Development of a biodegradable microfluidic paper-based device for blood-plasma separation integrated with non-enzymatic electrochemical detection of ascorbic acid.

In the present article, we developed an electrochemical microfluidic paper-based device (EµPAD) for the non-enzymatic detection of Ascorbic Acid (AA) concentration in plasma using whole human blood. We combined LF1 blood plasma separation membrane and Whatman grade 1 filter paper to separate plasma from whole blood through wax printing. A screen-printed electrode (SPE) was modified with spherical-shaped MgFe2O4 nanomaterial (n-MgF) to improve the catalytic properties of SPE. The n-MgF was prepared via hydrothermal method, and its material phase and morphology were confirmed via XRD, FTIR, TEM, SEM, and AFM analysis. The fabricated n-MgF/SPE/EµPAD exhibited detection of AA ranging from 0 to 80 µM. The obtained value of the detection limit, limit of quantification, sensitivity, and response time are 2.44 µM, 8.135 µM, 5.71 × 10-3 mA µM-1 cm-2, and 10 s, respectively. Our developed n-MgF/SPE/EµPAD shows marginal interference with the common analytes present in plasma, such as uric acid, glutamic acid, glucose, urea, lactic acid, and their mixtures. Overall, our low-cost, portable device with its user-friendly design and efficient plasma separation capability offers a practical and effective solution for estimating AA concentration from whole human blood in a single step.

Documento multidisciplinar de consenso sobre el manejo de la hemorragia masiva. Primera actualización 2023 (documento HEMOMAS-II) / Multidisciplinary consensus document on the management of massive haemorrhage. First update 2023 (document HEMOMAS-II)

El presente documento supone una puesta al día del documento multidisciplinar HEMOMAS, publicado en el año 2016 con el aval de las Sociedades Científicas Españolas de Anestesiología y Reanimación (SEDAR), Medicina Intensiva, Crítica y Unidades Coronarias (SEMICYUC) y de Trombosis y Hemostasia (SETH). El objetivo de este documento fue revisar y actualizar las recomendaciones existentes sobre el manejo de la hemorragia masiva (HM). Se siguió una metodología basada en elementos del método ADAPTE (búsqueda y adaptación de guías publicadas en el ámbito específico de la HM desde 2014, más búsqueda bibliográfica en PubMed y EMBASE desde enero-2014 hasta junio-2021). Tras la revisión de nueve guías y 207 artículos seleccionados, se actualizaron las 47 recomendaciones existentes en el artículo original, manteniendo, suprimiendo o modificando cada una de ellas y sus grados de recomendación y evidencia. Consensuadamente, los autores aprobaron la redacción final del artículo y las 41 recomendaciones resultantes.(AU)

This document is an update of the multidisciplinary document HEMOMAS, published in 2016 with the endorsement of the Spanish Scientific Societies of Anaesthesiology (SEDAR), Intensive Care (SEMICYUC) and Thrombosis and Haemostasis (SETH). The aim of this document was to review and update existing recommendations on the management of massive haemorrhage. The methodology of the update was based on several elements of the ADAPTE method by searching and adapting guidelines published in the specific field of massive bleeding since 2014, plus a literature search performed in PubMed and EMBASE from January 2014 to June 2021. Based on the review of 9 guidelines and 207 selected articles, the 47 recommendations in the original article were reviewed, maintaining, deleting, or modifying each of them and the accompanying grades of recommendation and evidence. Following a consensus process, the final wording of the article and the resulting 41 recommendations were approved by all authors.(AU)

Sampling Cerebrospinal Fluid and Blood from Lateral Tail Vein in Rats during EEG Recordings.

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many experimental contexts to measure molecules of interest, such as hormones, growth factors, proteins, or small non-coding RNAs. A specific example is the sampling of biological liquids in the research of biomarkers for epilepsy. In these studies, it is desirable to compare the levels of molecules in cerebrospinal fluid (CSF) and in plasma, by withdrawing CSF and plasma in parallel and considering the time distance of the sampling from and to seizures. The combined CSF and plasma sampling, coupled with video-EEG monitoring in epileptic animals, is a promising approach for the validation of putative diagnostic and prognostic biomarkers. Here, a procedure of combined CSF withdrawal from cisterna magna and blood sampling from the lateral tail vein in epileptic rats that are continuously video-EEG monitored is described. This procedure offers significant advantages over other commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, and reduced time of anesthesia. Additionally, it can be used to obtain CSF and plasma samples in both tethered and telemetry EEG recorded rats, and it may be used repeatedly across multiple days of experiment. By minimizing the stress due to sampling by shortening isoflurane anesthesia, measures are expected to reflect more accurately the true levels of investigated molecules in biofluids. Depending on the availability of an appropriate analytical assay, this technique may be used to measure the levels of multiple, different molecules while performing EEG recording at the same time.

Risk factors associated with prolonged intensive care unit stay following surgery for total anomalous pulmonary venous connection: a retrospective study.

BACKGROUND: Prolonged intensive care unit (ICU) stays consume medical resources and increase medical costs. This study identified risk factors associated with prolonged postoperative intensive care unit (ICU) stay in children with total anomalous pulmonary venous connection (TAPVC). METHODS: The medical records of 85 patients who underwent surgical repair of TAPVC were retrospectively analyzed. The patients were divided into prolonged-stay and standard-stay groups. The prolonged stay group included all patients who exceeded the 75th percentile of the ICU stay duration, and the standard stay group included all remaining patients. The effects of patient variables on ICU stay duration were investigated using univariate and logistic regression analyses. RESULTS: Patient median age was 41 (18-103) days, and median weight was 3.80 (3.30-5.35) kg.Postoperative duration of ICU stay was 11-68 days in the prolonged stay group (n = 23) and 2-10 days in the standard stay group (n = 62). Lower preoperative pulse oximetry saturation (SpO2), higher intraoperative plasma lactate levels, and prolonged postoperative mechanical ventilation were independent risk factors for prolonged ICU stay. Preoperative SpO2 < 88.5%, highest plasma lactate value > 4.15 mmol/L, and postoperative mechanical ventilation duration was longer than 53.5 h, were associated with increased risk of prolonged ICU stay. Young age, low body weight, subcardiac type, need for vasoactive drug support, emergency surgery, long anesthesia time, low SpO2 after anesthesia induction, long cardiopulmonary bypass (CPB) and aortic clamp times, high lactate level, low temperature, large volume of ultrafiltration during CPB, large amounts of chest drainage, large red blood cells (RBCs) and plasma transfusion, and postoperative cardiac dysfunction may be associated with prolonged ICU stay. CONCLUSIONS: Lower preoperative SpO2, higher intraoperative plasma lactate levels, and prolonged postoperative mechanical ventilation were independent risk factors for prolonged ICU stay in children with TAPVC. When SpO2 was lower than 88.5%, the highest plasma lactate value was more than 4.15 mmol/L, and the postoperative mechanical ventilator duration was longer than 53.5 h, the risk of prolonged ICU stay increased. Improved clinical management, including early diagnosis and timely surgical intervention to reduce hypoxia time and protect intraoperative cardiac function, may reduce ICU stay time.

Small RNA sequencing analysis of exosomes derived from umbilical plasma in IUGR lambs.

During the summer, pregnant ewes experience heat stress, leading to the occurrence of IUGR lambs. This study aims to explore the biomarkers of exosomal miRNAs derived from umbilical plasma in both IUGR and normal Hu lambs. We establish a heat-stressed Hu sheep model during mid-late gestation and selected IUGR and normal lambs for analysis. Exosomes from umbilical plasma were separated and small RNA sequencing is used to identify differentially expressed miRNAs. Next, we utilize MiRanda to predict the target genes of the differentially expressed miRNAs. To further understand the biological significance of these miRNAs, we conduct GO and KEGG pathway enrichment analysis for their target genes. The study's findings indicate that oar-miR-411a-5p is significantly downregulated in exosomes derived from umbilical plasma of IUGR lambs, while oar-miR-200c is significantly upregulated in the HS-IUGR group (P < 0.05). Furthermore, GO and KEGG enrichment analysis demonstrate that the target genes are involved in the Wnt, TGF-beta, and Rap1 signaling pathways. miRNAs found in exosomes have the potential to be utilized as biomarkers for both the diagnosis and treatment of IUGR fetuses.

Pharmacokinetics of Oral Minor Cannabinoids in Blood and Brain.

Introduction: Minor cannabinoids are increasingly being consumed in oral formulations (i.e., edibles, tinctures) for medical and nonmedical purposes. This study examined the pharmacokinetics (PKs) of cannabinoids tetrahydrocannabivarin (THCV), cannabichromene (CBC), cannabinol (CBN), and delta-8-tetrahydrocannabinol (D8-THC) after the first and last oral dose during a 14-day administration period. Materials and Methods: Sprague-Dawley rats (N=6 animals/dose, 50% female) were given an assigned dose of one of four cannabinoids (THCV=3.2-100 mg/kg, CBC=3.2-100 mg/kg, CBN=1-100 mg/kg, or D8-THC=0.32-10 mg/kg) or vehicle (medium-chain triglyceride oil) through oral gavage once daily for 14 days. Blood was collected 45 min and 1.5, 3, and 24 h following the first dose (day 1) and the last dose (day 14) of repeated oral cannabinoid treatment for PK analysis. Outcomes of interest included time to maximum concentration (Tmax), maximum concentration (Cmax), and area under the concentration versus time curve (AUClast). Dose-normalized (DN) Cmax and DN AUClast were also calculated. Brain tissue was collected 24 h post-administration of the first (day 1) and the last (day 14) dose of each cannabinoid to determine concentrations in brain. Results: All cannabinoids tested were detectable in plasma after single and 14-day repeated dosing. DN Cmax and DN AUClast were highest for D8-THC, followed by CBC, CBN, and THCV. There was no sex difference observed in cannabinoid kinetics. Accumulation of D8-THC in plasma was observed after 14 days of administration. THCV levels in plasma were lower on day 14 compared to day 1, indicating potential adaptation of metabolic pathways and increased drug elimination. Cannabinoids were detected in brain tissue 24 h post-administration of the first and the last dose of 17-100 mg/kg THCV, 3.2-100 mg/kg CBC, 10-100 mg/kg CBN, and 10 mg/kg D8-THC. Conclusions: THCV, CBC, CBN, and D8-THC produced detectable levels in plasma and translocated to brain tissue after the first dose (day 1) and the last dose (day 14) of repeated oral dosing. Examination of PKs of these minor cannabinoids in blood and brain provides a critical step for informing target dose ranges and dosing schedules in future studies that evaluate the potential effects of these compounds.

SIMOA-based analysis of plasma NFL levels in MCI and AD patients: a systematic review and meta-analysis.

BACKGROUND: The single-molecule array assay (SIMOA)-based detection of neurofilament light (NFL) chain could be useful in diagnosing mild cognitive impairment (MCI) and Alzheimer's disease (AD). This meta-analysis aimed to evaluate the circulating concentration of NFL in AD and MCI patients compared with healthy controls using the SIMOA technique. METHODS: To this end, Google Scholar, PubMed, Scopus, Web of Science, and the reference lists of relevant articles were systematically searched for studies reporting serum NFL chain levels in healthy controls, MCI, and AD patients. Appropriate statistical methods were employed to achieve the study purpose. RESULTS: Fifteen eligible studies including 3086 patients were pooled out of a total of 347 publications. Fixed effect model analysis showed that NFL chain level was significantly higher in the serum of patients with MCI (0.361 SMD, 95% CI, 0.286-0.435, p = 0.000, I2 = 49.179) and AD (0.808 SMD, 95% CI, 0.727-0.888, p = 0.000, I2 = 39.433) compared with healthy individuals. The analysis also showed that the NFL chain levels in plasma were significantly different between patients with MCI and AD (0.436 SMD, 95% CI, 0.359-0.513, p = 0.000, I2 = 37.44). The overall heterogeneity of the studies was modest. CONCLUSIONS: This study highlights the potential of serum NFL chain detected using SIMOA in differentiating MCI, AD, and healthy controls.

Head-to-Head Comparison of Two Plasma Phospho-tau Assays in Predicting Conversion of Mild Cognitive Impairment to Dementia.

BACKGROUND: Among blood biomarkers, phospho-tau181 (pTau181) is one of the most efficient in detecting Alzheimer disease across its continuum. However, transition from research to routine clinical use will require confirmation of clinical performance in prospective cohorts and evaluation of cofounding factors. METHODS: Here we tested the Lumipulse assay for plasma pTau181 in mild cognitive impairment (MCI) participants from the Baltazar prospective cohort. We compared the performance of this assay to the corresponding Simoa assay for the prediction of conversion to dementia. We also evaluated the association with various routine blood parameters indicative of comorbidities. RESULTS: Lumipulse and Simoa gave similar results overall, with hazard ratios for conversion to dementia of 3.48 (95% CI, 2.23-5.45) and 3.70 (95%CI, 2.39-5.87), respectively. However, the 2 tests differ somewhat in terms of the patients identified, suggesting that their use may be complementary. When combined with age, sex, and apolipoprotein E (APOE)ε4 status, areas under the curves for conversion detection were 0.736 (95% CI, 0.682-0.791) for Lumipulse and 0.733 (95% CI, 0.679-0.788) for Simoa. Plasma pTau181 was independently associated with renal dysfunction (assessed by creatinine and glomerular filtration) for both assays. Cardiovascular factors (adiponectin and cholesterol), nutritional, and inflammatory markers (total protein content, C-reactive protein) also impacted plasma pTau181 concentration, although more so with the Simoa than with the Lumipulse assay. CONCLUSIONS: Plasma pTau181 measured using the fully automated Lumipulse assay performs as well as the Simoa assay for detecting conversion to dementia of MCI patients within 3 years and Lumipulse is less affected by comorbidities. This study suggests a pathway to routine noninvasive in vitro diagnosis-approved testing to contribute to the management of Alzheimer disease. CLINICALTRIALS.GOV REGISTRATION NUMBER: NCT01315639.

Macrophage-Derived Chemokine MDC/CCL22: An Ambiguous Finding in COVID-19.

Macrophage-derived chemokine (MDC/CCL22) is a chemokine of the C-C subfamily. It is involved in T-cellular maturation and migration. Our previous research shows that plasma CCL22/MDC tends to show a statistically significant depletion of concentrations in acute patients and convalescents when compared to healthy donors. In the current work, we investigate existing views on MDC/CCL22 dynamics in association with various pathologies, including respiratory diseases and, specifically, COVID-19. Additionally, we present our explanations for the observed decrease in MDC/CCL22 concentrations in COVID-19. The first hypothesis we provide implies that viral products bind to MDC/CCL22 and block its activity. Another explanation for this phenomenon is based on dendritic cells population and the inhibition of their function.

Performance Evaluation of the VIDAS® D-Dimer Exclusion™ II Assay and the Beckman Coulter D-Dimer Assay.

BACKGROUND: The breakdown of fibrinogen and fibrin during the process of fibrinolysis creates D-dimer. A D-dimer analysis is crucial for the diagnosis of pulmonary embolism, deep vein thrombosis, and disseminated intravascular coagulation. The aim of this study is to evaluate the validity of two different D-dimer assays. METHODS: To analyze the D-dimer, venous blood taken in a vacuette containing 3.2% sodium citrate was used. Peripheral whole blood specimens were collected from 89 subjects, and plasma was separated from these specimens. VIDAS® D-dimer Exclusion™ II and the Beckman Coulter D-dimer assays were used for the quantitative detection of D-dimer levels. The analytical performance of the two different D-dimer assays was compared. RESULTS: The plasma values of D-dimer ranged from 89.2 to 7,452.9 ng/mL (FEU) when tested on the VIDAS® platform and from 20 to 7,770 ng/mL (FEU) when tested on the Beckman Coulter platform. Our results showed that the agreement between the two methods was acceptable and Pearson's correlation coefficient (r) was 0.93 (p < 0.001). CONCLUSIONS: A high correlation exists between quantitative D-dimer measurements conducted with the VIDAS® D-dimer Exclusion™ II and the Beckman Coulter D-dimer assays.

Lipid alternations in the plasma of COVID-19 patients with various clinical presentations.

Background: COVID-19 is a highly infectious respiratory disease that can manifest in various clinical presentations. Although many studies have reported the lipidomic signature of COVID-19, the molecular changes in asymptomatic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected individuals remain elusive. Methods: This study combined a comprehensive lipidomic analysis of 220 plasma samples from 166 subjects: 62 healthy controls, 16 asymptomatic infections, and 88 COVID-19 patients. We quantified 732 lipids separately in this cohort. We performed a difference analysis, validated with machine learning models, and also performed GO and KEGG pathway enrichment analysis using differential lipids from different control groups. Results: We found 175 differentially expressed lipids associated with SASR-CoV-2 infection, disease severity, and viral persistence in patients with COVID-19. PC (O-20:1/20:1), PC (O-20:1/20:0), and PC (O-18:0/18:1) better distinguished asymptomatic infected individuals from normal individuals. Furthermore, some patients tested positive for SARS-CoV-2 nucleic acid by RT-PCR but did not become negative for a longer period of time (≥60 days, designated here as long-term nucleic acid test positive, LTNP), whereas other patients became negative for viral nucleic acid in a shorter period of time (≤45 days, designated as short-term nucleic acid test positive, STNP). We have found that TG (14:1/14:1/18:2) and FFA (4:0) were differentially expressed in LTNP and STNP. Conclusion: In summary, the integration of lipid information can help us discover novel biomarkers to identify asymptomatic individuals and further deepen our understanding of the molecular pathogenesis of COVID-19.

Ex Vivo Placental Explant Flow Culture - Mimicking the Dynamic Conditions In Utero.

The existing ex vivo placental explant culture models are primarily grounded in static culture systems using well plates. However, these models inadequately reflect the dynamic in utero setting, where the placenta encounters constant slight shear stress due to plasma or blood flow. To address this limitation, a flow culture system has been devised to bring ex vivo placental explant cultivation closer to the in utero flow conditions experienced within the maternal body. Within this approach, placental explants are cultivated in a sequence of five interconnected flow chambers. This setting maintains physiological oxygen concentrations and a consistent flow rate. The collected data reveals that under flow conditions, the preservation of tissue morphology exhibits notable enhancement compared to conventional static methods. This innovative technique introduces a straightforward means of ex vivo placental explant culture, offering a more faithful representation of the dynamic in vivo environment. Moreover, this study introduces new possibilities for investigating the functional dynamics of the feto-maternal interface. By embracing feasible dynamic methodologies, a deeper comprehension of placental biology is facilitated, underscoring its relevance for maternal-fetal health.

A prospective evaluation of the diagnostic potential of EBV-DNA in plasma and whole blood.

BACKGROUND: Quantitative polymerase chain reaction (qPCR) for Epstein-Barr virus (EBV)-DNA is an important diagnostic tool for EBV-associated disease, but interpretation of its clinical significance is challenging. OBJECTIVES: We assessed the diagnostic and clinical performance of WHO-standardised qPCR for EBV-DNA (WHO EBV-qPCR) in plasma and whole blood (WB) for proven EBV disease in a prospectively accrued patient cohort. STUDY DESIGN: Central Denmark Region patients, tested with WHO EBV-qPCR from November 2017 to March 2019, were screened for EBV disease. Incidence (IR) was estimated by Poisson regression. Sensitivity, specificity, positive and negative predictive values (PPV, NPV) were calculated for EBV-qPCR in plasma and WB. Risk of diagnostic latency was compared between patients with EBV-positive and EBV-negative lymphomas. RESULTS: EBV disease was diagnosed in 95 of 1484 participants (IR: 16.3 per 1000 patientyears 95%CI; 13.3-19.9). Sensitivity and specificity of WHO EBV-qPCR in plasma was 82.4% (95% CI; 74.2-90.7%) and 87.8% (95% CI; 85.6-90%), yielding a PPV of 32.2% (95% CI; 24.9-39.5%) and NPV of 98.6% (95% CI; 97.7-99.5%) for proven EBV disease. Sensitivity and NPV were comparable in WB, while specificity and PPV decreased to 66.9% (95% CI; 60.6-73.1%) and 18.1% (95% CI; 7.5-28.7%). Risk of diagnostic latency was 2.3-fold (95% CI 1.4-4.1) higher for patients with EBV-positive compared with EBV-negative lymphomas. CONCLUSIONS: WHO EBV-qPCR in plasma and WB have a low PPV but a high NPV for proven EBV disease. Implementation of WHO EBV-qPCR could improve interpretation and facilitate EBV-positive lymphoma diagnosis.

Hydration Status of Geriatric Patients Is Associated with Changes in Plasma Proteome, Especially in Proteins Involved in Coagulation.

Due to multifactorial reasons, such as decreased thirst and decreased total body water, elderly patients are vulnerable to dehydration. The study aims to investigate whether moderate dehydration or hyperhydration affects the blood proteome. Blood samples, medication, and bioelectrical impedance analysis (BIA) details were collected from 131 geriatric patients (77 women and 54 men aged 81.1 ± 7.2 years). Based on an evaluation by Bioelectrical Impedance Vector Analyses (BIVAs) of this cohort, for each hydration status (dehydrated, hyperhydrated, and control), five appropriate blood plasma samples for both males and females were analyzed by liquid chromatography-mass spectrometry (LC-MS). Overall, 262 proteins for female patients and 293 proteins for male patients could be quantified. A total of 38 proteins had significantly different abundance, showing that hydration status does indeed affect the plasma proteome. Protein enrichment analysis of the affected proteins revealed "Wound Healing" and "Keratinization" as the two main biological processes being dysregulated. Proteins involved in clot formation are especially affected by hydration status.

Pre-analytical variables influence zinc measurement in blood samples.

Zinc deficiency continues to be a major concern for global public health. The zinc status of a target population is typically estimated by measuring circulating zinc levels, but the sampling procedures are not standardized and thus may result in analytical discrepancies. To examine this, we designed a study that controlled most of the technical parameters in order to focus on five pre-analytical variables reported to influence the measurement of zinc in blood samples, including (1) blood draw site (capillary or venous), (2) blood sample matrix (plasma or serum), (3) blood collection tube manufacturer (Becton, Dickinson and Company or Sarstedt AG & Co), (4) blood processing time (0, 4, or 24 hours), and (5) blood holding temperatures (4°C, 20°C, or 37°C). A diverse cohort of 60 healthy adults were recruited to provide sequential capillary and venous blood samples, which were carefully processed under a single chain of custody and measured for zinc content using inductively coupled plasma optical emission spectrometry. When comparing blood draw sites, the mean zinc content of capillary samples was 0.054 mg/L (8%; p<0.0001) higher than venous blood from the same donors. When comparing blood sample matrices, the mean zinc content of serum samples was 0.029 mg/L (5%; p<0.0001) higher than plasma samples from the same donors. When comparing blood collection tube manufacturer, the mean zinc content from venous blood samples did not differ between venders, but the mean zinc content from BD capillary plasma was 0.036 mg/L (6%; p<0.0001) higher than Sarstedt capillary plasma from the same donors. When comparing processing times, the mean zinc content of plasma and serum samples was 5-12% higher (p<0.0001) in samples processed 4-24 hour after collection. When comparing holding temperatures, the mean zinc content of plasma and serum samples was 0.5-7% higher (p = 0.0007 or p = 0.0061, respectively) in samples temporarily held at 20°C or 37°C after collection. Thus even with the same donors and blood draws, significant differences in zinc content were observed with different draw sites, tube types, and processing procedures, demonstrating that key pre-analytic variables can have an impact on zinc measurement, and subsequent classification of zinc status. Minimizing these pre-analytical variables is important for generating best practice guidelines for assessment of zinc status.

The Effect of Mycotoxins and Their Mixtures on Bovine Spermatozoa Characteristics.

There is growing concern about the effects of mycotoxins on mammalian reproduction. Although the effects of single mycotoxins have been well documented, the impact of their mixtures on spermatozoon quality is less known. Here, frozen-thawed semen (n = 6 bulls) was in-vitro-cultured (2 h) without (control) or with (i) a single mycotoxin [zearalenone (ZEN), ochratoxin A (OTA), toxin 2 (T2), and diacetoxyscirpenol (DAS)] in a dose-response manner; (ii) binary mixtures (OTA + T2, OTA + ZEN, OTA + DAS, ZEN + T2, DAS + T2 and ZEN + DAS); or (iii) ternary mixtures (OTA + DAS + T2, OTA + ZEN + T2, and ZEN + DAS + T2). Then, the spermatozoa quality was characterized according to its plasma- and acrosome-membrane integrity, mitochondrial membrane potential, and oxidation status by a flow cytometer. Exposure to single mycotoxins or binary mixtures did not affect the spermatozoa characteristics. However, exposure to the ternary mixtures, OTA + DAS + T2 and OTA + ZEN + T2, reduced (p < 0.05) the mitochondrial membrane potential relative to the control. In addition, OTA + ZEN + T2 increased (p < 0.05) the proportion of spermatozoa with reactive oxygen species relative to the control. The most suggested interaction effect between the mycotoxins was found to be an additive one. A synergistic interaction, mainly regarding the oxidation status of the spermatozoa, was also found between the mycotoxins. The current study sheds light on the potential risk of exposing spermatozoa to a mycotoxin mixture.

Impact of Nonthermal Plasma on Lipid Oxidation from the Perspective of Plasma Treatment Parameters and Plasma Species: Identification of Key Reactive Species.

Nonthermal plasma is a mild processing technology for food preservation. Its impact on lipid oxidation was investigated in this study. Stripped methylesters were considered as a basic lipid model system and were treated by a multihollow surface dielectric barrier discharge. In dry air plasma, O3, ·NO2, ·NO3, and 1O2 were identified as the main reactive species reaching the sample surface. Treatment time was the most prominent parameter affecting lipid oxidation, followed by the (specific) power input and the plasma-sample distance. In humid air plasma, less O3 was detected, but ONOOH and O2NOOH were generated and presumed to play a role in lipid oxidation. Ozone mainly resulted in the formation of carbonyl substances via the trioxolane pathway, while reactive nitrogen species (i.e., ·NO2, ·NO3, ONOOH, and O2NOOH) led to the formation of hydroperoxides. The impact of short-living radicals (e.g., ·O, ·N, ·OH, and ·OOH) was restricted in general, since they dissipated too fast to reach the sample.·NO, HNO3, H2O2, and UV radiation did not induce lipid oxidation. All the reactive species identified in this study were associated with the presence of O2 in the input gas.

Bifunctional Ti4+-modified paper for selective extraction or removal of phospholipids and paper spray mass spectrometry for bioanalysis in urine and plasma.

BACKGROUND: Phospholipids (PLs) are major constituents of cell membranes, play important roles in cell proliferation and death, as well as in signal transduction, and therefore are relevant biomarkers for different pathologies. On the other hand, when the analysis of small compounds, such as therapeutics in blood is desired, then phospholipids are part of the matrix and cause serious interference during analysis. Currently, both the analysis and removal of PLs from biological samples are limited by extensive sample preparation and instrumental separation. RESULTS: A fast and simple quantitative Ti4+-modified paper spray tandem mass spectrometric (TiPS-MS/MS) method was established in urine, where the enrichment of phospholipids was achieved, as well as reduction of matrix effects (primarily caused by high salt content) that ultimately led to improved sensitivity and selectivity. The method could achieve a physiologically relevant limit of detection (0.01-0.03 µg mL-1). Also, the usefulness of the Ti4+-modified paper was investigated in the opposite mode, namely for the selective removal of phospholipids from matrices such as plasma. Clonidine is used as model compound, as the detection of this compound is known to suffer from ion suppression by phospholipids. Compared with blank paper spray tandem mass spectrometry, the limit of detection could be improved from 0.3 µg mL-1 to 0.03 µg mL-1 by employing a Ti4+-modified paper on top of the spray tip to capture phospholipids from the sample. SIGNIFICANCE AND NOVELTY: A novel Ti4+-modified paper was developed to allow for rapid solid-phase extraction of phospholipids from urine or selective removal from plasma, followed by direct paper spray mass spectrometric detection as a fast and convenient sample preparation and analysis combination. The paper properties are based on the Ti4+ metal ion, which can selectively bind phosphate-containing compounds under acidic conditions, and its applicability was demonstrated in relevant biological matrices.

Bioinspired superhydrophobic SERS substrates for machine learning assisted miRNA detection in complex biomatrix below femtomolar limit.

Surface-enhanced Raman spectroscopy (SERS) is an analytical method with high potential in the field of medicine. The design of SERS substrates, based on specific morphology and/or chemical modification, allow the recognition of the presence of specific analytes with precision close to a single-molecule detection limit. However, the SERS analysis of real samples is significantly complicated by the presence of a large number of "minor" molecules that can shield the signal from the target analyte and make it impossible to determine it in practice. In this work, an advanced SERS approach was used for the detection of cancer-related miRNA-21 in blood plasma, used as a molecular model background. The approach was based on the combination of the biomimetic plasmon-active SERS substrate, its tuned surface chemistry and advanced SERS data analysis, making use of artificial machine learning. In the first step, biomimetic SERS substrates were created using a butterfly wing as a starting template. The substrates were covered by thin Au layer and covalently grafted with hydrophobic chemical moieties to introduce superhydrophobic and water-adhesive properties. The self-concentration of the analyte on the substrates was achieved by minimizing the contact area between the analyte drop and the substrate, which is facilitated by surface superhydrophobicity and additionally enhanced by drop evaporation on the flipped over substrate. Due to the presence of cancer miRNA and blood plasma background, the measured SERS spectra represent a complex of interfering peaks. Thus, their interpretation was carried out using a specially trained machine learning model. As a result, reliable and repeatable quantitative detection of miRNAs below the femtomolar level (up to 10-16 M) on the background of human blood plasma becomes possible.

Concordance of ctDNA and tissue mutations in NSCLC: A meta-analysis.

Systematic evaluation of the consistency between circulating tumor DNA (ctDNA) in plasma and tumor tissue samples in mutations in non-small cell lung cancer (NSCLC) patients. To collect publicly published literature from numerous important international medical databases through computer retrieval. To compare the differences in literature data related to gene mutations between plasma ctDNA and tumor tissue samples, a meta-analysis was performed using Stata 12.0 software while taking into account the inclusion and exclusion criteria. This article includes a total of 15 research data and collected data reports from 15 groups of NSCLC patients. The results are displayed using tissue samples as the gold standard. The Pearson correlation coefficients of sensitivity and specificity were used to calculate rho=0.044, and P=0.893. The Q-test found poor homogeneity and high heterogeneity in sensitivity and specificity among research data from various literature studies (I2>50%, P<0.1). The area under the SROC curve is 0.97 (95% CI: 0.96~0.99). The small sample subgroup has high heterogeneity, and the combined diagnosis effect size is 26[6~111], lower than the large sample subgroup 185320 [0~2.7×1012]. When taking 200 as the cut-off point, the combined effect size of the small sample subgroup is 46 [12~183], still lower than that of the large sample subgroup 429 [52~3574]. From this, it can be concluded that the consistency of small-sample studies is higher than the quality of large-sample studies, and the heterogeneity is relatively low. From the perspective of mutation types, the heterogeneity of literature with EGFR gene mutations alone is higher than that of literature with non-EGFR mutations alone, and the consistency is lower. The consistency of using plasma ctDNA to detect mutations in NSCLC patients with tumor tissue samples is influenced by the type of mutation gene and sample size measured by the patient, and there is a significant bias in related studies.