Synthesis and evaluation of antisense oligonucleotides prodrug with G-quadruplex assembly and lysosome escape capabilities for oncotherapy.

The applications of antisense oligonucleotides (ASOs) in rare or common diseases treatment have garnered great attention in recent years. Nevertheless, challenges associated with stability and bioavailability still persist, hampering the efficiency of ASOs. This work presents an ASO prodrug with parallel G-quadruplex assembly and lysosome escape capabilities for oncotherapy. Our findings revealed that the end-assembled quadruplex structure effectively shielded the ASO from enzymatic degradation. Meanwhile, the conjugation of maleimide within the quadruplex enhanced cellular uptake, potentially offering an alternative cell entry mechanism that circumvents lysosome involvement. Notably, an optimized molecule, Mal2-G4-ASO, exhibited remarkable therapeutic effects both in vitro and in vivo. This work presents a promising avenue for enhancing the activity of nucleic acid drugs in oncotherapy and potentially other disease contexts.

Pretreatment and analysis techniques development of TKIs in biological samples for pharmacokinetic studies and therapeutic drug monitoring.

Tyrosine kinase inhibitors (TKIs) have emerged as the first-line small molecule drugs in many cancer therapies, exerting their effects by impeding aberrant cell growth and proliferation through the modulation of tyrosine kinase-mediated signaling pathways. However, there exists a substantial inter-individual variability in the concentrations of certain TKIs and their metabolites, which may render patients with compromised immune function susceptible to diverse infections despite receiving theoretically efficacious anticancer treatments, alongside other potential side effects or adverse reactions. Therefore, an urgent need exists for an up-to-date review concerning the biological matrices relevant to bioanalysis and the sampling methods, clinical pharmacokinetics, and therapeutic drug monitoring of different TKIs. This paper provides a comprehensive overview of the advancements in pretreatment methods, such as protein precipitation (PPT), liquid-liquid extraction (LLE), solid-phase extraction (SPE), micro-SPE (µ-SPE), magnetic SPE (MSPE), and vortex-assisted dispersive SPE (VA-DSPE) achieved since 2017. It also highlights the latest analysis techniques such as newly developed high performance liquid chromatography (HPLC) and high-resolution mass spectrometry (HRMS) methods, capillary electrophoresis (CE), gas chromatography (GC), supercritical fluid chromatography (SFC) procedures, surface plasmon resonance (SPR) assays as well as novel nanoprobes-based biosensing techniques. In addition, a comparison is made between the advantages and disadvantages of different approaches while presenting critical challenges and prospects in pharmacokinetic studies and therapeutic drug monitoring.

Boron-Containing Mesoporous Silica Nanoparticles with Effective Delivery and Targeting of Liver Cancer Cells for Boron Neutron Capture Therapy.

Nanocarriers have been researched comprehensively for the development of novel boron-containing agents in boron neutron capture therapy (BNCT). We designed and synthesized a multifunctional mesoporous silica nanoparticle (MSN)-based boron-containing agent. The latter was coated with a lipid bilayer (LB) and decorated with SP94 peptide (SFSIIHTPILPL) on the surface as SP94-LB@BA-MSN. The latter incorporated boric acid (BA) into hydrophobic mesopores, coated with an LB, and modified with SP94 peptide on the LB. SP94-LB@BA-MSN enhanced nano interface tumor-targeting ability but also prevented the premature release of drugs, which is crucial for BNCT because adequate boron content in tumor sites is required. SP94-LB@BA-MSN showed excellent efficacy in the BNCT treatment of HepG-2 cells. In animal studies with tumor-bearing mice, SP94-LB@BA-MSN exhibited a satisfactory accumulation at the tumor site. The boron content reached 40.18 ± 5.41 ppm in the tumor site 4 h after injection, which was 8.12 and 15.51 times higher than those in mice treated with boronated phenylalanine and those treated with BA. For boron, the tumor-to-normal tissue ratio was 4.41 ± 1.13 and the tumor-to-blood ratio was 5.92 ± 0.45. These results indicated that nanoparticles delivered boron to the tumor site effectively while minimizing accumulation in normal tissues. In conclusion, this composite (SP94-LB@BA-MSN) shows great promise as a boron-containing delivery agent for the treatment of hepatocellular carcinoma using BNCT. These findings highlight the potential of MSNs in the field of BNCT.

Anisamide-conjugated hairpin antisense oligonucleotides prodrug co-delivering doxorubicin exhibited enhanced anticancer efficacy.

Antisense oligonucleotides (ASONs)-based therapeutics offers tremendous promise for the treatment of diverse diseases. However, there is still a need to develop ASONs with enhanced stability against enzymes, improved drug delivery, and enhanced biological potency. In this study, we propose a novel anisamide (AA)-conjugated hairpin oligonucleotide prodrug loading with chemotherapeutic agent (doxorubicin, DOX) (AA-loop-ASON/DOX) for oncotherapy. Results indicated that the introduction of a hairpin conformation and AA ligand in prodrug significantly improved the stability against enzymatic hydrolysis, as well as the cellar uptake of ASONs and DOX. The incorporation of disulfide bonds could trigger mechanical opening, resulting in the release of ASON and DOX in response to the intracellular glutathione (GSH) in tumors. Moreover, the composite of DOX-loading ASONs prodrug exhibited a robust and selective inhibition of tumor cell proliferation. This paper introduces a novel design concept for nucleic acid-based therapeutics, aiming to enhance the delivery of drug and improve biological effectiveness.

Psat1-generated α-ketoglutarate and glutamine promote muscle stem cell activation and regeneration.

By satisfying bioenergetic demands, generating biomass, and providing metabolites serving as cofactors for chromatin modifiers, metabolism regulates adult stem cell biology. Here, we report that a branch of glycolysis, the serine biosynthesis pathway (SBP), is activated in regenerating muscle stem cells (MuSCs). Gene inactivation and metabolomics revealed that Psat1, one of the three SBP enzymes, controls MuSC activation and expansion of myogenic progenitors through production of the metabolite α-ketoglutarate (α-KG) and α-KG-generated glutamine. Psat1 ablation resulted in defective expansion of MuSCs and impaired regeneration. Psat1, α-KG, and glutamine were reduced in MuSCs of old mice. α-KG or glutamine re-established appropriate muscle regeneration of adult conditional Psat1 -/- mice and of old mice. These findings contribute insights into the metabolic role of Psat1 during muscle regeneration and suggest α-KG and glutamine as potential therapeutic interventions to ameliorate muscle regeneration during aging.

Marine toxins in seafood: Recent updates on sample pretreatment and determination techniques.

Marine toxins can lead to varying degrees of human poisoning, often resulting in fatal symptoms and causing significant economic losses in seafood-producing regions. To gain a deeper comprehension of the role of marine toxins in seafood and their impact on the environment, it is imperative to develop rapid, cost-effective, environmentally friendly, and efficient methods for sample pretreatment and determination to mitigate adverse impacts of marine toxins. This review presents a comprehensive overview of advancements made in sample pretreatment and determination techniques for marine toxins since 2017. The advantages and disadvantages of various technologies were critically examined. Additionally, the current challenges and future development strategies for the analysis of marine toxins are provided.

Site-Specific Profiling of N-Glycans in Drosophila melanogaster.

BACKGROUND: Drosophila melanogaster is a well-studied and highly tractable genetic model system for deciphering the molecular mechanisms underlying various biological processes. Although being one of the most critical post-translational modifications of proteins, the understanding of glycosylation in Drosophila is still lagging behind compared with that of other model organisms. METHODS: In this study, we systematically investigated the site-specific N-glycan profile of Drosophila melanogaster using intact glycopeptide analysis technique. This approach identified the glycans, proteins, and their glycosites in Drosophila, as well as information on site-specific glycosylation, which allowed us to know which glycans are attached to which glycosylation sites. RESULTS: The results showed that the majority of N-glycans in Drosophila were high-mannose type (69.3%), consistent with reports in other insects. Meanwhile, fucosylated N-glycans were also highly abundant (22.7%), and the majority of them were mono-fucosylated. In addition, 24 different sialylated glycans attached with 16 glycoproteins were identified, and these proteins were mainly associated with developmental processes. Gene ontology analysis showed that N-glycosylated proteins in Drosophila were involved in multiple biological processes, such as axon guidance, N-linked glycosylation, cell migration, cell spreading, and tissue development. Interestingly, we found that seven glycosyltransferases and four glycosidases were N-glycosylated, which suggested that N-glycans may play a regulatory role in the synthesis and degradation of N-glycans and glycoproteins. CONCLUSIONS: To our knowledge, this work represents the first comprehensive analysis of site-specific N-glycosylation in Drosophila, thereby providing new perspectives for the understanding of biological functions of glycosylation in insects.

The environmental sources of benzophenones: Distribution, pretreatment, analysis and removal techniques.

Benzophenones (BPs) have wide practical applications in real human life due to its presence in personal care products, UV-filters, drugs, food packaging bags, etc. It enters the wastewater by daily routine activities such as showering, impacting the whole aquatic system, then posing a threat to human health. Due to this fact, the monitoring and removal of BPs in the environment is quite important. In the past decade, various novel analytical and removal techniques have been developed for the determination of BPs in environmental samples including wastewater, municipal landfill leachate, sewage sludge, and aquatic plants. This review provides a critical summary and comparison of the available cutting-edge pretreatment, determination and removal techniques of BPs in environment. It also focuses on novel materials and techniques in keeping with the concept of "green chemistry", and describes on challenges associated with the analysis of BPs, removal technologies, suggesting future development strategies.

Non-steroidal anti-inflammatory drugs (NSAIDs) in the environment: Updates on pretreatment and determination methods.

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used in human and animal health care to reduce persistent inflammation, pain and fever because of their anti-inflammatory, analgesic and antipyretic effects. However, the improper discharge and disposal make it becomes a major contaminant in the environment, which poses a big threat to the ecosystem. For this reason, accurate, sensitive, effective, green, and economic techniques are urgently required and have been rapidly developed in recent years. This review summarizes the advancement of sample preparation technologies for NSAIDs involving solid-phase extraction, solid-phase microextraction, liquid-phase microextraction, QuEChERS, and matrix solid-phase dispersion. Meanwhile, we overview and compare analytical technologies for NSAIDs, including liquid chromatography-based methods, gas chromatography-based methods, capillary electrophoresis, and sensors, particularly the development of liquid chromatography-based methods. Furthermore, we focus on their progress and conduct a comparison between their advantages and disadvantages.

Non-steroidal anti-inflammatory drugs (NSAIDs) in the environment: Recent updates on the occurrence, fate, hazards and removal technologies.

Non-steroidal Anti-inflammatory Drugs (NSAIDs) are extensively utilized pharmaceuticals worldwide. However, owing to the improper discharge and disposal practices, they have emerged as significant contaminants that are widely distributed in water, soils, and sewage sediments. This ubiquity poses a substantial threat to the ecosystem and human health. Consequently, it is imperative to develop rapid, cost-effective, efficient and reliable approaches for containing these substance in order to mitigate the deleterious impact of NSAIDs. This research provides a comprehensive review of the occurrence, fate, and hazards associated with NSAIDs in the general environment. Additionally, various removal technologies, including advanced oxidation processes, biodegradation, and adsorption, were systematically summarized. The study also presents a comparative analysis of the benefits and drawbacks of different removal technologies while interpreting challenges related to NSAIDs' removal and proposing strategies for future development.

Arriving late and lean at a stopover site is selected against in a declining migratory bird population.

Loss and/or deterioration of refuelling habitats have caused population declines in many migratory bird species but whether this results from unequal mortality among individuals varying in migration traits remains to be shown. Based on 13 years of body mass and size data of great knots (Calidris tenuirostris) at a stopover site of the Yellow Sea, combined with resightings of individuals marked at this stopover site along the East Asian-Australasian Flyway, we assessed year to year changes in annual apparent survival rates, and how apparent survival differed between migration phenotypes (i.e. migration timing and fuel stores). The measurements occurred over a period of habitat loss and/or deterioration in this flyway. We found that the annual apparent survival rates of great knots rapidly declined from 2006 to 2018, late-arriving individuals with small fuel stores exhibiting the lowest apparent survival rate. There was an advancement in mean arrival date and an increase in the mean fuel load of stopping birds over the study period. Our results suggest that late-arriving individuals with small fuel loads were selected against. Thus, habitat loss and/or deterioration at staging sites may cause changes in the composition of migratory phenotypes at the population-level.

Design, characterization and biological evaluation of a new chimeric 4A2-5-antisense prodrug combined with chemotherapy.

Issues surrounding rapid degradation and limited therapeutic efficacy still exist in the development of native antisense oligonucleotides (ASONs). In this paper, a novel strategy of chimeric 4A2-5-ASON prodrug combined with chemotherapy for oncotherapy was proposed. The self-assembled hairpin-end prodrug structure provided a DOX loading site, while enhancing stability against nuclease degradation. The disulfide led responsive drug release, and excellent therapeutic effects were achieved by the combined action of RNase H and RNase L recruitment, along with chemotherapy drug Doxorubicin (DOX), both in vitro and in vivo. This work provides evidence for the development of designing nucleic acid drugs with combined mechanisms.

Volatile Methylsiloxanes in Complex Samples: Recent Updates on Pretreatment and Analysis Methods.

Volatile methylsiloxanes (VMSs) are massively produced chemicals having applications in industry and home because of their physical and chemical characteristics. They are used in personal care products such as cosmetics, household coatings, cleaners, skin care products, and others. Resultantly, large number of VMSs are discharged into air where they can be subjected to atmospheric migrations over long distances causing toxic and estrogenic effects, persistence, and bioaccumulations. Many institutions have taken measures to control VMSs. They require accurate, rapid, and sensitive pretreatment and analysis methods for diverse samples. Herein, the pretreatment and determination methods of VMSs as reported in recent years are reviewed and summarized. Pretreatments include commonly methods such as membrane-assisted solvent extraction, liquid-liquid extraction, and others, while novel methods are solid phase extraction, solid phase microextraction, diverse liquid phase microextraction and others. Analyses are made through gas chromatography-based methods. In addition, the advantages, and disadvantages of techniques are compared, and the prospects of pretreatment and analysis methods are discussed.

Pros and cons of different pretreatment and detection methods for VMSs were discussed.Different novel pretreatment techniques for VMSs were discussed.High-resolution mass spectrometry (HRMS) for the determination of VMSs was discussed.

A Review on Pretreatment and Analysis Methods of Polyether Antibiotics in Complex Samples.

Polyether antibiotics (PAs) are the anti-coccidiosis drugs used for treating and preventing coccidiosis. Studies show the residues of these antibiotics in food cause adversities and threaten human health. PAs thus need robust, rugged, and accurate methods for their analysis. This review encompasses pretreatment and detection methods of PAs in diverse matrices since 2010. Both conventional and developed methods are part of the pretreatments, such as dispersive liquid-liquid microextraction, solid-phase extraction, solid-phase microextraction, solvent front position extraction, QuEChERS (Quick Easy Cheap Effective Rugged and Safe), supercritical fluid extraction, and others. The analysis methods involve liquid chromatography coupled with detectors, sensors, etc. The pros and cons of various techniques for PAs have been discussed and future tendencies are proposed.

Progress of pretreatment and analytical methods for PAs are summarized.Comparisons between different mass analyzers are discussed in detail.Novel materials in microextraction methods are depicted.

Trihalomethanes in water samples: Recent update on pretreatment and detection methods.

Trihalomethanes (THMs) are classified as volatile organic compounds, considered to be a disinfection by-product during water disinfection process. THMs have been shown to be cytotoxic, genotoxic and mutagenic, with a risk of cancer when they contact with people directly. To protect public health and monitor water quality, it is important to monitor and measure THMs in drinking water. Therefore, it is crucial to develop fast, accurate, highly sensitivity and green analysis methods of THMs in various complicated matrices. Here, this review presents an overall summary of the current state of the pretreatment and detection methods for THMs in various sample matrices since 2005. In addition to the traditionally used pretreatment methods for THMs (such as headspace extraction, microwave-assisted extraction, liquid-liquid extraction), the new-developed methods, including solid-phase extraction, QuEChERS and different microextraction methods, have been summarized. The detection methods include gas chromatography-based methods, sensors and several other approaches. Additionally, benefits and limitations of different techniques were also discussed and compared. This study is anticipated to offer fruitful insights into the further advancement and widespread applications of pretreatment and detection technologies for THMs as well as for related substances.

Zinc (â ¡) functionalized magnetic geopolymer as sorbents for rapid extraction of Fluoroquinolones in food prior to quantification by UHPLC-MS/MS.

A novel Zn@MGeo sorbent was easily constructed and can bind with FQs through the synergistic effect of electrostatic interaction and coordination. With the Zn@MGeo as sorbent, a MSPE-UHPLC-MS/MS method was established for simultaneous detection of FQs in complex matrices. The whole extraction process could be completed using 6.0 mg sorbent within 10 min under the optimal conditions. The established quantitative method obtained a wide linear range (0.01-200 µg/kg, R2 > 0.9987), high sensitivity (LODs: 0.005-0.05 µg/kg) and negligible matrix effect. The method was applied for analysis of real samples, with recoveries between 75.6% and 103.7%. In addition, the sorbent could be reused at least 9 times without reducing the adsorption performance. In general, the established method not only proposes a novel sorbent for FQs extraction, but also provides a powerful tool for rapid and sensitive detection of FQs in food matrices with practical application value.

Benzodiazepines in complex biological matrices: Recent updates on pretreatment and detection methods.

Benzodiazepines (BDZs) are used in clinics for anxiolysis, anticonvulsants, sedative hypnosis, and muscle relaxation. They have high consumptions worldwide because of their easy availability and potential addiction. They are often used for suicide or criminal practices such as abduction and drug-facilitated sexual assault. The pharmacological effects of using small doses of BDZs and their detections from complex biological matrices are challenging. Efficient pretreatment methods followed by accurate and sensitive detections are necessary. Herein, pretreatment methods for the extraction, enrichment, and preconcentration of BDZs as well as the strategies for their screening, identification, and quantitation developed in the past five years have been reviewed. Moreover, recent advances in various methods are summarized. Characteristics and advantages of each method are encompassed. Future directions of the pretreatment and detection methods for BDZs are also reviewed.

Synthesis, Biophysical Properties, and Antitumor Activity of Antisense Oligonucleotides Conjugated with Anisamide.

Antisense oligonucleotides (ASONs) have proven potential for the treatment of various diseases. However, their limited bioavailability restricts their clinical application. New structures with improved enzyme resistance stability and efficient drug delivery are needed. In this work, we propose a novel category of ASONs bearing anisamide conjugation at phosphorothioate sites for oncotherapy. ASONs can be conjugated with the ligand anisamide very efficiently and flexibly in a solution. The conjugation sites and the ligand amount both influence anti-enzymatic stability and cellular uptake, resulting in changes in antitumor activity that are detectable by cytotoxicity assay. The conjugate with double anisamide (T6) was identified as the optimal conjugate, and its antitumor activity and the underlying mechanism were examined further in vitro and in vivo. This paper presents a new strategy for the design of nucleic acid-based therapeutics with improved drug delivery and biophysical and biological efficacy.

Diuretics in Different Samples: Update on the Pretreatment and Analysis Techniques.

Diuretics are drugs that promote the excretion of water and electrolytes in the body and produce diuretic effects. Clinically, they are often used in the treatment of edema caused by various reasons and hypertension. In sports, diuretics are banned by the World Anti-Doping Agency (WADA). Therefore, in order to monitor blood drug concentration, identify drug quality and maintain the fairness of sports competition, accurate, rapid, highly selective and sensitive detection methods are essential. This review provides a comprehensive summary of the pretreatment and detection of diuretics in various samples since 2015. Commonly used techniques to extract diuretics include liquid-liquid extraction, liquid-phase microextraction, solid-phase extraction, solid-phase microextraction, among others. Determination methods include methods based on liquid chromatography, fluorescent spectroscopy, electrochemical sensor method, capillary electrophoresis and so on. The advantages and disadvantages of various pretreatment and analytical techniques are elaborated. In addition, future development prospects of these techniques are discussed.

HIGHLIGHTSPretreatment and determination methods of diuretics in diverse samples are reviewed.Applications of novel materials and technologies for SPE and sensors are highlighted.Pros and cons of recent pretreatment and analysis techniques used for diuretics are discussed.Applications of high-resolution mass spectrometry are described in detail.

Natural Compounds, Optimal Combination of Brusatol and Polydatin Promote Anti-Tumor Effect in Breast Cancer by Targeting Nrf2 Signaling Pathway.

Triple-negative breast cancer (TNBC) has been clearly recognized as a heterogeneous tumor with the worst prognosis among the subtypes of breast cancer (BC). The advent and application of current small-molecule drugs for treating TNBC, as well as other novel inhibitors, among others, have made treatment options for TNBC more selective. However, there are still problems, such as poor patient tolerance, large administration doses, high dosing frequency, and toxic side effects, necessitating the development of more efficient and less toxic treatment strategies. High expression of Nrf2, a vital antioxidant transcription factor, often promotes tumor progression, and it is also one of the most effective targets in BC therapy. We found that in MDA-MB-231 cells and SUM159 cells, brusatol (BRU) combined with polydatin (PD) could significantly inhibit cell proliferation in vitro, significantly downregulate the expression of Nrf2 protein as well as the expression of downstream related target genes Heme Oxygenase-1 (HO-1) and NAD(P)H dehydrogenase, quinone 1 (NQO1), and promote reactive oxygen species (ROS) levels to further strengthen the anti-tumor effect. Furthermore, we discovered in our in vivo experiments that by reducing the drug dosage three times, we could significantly reduce tumor cell growth while avoiding toxic side effects, providing a treatment method with greater clinical application value for TNBC treatment.