Evaluation of a Novel Fixative Solution for Liquid-Based Cytology in Diagnostic Cytopathology.

Since its introduction in the early 2000s, liquid-based cytology (LBC) has been increasingly used for gynecologic and non-gynecologic cytology, and its multiple advantages have been widely recognized. The aim of this study was to investigate the use of a new fixative and pre-analytical method for morphological diagnosis in cytological samples. In particular, we evaluated the effect of a novel preservative solution on the preparation of diagnostic slides by comparing it with the standard reference used globally in cytology laboratories. This study included both gynecological (n = 139) and non-gynecological (n = 183) samples. Several morphologic variables were then identified and evaluated. Using this approach, we were then able to demonstrate the suitability of the new system, with improved safety, to be integrated within current pathology clinical practice. Overall, using a safer preservative solution, the study shows no statistical difference (and then non-inferiority) in the new fixation protocol compared with the standard reference used in routine practice in terms of diagnostic adequacy, evaluated both in clinically relevant gyn and non-gyn datasets.

Expanded biochemical analyses of human tear fluid: Polyvalent faces of the schirmer strip.

The tear film forms a protective barrier between the ocular surface and the external environment. Despite its small volume, recent advancements in preanalytical and analytical procedures have enabled its in-depth analysis using multiple approaches. However, the diversity of tear film collection methods and the lack of standardization in pre-analytical methods represent the main obstacles to reproducible results and comparison among different studies. In this study, we first improved the pre-analytical procedures for the extraction of various molecular entities from Schirmer strips (ScS). Subsequently, our investigation focused on analyzing the molecular variances that might occur between two primary tear collection methods: capillary tube (CT) and ScS. Additionally, we examined different parts of the ScS to underscore these variations, which could serve as crucial factors for developing a standardized, optimized protocol for sample processing. Our results show that the inclusion of surfactants in the extraction process enhanced both the yield of protein extraction and the number of proteins identified in ScS, by effectively lysing the cells and improving the solubility of several intracellular proteins. In addition to proteins, nucleic acids could also be recovered for gene expression analyses, particularly from the bulb region of the ScS which is placed in the cul-de-sac. Despite their diluted nature, extracts from ScS remain a suitable material for retrieving tear proteins such as IL-17A at levels as low as the fg/mL range, thanks to highly sensitive immunoassays. Collection methods can affect measured tear protein levels. Lactoferrin is found in higher percentages in capillary electrophoresis analysis of tears collected using ScS compared to tears collected by CT (39.6 ± 4.8% versus 31 ± 4.4%).

Recent advances in nanomaterials-based electrochemical sensors for tramadol analysis.

Tramadol is a centrally-acting analgesic used for treating moderate to severe acute and chronic pain. Pain is an unpleasant sensation that occurs most commonly as a result of tissue injury. Tramadol possesses agonist actions at the µ-opioid receptor and effects reuptake at the noradrenergic and serotonergic systems. In the last years, several analytical procedures have been published in the literature for the determination of tramadol from pharmaceutical formulations and biological matrices. Electrochemical methods have attracted tremendous attention for the quantification of this drug owing to their demonstrated potential for quick response, real-time measurements, elevated selectivity and sensitivity. In this review, we highlighted the recent advances and applications of nanomaterials-based electrochemical sensors for the analysis and detection of tramadol, which is extremely important for the indication of effective diagnoses and for quality control analyses in order to protect human health. Also, the main challenges in developing nanomaterials-based electrochemical sensors for the determination of tramadol will be discussed. At last, this review offers prospects for the future research and development needed for modified electrode sensing technology for the detection of tramadol.

The Art of Stem Cell-Based Therapy.

Potency assays represent crucial experiments at the hub of the comprehensive complexity surrounding cell therapy. Moreover, numerous factors beyond biological and scientific considerations are involved in achieving successful potency assays that fulfil regulatory authority approval for a new advanced therapy medicinal product. Though this can mean a frustratingly long period of discovery and development, progress in cell therapy is nowadays proceeding remarkably quickly, assisted by the potency assay rigorously placing emphasis on the need to critically analyse the key factor/s responsible for the therapeutic mechanism of action. History has shown that it can take many decades for there to be an improved understanding of a mechanism of action. Yet the chasing of precise targets has revolutionised medicine, with no clearer example than approaches to viral pandemics. The centuries involved in the eradication of smallpox have paved the way for an unprecedented pace of vaccine development for the Covid-19 pandemic. Such extraordinary accomplishments foster encouragement that similarly for stem cell-based therapy, our scientific knowledge will continue to improve apace. This chapter focuses on the art of experimentation and discovery, introducing potency assay requisites and numerous factors that can influence potency assay outcomes. A comprehensive understanding of potency assays and their development can hasten the provision of new cell therapies to help resolve burdensome diseases of unmet medical need.

Measurement Uncertainty and Conformity Assessment Applied to Drug and Medicine Analyses - A Review.

Analytical results are often used in scientific research, industrial and clinical applications to support decision making. Despite all efforts to ensure the reliability of analytical results (including method validation, internal quality control, use of certified reference materials, proficiency tests, and ISO 17025 accreditation), there will always be an uncertainty associated with the measured value. The measurement uncertainty expresses the quality of the analytical result and allows the comparability between analytical results or between the measured value and the specification limit(s). This work discusses the importance of measurement uncertainty, including the steps involved in the measurement uncertainty evaluation, the bottom-up and top-down approaches used in measurement uncertainty calculation, the measurement uncertainty evaluation in drug and medicine analyses, and the application of measurement uncertainty in conformity assessment for quality control, stability studies, and pharmaceutical equivalence.

Impact of the pre-examination phase on multicenter metabolomic studies.

The development of metabolomics in clinical applications has been limited by the lack of validation in large multicenter studies. Large population cohorts and their biobanks are a valuable resource for acquiring insights into molecular disease mechanisms. Nevertheless, most of their collections are not tailored for metabolomics and have been created without specific attention to the pre-analytical requirements for high-quality metabolome assessment. Thus, comparing samples obtained by different pre-analytical procedures remains a major challenge. Here, 1H NMR-based analyses are used to demonstrate how human serum and plasma samples collected with different operating procedures within several large European cohort studies from the Biobanking and Biomolecular Resources Infrastructure - Large Prospective Cohorts (BBMRI-LPC) consortium can be easily revealed by supervised multivariate statistical analyses at the initial stages of the process, to avoid biases in the downstream analysis. The inter-biobank differences are discussed in terms of deviations from the validated CEN/TS 16945:2016 / ISO 23118:2021 norms. It clearly emerges that biobanks must adhere to the evidence-based guidelines in order to support wider-scale application of metabolomics in biomedicine, and that NMR spectroscopy is informative in comparing the quality of different sample sources in multi cohort/center studies.

Key factors influencing metal concentrations in sediments along Western European Rivers: A long-term monitoring study (1945-2020).

Since 1945, a large amount of heterogeneous data has been acquired to survey river sediment quality, especially concerning regulatory metals such as Cd, Cr, Cu, Hg, Ni, Pb, and Zn. Large-scale syntheses are critical to assess the effectiveness of public regulations and the resiliency of the river systems. Accordingly, this data synthesis proposes a first attempt to decipher spatio-temporal trends of metal contamination along seven major continental rivers in Western Europe (France, Belgium, Germany, and the Netherlands). A large dataset (>12,000 samples) from various sediment matrices (bed and flood deposits - BFD, suspended particulate matter - SPM, dated sediment cores - DSC) was set up based on monitoring and scientific research from the 1950s to the 2010s. This work investigates the impact of analytical protocols (matrix sampling, fractionation, extraction), location and time factors (related to geology and anthropogenic activities) on metal concentration trends. Statistical analyses highlight crossed-interactions in space and time, as well as between sediment matrices (metal concentrations in SPM ≃ DSC > BFD) and extraction procedures (also related to river lithology). Major spatio-temporal trends are found along several rivers such as (i) an increase of metal concentrations downstream of the main urban industrial areas (e.g. Paris-Rouen corridor on the Seine River, Bonn-Duisburg corridor on the Rhine River), (ii) a long-term influence of former mining areas located in crystalline zones, releasing heavily contaminated sediments for decades (Upper Loire River, Middle Meuse section), (iii) a decrease of metal concentrations since the 1970s (except for Cr and Ni, rather low and stable over time). The improvement of sediment quality in the most recent years in Europe reflects a decisive role of environment policies, such as more efficient wastewater treatments, local applications of the Water Framework Directive and urban industrial changes in the river valleys.

Multicriteria Decision Analysis and Grouping of Analytical Procedures for Phthalates Determination in Disposable Baby Diapers.

This study presents the application of one of the tools from the multicriteria decision analysis set (MCDA), the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS). Selected green analytical chemistry metrics were used to rank analytical procedures for the phthalate determination in disposable baby diapers. Nine analytical procedures were assessed in order to find one that has the lowest environmental impact and the best analytical figures of merit. Nine different criteria, where weighting was based on the experts' evaluation, were used in the procedures' assessment. With the use of TOPSIS, an easy and straightforward technique, selection of the most appropriate procedure was made.

Bio Discarded from Waste to Resource.

The modern linear agricultural production system allows the production of large quantities of food for an ever-growing population. However, it leads to large quantities of agricultural waste either being disposed of or treated for the purpose of reintroduction into the production chain with a new use. Various approaches in food waste management were explored to achieve social benefits and applications. The extraction of natural bioactive molecules (such as fibers and antioxidants) through innovative technologies represents a means of obtaining value-added products and an excellent measure to reduce the environmental impact. Cosmetic, pharmaceutical, and nutraceutical industries can use natural bioactive molecules as supplements and the food industry as feed and food additives. The bioactivities of phytochemicals contained in biowaste, their potential economic impact, and analytical procedures that allow their recovery are summarized in this study. Our results showed that although the recovery of bioactive molecules represents a sustainable means of achieving both waste reduction and resource utilization, further research is needed to optimize the valuable process for industrial-scale recovery.

Understanding Quality Paradigm Shifts in the Evolving Pharmaceutical Landscape: Perspectives from the USP Quality Advisory Group.

Recent changes in the pharmaceutical industry have led to significant paradigm shifts in the pharmaceutical quality environment. Globalization of the pharmaceutical industry, increasingly rapid development of novel therapies, and adoption of new manufacturing techniques have presented numerous challenges for the established regulatory framework and quality environment and are impacting the approaches utilized to ensure the quality of pharmaceutical products. Regulators, industry, and standards-setting organizations have begun to recognize the need to rely more on integrated risk-based approaches and to create more nimble and flexible standards to complement these efforts. They also increasingly have recognized that quality needs to be built into systems and processes throughout the lifecycle of the product. Moreover, the recent COVID-19 crisis has emphasized the need to adopt practices that better promote global supply chain resilience. In this paper, the USP Quality Advisory Group explores the various paradigm shifts currently impacting pharmaceutical quality and the approaches that are being taken to adapt to this new environment. Broad adoption of the Analytical Procedure Lifecycle approach, improved data management, and utilization of digital technologies are identified as potential solutions that can help meet the challenges of these quality paradigm shifts. Further discussion and collaboration among stakeholders are needed to pursue these and other solutions that can ensure a continued focus on quality while facilitating pharmaceutical innovation and development.

Amygdalin: Toxicity, Anticancer Activity and Analytical Procedures for Its Determination in Plant Seeds.

Amygdalin (d-Mandelonitrile 6-O-ß-d-glucosido-ß-d-glucoside) is a natural cyanogenic glycoside occurring in the seeds of some edible plants, such as bitter almonds and peaches. It is a medically interesting but controversial compound as it has anticancer activity on one hand and can be toxic via enzymatic degradation and production of hydrogen cyanide on the other hand. Despite numerous contributions on cancer cell lines, the clinical evidence for the anticancer activity of amygdalin is not fully confirmed. Moreover, high dose exposures to amygdalin can produce cyanide toxicity. The aim of this review is to present the current state of knowledge on the sources, toxicity and anticancer properties of amygdalin, and analytical methods for its determination in plant seeds.

Attribution Markers and Data Mining in Art Authentication.

Today's global art market is a billion-dollar business, attracting not only investors but also forgers. The high number of forged works requires reliable authentication procedures to mitigate the risk of investments. However, with the developments in the methodology, continuous time pressure and the threat of litigation, authenticating artwork is becoming increasingly complex. In this paper, we examined whether the decision process involved in the authenticity examination may be supported by machine learning algorithms. The idea is motivated by existing clinical decision support systems. We used a set of 55 artworks (including 12 forged ones) with determined attribution markers to train a decision tree model. From our preliminary results, it follows that it is a very promising technique able to support art experts. Decision trees are able to summarize the existing knowledge about all investigations and may also be used as a classifier for new paintings with known markers. However, larger datasets with artworks of known provenance are needed to build robust classification models. The method can also utilize the most important markers and, consequently, reduce the costs of investigations.

[Estimating Postmortem Interval by Protein Chip Detection Technology Combined with Multidimensional Statistics].

ABSTRACT: Objective To obtain the protein expression profile of rat liver tissue after death by the 2100 bioanalyzer combined with protein chip, and infer the relationship between protein expression profile and postmortem interval. Methods Rats were killed by abdominal anesthesia and placed at 16 ℃. Water-soluble proteins in liver tissues were extracted at 14 time points after death. The expression profile data of proteins with relative molecular weight of 14 000-230 000 were obtained using protein chip, and principal component analysis （PCA）, partial least squares-discriminant analysis （PLS-DA） and Fisher discriminant were used to analyze the data. Results According to the changes of protein expression profile, the postmortem interval was divided into group A （0 d）, group B （1-9 d）, group C （12-30 d） according to the result of PLS-DA. The prediction accuracy of the training set and test set of the model were all 100.0%, and the internal cross-validation of the training set was 100.0% according to Fisher discriminant. The Fisher discriminant model at each time point of group B and C was established to narrow the time window of postmortem interval estimation. The prediction accuracy of the training set and test set were all 100.0%, and the internal cross-validation accuracy of the training set was 100.0% in group B. The prediction accuracy of the training set and test set were respectively 95.2% and 78.6% in group C, and the internal cross-validation of the training set was 88.1%. Conclusion Protein chip detection technology can quickly and easily obtain the expression profile of water-soluble proteins of rat liver tissue with a relative molecular weight of 14 000-230 000 at different time points after death. PLS-DA and Fisher discriminant models are established to classify and predict the postmortem interval, in order to provide new ideas and methods for postmortem interval estimation.

Estimation of an Elite Road Cyclist Performance in Different Positions Based on Numerical Simulations and Analytical Procedures.

The aim of this study was to use numerical simulations and analytical procedures to compare a cyclist's performance in three different cycling positions. An elite level road cyclist competing at a national level was recruited for this research. The bicycle was 7 kg and the cyclist 55 kg. A 3D scan was taken of the subject on the competition bicycle, wearing race gear and helmet in the upright position, in the handlebar drops (dropped position) and leaning on the elbows (elbows position). Numerical simulations by computer fluid dynamics in Fluent CFD code assessed the coefficient of drag at 11.11 m/s. Following that, a set of assumptions were employed to assess cycling performance from 1 to 22 m/s. Drag values ranged between 0.16 and 99.51 N across the different speeds and positions. The cyclist mechanical power in the elbows position differed from the upright position between 0 and 23% and from the dropped position from 0 to 21%. The cyclist's energy cost in the upright position differed 2 to 16% in comparison to the elbows position and the elbows position had less 2 to 14% energy cost in comparison to the dropped position. The estimated time of arrival was computed for a 220,000 m distance and it varied between 7,715.03 s (2 h:8 min:24 s) and 220,000 s (61 h:6 min:40 s) across the different speeds and positions. In the elbows position, is expected that a cyclist may improve the winning time up to 23% in comparison to he upright and dropped position across the studied speeds.

Analysis of Cyclist's Drag on the Aero Position Using Numerical Simulations and Analytical Procedures: A Case Study.

BACKGROUND: Resistance acting on a cyclist is a major concern among the cycling fraternity. Most of the testing methods require previous training or expensive equipment and time-consuming set-ups. By contrast, analytical procedures are more affordable and numerical simulations are perfect for manipulating and controlling inputs. The aim of this case study was to compare the drag of a cyclist in the aero position as measured using numerical simulation and analytical procedures. METHODS: An elite male cyclist (65 kg in mass and 1.72 m in height) volunteered to take part in this research. The cyclist was wearing his competition gear, helmet and bicycle. A three-dimensional model of the bicycle and cyclist in the aero position was obtained to run the numerical simulations. Computational fluid dynamics (CFD) and a set of analytical procedures were carried out to assess drag, frontal area and drag coefficient, between 1 m/s and 22 m/s, with increments of 1 m/s. The t-test paired samples and linear regression were selected to compare, correlate and assess the methods agreement. RESULTS: No significant differences (t = 2.826; p = 0.275) between CFD and analytical procedures were found. The linear regression showed a very high adjustment for drag (R2 = 0.995; p < 0.001). However, the drag values obtained by the analytical procedures seemed to be overestimated, even though without effect (d = 0.11). CONCLUSIONS: These findings suggest that drag might be assessed using both a set of analytical procedures and CFD.

[Expression and bioinformatics analysis of long-chain non-coding RNA PVT1 in tumors].

Objective: To study the expression and mechanism of long-chain non-coding RNA PVT1 in tumor by bioinformatics analysis and experimental verification, and to provide new ideas for the study of the pathogenesis of tumors. Methods: The expression of PVT1 in 14 common tumors was downloaded from starBase v2.0 public database, which also was verified by PVT1 RNA-in situ hybridization.The upstream transcription factors, the downstream target microRNA(miRNA) for PVT1 and the target genes for the target miRNAs were predicted and analyzed by using bioinformatics based on the database of UCSC Genome Browser, HMDD v2.0, miRTar Base, JASPAR databases. Results: StarBase database analysis and RNA in situ hybridization showed that PVT1 was highly expressed in kidney clear cell carcinoma and colon and rectal adenocarcinoma. PVT1 was regulated by the upstream transcription factors CREB1, Atf1, SP1, KLF5, STAT3, while it could control the expression of the downstream target miR-16. bcl-2, VEGFA, CCNE1, CCND1 and SHOC2 showed an interaction with the transcription factor of PVT1, which formed a feedback regulatory pathway. Conclusions: PVT1 is highly expressed in kidney clear cell carcinoma and colon and rectal adenocarcinoma.The predictive analysis of bioinformatics demonstrates that transcription factor/PVT1/miR-16/target gene signal axon may be an important molecular mechanism, which provide a valuable clue for further functional mechanism research of long-chain non-coding RNA.

Multielemental speciation analysis by advanced hyphenated technique - HPLC/ICP-MS: A review.

Speciation analysis has become an invaluable tool in human health risk assessment, environmental monitoring or food quality control. Another step is to develop reliable multielemental speciation methodologies, to reduce costs, waste and time needed for the analysis. Separation and detection of species of several elements in a single analytical run can be accomplished by high performance liquid chromatography hyphenated to inductively coupled plasma mass spectrometry (HPLC/ICP-MS). Our review assembles articles concerning multielemental speciation determination of: As, Se, Cr, Sb, I, Br, Pb, Hg, V, Mo, Te, Tl, Cd and W in environmental, biological, food and clinical samples analyzed with HPLC/ICP-MS. It addresses the procedures in terms of following issues: sample collection and pretreatment, selection of optimal conditions for elements species separation by HPLC and determination using ICP-MS as well as metrological approach. The presented work is the first review article concerning multielemental speciation analysis by advanced hyphenated technique HPLC/ICP-MS.

Microfluidic solutions enabling continuous processing and monitoring of biological samples: A review.

The last decade has witnessed tremendous advances in employing microfluidic solutions enabling Continuous Processing and Monitoring of Biological Samples (CPMBS), which is an essential requirement for the control of bio-processes. The microfluidic systems are superior to the traditional inline sensors due to their ability to implement complex analytical procedures, such as multi-step sample preparation, and enabling the online measurement of parameters. This manuscript provides a backgound review of microfluidic approaches employing laminar flow, hydrodynamic separation, acoustophoresis, electrophoresis, dielectrophoresis, magnetophoresis and segmented flow for the continuous processing and monitoring of biological samples. The principles, advantages and limitations of each microfluidic approach are described along with its potential applications. The challenges in the field and the future directions are also provided.

Procedimientos analítico-racionales en la adaptación de tests. Adaptación al español de la batería de pruebas de razonamiento: Adaptation to Spanish of the Reasoning Tests Battery / Judgmental-analytical procedures for adapting tests

Adaptar un test entre culturas o idiomas requiere considerar aspectos legales, culturales, lingüísticos, métricos y de uso. La atención prestada por la literatura especializada a los aspectos metodológicos implicados en el estudio de la equivalencia métrica ha sido significativamente mayor que la dedicada a los procedimientos analítico-racionales previos a la fase de confirmación empírica. Sin embargo, la consideración de estos últimos es crucial en el proceso de adaptación. En esta línea, el objetivo de este trabajo es incidir en la relevancia de las etapas previas y describirlas, ofreciendo una sistematización que considera diez apartados. Todos ellos contribuyen a garantizar y favorecer la construcción de un test adaptado y equivalente, en la medida de lo posible, al test original. El proceso es ejemplificado con la adaptación al español de una prueba cognitiva construida en portugués para la población portuguesa, la Bateria de Provas de Raciocínio.

Adapting a test between cultures or languages requires taking into account legal, linguistic, metric, and use-related considerations. Significantly more attention has been paid to the methodological aspects involved in the study of metric equivalence than to judgmental-analytical procedures prior to the empirical confirmation stage. However, considering the latter is crucial in the adaptation process. Along these lines, this paper seeks to describe and focus on the relevance of the previous stages, thereby offering a systematization process that comprises ten sections. This approach contributes to ensuring the construction of a test adapted and equivalent in as much as possible to the original. This process is exemplified by means of a Spanish language adaptation of a cognitive test originally designed in Portuguese for the Portuguese population, the Reasoning Test Battery.

Analytical procedures for the determination of fuel combustion products, anti-corrosive compounds, and de-icing compounds in airport runoff water samples.

The purpose of this study is to propose and evaluate new procedures for determination of fuel combustion products, anti-corrosive and de-icing compounds in runoff water samples collected from the airports located in different regions and characterized by different levels of the activity expressed by the number of flights and the number of passengers (per year). The most difficult step in the analytical procedure used for the determination of PAHs, benzotriazoles and glycols is sample preparation stage, due to diverse matrix composition, the possibility of interference associated with the presence of components with similar physicochemical properties. In this study, five different versions of sample preparation using extraction techniques, such as: LLE and SPE, were tested. In all examined runoff water samples collected from the airports, the presence of PAH compounds and glycols was observed. In majority of the samples, BT compounds were determined. Runoff water samples collected from the areas of Polish and British international airports as well as local airports had similar qualitative composition, but quantitative composition of the analytes was very diverse. New and validated analytical methodologies ensure that the necessary information for assessing the negative impact of airport activities on the environment can be obtained.