Valorization of pawpaw (Carica papaya L.) leaves as a source of polyphenols by ionic liquid-based microwave-assisted extraction: Comparison with other extraction methods and bioactivity evaluation.

This study aimed to valorize pawpaw (Carica papaya L.) leaves as a rich source of polyphenols through the application of ionic liquid-based microwave-assisted extraction (ILMAE). Initially, the ILMAE process was optimized using response surface methodology (RSM), resulting in a total polyphenols yield of 27.84 ± 0.33 mg GAE/g DW under the optimal conditions: [BMIM]Br concentration of 0.57 mol/L, extraction time of 14 min, microwave power of 460 W, extraction temperature of 77 °C, solvent-to-material ratio of 30 mL/g, and three extraction cycles. Compared to conventional methods such as maceration extraction (ME), heat reflux extraction (HRE), and microwave-assisted extraction (MAE), the ILMAE method exhibited a significantly higher PLTP yield. Furthermore, the PLTP extracts demonstrated strong antioxidant activity against DPPH• and ABTS+• radicals, as well as a significant inhibitory effect on α-glucosidase activity. This work demonstrates that ILMAE is a green and efficient strategy for the valorization of pawpaw leaves.

Dynamic Nomogram for Subsyndromal Delirium in Adult Intensive Care Unit: A Prospective Cohort Study.

Purpose: To develop a dynamic nomogram of subsyndromal delirium (SSD) in intensive care unit (ICU) patients and internally validate its efficacy in predicting SSD. Patients and Methods: Patients who met the inclusion and exclusion criteria in the ICU of a tertiary hospital in Zhejiang from September 2021 to June 2022 were selected as the research objects. The patient data were randomly divided into the training set and validation set according to the ratio of 7:3. The least absolute shrinkage and selection operator (LASSO) and multivariate logistic regression were used to screen the predictors of SSD, and R software was used to construct a dynamic nomogram. Receiver operating characteristic (ROC) curve, calibration band and decision curve were used to evaluate the discrimination, calibration and clinical effectiveness of the model. Results: A total of 1000 eligible patients were included, including 700 in the training set and 300 in the validation set. Age, drinking history, C reactive protein level, APACHE II, indwelling urinary catheter, mechanical ventilation, cerebrovascular disease, respiratory failure, constraint, dexmedetomidine, and propofol were predictors of SSD in ICU patients. The ROC curve values of the training set was 0.902 (95% confidence interval: 0.879-0.925), the best cutoff value was 0.264, the specificity was 78.4%, and the sensitivity was 88.0%. The ROC curve values of the validation set was 0.888 (95% confidence interval: 0.850-0.930), the best cutoff value was 0.543, the specificity was 94.9%, and the sensitivity was 70.9%. The calibration band showed good calibration in the training and validation set. Decision curve analysis showed that the net benefit in the model was significantly high. Conclusion: The dynamic nomogram has good predictive performance, so it is a precise and effective tool for medical staff to predict and manage SSD in the early stage.

The application of Aptamer in biomarker discovery.

Biomarkers are detectable molecules that can reflect specific physiological states of cells, organs, and organisms and therefore be regarded as indicators for specific diseases. And the discovery of biomarkers plays an essential role in cancer management from the initial diagnosis to the final treatment regime. Practically, reliable clinical biomarkers are still limited, restricted by the suboptimal methods in biomarker discovery. Nucleic acid aptamers nowadays could be used as a powerful tool in the discovery of protein biomarkers. Nucleic acid aptamers are single-strand oligonucleotides that can specifically bind to various targets with high affinity. As artificial ssDNA or RNA, aptamers possess unique advantages compared to conventional antibodies. They can be flexible in design, low immunogenicity, relative chemical/thermos stability, as well as modifying convenience. Several SELEX (Systematic Evolution of Ligands by Exponential Enrichment) based methods have been generated recently to construct aptamers for discovering new biomarkers in different cell locations. Secretome SELEX-based aptamers selection can facilitate the identification of secreted protein biomarkers. The aptamers developed by cell-SELEX can be used to unveil those biomarkers presented on the cell surface. The aptamers from tissue-SELEX could target intracellular biomarkers. And as a multiplexed protein biomarker detection technology, aptamer-based SOMAScan can analyze thousands of proteins in a single run. In this review, we will introduce the principle and workflow of variations of SELEX-based methods, including secretome SELEX, ADAPT, Cell-SELEX and tissue SELEX. Another powerful proteome analyzing tool, SOMAScan, will also be covered. In the second half of this review, how these methods accelerate biomarker discovery in various diseases, including cardiovascular diseases, cancer and neurodegenerative diseases, will be discussed.

Family members' perceptions of surrogate decision-making in the intensive care unit: A systematic review.

BACKGROUND: A better understanding of the perceptions of family members in making surrogate decisions for loved ones during intensive care is needed to inform the development of targeted supportive interventions. OBJECTIVE: To examine and synthesize qualitative data on family members' perceptions of surrogate decision-making in the intensive care unit. DESIGN: We conducted a systematic review and qualitative data synthesis. Eligible studies contained family members' quotes about surrogate decision-making experiences and perceptions in adult intensive care units, published in English or Chinese, in a peer-reviewed journal up to February 2022. Data sources included Embase, PubMed, ISI Web of Science, PsychINFO, CINAHL, Biomedical Literature Service System, China National Knowledge Infrastructure (CNKI), Wanfang Data, and VIP Journal. METHODS: The searches yielded 5974 identified articles, of which 23 studies were included. At least two different reviewers independently assessed the study quality and extracted data into a Microsoft Excel spreadsheet. A thematic synthesis was performed by classifying all text units into one of the broad themes and subsequently analyzed to inductively develop the first-, second-, and third-order themes. Six family members with experience in intensive care unit surrogate decision-making contributed to the analysis. RESULTS: The qualitative data synthesis resulted in five major themes. The following key new insights into family members' perceptions of surrogate decision-making in the intensive care unit were obtained: in individual systems, family members suffered from emotional distress and psychological stress; different cognitive styles emerged; some family members reshaped a new order of life in the disruption; in family systems, the family as a whole was closely connected with each other; and in medical systems families perceived asymmetry in relationships with clinicians, many factors influencing trust, the necessity for role-specific mediators and issues with operations and environments not being sufficiently humanized. CONCLUSION: This qualitative synthesis showed that individuals' emotions and cognition underwent complex processes during surrogate decision-making. The family as a whole, with disparate functional states, also faced different processes and outcomes under the crisis situation. At a broader level, the decision-making process reflected society's perceptions of the medical system. Future studies should use these insights to further explore and optimize the many aspects of surrogate decision support measures for families of critically ill patients and include the measurement of outcomes after interventions at multiple layers of the individual, family, and medical systems. REGISTRATION NUMBER: The protocol was prospectively published on International Prospective Register of Systematic Reviews (PROSPERO)-CRD42022316687. TWEETABLE ABSTRACT: Families of critically ill patients undergo a complex interactional process within the individual, family, and medical systems during surrogate decision-making.

Aptamer nucleotide analog drug conjugates in the targeting therapy of cancers.

Aptamers are short single-strand oligonucleotides that can form secondary and tertiary structures, fitting targets with high affinity and specificity. They are so-called "chemical antibodies" and can target specific biomarkers in both diagnostic and therapeutic applications. Systematic evolution of ligands by exponential enrichment (SELEX) is usually used for the enrichment and selection of aptamers, and the targets could be metal ions, small molecules, nucleotides, proteins, cells, or even tissues or organs. Due to the high specificity and distinctive binding affinity of aptamers, aptamer-drug conjugates (ApDCs) have demonstrated their potential role in drug delivery for cancer-targeting therapies. Compared with antibodies which are produced by a cell-based bioreactor, aptamers are chemically synthesized molecules that can be easily conjugated to drugs and modified; however, the conventional ApDCs conjugate the aptamer with an active drug using a linker which may add more concerns to the stability of the ApDC, the drug-releasing efficiency, and the drug-loading capacity. The function of aptamer in conventional ApDC is just as a targeting moiety which could not fully perform the advantages of aptamers. To address these drawbacks, scientists have started using active nucleotide analogs as the cargoes of ApDCs, such as clofarabine, ara-guanosine, gemcitabine, and floxuridine, to replace all or part of the natural nucleotides in aptamer sequences. In turn, these new types of ApDCs, aptamer nucleotide analog drug conjugates, show the strength for targeting efficacy but avoid the complex drug linker designation and improve the synthetic efficiency. More importantly, these classic nucleotide analog drugs have been used for many years, and aptamer nucleotide analog drug conjugates would not increase any unknown druggability risk but improve the target tumor accumulation. In this review, we mainly summarized aptamer-conjugated nucleotide analog drugs in cancer-targeting therapies.

Accurate quantification of SARS-CoV-2 RNA by isotope dilution mass spectrometry and providing a correction of reverse transcription efficiency in droplet digital PCR.

The novel coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected more than 505 million confirmed cases, including over 6 million deaths. Reference materials (RMs) of SARS-CoV-2 RNA played a crucial role in performance evaluation and quality control of testing laboratories. As the potential primary characterization method of RMs, reverse transcription digital PCR (RT-dPCR) measures the copy number of RNA, but the accuracy of reverse transcription (RT) efficiency has yet to be confirmed. This study established a method of enzymatic digestion followed by isotope dilution mass spectrometry (IDMS), which does not require an RT reaction, to quantify in vitro-transcribed SARS-CoV-2 RNA. RNA was digested to nucleotide monophosphate (NMP) within 15 min and analyzed by IDMS within 5 min. The consistency among the results of four different NMPs demonstrated the reliability of the proposed method. Compared to IDMS, the quantitative result of RT-dPCR turned out to be about 10% lower, possibly attributed to the incompleteness of the reverse transcription process. Therefore, the proposed approach could be valuable and reliable for quantifying RNA molecules and evaluating the RT efficiency of RT-based methods.

Urinary analysis reveals high Alternaria mycotoxins exposure in the general population from Beijing, China.

Alternaria mycotoxins are of concern due to its adverse health effect, they affect various cereal crops and grain-based food along with modified forms that contribute to overall exposure. This study aimed to determine the frequency and level of exposure to Alternaria mycotoxins (tenuazonic acid, TeA; alternariol, AOH; alternariol monomethyl ether, AME; tentoxin, TEN; and altenuene, ALT) in human urine from Beijing adults. A total of 2212 urine samples were collected and analyzed for five mycotoxins using LC-ESI-MS/MS. More than 98% of the samples had at least one Alternaria mycotoxin detected. Among the mycotoxins, AME had the highest detection rate (96.0%), followed by TeA (70.5%). The calculated average daily intake values of AME (12.5 ng/kg b.w.) was 5 times the TTC value (2.5 ng/kg b.w.) set by the EFSA, indicating the potential health risks associated with mycotoxins. Immediate attention and subsequent actions should be taken to identify the sources of mycotoxins and the corresponding exposure pathways to humans in the investigated regions.

Development of a simple and rapid LC-MS/MS method for the simultaneous quantification of five Alternaria mycotoxins in human urine.

Alternaria mycotoxins, such as tenuazonic acid (TeA), altenuene (ALT), alternariol (AOH), tentoxin (TEN) and alternariol monomethyl ether (AME) are frequently found in foods and may pose a potential risk to human health. Human biomonitoring can help measure our exposure to these mycotoxins, and help us determine if the exposure is changing over time. In this study, a simple liquid-liquid extraction sample preparation procedure followed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was developed for the simultaneous analysis of five Alternaria mycotoxins in human urine. High recoveries (92.7-103.2%) were obtained for all the tested mycotoxins with relative standard deviations (RSDs, %) of less than 6.4%. The limits of quantification (LOQs) for the analytes in urine ranged from 0.001 to 0.05 ng/mL. The method was successfully applied to investigate the levels of five Alternaria mycotoxins from 135 volunteers. In all of the samples, at least one Alternaria mycotoxin was detected. TeA, AME and AOH were the predominant Alternaria mycotoxins, and the detection rates were 85.9%, 96.3% and 51.9%, respectively.

Determination of Alternaria Mycotoxins in Fresh Sweet Cherries and Cherry-Based Products: Method Validation and Occurrence.

Sweet cherry is susceptible to disease caused by the Alternaria species and produces various Alternaria mycotoxins. Analytical methodologies based on solid-phase extraction (SPE) and LC-MS/MS to simultaneously determine five main Alternaria mycotoxins (tenuazonic acid, 1; alternariol, 2; alternariol methyl ether, 3; altenuene, 4; and tentoxin, 5) in fresh sweet cherries and cherry products were developed and validated. The limits of quantitation (LOQ) of the analytes ranged from 0.002-0.066 µg/kg. The method was successfully applied to 83 fresh cherry and cherry-related product samples. 1 and 5 were the predominant toxins with detection frequencies >50%, followed by 3 (42%), 2 (35%), and 4 (31%). Daily intakes of Alternaria mycotoxins via fresh sweet cherries were assessed preliminarily using the measured concentrations, and consumption data were obtained from a web-based dietary questionnaire ( n = 476). The maximum exposure of 1 and 3 were 4.6 and 16.7 times the threshold of the toxicological concern (TTC) value, respectively.

Toxicological evaluation of advanced glycation end product Nε-(carboxymethyl)lysine: Acute and subacute oral toxicity studies.

Nε-(carboxymethyl)lysine (CML) as a novel potential noxious compound in various food products has aroused extensive concern in recent years. This study aimed to investigate the oral acute and subacute toxicity of CML in mice as per OECD 420 and 407 guidelines. Acute administration of 2000 and 5000 mg/kg CML did not induce any mortality within 14 days, nevertheless some toxicological symptoms and histopathological changes were observed. The estimated LD50 of CML was >5000 mg/kg. In subacute toxicity test, CML was dosed at 200, 500 and 1000 mg/kg in both genders for 28 days. The body weights reduced which was accompanied with the decrease of food consumptions. Hematology parameters viz. RBC, HGB and MCH showed minor alteration but these were still within normal range. Biochemical analysis of hepatic and renal function markers showed significant elevating in AST, ALT, Cr and BUN etc. Histopathological alterations were observed in lung, liver, kidney and spleen. Subacute toxicity of CML involved oxidative stress caused by reducing antioxidant enzyme (SOD and GSH-Px) activities, and significantly increasing lipid peroxide (MDA) level. In conclusion, CML was unlikely to present an acute hazard, but repeated administration could produce deleterious effects on mice especially inducing liver and kidney damage through oxidative stress.