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Enhanced mixing in microfluidic systems is necessary in many applications such as chemical processing, biological assays, and diagnosis. We are developing a microfluidic system to efficiently mix minute reagents (down to several microliters) using vibration-induced flow (VIF), in which a net flow is generated around a micropillar by applying periodic vibration. In this study, we numerically investigate the enhancement in chaotic mixing using the VIF technique and periodic switching of vibrations. By extending our previous numerical simulation model, we investigate the flow field and trajectories of fluid particles in three-dimensional space. We demonstrate that chaotic advection characteristics can be observed by periodically switching the vibrational direction of a substrate using simple cylindrical pillars. In addition, using an appropriate interval for switching the vibration axes yields better mixing performance. The extent of chaotic advection is evaluated quantitatively using the Lyapunov exponent considering various vibration parameters, such as the vibration amplitude, separation distance between each pillar and pillar shape. The flow field induced by a large-amplitude and sharp-edged wall pillar provides excellent mixing results. Thus, VIF is successfully applied to obtain an efficient mixing strategy with the aid of the chaotic theory. [ABSTRACT FROM AUTHOR]

Detection sensitivity of aquatic species using environmental DNA (eDNA) generally decreases in turbid water but is poorly characterized. In this study, eDNA detection targeted delta smelt (Hypomesus transpacificus), a critically endangered estuarine fish associated with turbid water. eDNA sampling in the field was first paired with a trawl survey. Species-specific detection using a Taqman qPCR assay showed concordance between the methods, but a weak eDNA signal. Informed by the results of field sampling, an experiment was designed to assess how turbidity and filtration methods influence detection of a rare target. Water from non-turbid (5 NTU) and turbid (50 NTU) estuarine sites was spiked with small volumes (0.5 and 1 mL) of water from a delta smelt tank to generate low eDNA concentrations. Samples were filtered using four filter types: cartridge filters (pore size 0.45 μm) and 47 mm filters (glass fiber, pore size 1.6 μm and polycarbonate, pore sizes 5 and 10 μm). Prefiltration was also tested as an addition to the filtration protocol for turbid water samples. eDNA copy numbers were analyzed using a censored data method for qPCR data. The assay limits and lack of PCR inhibition indicated an optimized assay. Glass fiber filters yielded the highest detection rates and eDNA copies in non-turbid and turbid water. Prefiltration improved detection in turbid water only when used with cartridge and polycarbonate filters. Statistical analysis identified turbidity as a significant effect on detection probability and eDNA copies detected; filter type and an interaction between filter type and prefilter were significant effects on eDNA copies detected, suggesting that particulate-filter interactions can affect detection sensitivity. Pilot experiments and transparent criteria for positive detection could improve eDNA surveys of rare species in turbid environments.

mRNA measurement is dominated by RT-PCR, which requires expensive laboratory equipment and personnel with advanced degrees. Loop-mediated isothermal amplification (LAMP) is a versatile technique for detecting target DNA and RNA. The sensitivity of LAMP in early reports has been below that of the standard RT-PCR tests. Here, we report the use of a fluorescence-based RT-LAMP protocol to measure CDX2 expression patterns, which match extremely well to the standards of sophisticated RT-PCR techniques (r = 0.99, p

We conducted a study to investigate the potential effects of nutrients released from a fish farm, which fell within the typical range found in oligotrophic offshore waters of the Aegean Sea, on phytoplankton growth. We designed an in situ bioassay experiment at a fish farm and incubated natural phytoplankton assemblages inside dialysis membrane bags for six days. Changes in phytoplankton growth in samples of ambient seawater collected throughout the experiment served as controls and were considered indicative of the net population change rates. Half of the bags were filled with seawater filtered through a 150-µm mesh, while the other half contained unfiltered seawater. The growth rates, estimated based on chlorophyll a concentrations and phytoplankton cell numbers inside the filtered and unfiltered bags, showed no significant differences. While no detectable net phytoplankton growth occurred in the ambient seawater, there was an exponential increase in chlorophyll a content and cell numbers within the bags. Moreover, the species richness within the bags gradually declined throughout the experiment. The findings of the study confirm that continuous nutrient releases from fish farms can promote high population growth rates in oligotrophic environments, provided that phytoplankton losses due to grazing, advection, and sinking are minimized or eliminated. [ABSTRACT FROM AUTHOR]

Background: Green synthesized nanoparticles have recently gained significant medicinal applications and oftentimes outperform their green sources. Selenium is of fundamental importance to human health, stemming from its distinctive physicochemical properties, such as antioxidant activity, inhibition of Lipid peroxidation, stabilization of membrane proteins, maintenance of membrane fluidity and modulation of cell signaling. Though reports have shown some therapeutic potential of Ackee plant parts such as antioxidant, anti-inflammatory, antimicrobial, neuroprotective, very few scientific proofs still exist in support of these effects. Methods: This study synthesized selenium nanoparticles (Se-NPs) from crude methanolic extracts of Ackee leaves (AKL) and Ackee arils (AKA), examined the photo-physical characteristics of the Se-NPs and determined the in-vitro antioxidant, antidiabetic, and anti-inflammatory potentials of AKL, AKA, and their Se-NPs using established protocols. Results: In both leaves and arils Se-NPs: UV spectroscopy revealed a qualitative absorbance at 310 nm; FTIR indicated multiple vibrations around 4000 cm−1- 400 cm−1; SEM images of 5 µm principally showed consistent size distribution of amorphous and granular shape at a magnification of 10,000X; while EDS spectra strongly confirm the presence of atomic Se compound at 30 kV. Various antioxidant activities assays carried out showed a range of approximately 4 to 60 times higher activities of the AKL, AKA, and Se-NPs than Ascorbic acid—the standard drug used. Furthermore, appreciable activities of more than 50% were obtained for alpha-amylase and alpha-glucosidase inhibitory activities, along with highly significant activities of haemoglobin glycosylation, glucose uptake, membrane stabilization, anti-arthritic, anti-haemolysis activities, when AKL, AKA, and Se-NPs were compared with standard drugs. Conclusion: Encouraging the development and utilization of AKL, AKA, and Se-NPs will provide tremendous therapeutic efficacy and bioavailability approaches towards the management of diabetes, inflammation, and other oxidative stress-related diseases. [ABSTRACT FROM AUTHOR]

This study explores for the first time the impact of chitosan (CS) with varying molecular weights (MW), orange peel extract concentration, and hydroxypropyl methylcellulose (HPMC) content on the formulation of buccal tablets for treating oral infections. Utilizing a statistical design of experiments (DoE), nine different formulations were evaluated for mechanical properties, dissolution behavior, mucoadhesion, and biological activity. A formulation with high CS MW, 60% orange peel extract, and 8% HPMC, emerged as the optimal formulation, demonstrating superior tabletability, compressibility, and compactibility. Dissolution studies indicated that hesperidin release followed the Higuchi model, with higher extract content enhancing this phenomenon. Mucoadhesion improved with increased HPMC and CS concentrations, although higher extract content reduced bioadhesion. Biological assays showed that higher extract levels boosted antioxidant activity, while CS primarily contributed to anti-inflammatory effects. The optimized formulation exhibited broad antimicrobial activity against key oral pathogens, surpassing the effectiveness of the individual components. Principal component analysis (PCA) further confirmed the significant influence of extract content on tablet properties. These findings suggest that the optimized tablet formulation holds promise for effective buccal delivery in the treatment of oral infections, warranting further investigation in clinical settings. [ABSTRACT FROM AUTHOR]

Alzheimer's disease, characterized by cognitive decline and memory loss, is associated with decreased acetylcholine levels due to acetylcholinesterase (AChE) activity. Compounds containing a coumarin heterocyclic core coupled with thiazole exhibit excellent acetylcholinesterase inhibitory activity. In this work, we designed and synthesized a series of 4-(benzo[d]thiazole-2-yl) phenols based on 4-hydroxycoumarin. The compounds were synthesized and their inhibitory activities were evaluated through in vitro biological assays. Of the compounds investigated, 3i exhibited the strongest inhibitory activity, with an IC50 value of 2.7 µM. Molecular docking and molecular dynamics simulations were employed to elucidate the binding interactions and stability of the synthesized compounds with AChE. The results demonstrated promising inhibitory activity, suggesting potential therapeutic applications for Alzheimer's disease. This research contributes to the development of coumarin-based heterocyclic compounds as effective AChE inhibitors. [ABSTRACT FROM AUTHOR]

A series of new 5-(trifluoromethyl)-benzo[d]imidazole linked N-phenylamide compounds was designed and synthesized using 1-chloro-2-nitro-4-(trifluoromethyl)benzene as raw material via five steps. Their structures were confirmed by 1H NMR, 13C NMR and HRMS. The bioassay results indicated that some of them exhibited moderate insecticidal activity (>50% inhibition) against Mythimna separata and Nilaparvata lugens at 500 μg/mL. [ABSTRACT FROM AUTHOR]

• The study introduces a novel method for identifying DNA-binding sites using amino acid sequences. • DeepDBS uses deep learning techniques to generate representations which train a Random Forest classifier. • The study achieves high performance in accuracy (99%) in cross-validations and jackknife testing outperforming existing methods. Interactions of biological molecules in organisms are considered to be primary factors for the lifecycle of that organism. Various important biological functions are dependent on such interactions and among different kinds of interactions, the protein DNA interactions are very important for the processes of transcription, regulation of gene expression, DNA repairing and packaging. Thus, keeping the knowledge of such interactions and the sites of those interactions is necessary to study the mechanism of various biological processes. As experimental identification through biological assays is quite resource-demanding, costly and error-prone, scientists opt for the computational methods for efficient and accurate identification of such DNA-protein interaction sites. Thus, herein, we propose a novel and accurate method namely DeepDBS for the identification of DNA-binding sites in proteins, using primary amino acid sequences of proteins under study. From protein sequences, deep representations were computed through a one-dimensional convolution neural network (1D-CNN), recurrent neural network (RNN) and long short-term memory (LSTM) network and were further used to train a Random Forest classifier. Random Forest with LSTM-based features outperformed the other models, as well as the existing state-of-the-art methods with an accuracy score of 0.99 for self-consistency test, 10-fold cross-validation, 5-fold cross-validation, and jackknife validation while 0.92 for independent dataset testing. It is concluded based on results that the DeepDBS can help accurate and efficient identification of DNA binding sites (DBS) in proteins. [ABSTRACT FROM AUTHOR]

In-vitro protein refolding is a major bottleneck step in the large-scale manufacturing of antibody fragments expressed in a microbial host system. The formation of an inter-chain disulfide bond is a key rate-limiting step during in-vitro refolding of antibody fragment therapeutics. In this investigation, we report the use of two redox mutant strains of E. coli viz., SHuffle® T7 (DE3) and SHuffle® T7 Express (DE3) possessing an oxidizing cytoplasm for the soluble expression of a refolded biosimilar antibody fragment. The effect of various critical process parameters on antibody fragment refolding yield was studied using a Design of Experiment (DoE) approach. The optimized upstream processing led to the expression of 167 ± 2.53 mg/L and 95 ± 1.44 mg/L of soluble refolded biosimilar Ranibizumab using the SHuffle® T7 (DE3) and SHuffle® T7 Express (DE3) strains, respectively, at the bioreactor scale. Physicochemical characteristics of the in-vivo refolded antibody fragment were studied and compared with an innovator molecule using various orthogonal analytical methods. A biological activity study of in-vivo refolded Ranibizumab using HUVEC cell-based bioassay proved that cell proliferation inhibition is comparable to the innovator Ranibizumab. The proposed strategy offers a time and cost-effective manufacturing platform for antibody fragments. [Display omitted] • Efficient soluble expression of in-vivo refolded antibody fragment. • Elimination of in-vitro refolding step. • DoE-based optimization of process parameters to enhance soluble target protein yield. • Novel downstream processing platform for the purification of antibody fragments. • Reduction in overall manufacturing time from nine days to three days. [ABSTRACT FROM AUTHOR]

The human epidermal growth factor (hEGF) is a powerful mitogen used in medical treatment and cosmetics. However, the recovery of hEGF from natural sources is insufficient to meet market demands, thus recombinant protein production technology for recombinant hEGF (rhEGF) was developed. Among the heterologous expression systems used for rhEGF production, Escherichia coli is preferred due to its well-established advantages. In this review, we focus on the development of two rhEGF expression strategies (soluble rhEGF and rhEGF inclusion bodies), and discuss the design of fusion rhEGF protein and its cultivation strategies. We also explore downstream processes for rhEGF isolation and purification methods (with or without a column chromatography system), and the recovery of rhEGF from inclusion bodies through solubilisation and refolding. Lastly, methods of quantification, characterisation, and biological activity assays used to quantify and validate purified rhEGF are also listed. [Display omitted] • Recombinant human epidermal growth factor (rhEGF) is used for ulcer treatment and cosmetic applications. • rhEGF can be produced via soluble or insoluble (inclusion bodies) protein strategies. • Exploration on differences in fusion protein, cultivation conditions and purification for both rhEGF production strategies. • Method of analysis of produced rhEGF. [ABSTRACT FROM AUTHOR]

Astragalus membranaceus is a traditional Chinese medicine with multiple pharmacological activities. Modern pharmacological research has found that Astragalus membranaceus extract has an inhibitory effect on α-glucosidase, however, which component can inhibit the activity of α-glucosidase and its degree of inhibition are unknown. To address this issue, this study used affinity ultrafiltration screening combined with UPLC-ESI-Orbitrap-MS technology to screen α-glucosidase inhibitors in Astragalus membranaceus. Using affinity ultrafiltration technology, we obtained the active components, and using UPLC-ESI-Orbitrap-MS technology, we quickly analyzed and identified them. As a result, a total of 8 ingredients were selected as α-glucosidase inhibitors. [ABSTRACT FROM AUTHOR]

One promising approach to overcome drug resistance in asthma treatments involves dual-target therapy, specifically targeting the β2 adrenergic receptor (β2-AR) and muscarinic-3 acetylcholine receptor (M3R). This study investigated the anti-asthma effects and dual-target mechanisms of glycyrrhizic acid, hesperidin, and platycodin D (GHP) from Zhisou San. GHP administration effectively attenuated OVA-induced inflammatory infiltration and overproduction of mucus in asthmatic mice. Additionally, GHP treatment significantly suppressed M3R and promoted β2-AR activation, resulting in the relaxation of tracheal smooth muscle. These findings concluded that GHP mitigated asthma by targeting β2-AR and M3R to ameliorate airway inflammation and modulate airway smooth muscle relaxation. [ABSTRACT FROM AUTHOR]

Two new cucurbitane-type triterpenoid saponins, 2,20β,22β-trihydroxy-16α,23(R)-epoxycucurbita-1,5,24-triene-3,11-dione 2-O-β-D-glucopyranoside (1), 2,20β,22α-trihydroxy-16α,23(S)-epoxycucurbita-1,5,11,24-tetraene-3-one 2-O-β-D-glucopyranoside (2) were isolated from the fruit of Citrullus colocynthis (L.) Schrad. Their structures were elucidated by mass spectrometry, IR, 1D, and 2D NMR spectroscopy, etc. Besides, both of the compounds showed significant hepatoprotective activities at 10 μM against paracetamol-induced HepG2 cell damage. [ABSTRACT FROM AUTHOR]

BACKGROUND: Evaluation of herbicidal activity and identification of active compounds are important bases for the development of new botanical herbicides. RESULTS: This study confirmed that Symphoricarpos orbiculatus has high herbicidal activities against mono‐dicotyledonous weeds, including Echinochloa crusgalli, Digitaria sanguinalis, Amaranthus retroflexus and Portulaca oleracea. By bioassay‐guided isolation, 12 compounds were isolated and identified from S. orbiculatus for the first time, including iridoids: naucledal (K1), loganin (K2), loganigenin (K3), loganin acid (K4), glucologanin (K5) and vogeloside (K6), as well as flavonoids: quercetine (K7), luteolin (K8), nobiletin (K9), astragalin (K10), isorhamnetin 3‐d‐glucoside (K11) and rutin (K12). Biological assays showed that iridoids are the main active ingredients of S. orbiculatus. The compounds of K5 and K6 could inhibit both the root (IC50 = 37.54 and 38.91 μg mL−1, respectively) and shoot (IC50 = 42.78 and 45.72 μg mL−1, respectively) of Portulaca oleracea, which have a weeding toxicity similar to that of the commercialized plant‐based herbicide pelargonic acid. In addition, the results of pot culture assay showed that S. orbiculatus ethanol extracts had high fresh weight control effect against Digitaria sanguinalis and P. oleracea at the concentration of 40 g L−1. After 7 days, both the soil treatment and the stem and leaf spray method resulted in severe leaf necrosis and significant leaf etiolation. CONCLUSION: Symphoricarpos orbiculatus and its herbicidal active compounds have the potential to develop into botanical herbicides, and are first reported in the present study. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The incidence of prostate cancer is increasing, and its poor prognosis is associated with metadherin overexpression; however, its molecular mechanisms are unclear. This study used Western blot, quantitative real-time polymerase chain reaction, immunofluorescence, and tumorigenicity assays to investigate metadherin and epithelial-mesenchymal transition in prostate cancer. We found that metadherin was highly expressed in prostate cancer tissues, especially PC-3 cells. Metadherin overexpression promoted t epithelial-mesenchymal transition and tumor growth. Bioinformatics analysis and assays revealed that lymphoid enhancer-binding factor 1 directly activates the metadherin promoter. The epithelial- mesenchymal transition and tumorigenicity were suppressed by the silencing of lymphoid enhancer-binding factor 1; however, these effects were reversed by metadherin overexpression. Vitamin D treatment downregulated metadherin and lymphoid enhancer-binding factor 1, counteracting lymphoid enhancer-binding factor 1-induced metadherin upregulation. Our findings indicate that metadherin enhances epithelial-mesenchymal transition and tumorigenicity through lymphoid enhancer-binding factor 1, with vitamin D modulating this interaction in prostate cancer. [ABSTRACT FROM AUTHOR]

Pharmacogenomic (PGx) testing is a growing area of personalized medicine with demonstrated clinical utility in improving patient outcomes in oncology. PGx testing of pharmacogenes affecting drug pharmacokinetics, pharmacodynamics, and response can help inform drug selection and dosing of several anticancer therapies and supportive care medications. Several PGx testing techniques exist including polymerase chain reaction (PCR), MassARRAY, microarray, and sequencing. This review article provides a clinician-friendly guide of these techniques. Understanding the advantages, limitations, ideal use, and potential clinical applications of each platform can help clinicians choose the appropriate PGx testing platform for specific use cases. A clinician-friendly guide to available pharmacogenomic testing techniques and their use in oncology. [ABSTRACT FROM AUTHOR]

BACKGROUND: Elucidating fitness cost associated with field‐evolved insect resistance to insecticide is of particular importance to current sustainable pest control. The global pest whitefly Bemisia tabaci has developed resistance to many members of neonicotinoids, but little is known about whitefly resistance to neonicotinoid nitenpyram and its associated fitness cost. Using insecticide bioassay and life‐table approach, this study aims to investigate nitenpyram resistance status in field‐collected whitefly populations, and to explore whether such resistance is accompanied by a fitness cost. RESULTS: The bioassay results revealed that 14 of 29 whitefly populations displayed moderate to extremely high resistance to nitenpyram, demonstrating a widespread field‐evolved resistance to nitenpyram. This field‐evolved resistance in the whitefly has increased gradually over the past 3 years from 2021 to 2023. Further life‐table study showed that two resistant whitefly populations exhibited longer developmental time, shorter lifespans of adult, and lower fecundity compared with the most susceptible population. The relative fitness cost of the two resistant populations was calculated as 0.69 and 0.56 by using net productive rate R0, which suggests that nitenpyram resistance comes with fitness cost in the whitefly, especially on reproduction. CONCLUSION: Overall, these results represent field‐evolved high resistance to nitenpyram in the whitefly. The existing fitness costs associated with nitenpyram resistance are helpful to propose a suitable strategy for sustainable control of whiteflies by rotation or mixture of insecticide with different modes of action. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Land managers, researchers, and regulators increasingly utilize environmental DNA (eDNA) techniques to monitor species richness, presence, and absence. In order to properly develop a biological assay for eDNA metabarcoding or quantitative PCR, scientists must be able to find not only reference sequences (previously identified sequences in a genomics database) that match their target taxa but also reference sequences that match non-target taxa. Determining which taxa have publicly available sequences in a time-efficient and accurate manner currently requires computational skills to search, manipulate, and parse multiple unconnected DNA sequence databases. Our team iteratively designed a Graphic User Interface (GUI) Shiny application called the Reference Sequence Browser (RSB) that provides users efficient and intuitive access to multiple genetic databases regardless of computer programming expertise. The application returns the number of publicly accessible barcode markers per organism in the NCBI Nucleotide, BOLD, or CALeDNA CRUX Metabarcoding Reference Databases. Depending on the database, we offer various search filters such as min and max sequence length or country of origin. Users can then download the FASTA/GenBank files from the RSB web tool, view statistics about the data, and explore results to determine details about the availability or absence of reference sequences. [ABSTRACT FROM AUTHOR]

Background: Lactoferrin (LF) in human milk has various biological properties and contributes to the prevention of preterm birth complications. Enzyme-linked immunosorbent assay (ELISA) is one of the most commonly used methods to measure LF in human milk, but this method is time-consuming and laborious. In Japanese human milk banks, the concentration of LF in donor human milk (DHM) is measured routinely. Here, we reported a rapid, simple, and accurate method for determining LF in human milk using a new reagent based on a latex agglutination assay. Methods: We obtained 208 human milk pools from 148 mothers, and samples were collected before and after Holder pasteurization. Milk samples were diluted 100- or 200-fold and LF concentrations were measured by a latex agglutination assay using an automated analyzer. The reagent was validated in terms of repeatability, linearity, detection limit, recovery, and comparison with ELISA. Results: The coefficient of variation (CV) for intra-assay precision ranged from 0.6 to 5.0% in human milk with high, medium, and low LF concentrations. The linearity was also tested by serial sample dilution and was confirmed up to 16 µg/mL with a detection limit of 0.2 µg/mL. The recovery rates in a spiked recovery test were ranged from 90 to 120% at high, medium, and low concentrations of LF. Furthermore, a strong correlation was observed between LF levels determined by the latex agglutination assay and ELISA (r = 0.978, p < 0.001, n = 255). The regression equation was y = 0.991x + 0.545 (r2 = 0.974, p < 0.001). Compared with ELISA, the latex agglutination assay reduces the measurement time by 160 min and the cost by 55%. Conclusions: The latex agglutination assay used to determine LF in human milk is rapid, simple, and accurate enough to be used routinely. Its use may contribute to the quick and easy provision of appropriate DHM to preterm infants. [ABSTRACT FROM AUTHOR]

Background/objective: To examine the cognitive benefits of 6  months of prescribed intermittent exercise (10-min bouts totaling 150 weekly minutes) in community-dwelling older adults, comparing effects of low-intensity movement (LIM) and moderate-intensity aerobic exercise (aerobic exercise; AE) training; and exploring biological mechanisms of exercise-related cognitive improvement. Method: Twenty-five adults (>60  years old) participated in a 6-month controlled trial and were randomized into LIM or AE intermittent training. Cognition was assessed using a neuropsychological test battery including the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS), California Verbal Learning Test, 2nd Edition (CVLT-II), and Delis-Kaplan Executive Function System (D-KEFS). Neuroimaging measures were collected using a 7  T human MRI scanner. Serologic neurotrophic and inflammatory factors were analyzed using Luminex multiplex assays [brain derived neurotrophic factor (BDNF); vascular endothelial growth factor (VEGF)]; interleukin-6 (IL-6), C-reactive protein (CRP), plasminogen activator inhibitor (PAI-1). Results: LIM and AE intermittent training had dissociable effects on cognition, with LIM resulting in improved learning and memory and AE resulting in improved executive functioning. Intervention groups differed on change in cognitive performance on CVLT-II learning and D-KEFS trail making test. Increase in right dorsolateral prefrontal cortex (DLPFC) surface area was linked to executive improvement (i.e., phonemic fluency) regardless of intervention group. A decline in circulating PAI-1 was linked to learning and memory improvement in response to LIM over 6 months. Conclusion: Moderate-intensity AE and LIM intermittent training likely have distinct cognitive benefits, though low-intensity activity is often included as a control group in exercise trials in aging. [ABSTRACT FROM AUTHOR]

Introduction:Vernonia amygdalina Delile (VAD), also known as bitter leaf, is widely utilized in traditional medicine for the treatment of various ailments, including cancer. The presence of bioactive compounds in VAD is believed to be responsible for its characteristic bitterness. In Ghana, it is a common practice to mitigate the bitterness of VAD by combining it with Citrus aurantifolia (Christm.) Swingle (lime) juice extracts, although this method lacks scientific evidence and documentation. Therefore, the antioxidant and anticancer activities of VAD and lime juice extracts (V5) and their combined effects were evaluated in vitro. Method: The antioxidant activity and cytotoxic effects of VAD extracts were determined against Jurkat, MCF‐7, HepG2, and PNT2 cells using the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) assay to quantify antioxidant activity and the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay to assess cytotoxicity. The statistical analysis of the data was conducted using Microsoft Excel and GraphPad Prism 8.0. Linear regression was employed to determine the correlation between the concentration and the percentage of antioxidant activity, while p values were calculated using Student's t‐test. Results: The laboratory analysis focused on the extracts V1, V2, V3, V4, and V5. Briefly, V1 and V2 contained equal amounts of saponins and terpenoids. Among these, V2 exhibited the highest free radical scavenging activity, as indicated by an EC50 value of 2.14 ± 0.06 mg/mL. V2 also demonstrated cytotoxicity against the MCF‐7, HepG2, Jurkat, and PNT2 cell lines. On the other hand, V3 and V4 did not show any cytotoxic effects across all tested cell lines. In contrast, V5 was toxic to HepG2 and MCF‐7 cells but had no cytotoxic effect on Jurkat cell lines. V2 exhibited dose‐dependent cytotoxicity (0–1000 μg/mL), with the strongest inhibition observed against Jurkat cells (IC50 value = 96.341 μg/mL) and a selective index of 3.567. The difference in activity between the extracts from different parts of the plant and the extract combined with lime juice was significant (p < 0.05), indicating a synergistic effect of the phytochemicals in both VAD and lime juice. Conclusion: V2 and V5 demonstrated a remarkable antioxidant property, and they are effective in inhibiting cancer cell lines, respectively. [ABSTRACT FROM AUTHOR]

Background: The present study aimed to investigate the effects of infrared diode laser irradiation on the proliferation and differentiation capacity of periodontal ligament stem cells (hPDLSCs), which are optimal cell sources for periodontal regeneration. Methods: hPDLSCs were isolated and characterized by flow cytometric analysis of mesenchymal stem cell markers, and their trilineage differentiation capacity was tested. hPDLSCs were then cultured and irradiated with infrared diode laser (970 nm) at a power of 200 mW and a fluence of 4 J/cm2 for 3 s. MTT assay was performed to assess cellular proliferation. Cell cycle analysis was performed, and the impact of infrared diode laser irradiation on the stemness and osteogenic differentiation potential of hPDLSCs was evaluated via RT‒PCR. Results: Infrared diode laser application enhanced the stemness, viability, proliferation, and differentiation of PDLSCs. Stem cell markers (OCT4, SOX2, and NANOG) were significantly upregulated in hPDLSCs exposed to laser irradiation. There was significant overexpression of RUNX2, ALP, OPN, and OCN on day 14 after laser application. Conclusions: These findings provide valuable insights into the specific applications of infrared diode lasers to effectively regenerate periodontal tissues. The results can aid in the development of precise clinical protocols aimed at enhancing osseointegration and promoting tissue regeneration. Ultimately, the combination of infrared diode laser with hPDLSCs is promising for stimulating periodontal regeneration. [ABSTRACT FROM AUTHOR]

Cerium oxide (CeO2) and yttrium oxide (Y2O3) nanoparticles possess interesting surface properties and interfacial interactions that make them attractive candidates for various applications. This study presents a comprehensive investigation into the synthesis and toxicity assessment of yttrium-doped cerium oxide (Ce-Y) nanocomposite using a combination of analytical techniques and biological assays. The nanocomposite was characterized using UV–Vis spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), electron microscopy imaging, dynamic light scattering (DLS), and zeta potential measurements to elucidate their structural, optical and physicochemical properties. The synthesized nanocomposite exhibited distinctive absorption spectra and precise alignment of diffraction peaks, confirming the successful incorporation of yttrium ions into the cerium oxide lattice. Electron microscopy images revealed well-dispersed yttrium particles within the ceria matrix, indicating uniform distribution and morphology. DLS and zeta potential analysis provided insights into the size distribution and stability of the nanocomposite. Furthermore, in vivo toxicity assessment using Drosophila melanogaster model revealed no significant toxicity of Ce:Y nanocomposite, as evidenced by survival assay and behavioral assays. Superoxide dismutase (SOD) activity and reactive oxygen species (ROS) levels were also evaluated, demonstrating no discernible nanoparticle-induced toxicity. Overall, this study highlights the potential applications of Ce:Y nanocomposite and underscores the importance of comprehensive toxicity evaluation in nanomaterial development. [ABSTRACT FROM AUTHOR]

Objective: Our study focused on the effects and molecular mechanisms of kaempferol, a major active component of Eucommia ulmoides Oliver (EUO), on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods: Target molecules for EUO, osteoarthritis, and osteogenic differentiation were identified through network pharmacology analysis. BMSCs were isolated and treated with various concentrations of kaempferol. Optimal concentration was determined through MTT assays. Osteogenic differentiation was assessed using alkaline phosphatase (ALP) and Alizarin Red S staining, while osteogenic markers (Collagen I, RUNX2, and OPN) and CAV-1 expression were analyzed using RT-qPCR and Western blot. The effects of combined treatment with kaempferol and an overexpression vector for CAV-1 (oe-CAV-1) on osteogenic differentiation were also observed. Results: Network pharmacology analysis identified kaempferol as the primary active component influencing CAV-1 targeted in subsequent experiments. It was found that 10 µM kaempferol was optimal for treating BMSCs. Post-treatment, significant increases in ALP activity and calcium deposition were observed, along with elevated expression of osteogenic markers, and decreased CAV-1. Overexpression of CAV-1 significantly reversed the promotive effects of kaempferol on BMSC osteogenic differentiation, effectively inhibiting the process. Conclusion: Collectively, kaempferol promotes osteogenic differentiation in BMSCs by inhibiting CAV-1 expression. [ABSTRACT FROM AUTHOR]

Background: Screening for anaplastic lymphoma kinase (ALK) rearranged non‐small cell lung cancer (NSCLC) is crucial for identifying patients eligible for targeted therapy. The FDA‐approved ALK (D5F3) immunohistochemistry (IHC) assay, used with the OptiView Amplification Kit, demonstrates excellent sensitivity and specificity in detecting these patients. However, the clinical significance of resulting focal positivity remains unclear, and ALK (D5F3) expression unrelated to ALK fusion is observed in some cases of neuroendocrine differentiation. This study aims to validate these findings with molecular testing and contribute to the accurate interpretation of ALK (D5F3) IHC results. Methods: A total of 1619 patients diagnosed with NSCLC and neuroendocrine carcinoma were evaluated using ALK (D5F3) IHC. For cases with strong but focal expression and those with diffuse strong positivity in neuroendocrine differentiation, ALK fluorescence in situ hybridization (FISH) and/or next‐generation sequencing (NGS) tests were performed. Results: Seven out of 1109 adenocarcinomas (0.6%) and six out of 289 squamous cell carcinomas (2.1%) exhibited strong focal ALK (D5F3) expression. Nine out of 209 neuroendocrine carcinomas (4.3%) showed homogeneously strong ALK (D5F3) expression. All these cases, including adenocarcinoma with neuroendocrine differentiation and combined small cell carcinoma, were negative for ALK fusions by FISH and/or NGS. Conclusion: This study demonstrates that strong but focal ALK (D5F3) immunostaining and strong expression in neuroendocrine differentiation may not indicate ALK fusion. By considering these findings, we can improve the accuracy of patient selection for targeted therapy by minimizing false‐positive interpretations of ALK (D5F3) staining. [ABSTRACT FROM AUTHOR]

The translated protein undergoes a specific modification process, which involves the formation of covalent bonds on lysine residues and the attachment of small chemical moieties. The protein's fundamental physicochemical properties undergo a significant alteration. The change significantly alters the proteins' 3D structure and activity, enabling them to modulate key physiological processes. The modulation encompasses inhibiting cancer cell growth, delaying ovarian aging, regulating metabolic diseases, and ameliorating depression. Consequently, the identification and comprehension of post-translational lysine modifications hold substantial value in the realms of biological research and drug development. Post-translational modifications (PTMs) at lysine (K) sites are among the most common protein modifications. However, research on K-PTMs has been largely centered on identifying individual modification types, with a relative scarcity of balanced data analysis techniques. In this study, a classification system is developed for the prediction of concurrent multiple modifications at a single lysine residue. Initially, a well-established multi-label position-specific triad amino acid propensity algorithm is utilized for feature encoding. Subsequently, PreMLS: a novel ClusterCentroids undersampling algorithm based on MiniBatchKmeans was introduced to eliminate redundant or similar major class samples, thereby mitigating the issue of class imbalance. A convolutional neural network architecture was specifically constructed for the analysis of biological sequences to predict multiple lysine modification sites. The model, evaluated through five-fold cross-validation and independent testing, was found to significantly outperform existing models such as iMul-kSite and predML-Site. The results presented here aid in prioritizing potential lysine modification sites, facilitating subsequent biological assays and advancing pharmaceutical research. To enhance accessibility, an open-access predictive script has been crafted for the multi-label predictive model developed in this study. Author summary: Proteins undergo a variety of post-translational modifications (PTMs) after synthesis, such as lysine modifications, which significantly influence their structure and function. These modifications of lysine are known to regulate physiological processes, including the inhibition of cancer cell growth, the delay of aging, the regulation of metabolic diseases, and the improvement of depressive disorders. Abnormal modifications are closely associated with the occurrence and progression of a multitude of diseases. Therefore, the identification and comprehension of these modifications are of paramount importance for biological research and drug development. A multitude of studies have focused on a single type of lysine modification, with prediction methods for multiple lysine modification sites being relatively scarce. In this research, a novel multi-label prediction model named PreMLS has been developed for the simultaneous identification of four lysine modifications: methylation, acetylation, crotonylation, and succinylation. The imbalance issue in the dataset was addressed utilizing the ClusterCentroids undersampling algorithm, following which a predictive model, PreMLS, was constructed using a CNN to forecast multiple lysine modification sites. Compared to existing models, this new approach has significantly enhanced the accuracy and reliability of the predictions. [ABSTRACT FROM AUTHOR]

Numerous clinical disorders have been linked to the etiology of dysregulated NLRP3 (NACHT, LRR, and PYD domain‐containing protein 3) inflammasome activation. Despite its potential as a pharmacological target, modulation of NLRP3 activity remains challenging. Only a sparse number of compounds have been reported that can modulate NLRP3 and none of them have been developed into a commercially available drug. In this research, we identified three potent NLRP3 inflammasome inhibitors, gymnoasins A‐C (1–3), with unprecedented pentacyclic scaffolds, from an Antarctic fungus Pseudogymnoascus sp. HDN17‐895, which represent the first naturally occurring naphthopyrone‐macrolide hybrids. Additionally, biomimetic synthesis of gymnoasin A (1) was also achieved validating the chemical structure and affording ample amounts of material for exhaustive bioactivity assessments. Biological assays indicated that 1 could significantly inhibited in vitro NLRP3 inflammasome activation and in vivo pro‐inflammatory cytokine IL‐1β release, representing a valuable new lead compound for the development of novel therapeutics with the potential to inhibit the NLRP3 inflammasome. [ABSTRACT FROM AUTHOR]

Currently, research in the development of high-performance sensing platforms has increased to fulfill the needs of analysis and detection. In this study, we developed a novel type of surface-enhanced Raman scattering (SERS) chip composed of a covalent organic framework (COF)-silver nanoparticles (AgNPs) nanocomposite, and this nanocomposite was fabricated by a one-step method of ultrasonically mixing the obtained COF and AgNPs. The fabricated chip exhibited high sensitivity and repeatable SERS effects. Practical application results showed that the chip was highly sensitive and reliable and capable of detecting DNA bases (adenine) to fit an equation in the range from 0.01 pM to 1 nM, with an R-square of 0.97253 and a detection limit of ~0.026 pM (signal-to-noise ratio (S/N) = 3). Therefore, the proposed SERS system has potential applications in biological assays. Schematic of AgNP@COF as SERS substrates for the detection of DNA bases. [ABSTRACT FROM AUTHOR]

Background: The fabrication of anti-Toxoplasma drugs with less side effects and desirable efficacy is one of the important research goals facing with toxoplasmosis. This study aimed to determine the anti-Toxoplasma effects of Cinnamon zeylanicum (CZ), Moringa oleifera (MO) oil encapsulated into solid lipid nanoparticles (SLNs). Methods: Vero cells were cultured with serial concentrations (1 mg/mL to 100 µg/mL) of CZ-SLNs and MO-SLNs in DMEM culture medium. The morphological, physical, and chemical features of nanoparticles were calculated. The cell viability assays and anti-T. gondii effects of CZ-SLNs and MO-SLNs were evaluated. The CC50 and IC50 indices of SLNs-enveloped extracts were calculated. Results: The particle sizes of MO-SLNs and CZ-SLNs were 411.5 and 365 nm, while PDI indices were 0.53 and 0.7, respectively. Transmission electron microscopy (TEM) showed that both MO-SLNs and CZ-SLNs were smoothed spherical nanoparticles with rounded edges. The cytotoxicity assay showed the CC50 value of MO-SLNs at concentrations of ˃10 mg/mL. In addition, 60% of T. gondii-infected Vero cells remained alive at the concentrations ≤ 1 mg/ml, while the MO-SLNs killed at least 90% of T. gondii tachyzoites with an IC50 > 1 µg/ml. The cytotoxicity of CZ-SLNs extract showed the CC50 at the concentration 0.1 mg/mL. More than 50% of Vero cells, infected with T. gondii tachyzoites, survived at a concentration less than 0.1 mg/mL (IC50 ˂ 0.1 mg/mL), while the CZ-SLNs killed at least 85% of T. gondii tachyzoites in all concentrations. Conclusion: The current results represented that the use of SLNs as a nano-carrier for M. oleifera and C. zeylanicum could kill T. gondii tachyzoites with low cytotoxicity, suggesting the effectiveness of these nano-emulsions along with the chemical agents in the treatment of Toxoplasma. [ABSTRACT FROM AUTHOR]

Arterial thrombosis is a leading cause of death and disability worldwide with no effective bioassay for clinical prediction. As a symbolic feature of arterial thrombosis, severe stenosis in the blood vessel creates a high-shear, high-gradient flow environment that facilitates platelet aggregation towards vessel occlusion. Here, we present a thrombus profiling assay that monitors the multi-dimensional attributes of thrombi forming in such biomechanical conditions. Using this assay, we demonstrate that different receptor–ligand interactions contribute distinctively to the composition and activation status of the thrombus. Our investigation into hypertensive and older individuals reveals intensified biomechanical thrombogenesis and multi-dimensional thrombus profile abnormalities, endorsing the diagnostic potential of the assay. Furthermore, we identify the hyperactivity of GPIbα-integrin αIIbβ3 mechanosensing axis as a molecular mechanism that contributes to hypertension-associated arterial thrombosis. By studying drug-disease interactions and inter-individual variability, our work reveals a need for personalized anti-thrombotic drug selection that accommodates each patient's pathological profile. Arterial thrombosis is a leading cause of death and disability with no clinical bioassay for prediction. Here, the authors develop an assay that monitors the multi-dimensional attributes of thrombi forming in its physiologically mimicking rheological condition to predict arterial thrombotic risks [ABSTRACT FROM AUTHOR]

Introduction: Prostate cancer (PCa) is the second most frequent cancer diagnosis in men and the fifth leading cause of death worldwide. Prostate Specific Membrane Antigen (PSMA) and Gastrin Releasing Peptide (GRP) receptors are overexpressed in PCa. In this study, we have developed iron oxide nanoparticles (IONs) functionalized with the Prostate Specific Membrane Antigen (PSMA) and Gastrin Releasing Peptide (GRP) ligands for dual targeting of Prostate cancer. Methods: IONs were developed with a thin silica layer on their surface with MPTES (carrying -SH groups, IONs-SH), and they were coupled either with a pharmacophore targeting PSMA (IONs-PSMA) or with bombesin peptide (IONs-BN), targeting GRP receptors, or with both (IONs-PSMA/BN). The functionalized IONs were characterized for their size, zeta potential, and efficiency of functionalization using dynamic light scattering (DLS) and Fourier-Transform Infrared Spectroscopy (FT-IR). All the aforementioned types of IONs were radiolabeled directly with Technetium-99m (99mTc) and evaluated for their radiolabeling efficiency, stability, and binding ability on two different PCa cell lines (PC3 and LNCaP). Results and Discussion: The MTT assay demonstrated low toxicity of the IONs against PC3 and LNCaP cells, while the performed wound-healing assay further proved that these nanostructures did not affect cellular growth mechanisms. The observed hemolysis ratio after co-incubation with red blood cells was extremely low. Furthermore, the 99mTc-radiolabeled IONs showed good stability in human serum, DTPA, and histidine, and high specific binding rates in cancer cells, supporting their future utilization as potential diagnostic tools for PCa with Single Photon Emission Computed Tomography (SPECT) imaging. [ABSTRACT FROM AUTHOR]

Paclobutrazol, a fungicide of the triazole class, is widely used as an inducer of early flowering and fruiting by inhibiting gibberellin formation. However, biological assays using model organisms to evaluate their cytogenotoxic and mutagenic potential are still scarce. Therefore, this study aimed to investigate the effects of the commercial product Cultar® 250 SC (CP) and the pure substance (PBZ) on the germination and initial seedling development of Lactuca sativa L. (lettuce), in addition to evaluating the effects of CP on the mitotic activity and DNA, as we believe that PBZ has a greater toxic potential than CP on seed germination, and that the latter has cytogenotoxic and mutagenic effects on L. sativa. Lettuce seeds treated with CP and with PBZ in the doses of 0.25, 0.50, 1, 1.5, and 2 g L−1 showed significant reductions in germination rate, as well the CP reduced the root and initial development seedling development. PBZ showed greater inhibition of germination compared to CP. In direct exposure to PBZ, there was not sufficient seedling development for analysis, while in discontinuous treatment, there was inhibition of root growth (except for doses of 0.25 and 0.50 g L−1) and in the development of the aerial part. While no mitodepressive effect was observed in meristematic cells treated with CP, increased frequencies of chromosomal alterations, including condensed nuclei and micronuclei, were evident in both meristematic cells and the F1 region. The Comet assay further demonstrated higher levels of DNA damage at higher paclobutrazol doses, supporting the findings of increased micronucleus frequencies. Consequently, it can be concluded that the CP exhibits greater toxicity towards seed germination compared to lettuce seedlings, and PBZ has a greater toxic potential than CP in relation to these parameters. However, the impact of CP on seedlings was relatively minimal, as evidenced by their limited effects on development, cell proliferation, and DNA, suggesting a slight toxicity of this agent. Therefore, we infer that Cultar® 250 SC should be used with caution. Thus, this study emphasizes the importance of employing joint analyses to better elucidate and correlate the mechanisms of action of potentially toxic substances. Furthermore, it provides a basis for discussing the application of Cultar® 250 SC and seeking more sustainable alternatives in food production. [ABSTRACT FROM AUTHOR]

Image-based models that use features extracted from cell microscopy images can estimate the activity of small molecules in various biological assays. Typically, models are trained on images stained by an optimized protocol (e.g. Cell Painting) after exposure to a fairly high small molecule concentration (referred to as 'image concentration') of 10 μ M or higher. Low concentration images (e.g. 0.16 μM, 0.8 μM, 4 μM) tend to yield models with worse performance. In this work, we nevertheless report a practical use for low image concentration data. We propose the combination of well-performing models trained at higher image concentrations, with lower image concentration for inference to identify more potent compounds. We show that this approach improves on the conventional method (directly training a high-potency model) in 65 % of assays investigated in terms of AUC-ROC, and 75 % of assays in terms of RIPtoP-corrected AUC-PR. [ABSTRACT FROM AUTHOR]

Simple Summary: The frequent activation of the PI3K/AKT/mTOR (PAM) pathway makes it an attractive therapeutic target in gynecological cancers. Many PAM inhibitors selectively target single PAM pathway nodes, which can lead to reduced efficacy and increased drug resistance. In addition, compensatory pathways can be activated when only the PAM pathway is inhibited. Here, we show that gedatolisib, a PAM inhibitor targeting multiple PAM pathway nodes, exerted greater growth-inhibitory effects relative to single-node PAM inhibitors in gynecologic cancer cell models. In addition, gedatolisib combined with inhibitors of compensatory pathways involved in the estrogen response and cell cycle progression inhibited tumor growth in endometrial and ovarian cancer mouse models. Gedatolisib in combination with other therapies has previously shown promising preliminary clinical efficacy and safety in various solid tumor types. The non-clinical data presented here support the development of gedatolisib in combination with hormonal therapy and/or cell cycle inhibitors for gynecologic cancer treatment. Background/Objectives: The PI3K/AKT/mTOR (PAM) pathway is frequently activated in gynecological cancers. Many PAM inhibitors selectively target single PAM pathway nodes, which can lead to reduced efficacy and increased drug resistance. To address these limitations, multiple PAM pathway nodes may need to be inhibited. Gedatolisib, a well-tolerated panPI3K/mTOR inhibitor targeting all Class I PI3K isoforms, mTORC1 and mTORC2, could represent an effective treatment option for patients with gynecologic cancers. Methods: Gedatolisib and other PAM inhibitors (e.g., alpelisib, capivasertib, and everolimus) were tested in endometrial, ovarian, and cervical cancer cell lines by using cell viability, cell proliferation, and flow cytometry assays. Xenograft studies evaluated gedatolisib in combination with a CDK4/6 inhibitor (palbociclib) or an anti-estrogen (fulvestrant). A pseudo-temporal transcriptomic trajectory of endometrial cancer clinical progression was computationally modeled employing data from 554 patients to correlate non-clinical studies with a potential patient group. Results: Gedatolisib induced a substantial decrease in PAM pathway activity in association with the inhibition of cell cycle progression and the decreased cell viability in vitro. Compared to single-node PAM inhibitors, gedatolisib exhibited greater growth-inhibitory effects in almost all cell lines, regardless of the PAM pathway mutations. Gedatolisib combined with either fulvestrant or palbociclib inhibited tumor growth in endometrial and ovarian cancer xenograft models. Conclusions: Gedatolisib in combination with other therapies has shown an acceptable safety profile and promising preliminary efficacy in clinical studies with various solid tumor types. The non-clinical data presented here support the development of gedatolisib combined with CDK4/6 inhibitors and/or hormonal therapy for gynecologic cancer treatment. [ABSTRACT FROM AUTHOR]

Simple Summary: Treatments for colorectal cancer (CRC) largely rely on chemotherapeutics. For CRC treatment, irinotecan is widely used in various combinations with other drugs. Despite being an effective drug, irinotecan treatment often leads to drug resistance and tumor relapse. Mechanistically, it inhibits topoisomerase I and induces double-strand DNA breaks, which result in p53-dependent apoptosis. In the current study, we explored irinotecan-mediated, p53-independent pro- and anti-apoptotic effects. Our data show that irinotecan induces cyclin-dependent kinase inhibitors as well as anti-apoptotic proteins [osteopontin (OPN), survivin, and ISG15], the immunomodulatory molecule PD-L1, immune modulation mediated by proliferative NF-κB signaling, and metastatic markers (CD44, Sox2, Oct4, Snail, and Slug). Inhibition of OPN and/or NF-κB signaling potentiates irinotecan efficacy. The combination of OPN and/or an NF-κB inhibitor with irinotecan may lead to increased efficacy with reduced drug resistance. Among the most popular chemotherapeutic agents, irinotecan, regarded as a prodrug belonging to the camptothecin family that inhibits topoisomerase I, is widely used to treat metastatic colorectal cancer (CRC). Although immunotherapy is promising for several cancer types, only microsatellite-instable (~7%) and not microsatellite-stable CRCs are responsive to it. Therefore, it is important to investigate the mechanism of irinotecan function to identify cellular proteins and/or pathways that could be targeted for combination therapy. Here, we have determined the effect of irinotecan treatment on the expression/activation of tumor suppressor genes (including p15Ink4b, p21Cip1, p27Kip1, and p53) and oncogenes (including OPN, IL8, PD-L1, NF-κB, ISG15, Cyclin D1, and c-Myc) using qRT-PCR, Western blotting, immunofluorescence (IF), and RNA sequencing of tumor specimens. We employed stable knockdown, neutralizing antibodies (Abs), and inhibitors of OPN, p53, and NF-κB to establish downstream signaling and sensitivity/resistance to the cytotoxic activities of irinotecan. Suppression of secretory OPN and NF-κB sensitized colon cancer cells to irinotecan. p53 inhibition or knockdown was not sufficient to block or potentiate SN38-regulated signaling, suggesting p53-independent effects. Irinotecan treatment inhibited tumor growth in syngeneic mice. Analyses of allograft tumors from irinotecan-treated mice validated the cell culture results. RNA-seq data suggested that irinotecan-mediated activation of NF-κB signaling modulated immune and inflammatory genes in mice, which may compromise drug efficacy and promote resistance. In sum, these results suggest that, for CRCs, targeting OPN, NF-κB, PD-L1, and/or ISG15 signaling may provide a potential strategy to overcome resistance to irinotecan-based chemotherapy. [ABSTRACT FROM AUTHOR]

Simple Summary: Cervical cancer is the fourth most common cancer in women worldwide, with HPV being a prevalent cause. Recent automated signal detection methods in ISH assays have shown promise for detecting HPV DNA in cervical tissue. This study compared an ISH probe (Inform HPV II and III) to PCR assays in cervical tissue samples with varying degrees of cervical intraepithelial neoplasia (CIN) and normal cervix tissue. Findings indicated a significant association between ISH III levels and HPV outcomes; patients with positive outcomes had lower ISH III levels (MD = −7961.82, p = 0.005). However, automatic signal detection for ISH is limited in cervical tissue, making genotyping-based HPV testing more effective, as it allows for larger sample collection. ISH results should be interpreted alongside clinical history and morphology. Background: Cervical cancer is fourth the most common cancer in women worldwide. Due to the prevalence of human papillomavirus (HPV) in the population (80–90%), scientists are likely to discover even more associations of this pathogen with other diseases in the future. In recent years, In Situ Hybridization (ISH) assays that use automated signal-detecting methods in formalin-fixed, paraffin-embedded (FFPE) cervical tissue, such as the enzyme-categorized signal-detecting system, have shown a higher sensitivity. Objectives and Methods: To evaluate automatic signal detection of ISH assay for detecting HPV DNA, we compared the ability of an ISH probe, Inform HPV II and III (Ventana Medical Systems, Tucson, AZ), to that of PCR assays to detect HPV DNA in cervical tissue specimens with cervical intraepithelial neoplasia (CIN; CIN 1, 28 cases; CIN 2, 22 cases; and CIN 3, 20 cases) and normal cervix (2 cases). Results: Our findings showed a significant relation was confirmed between ISH III level and HPV outcome (positive/negative). Patients with positive HPV outcomes had significantly lower ISH III levels, MD = −7961.82 CI95 [−17,230.00; −199.21], p = 0.005. Conclusions: Automatic signal detection of ISH assay is not particularly applicable to cervical tissue material. A more useful method of confirming the presence of HPV in the cervix is the HPV test with genotyping, as it allows for collecting a larger amount of material from the cervical disc and canal. The interpretation of a positive or negative ISH test must be guided in the context of clinical history and morphology. [ABSTRACT FROM AUTHOR]

The search and development of effective sirtuin small molecule inhibitors (SIRTIs) continues to draw great attention due to their wide range of pharmacological applications. Based on SIRTs' involvement in different biological pathways, their ligands were investigated for many diseases, such as cancer, neurodegenerative disorders, diabetes, cardiovascular diseases and autoimmune diseases. The elucidation of a substantial number of SIRT2–ligand complexes is steering the identification of novel and more selective modulators. Among them, SIRT2 in the presence of the SirReal2 analog series was the most studied. On this basis, we recently reported structure-based analyses leading to the discovery of thiazole-based compounds acting as SIRT2 inhibitors (T1, SIRT2 IC50 = 17.3 µM). Herein, ligand-based approaches followed by molecular docking simulations allowed us to evaluate in silico a novel small series of thiazoles (3a–3d and 5a, 5d) as putative SIRT2 inhibitors. Results from the computational studies revealed comparable molecular interaction fields (MIFs) and docking positionings of most of these compounds with respect to reference SIRT2Is. Biochemical and biological assays validated this study and pointed to compound 5a (SIRT2 IC50 = 9.0 µM) as the most interesting SIRT2I that was worthy of further development as an anticancer agent. [ABSTRACT FROM AUTHOR]

SUMMARY: Presilphiperfolane‐type sesquiterpenes represent a unique group of atypical sesquiterpenoids characterized by their distinctive tricyclic structure. They have significant potential as lead compounds for pharmaceutical and agrochemical development. Herein, we utilized a transcriptomic approach to identify a terpene synthase (TPS) gene responsible for the biosynthesis of rare presilphiperfolane‐type sesquiterpenes in Inula lineariifolia, designated as IlTPS1. Through phylogenetic analysis, we have identified the evolutionary conservation of key motifs, including RR(x)8W, DDxxD, and NSE/DTE in IlTPS1, which are shared with other tricyclic sesquiterpene synthases in the TPS‐a subfamily of Asteraceae plants. Subsequent biochemical characterization of recombinant IlTPS1 revealed it to be a multiproduct enzyme responsible for the synthesis of various tricyclic sesquiterpene alcohols from farnesyl diphosphate (FPP), resulting in production of seven distinct sesquiterpenes. Mass spectrometry and nuclear magnetic resonance (NMR) spectrometry identified presilphiperfolan‐8β‐ol and presilphiperfol‐7‐ene as predominant products. Furthermore, biological activity assays revealed that the products from IlTPS1 exhibited a potent antifungal activity against Nigrospora oryzae. Our study represents a significant advancement as it not only functionally identifies the first step enzyme in presilphiperfolane biosynthesis but also establishes the heterologous bioproduction of these unique sesquiterpenes. Significance Statement: Our research has uncovered the molecular mechanism responsible for the synthesis of rare, bridged tricyclic sesquiterpenes by IlTPS1. Furthermore, we have established their bioproduction and assessed their antimicrobial properties. This study serves as a promising example of how unique sesquiterpene natural products can be harnessed and leveraged in the pharmaceutical and agrochemical sectors for various applications. [ABSTRACT FROM AUTHOR]

In this study, new peripherally substituted symmetric zinc and magnesium phthalocyanines (4 and 5) were successfully prepared by cyclotetramerization of the tetrahydropyrimidone (THPM)-linked phthalonitrile 3. The identity of the compounds were confirmed primarily through spectroscopic analysis including NMR, FT-IR, UV-Vis and MALDI-TOF mass spectroscopy. The photophysical and photochemical properties of the synthesized phthalocyanines (Pcs) were examined using UV–Vis absorption and fluorescence emission spectroscopy techniques. The quantum yields of singlet oxygen were found to be 0.50 and 0.33 for compounds 4 and 5 in DMSO, respectively. In addition to photo-physicochemical properties, the enhanced biological activities of compounds 4 and 5 were investigated using a range of biological assays, namely, antibiofilm, microbial cell viability, antioxidant, DNA cleavage, antimicrobial and photodynamic antimicrobial assays. The maximum DPPH inhibition of 4 and 5 was detected as 40.46% and 25.76% at 100 mg L−1, respectively. Fragmentation of the DNA molecule was observed at concentrations of 25 mg L−1, 50 mg L−1 and 100 mg L−1 for 4 and 5. Additionally, effective inhibition of microbial cell viability was observed with the targeted Pcs. The antibiofilm properties of these compounds were found to be concentration-dependent. The biofilm inhibition activities of 4 and 5 were found to be 96.01% and 92.04% for S. aureus, while they were 95.42% and 91.27%, for P. aeruginosa, respectively. The antimicrobial activities of 4 and 5 on different microorganisms were evaluated using the microdilution assay. In the case of photodynamic antimicrobial treatment, the newly synthesized Pcs showed more effective antimicrobial inhibition compared to the control. These findings suggest that compounds 4 and 5 can be used as promising photodynamic antimicrobial agents for the treatment of many diseases, particularly infectious diseases. [ABSTRACT FROM AUTHOR]

ABSTRACT A thiophosphoric triamide (SPT) ligand P(S)[NH‐2Py]3 was synthesized and utilized as a ligand to make novel binuclear and mononuclear Co2+ and Cd2+ complexes. The compounds are the first examples of discrete chelate SPT complexes with a [N]3P(S)‐based backbone in which the SPT acts as a flexible tridentate N,N,N‐donor or N,N,O‐donor ligand. The metal cation features a hexacoordinate environment M(N)3(Cl)3/M(N)4(O)2 adopting a distorted octahedral geometry. Hirshfeld surface analysis affirms that the pyridinyl nitrogen is a more dominant H‐bond acceptor than the thiophosphoryl sulfur atom. The inhibitory activity against the MDA‐MB‐231 cancer cell line in vitro indicated a good inhibitory effect for all compounds. The mononuclear Co2+ complex showed a stronger inhibitory activity against cancer cells than other studied compounds with an IC50 of 62 ± 1.4 nM. Antioxidant and antihemolytic activity of the compounds showed appreciable performance with the highest activities for the Cd2+ complex. A computational DNA binding study by molecular docking indicated a good affinity of the compounds with the DNA receptor by a mixed mode of interactions. Docking was also performed on SARS‐CoV‐2 and monkeypox (Mpox), resulting in binding energies (−5.5 to −8.9 kcal/mol) that are competitive with current effective drugs. [ABSTRACT FROM AUTHOR]

The escalating incidence of hospital infections due to antibiotic resistance necessitates the identification of alternative therapeutic agents such as probiotics. This study was designed to isolate and evaluate the efficacy of probiotics against Staphylococcus saprophyticus, a prevalent etiological agent of urinary tract infections (UTIs). A total of 100 S. saprophyticus strains were isolated from clinical samples and subjected to antibiotic susceptibility testing via the disc diffusion method. Concurrently, probiotic bacteria were isolated from Bulgarian cheese and shallot yogurt, and their antibacterial activity against S. saprophyticus strains was assessed. The inhibitory potential of probiotic supernatants was evaluated using microtiter plate assays, with the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) determined at a 1/2 dilution. Cytotoxicity was evaluated using the MTT assay, and high‐performance liquid chromatography (HPLC) was employed to analyze the concentrations of organic acids produced by the probiotics. The results revealed that all S. saprophyticus strains were resistant to tetracycline and doxycycline but susceptible to other antibiotics. Lactobacillus rhamnosus strains M and B demonstrated notable antibacterial and antibiofilm activity against S. saprophyticus isolates. These probiotics exhibited susceptibility to most antibiotics and lacked virulence factors, suggesting their safety for therapeutic use. The organic acids produced by the probiotics were identified as lactic acid, acetic acid, and formic acid. In conclusion, L. rhamnosus strains M and B exhibit potent antimicrobial properties against S. saprophyticus, indicating their potential as therapeutic agents for UTIs. Further research is warranted to validate these findings and explore the possibility of these probiotics in clinical applications. [ABSTRACT FROM AUTHOR]

ABSTRACT Aromia bungii is a serious wood borer damaging Prunus persica and other numerous economic tree species. Host plant volatiles play a key role in pest population monitoring and trapping. To ascertain the influence of volatile organic compounds emitted by P. persica at varying physiological statuses on the host‐searching behaviour of A. bungii adults, we initially collected and identified the volatiles emitted by P. persica leaves infested with A. bungii using gas chromatography–mass spectrometry. Subsequently, we evaluated the behavioural response of A. bungii adults to the crude extracts, single compounds and blends of these compounds by Y‐tube olfactometer bioassay. Results showed that volatiles emitted from P. persica infested with A. bungii larvae at Grade I were more attractive to females, but males were more attracted to volatiles from P. persica infested with A. bungii larvae at Grade IV. Five compounds were identified and showed relatively high levels from both infested plants at Grade I and Grade IV. Among them, 3‐carene, cis‐3‐hexen‐1‐ol and cis‐3‐hexenyl acetate were attractive to females, and a blend of the three volatiles in a ratio of 3:91:6 for Grade I infestation was more attractive to females than a ratio of 3:83:14 from Grade IV infestation. The compounds, 3‐carene, cis‐3‐hexenyl acetate, cis‐3‐hexene‐1‐ol and benzaldehyde, were attractive to males and a blend in a ratio of 3:81:13:3 from Grade IV infestation was more attractive to males than a ratio of 3:89:6:2 from Grade I infestation. The results show that the kinds and ratios of volatiles emitted from P. persica infested with A. bungii larvae play important roles in host‐searching behaviour of A. bungii adults. [ABSTRACT FROM AUTHOR]

Anthracnose of the tea plant (Camellia sinensis), caused by Colletotrichum spp., poses a significant threat to both the yield and quality of tea production. To address this challenge, researchers have looked to the application of endophytic bacteria as a natural alternative to the use chemical pesticides, offering potential for enhancing disease resistance and abiotic stress tolerance in tea plants. This study focused on identifying effective microbial agents to combat tea anthracnose caused by Colletotrichum fructicola. A total of 38 Bacillus-like strains were isolated from the tea rhizosphere, with 8 isolates showing substantial inhibitory effects against the mycelial growth of C. fructicola, achieving an average inhibition rate of 60.68%. Among these, strain T3 was particularly effective, with a 69.86% inhibition rate. Through morphological, physiological, and biochemical characterization, along with 16S rRNA gene phylogenetics analysis, these strains were identified as B. inaquosorum (T1 and T2), B. tequilensis (T3, T5, T7, T8, and T19), and B. spizizenii (T6). Biological and molecular assays confirmed that these strains could induce the expression of genes associated with antimicrobial compounds like iturin, fengycin, subtilosin, and alkaline protease, which effectively reduced the disease index of tea anthracnose and enhanced tea plant growth. In conclusion, this study demonstrates that B. inaquosorum, B. tequilensis, and B. spizizenii strains are promising biocontrol agents for managing tea anthracnose. [ABSTRACT FROM AUTHOR]

Background: Neuroinflammation is widely recognized as a key factor in the pathogenesis of Alzheimer's disease (AD), alongside ß-amyloid deposition and the formation of neurofibrillary tangles. The NLR family pyrin domain containing 3 (NLRP3) inflammasome, part of the innate immune system, has been implicated in the neuropathology of both preclinical amyloid and tau transgenic models. Activation of the NLRP3 pathway involves an initial priming step, which increases the expression of Nlrp3 and interleukin (IL)-1ß, followed by the assembly of the NLRP3 inflammasome complex, comprising NLRP3, ASC, and caspase-1. This assembly leads to the proteolytic maturation of the pro-inflammatory cytokines IL-1ß and IL-18. Additionally, the NLRP3 inflammasome induces Gasdermin D (GSDMD) cleavage, forming membrane pores through which IL-1ß and IL-18 are secreted. Inhibition of NLRP3 has been shown to enhance plaque clearance by modulating microglial activation. Furthermore, blocking NLRP3 in tau transgenic mice has been found to reduce tau phosphorylation by affecting the activity of certain tau kinases and phosphatases. Methods: In this study, organotypic brain slice cultures from P301S transgenic mice were treated with lipopolysaccharide (LPS) plus nigericin as a positive control or exposed to tau seeds (K18) to evaluate NLRP3 inflammasome activation. The effect of tau seeding on NLRP3 activity was further examined using Meso Scale Discovery (MSD) assays to measure IL1ß secretion levels in the presence and absence of NLRP3 inhibitors. The role of NLRP3 activity was investigated in fullbody Nlrp3 knockout mice crossbred with the tau transgenic P301S model. Additionally, full-body and microglia-selective Gsdmd knockout mice were crossbred with P301S mice, and tau pathology and neurodegeneration were evaluated at early and late stages of the disease using immunohistochemistry and biochemical assays. Results: Activation of the NLRP3 pathway was observed in the mouse organotypic slice culture (OSC) model following stimulation with LPS and nigericin or exposure to tau seeds. However, Nlrp3 deficiency did not mitigate tauopathy or neurodegeneration in P301S mice in vivo, showing only a minor effect on plasma neurofilament (NF-L) levels. Consistently, Gsdmd deficiency did not alter tau pathology in P301S mice. Furthermore, neither full-body nor microglia-selective Gsdmd deletion had an impact on neuronal pathology or the release of pro-inflammatory cytokines. Conclusion: The absence of key components of the NLRP3 inflammasome pathway did not yield a beneficial effect on tau pathology or neurodegeneration in the preclinical Tau-P301S mouse model of AD. Nonetheless, organotypic slice cultures could serve as a valuable ex vivo mechanistic model for evaluating NLRP3 pathway activation and pharmacological inhibitors. [ABSTRACT FROM AUTHOR]

This study explores and verifies potential molecular targets through which KRAS mutations regulate the colonization of Fusobacterium nucleatum (FN) in colorectal cancer (CRC). This study combined multiple bioinformatics methods and biological assays. Through The Cancer Genome Atlas, Gene Expression Omnibus, Human Protein Atlas, immunohistochemistry, and co‐culture assays, we further confirmed the differential expression of SERTAD4 in CRC. We delved deeper into examining how expression of SERTAD4 is linked with immune cell infiltration and the enrichment of potential pathways. Lastly, through bacterial phenotypic assays, we validated the function of SERTAD4. As a molecule associated with KRAS mutations and FN infection, the expression levels of SERTAD4 were downregulated in CRC. The diagnostic efficacy of SERTAD4 for CRC is not inferior to that of CEA. Low expression of SERTAD4 is associated with poorer overall survival in CRC. Correlation analysis found that increased expression of SERTAD4 is associated with various immune cell infiltrations and immune checkpoint genes. Finally, bacterial adhesion and invasion assays verify that SERTAD4 inhibits the adhesion and invasion abilities of FN in CRC. This study demonstrates that SERTAD4 exerts a protective role in CRC by inhibiting the colonization of FN. [ABSTRACT FROM AUTHOR]

Beech mushrooms (Hypsizygus marmoreus) are edible mushrooms commercially used in South Korea. They can be classified into white and brown according to their pigmentation. This study analyzed the metabolites and biological activities of these mushrooms. Specifically, 42 metabolites (37 volatiles, two phenolics, and three carbohydrates) were quantified in white beech mushrooms, and 47 (42 volatiles, two phenolics, and three carbohydrates) were detected in brown mushrooms. The major volatiles detected were hexanal, pentanal, 1-hexanol, and 1-pentanol. Brown mushrooms contained higher levels of hexanal (64%) than white mushrooms (35%), whereas white mushrooms had higher levels of pentanal (11%) and 1-pentanol (3%). Most volatiles were more abundant in white mushrooms than in brown mushrooms. Furthermore, brown beech mushrooms had a higher phenolic content than white mushrooms. Biological assays revealed that both types of mushroom demonstrated anti-microbial activities against bacterial and yeast pathogens and weak DPPH scavenging activity. The extracts from both mushrooms (50 μg/mL) also exhibited strong anti-inflammatory properties. Brown mushroom extracts showed higher antioxidant, anti-microbial, and anti-inflammatory properties than white mushroom extracts. This study reported that the differences in phenotype, taste, and odor were consistent with the metabolite differences between white and brown beech mushrooms, which have high nutritional and biofunctional values. [ABSTRACT FROM AUTHOR]

Iron (Fe) is a key limiting nutrient for Southern Ocean phytoplankton. Input of Fe into the Southern Ocean is projected to change due to global warming, yet the combined effects of a concurrent increase in temperature with dissolved Fe (dFe) addition on phytoplankton growth and community composition have not been extensively studied. To improve our understanding of how Antarctic phytoplankton communities respond to Fe and enhanced temperature, we performed four full factorial onboard bioassays under trace-metal-clean conditions with phytoplankton communities from different regions of the Weddell Sea and the Amundsen Sea in the Southern Ocean. Treatments consisted of 2 nM Fe addition with 2 °C warming (TF), Fe addition at in situ temperature (F) + 2 °C warming with no Fe addition (T) and a control at in situ temperature with no Fe addition (control, C). Temperature had a limited effect by itself but boosted the positive response of the phytoplankton to Fe addition. Photosynthetic efficiency, phytoplankton abundances and chlorophyll a concentrations typically increased (significantly) with Fe addition (F and/or TF treatment), and the phytoplankton community generally shifted from haptophytes to diatoms upon Fe addition. The < 20 µ m phytoplankton fraction displayed population-specific growth responses, resulting in a pronounced shift in community composition and size distribution (mainly towards larger-sized phytoplankton) for the F and TF treatments. Such a distinct enhanced impact of dFe supply with warming on Antarctic phytoplankton size, growth and composition will likely affect trophic transfer efficiency and ecosystem structure, with potential significance for the biological carbon pump. [ABSTRACT FROM AUTHOR]

Background: Many diseases are increasingly recognized as public health concerns worldwide because of the increasing incidence of multidrug-resistant bacteria. Recently, interest in the use of indigenous medicinal plants to treat infectious illnesses has increased, highlighting the need to find new bioactive phytochemicals. Ajuga integrifolia is a plant commonly utilized in traditional drugs to treat a wide range of diseases, although its effectiveness has not been scientifically validated. The present study aimed to evaluate the total phenolic and flavonoid contents and assess the biological activities of A. integrifolia leaf extracts produced via different solvent systems. Methods: Soxhlet extraction was employed to obtain crude extracts from different solvents (methanol, ethanol and water). The 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric-reducing power assays were used to measure the antioxidant activity, and the antibacterial activity of the extract was evaluated on the basis of its minimum inhibitory concentration (MIC) against two gram-negative bacteria (Escherichia coli (ATCC-25922) and Pseudomonas aeruginosa (ATCC-43495)) and two gram-positive bacteria (Staphylococcus aureus (ATCC-25923) and Streptococcus pyogenes (ATCC-19615)) via the agar disk-diffusion technique. Results: A significant amount of total phenolic content (TPC) and flavonoid content (TFC) were present in all the extracts. The extracts presented powerful antioxidant activity in all the assays. The disc diffusion and MIC results revealed the ability of the methanol and ethanol extracts of A. integrifolia leaves to inhibit S. aureus growth at a concentration of 3.125 mg/mL. However, the water extracts were ineffective against E. coli and P. aeruginosa. Conclusions: These findings indicate that A. integrifolia leaf extracts have reasonable biological activities. These findings underscore the importance of A. integrifolia leaves as a source of health benefits. [ABSTRACT FROM AUTHOR]