in Alzheimer's disease fluid biomarkers.

Detecting schizophrenia, bipolar disorder, psychosis vulnerability and major depressive disorder from 5 minutes of online-collected speech.

Psychosis poses substantial social and healthcare burdens. The analysis of speech is a promising approach for the diagnosis and monitoring of psychosis, capturing symptoms like thought disorder and flattened affect. Recent advancements in Natural Language Processing (NLP) methodologies enable the automated extraction of informative speech features, which has been leveraged for early psychosis detection and assessment of symptomology. However, critical gaps persist, including the absence of standardized sample collection protocols, small sample sizes, and a lack of multi-illness classification, limiting clinical applicability. Our study aimed to (1) identify an optimal assessment approach for the online and remote collection of speech, in the context of assessing the psychosis spectrum and evaluate whether a fully automated, speech-based machine learning (ML) pipeline can discriminate among different conditions on the schizophrenia-bipolar spectrum (SSD-BD-SPE), help-seeking comparison subjects (MDD), and healthy controls (HC) at varying layers of analysis and diagnostic complexity. We adopted online data collection methods to collect 20 minutes of speech and demographic information from individuals. Participants were categorized as "healthy" help-seekers (HC), having a schizophrenia-spectrum disorder (SSD), bipolar disorder (BD), major depressive disorder (MDD), or being on the psychosis spectrum with sub-clinical psychotic experiences (SPE). SPE status was determined based on self-reported clinical diagnosis and responses to the PHQ-8 and PQ-16 screening questionnaires, while other diagnoses were determined based on self-report from participants. Linguistic and paralinguistic features were extracted and ensemble learning algorithms (e.g., XGBoost) were used to train models. A 70%-30% train-test split and 30-fold cross-validation was used to validate the model performance. The final analysis sample included 1140 individuals and 22,650 minutes of speech. Using 5-minutes of speech, our model could discriminate between HC and those with a serious mental illness (SSD or BD) with 86% accuracy (AUC = 0.91, Recall = 0.7, Precision = 0.98). Furthermore, our model could discern among HC, SPE, BD and SSD groups with 86% accuracy (F1 macro = 0.855, Recall Macro = 0.86, Precision Macro = 0.86). Finally, in a 5-class discrimination task including individuals with MDD, our model had 76% accuracy (F1 macro = 0.757, Recall Macro = 0.758, Precision Macro = 0.766). Our ML pipeline demonstrated disorder-specific learning, achieving excellent or good accuracy across several classification tasks. We demonstrated that the screening of mental disorders is possible via a fully automated, remote speech assessment pipeline. We tested our model on relatively high number conditions (5 classes) in the literature and in a stratified sample of psychosis spectrum, including HC, SPE, SSD and BD (4 classes). We tested our model on a large sample (N = 1150) and demonstrated best-in-class accuracy with remotely collected speech data in the psychosis spectrum, however, further clinical validation is needed to test the reliability of model performance.

Implementation of a Microbiological Surveillance Protocol in a Portuguese Tertiary Care Academic Endoscopy Unit

Introduction: International societies recommend microbiological surveillance of endoscopes to reduce the incidence of endoscope-associated infections, particularly for duodenoscopes. However, surveillance protocols are not internationally standardized, both regarding sample collection, processing, and culture. This study aims to provide a framework protocol encompassing the experience of a tertiary large volume endoscopy center and the microbiology laboratory for collecting and culturing of endoscope samples for microbiological surveillance. Methods: A sample collection and processing protocol was designed as a result of a cooperation between the Endoscopy Center of the Gastroenterology Department and the Microbiology Laboratory of the Department of Clinical Pathology. This protocol reflects international recommendations in this topic and the human and technological resources of the involved departments. Results: The established protocol for collecting samples varies according to the type and model of endoscope. The specimens are collected as sterile saline liquid samples, as well as swabs (with and without transport media). Together with the collection of samples from the endoscope, samples from the final rinse water as well as the water bottle are also collected. For duodenoscopes and curvilinear echoendoscopes, we perform microbiological surveillance every 3 months; for gastroscopes and colonoscopes, at least, once a year; and for specific endoscopes, such as the pediatric or dual-channel therapeutic endoscopes, enteroscopes, or radial echoendoscopes, every 6 months. Conclusion: Endoscopy units should have detailed protocols for microbiological surveillance of endoscopes. These protocols should be drawn up by a multidisciplinary team that includes endoscopy nurses, gastroenterologists, microbiologists, and the antimicrobial stewardship team, following international recommendations, adapted to each institution resources.

BIOM-09. TOWARDS CLINICAL IMPLEMENTATION OF LIQUID BIOPSY ANALYSIS FOR PAEDIATRIC BRAIN TUMOUR PATIENTS – A PAN-EUROPEAN EFFORT FOR STANDARDIZATION

Abstract BACKGROUND Liquid biopsies (LBs) allow for analysis of tumour-derived molecules, such as cell-free DNA (cfDNA) and microRNAs (miRNAs), complementing cancer diagnostics, therapy monitoring, and minimal residual disease detection. While circulation-based LB has benefit for extracranial tumour entities, peripheral blood has not historically been a reliable source for diagnosis and disease-monitoring in brain tumours. For these tumours, cerebrospinal fluid (CSF) analysis is better suited for detection despite being a more invasive procedure. However, standardized protocols for sample collection and pre-analytical parameters for these LBs currently are limited. AIM We aim to establish an evidence-based gold standard protocol for cfDNA and miRNA analyses from CSF in paediatric brain tumour patients. In addition to higher compliance in trial-related CSF sampling, this initiative seeks to facilitate identification of clinically relevant indications and time-points for collection, thereby paving the way for well-informed, routine CSF molecular diagnostics. METHODS AND RESULTS We have initiated the Liquid Biopsy Working Group within the framework of the International Society of Paediatric Oncology Europe Brain Tumour Group (SIOPE-BTG). Available literature on LB in brain tumours was critically assessed regarding pre-analytical parameters, downstream analyses and LB assay performance. To directly evaluate confounding by pre-analytical factors, we experimentally compared the impact of CSF sampling sites, storage conditions, and different pre-processing protocols. cfDNA and miRNA yields, purity, and the fraction of tumour-derived material were selected as read-outs and correlated with tumour tissue information. To prepare for clinical translation, we consulted clinical paediatric brain tumour trial coordinators to identify major bottlenecks for CSF sampling in routine clinical practice. In addition, clinical practitioners and patient/ parent representatives have been consulted to comprehensively represent the perspectives and needs of all stakeholders. Together, results of this collaborative effort will be fundamental to the successful clinical integration of LBs to improve the future management of paediatric brain tumour patients.

Tongue depressor (bio)sensors: A fast decentralized self-testing of salivary biomarkers for personalized medicine

The clinical tests usually make use of blood samples, requiring invasive sample collection protocols, which are not appropriate for decentralized, frequent, and point-of-care (POC) applications. Here, we report disposable (bio)sensors combining electrochemical and colorimetric technologies on an accessible and lightweight wooden tongue depressor platform. The array of (bio)sensors on this commonly used medical tool enables rapid, and safe saliva sampling and showed promise to be used for frequent self-testing of four clinically relevant biomarkers (glucose, uric acid, nitrite, and thiocyanate) in saliva. The electrochemical system was designed to detect uric acid and glucose by chronoamperometric measurements in clinically relevant concentrations ranging from 50 to 500 µmol L−1 and from 10 to 5000 µmol L−1, respectively. Meanwhile, the colorimetric paper-based spot tests were optimized to detect nitrite and thiocyanate in concentrations ranging from 5 to 200 µmol L−1 and 0.5 to 2.1 mmol L−1, respectively, using digital image colorimetry. Our affordable tongue depressor (bio)sensors provide results within 5 min and achieve limits of detection in the µmol L−1 range. Importantly, the results provided by our tests when applied to human saliva samples presented excellent concordance with the traditional spectrophotometric method (at a 95 % confidence level), demonstrating the reliability and robustness of our method. In addition, the toxicity and biocompatibility studies of the tongue depressor (bio)sensors were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Calcein acetoxymethyl ester (Calcein AM), and Calcein Propidium Iodide Hoechst (Calcein PI) Analysis, confirming the safety use of the test. Therefore, our tongue depressor (bio)sensor facilitates frequent testing of several biomarkers and can contribute to personalized medicine.

Temporal dynamics of fecal microbiota community succession in broiler chickens, calves, and piglets under aerobic exposure.

Researchers have extensively studied the effect of oxygen on the growth and survival of bacteria. However, the impact of oxygen on bacterial community structure, particularly its ability to select for taxa within the context of a complex microbial community, is still unclear. In a 21-day microcosm experiment, we investigated the effect of aerobic exposure on the fecal community structure and succession pattern in broiler, calf, and piglet feces (n = 10 for each feces type). Bacterial diversity decreased and community structure changed rapidly in the broiler microbiome (P < 0.001), while the fecal community of calves and piglets, which have higher initial diversity, was stable after initial exposure but decreased in diversity after 3 days (P < 0.001). The response to aerobic exposure was host animal specific, but in all three animals, the change in community structure was driven by a decrease in anaerobic species, primarily belonging to Firmicutes and Bacteroidetes (except in broilers where Bacteroidetes increased), along with an increase in aerobic species belonging to Proteobacteria and Actinobacteria. Using random forest regression, we identified microbial features that predict aerobic exposure. In all three animals, host-beneficial Prevotella-related ASVs decreased after exposure, while ASVs belonging to Acinetobacter, Corynbacterium, and Tissierella were increased. The decrease of Prevotella was rapid in broilers but delayed in calves and piglets. Knowing when these pathobionts increase in abundance after aerobic exposure could inform farm sanitation practices and could be important in designing animal experiments that modulate the microbiome.IMPORTANCEThe fecal microbial community is contained within a dynamic ecosystem of interacting microbes that varies in biotic and abiotic components across different animal species. Although oxygen affects bacterial growth, its specific impact on the structure of complex communities, such as those found in feces, and how these effects vary between different animal species are poorly understood. In this study, we demonstrate that the effect of aerobic exposure on the fecal microbiota was host-animal-specific, primarily driven by a decrease in Firmicutes and Bacteroidetes, but accompanied by an increase in Actinobacteria, Proteobacteria, and other pathobionts. Interestingly, we observed that more complex communities from pig and cattle exhibited initial resilience, while a less diverse community from broilers displayed a rapid response to aerobic exposure. Our findings offer insights that can inform farm sanitation practices, as well as experimental design, sample collection, and processing protocols for microbiome studies across various animal species.

Operational Procedures of Forensic Science Laboratories and Protocols for Sample Collection

This research paper delves into the operational procedures employed within forensic science laboratories, with a specific focus on the protocols governing sample collection. Forensic science plays a crucial role in the criminal justice system, providing evidence and expert analysis to aid investigations and legal proceedings. Effective operational procedures and rigorous protocols for sample collection are essential to maintain the integrity and reliability of forensic evidence. Through an in-depth examination of existing literature, case studies, and expert insights, this paper aims to provide a comprehensive understanding of the methodologies and guidelines employed in forensic laboratories worldwide. Additionally, it explores the challenges faced by forensic scientists in sample collection and processing, along with recommendations for improving practices and enhancing the quality of forensic evidence.

Ensuring Accuracy: Critical Validation Techniques in Geochemical Analysis for Sustainable Geothermal Energy Development

Geochemical analysis is a critical tool in geothermal exploration, providing valuable insights into reservoir characteristics. However, obtaining accurate and reliable geochemical data requires rigorous validation techniques. This review examines key factors affecting the accuracy of geochemical data and discusses best practices for ensuring quality. Proper sampling methods, including selection of representative locations, use of appropriate equipment, and adherence to robust protocols for sample collection, filtration, preservation, and storage, are essential for maintaining integrity. Analytical techniques must be carefully selected, with regular calibration and standardization of instruments using certified reference materials. Implementing comprehensive quality assurance and quality control procedures, such as analyzing blanks, duplicates, and spike samples, helps monitor precision and accuracy. Data interpretation should consider the complexities of the geological and hydrological settings, integrating multiple lines of evidence. By following established guidelines and continuously updating methods based on emerging technologies and inter-laboratory comparisons, geothermal teams can optimize the reliability of their geochemical data. Accurate and precise geochemical information, when combined with geological, geophysical, and hydrological data, enables informed decision-making and enhances the success of geothermal projects. As geothermal energy gains importance in the transition to sustainable resources, ensuring the accuracy of geochemical analysis will be crucial for effective exploration and development.

Challenges in the Metabolomics-Based Biomarker Validation Pipeline.

As end-products of the intersection between the genome and environmental influences, metabolites represent a promising approach to the discovery of novel biomarkers for diseases. However, many potential biomarker candidates identified by metabolomics studies fail to progress beyond analytical validation for routine implementation in clinics. Awareness of the challenges present can facilitate the development and advancement of innovative strategies that allow improved and more efficient applications of metabolite-based markers in clinical settings. This minireview provides a comprehensive summary of the pre-analytical factors, required analytical validation studies, and kit development challenges that must be resolved before the successful translation of novel metabolite biomarkers originating from research. We discuss the necessity for strict protocols for sample collection, storage, and the regulatory requirements to be fulfilled for a bioanalytical method to be considered as analytically validated. We focus especially on the blood as a biological matrix and liquid chromatography coupled with tandem mass spectrometry as the analytical platform for biomarker validation. Furthermore, we examine the challenges of developing a commercially viable metabolomics kit for distribution. To bridge the gap between the research lab and clinical implementation and utility of relevant metabolites, the understanding of the translational challenges for a biomarker panel is crucial for more efficient development of metabolomics-based precision medicine.

Determination of metals in water samples within the irrigation area in Telo District, Kaffa Zone, South Western Ethiopia

Water contamination due to the accumulation of metal is not only an environmental problem, but it is also a global issue. The river flows alongside the town in its downward direction, where runoff from the uplands and municipal trash discharge during floods may contribute to the contamination of the river. Despite the fact that this river is the area's primary source of water, the water near the effluent release point is extremely corrosive. The nearby settlements depend on this untreated river water for drinking, irrigation, and other domestic uses since they have no access to a municipal water supply. The aim of the present study was to evaluate the degree of metal contamination in the Obasho river water samples collected from irrigation water sites in the Gurguba region. Three samples were collected from each of four irrigation water sites, which were 1.5 km away from each other. A total of 12 water samples were collected by following the standard water sample collection protocol. 100 mL of the irrigation water sample was digested in aquaragia, and AAS was used to determine the concentration of metals in the water samples. The concentration ranges of some metals were: Ca (1.84387–2.810824 mg/L), and its maximum limit in FAO is 20.0 mg/L; Mg (3.176942–4.543031 mg/L), and its maximum limit in FAO is 5.0 mg/L; Cr (0.039227–0.047872 mg/L), and its WHO/FAO permissible level is 0.1 mg/L; Co (0.036703–0.057218 mg/L), and its permissibility level is 0.05 mg/L WHO/FAO, Cd (0.006198–0.02856 mg/L), whose maximum limit in FAO is 0.01 mg/L, and Pb (0.065138–0.091131 mg/L) WHO/FAO permissible level is 5.0 mg/L. The mean concentrations of metals (Ca, Mg, Co, Cd, Pb, and Cr) in all study sites were below the regulatory limits except for Co and Cd; no water contamination was caused by these metals at the study sites. However, the mean concentration levels of Co in all study sites were above regulatory limits. The mean concentration of Co at Konit Kochito and Cd at Geremew Mamo, Konit Kochito, and Gereno Gebito is also greater than regulatory limits. A pair-wise comparison of some metals in study sites was carried out, thus the mean concentration of the irrigation water sites sample was significantly different from some metal concentration (p < 0.05) at the 95% confidence level. Generally, there was no cumulative effect of water contamination caused by Ca, Mg, Co, Cd, Pb, and Cr in Gurguba Kebele study sites, and it may not appear to pose very serious environmental problems at this moment. Thus, determining the metal content in irrigation water samples is crucial for ensuring the safety and sustainability of agricultural practices. It raises awareness regarding the contents of heavy metal contamination and sets limits for acceptable metal concentrations. It has also vital role in maintaining soil health.

Clinical laboratory strategies for managing pandemic outbreaks

This review article comprehensively examines clinical laboratory strategies for managing pandemic outbreaks, with a particular focus on lessons learned from the COVID-19 pandemic. Drawing upon an extensive analysis of existing literature from databases such as PubMed, Web of Science, and Cochrane, as well as manual searches on Google Scholar, the study aims to provide insights into key aspects of pandemic preparedness and response. The literature review encompasses diverse combinations of medical terminology, emphasizing the role and response of clinical laboratories during pandemics. The primary focus areas include rapid test development and deployment, increased testing capacity, sample collection, and transportation protocols, data management, personnel safety, resource allocation, adaptability, and quality control measures. The inclusion of articles in this study adheres to rigorous criteria, ensuring a thorough and robust review process. The findings highlight the critical importance of rapid and accurate diagnostic testing, especially in the early stages of a pandemic, to facilitate timely interventions and containment. Efficient sample collection and transportation systems are crucial for maintaining the integrity of specimens, while advanced data management and reporting systems contribute to informed decision-making. Adaptability and flexibility are identified as indispensable traits for navigating the dynamic challenges of a pandemic, and stringent quality control measures are foundational for upholding the accuracy of diagnostic testing. The review underscores the collaborative efforts needed among clinical laboratories, public health agencies, and research institutions to strengthen global pandemic preparedness and response strategies.

Modification of sensitive pyridine-pyrazolone method for determination of blood cyanide and its possible applications in fatal aircraft accidents

BackgroundFatal aviation accidents are a critical concern for armed forces, necessitating precise toxicological analysis of postmortem blood samples. This paper introduces a novel spectrophotometric method for quantifying cyanide in blood samples with an aim to aid in aircraft accident reconstruction in the future. MethodsBlood samples were collected from healthy non-smokers aged 20–45 years, excluding individuals with hepatic impairment, or those using drugs affecting blood cyanide levels. The spectrophotometric method involved cyanide adsorption in a sodium hydroxide solution using microdiffusion discs and chemical reactions resulting in a pinkish-purple end product. The optimal absorption wavelength (λmax) was determined, and the lower limits of detection and quantitation were established. The method underwent validation for precision, sensitivity, specificity, linearity, and accuracy. ResultsThe procedure yielded a pinkish-purple compound with an optimal absorption wavelength (λmax) of 531 nm. The established limits of detection (LoD) and quantitation (LoQ) were 0.0625 μg/ml and 0.125 μg/ml, respectively. The method demonstrated strong linearity up to cyanide concentrations of 4 μg/ml. It exhibited a sensitivity of 99.7% and accuracy of 85.92%. The intraday and inter-day precision were determined as 9.04% and 4.27%, respectively. The method was highly specific, showing no interference with blanks at the peak wavelength with a working range of 0.125–4 μg/ml. ConclusionThe study describes developing and validating a sensitive method for quantitatively estimating cyanide in blood samples. Incorporating this method into the existing postmortem toxicological sample collection and investigation protocol will enhance the analysis capabilities and improve aircraft accident investigation outcomes within the country.

A Clinical Breathomics Dataset

This study entailed a comprehensive GC‒MS analysis conducted on 121 patient samples to generate a clinical breathomics dataset. Breath molecules, indicative of diverse conditions such as psychological and pathological states and the microbiome, were of particular interest due to their non-invasive nature. The highlighted noninvasive approach for detecting these breath molecules significantly enhances diagnostic and monitoring capacities. This dataset cataloged volatile organic compounds (VOCs) from the breath of individuals with asthma, bronchiectasis, and chronic obstructive pulmonary disease. Uniform and consistent sample collection protocols were strictly adhered to during the accumulation of this extensive dataset, ensuring its reliability. It encapsulates extensive human clinical breath molecule data pertinent to three specific diseases. This consequential clinical breathomics dataset is a crucial resource for researchers and clinicians in identifying and exploring important compounds within the patient’s breath, thereby augmenting future diagnostic and therapeutic initiatives.

Prevalence of Enterococcus faecalis and Candida albicans in endodontic retreatment Cases: A comprehensive study

IntroductionEndodontic treatment failures often stem from the presence of microbial pathogens, particularly Enterococcus faecalis and Candida albicans. This study systematically assesses the prevalence of E. faecalis and C. albicans in endodontic retreatment cases, aiming to explore their impact on treatment outcomes. MethodsEmploying a systematic sampling approach, 30 patients with a history of previous endodontic treatment were selected. Rigorous clinical and radiographic assessments were conducted, following standardized protocols for root canal sample collection. Microbiological analysis, utilizing selective culture media, was employed to identify and quantify E. faecalis and C. albicans. Statistical analyses, including chi-square and logistic regression tests, were performed. ResultsThe study involved 30 patients undergoing endodontic retreatment, with comprehensive clinical and radiographic evaluations for cases with and without periradicular lesions. Microbiological analysis unveiled a significant prevalence of E. faecalis and C. albicans, establishing a robust association between these pathogens and retreatment failure. These findings underscore the critical need for targeted antimicrobial interventions to enhance the overall success rates of endodontic retreatment procedures. ConclusionThis study highlights the substantial prevalence of E. faecalis and C. albicans in endodontic retreatment cases, emphasizing the importance of identifying and effectively managing these pathogens for successful treatment outcomes. The notable association between these microbial agents and retreatment failure underscores the imperative for tailored antimicrobial strategies to enhance the efficacy of endodontic retreatment procedures.

Biomedical SERS - the current state and future trends.

Surface enhanced Raman spectroscopy (SERS) is meeting the requirements in biomedical science being a highly sensitive and specific analytical tool. By employing portable Raman systems in combination with customized sample pre-treatment, point-of-care-testing (POCT) becomes feasible. Powerful SERS-active sensing surfaces with high stability and modification layers if required are available for testing and application in complex biological matrices such as body fluids, cells or tissues. This review summarizes the current state in sample collection and pretreatment in SERS detection protocols, SERS detection schemes, i.e. direct and indirect SERS as well as targeted and non-targeted SERS, and SERS-active sensing surfaces. Moreover, the recent developments and advances of SERS in biomedical application scenarios, such as infectious diseases, cancer diagnostics and therapeutic drug monitoring is given, which enables the readers to identify the sample collection and preparation protocols, SERS substrates and detection strategies that are best-suited for their specific applications in biomedicine.

Activation of Dendritic Cells Isolated from the Blood of Patients with Prostate Cancer by Ex Vivo Fluid Shear Stress Stimulation.

Prostate cancer is one of the most common cancers among men in the United States and a leading cause of cancer-related death in men. Treatment options for patients with advanced prostate cancer include hormone therapies, chemotherapies, radioligand therapies, and immunotherapies. Provenge (sipuleucel-T) is an autologous cancer-vaccine-based immunotherapy approved for men with asymptomatic or minimally symptomatic metastatic castration-resistant prostate cancer (mCRPC). Administration of sipuleucel-T involves leukapheresis of patient blood to isolate antigen-presenting cells (APCs), including dendritic cells (DCs), and subsequent incubation of isolated APCs with both an antigen, prostatic acid phosphatase (PAP), and granulocyte macrophage-colony stimulating factor (GM-CSF) before their infusion back into the patient. Although sipuleucel-T has been shown to improve overall survival, other meaningful outcomes, such as prostate-specific antigen (PSA) levels and radiographic response, are inconsistent. This lack of robust response may be due to limited ex vivo activation of DCs using current protocols. Earlier studies have shown that many cell types can be activated ex vivo by external forces such as fluid shear stress (FSS). We hypothesize that novel fluid shear stress technologies and methods can be used to improve ex vivo efficacy of prostate cancer DC activation in prostate cancer. Herein, we report a new protocol for activating DCs from patients with prostate cancer using ex vivo fluid shear stress. Ultimately, the goal of these studies is to improve DC activation to expand the efficacy of therapies such as sipuleucel-T. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Sample collection and DC isolation Basic Protocol 2: Determination and application of fluid shear stress Basic Protocol 3: Flow cytometry analysis of DCs after FSS stimulation.