An RNA Helicase DHX33 Inhibitor Shows Broad Anticancer Activity via Inducing Ferroptosis in Cancer Cells.

RNA helicase DHX33 has been identified as a critical factor promoting cancer development. In the present study, a previously developed small molecule inhibitor for DHX33, KY386, was found to robustly kill cancer cells via a new path, the ferroptosis pathway. Mechanistically, DHX33 promotes the expression of critical players in lipid metabolism including FADS1, FADS2, and SCD1 genes, thereby sensitizing cancer cells to ferroptosis mediated cell death. Our study reveals a novel mechanism of DHX33 in promoting tumorigenesis and highlights that pharmacological targeting DHX33 can be a feasible option in human cancers. Normally differentiated cells are insensitive to DHX33 inhibition, and DHX33 inhibitors have little cellular toxicity in vitro and in vivo. Our studies demonstrated that DHX33 inhibitors can be promising anticancer agents with great potential for cancer treatment.

Exogenous iron caused osteocyte apoptosis, increased RANKL production, and stimulated bone resorption through oxidative stress in a murine model.

Iron overload is a risk factor for osteoporosis due to its oxidative toxicity. Previous studies have demonstrated that an excessive amount of iron increases osteocyte apoptosis and receptor activator of nuclear factor κ-B ligand (RANKL) production, which stimulates osteoclast differentiation in vitro. However, the effects of exogenous iron supplementation-induced iron overload on osteocytes in vivo and its role in iron overload-induced bone loss are unknown. This work aimed to develop an iron overloaded murine model of C57BL/6 mice by intraperitoneal administration of iron dextran for two months. The iron levels in various organs, bone, and serum, as well as the microstructure and strength of bone, apoptosis of osteocytes, oxidative stress in bone tissue, and bone formation and resorption, were assessed. The results showed that 2 months of exogenous iron supplementation significantly increased iron levels in the liver, spleen, kidney, bone tissue, and serum. Iron overload negatively affected bone microstructure and strength. Osteocyte apoptosis and empty lacunae rate were elevated by exogenous iron. Iron overload upregulated RANKL expression but had no significant impact on osteoprotegerin (OPG) and sclerostin levels. Static and dynamic histologic analyses and serum biochemical assay showed that iron overload increased bone resorption without significantly affecting bone formation. Exogenous iron promoted oxidative stress in osteocytes in vivo and in vitro. Additional supplementation of iron chelator (deferoxamine) or N-acetyl-L-cysteine (NAC) partially alleviated bone loss, osteocyte apoptosis, osteoclast formation, and oxidative stress due to iron overload. These findings, in line with prior in vitro studies, suggest that exogenous iron supplementation induces osteoclastogenesis and osteoporosis by promoting osteocyte apoptosis and RANKL production via oxidative stress.

Development and clinical validation of a dual ddPCR assay for detecting carbapenem-resistant Acinetobacter baumannii in bloodstream infections.

Objective: Acinetobacter baumannii (A. baumannii, AB) represents a major species of Gram-negative bacteria involved in bloodstream infections (BSIs) and shows a high capability of developing antibiotic resistance. Especially, carbapenem-resistant Acinetobacter baumannii (CRAB) becomes more and more prevalent in BSIs. Hence, a rapid and sensitive CRAB detection method is of urgent need to reduce the morbidity and mortality due to CRAB-associated BSIs. Methods: A dual droplet digital PCR (ddPCR) reaction system was designed for detecting the antibiotic resistance gene OXA-23 and AB-specific gene gltA. Then, the specificity of the primers and probes, limit of detection (LOD), linear range, and accuracy of the assay were evaluated. Furthermore, the established assay approach was validated on 37 clinical isolates and compared with blood culture and drug sensitivity tests. Results: The dual ddPCR method established in this study demonstrated strong primer and probe specificity, distinguishing CRAB among 21 common clinical pathogens. The method showed excellent precision (3 × 10-4 ng/µL, CV < 25%) and linearity (OXA-23: y = 1.4558x + 4.0981, R2 = 0.9976; gltA: y = 1.2716x + 3.6092, R2 = 0.9949). While the dual qPCR LOD is 3 × 10-3 ng/µL, the dual ddPCR's LOD stands at 3 × 10-4 ng/µL, indicating a higher sensitivity in the latter. When applied to detect 35 patients with BSIs of AB, the results were consistent with clinical blood culture identification and drug sensitivity tests. Conclusion: The dual ddPCR detection method for OXA-23 and gltA developed in this study exhibits good specificity, excellent linearity, and a higher LOD than qPCR. It demonstrates reproducibility even for minute samples, making it suitable for rapid diagnosis and precision treatment of CRAB in BSIs.

Lead Optimization of Butyrolactone I as an Orally Bioavailable Antiallergic Agent Targeting FcγRIIB.

Food allergy (FA) poses a growing global food safety concern, yet no effective cure exists in clinics. Previously, we discovered a potent antifood allergy compound, butyrolactone I (BTL-I, 1), from the deep sea. Unfortunately, it has a very low exposure and poor pharmacokinetic (PK) profile in rats. Therefore, a series of structural optimizations toward the metabolic pathways of BTL-I were conducted to provide 18 derives (2-19). Among them, BTL-MK (19) showed superior antiallergic activity and favorable pharmacokinetics compared to BTL-I, being twice as potent with a clearance (CL) rate of only 0.5% that of BTL-I. By oral administration, Cmax and area under the concentration-time curve (AUC0-∞) were 565 and 204 times higher than those of BTL-I, respectively. These findings suggest that butyrolactone methyl ketone (BTL-BK) could serve as a drug candidate for the treatment of FAs and offer valuable insights into optimizing the druggability of lead compounds.

A new AI-based approach for automatic identification of tea leaf disease using deep neural network based on hybrid pooling.

The degree of production efficiency and the quality of the commodities produced may both be directly impacted by the presence of illnesses in tea leaves. These days, this procedure may be automated with the use of artificial intelligence tools, and a number of approaches have been put out to satisfy these needs. Nonetheless, current research efforts have focused on improving diagnosis accuracy and expanding the variety of illnesses that might affect tea leaves. In this article, a new method is proposed for accurately diagnosing tea leaf diseases using artificial intelligence techniques. In the proposed method, the input images are preprocessed to remove redundant information. Then, a hybrid pooling-based Convolutional Neural Network (CNN) is employed to extract image features. In this method, the pooling layers of the CNN model are randomly adjusted based on either max pooling or average pooling functions. This strategy can enhance the efficiency of the CNN-based feature extraction model. In this method, the pooling layers of the CNN model are randomly adjusted based on either max pooling or average pooling functions. This strategy can enhance the efficiency of the CNN-based feature extraction model. After feature extraction, a weighted Random Forest (WRF) model is used for the detection of tea leaf diseases. The outputs of the decision tree models and their corresponding weights are used to identify tea leaf illnesses in this classification model, where each tree in the random forest is given a weight depending on how well it performs. The Cuckoo Search Optimization (CSO) method is used in the proposed classification model to give a weight to each tree. Tea Sickness Dataset (TSD) has been used as the basis for evaluating the suggested method's effectiveness. The findings show that the suggested approach has an average accuracy of 92.47% in identifying seven different forms of tea leaf illnesses. Additionally, the recall and accuracy metrics indicate results of 92.35 and 92.26, respectively, indicating improvements over earlier techniques.

Preparation, characterization, antioxidant, and antifungal activity of phenyl/indolyl-acyl chitooligosaccharides.

In this study, carboxylic acids compounds were grafted onto chitooligosaccharides to prepare seven phenyl/indolyl-acyl chitooligosaccharides derivatives. The structures of the derivatives were characterized by IR spectroscopy, 13C NMR and elemental analysis. Meanwhile, antioxidant activities in vitro of the novel derivatives were analyzed. Compared to COS and carboxylic acid, the derivatives showed higher scavenging capacity for superoxide anion and DPPH radicals, with scavenging rates of 59.39% and 94.86%, respectively. The hydroxyl radical scavenging ability of the derivatives was only 18.89%. The antifungal activities of chitooligosaccharide derivatives against Diaporthe batatas and Phytophthora capsici were studied by the growth rate method. Compared with chitooligosaccharide itself, derivatives were inhibited by 97.77% and 100%. The above results showed that chitooligosaccharide derivatives have good biocompatibility and can be used in food, agriculture and medicine.

Poly-ß-cyclodextrin strengthen Pr6O11 porous oxidase mimic for dual-channel visual recognition of bioactive cysteine and Fe2.

To conveniently monitor bioactive cysteine (Cys) and Fe2+ in practice, a kind of poly-ß-cyclodextrin strengthen praseodymium oxide (Pr6O11) porous oxidase mimic (p-ß-CD@Pr6O11) was constructed by virtue of the strong coordination between nano Pr6O11 and poly-ß-cyclodextrin substrate. After its microstructure and physicochemical property were characterized in detail, it was noted that porous p-ß-CD@Pr6O11 exhibited excellent enzyme-like catalytic activity to accelerate the oxidation of 3,3',5,5,'-tetramethylbanzidine (TMB) and 2,2'-azinobis (3-ethylbenzo-thiazoline-6-sulfonic acid) ammonium salt (ABTS) with significant color-enhancement effect in the air. Based on the signal amplification, trace Cys could exclusively deteriorate the UV-vis absorbance at 653 nm of p-ß-CD@Pr6O11-TMB and Fe2+ alter the one at 729 nm of p-ß-CD@Pr6O11-ABTS with visual color changes. Under the optimized conditions, the proposed p-ß-CD@Pr6O11-TMB and p-ß-CD@Pr6O11-ABTS systems were successfully applied for dual-channel monitoring of Cys in Cys capsules and fetal bovine serum and Fe2+ in agricultural products with quite low detection limits, i.e., 7.8×10-9 mol·L-1 for Cys and 6.93×10-8 mol·L-1 (S/N=3) for Fe2+, respectively. The synergetic-enhancement detection mechanisms to Cys and Fe2+ were also proposed.

Iron-Catalyzed C(sp3)-C(sp3) Coupling to Construct Quaternary Carbon Centers.

The construction of quaternary carbon centers via C-C coupling protocols remains challenging. The coupling of tertiary C(sp3) with secondary or tertiary C(sp3) counterparts has been hindered by pronounced steric clashes and many side reactions. Herein, we have successfully developed a type of bisphosphine ligand iron complex-catalyzed coupling reactions of tertiary alkyl halides with secondary alkyl zinc reagents and efficiently realized the coupling reaction between tertiary C(sp3) and secondary C(sp3) with high selectivity for the initial instance, which provided an efficient method for the construction of quaternary carbon centers with high steric hindrance. The combination of an iron catalyst and directing group of the substrate makes the great challenging transformation possible.

Gain-of-Function KIDINS220 Variants Disrupt Neuronal Development and Cause Cerebral Palsy.

BACKGROUND: Kinase D-interacting substrate of 220 kDa (KIDINS220) is a multifunctional scaffolding protein essential for neuronal development. It has been implicated in neurological diseases with either autosomal dominant (AD) or autosomal recessive (AR) inheritance patterns. The molecular mechanisms underlying the AR/AD dual nature of KIDINS220 remain elusive, posing challenges to genetic interpretation and clinical interventions. Moreover, increased KIDINS220 exhibited neurotoxicity, but its role in neurodevelopment remains unclear. OBJECTIVE: The aim was to investigate the genotype-phenotype correlations of KIDINS220 and elucidate its pathophysiological role in neuronal development. METHODS: Whole-exome sequencing was performed in a four-generation family with cerebral palsy. CRISPR/Cas9 was used to generate KIDINS220 mutant cell lines. In utero electroporation was employed to investigate the effect of KIDINS220 variants on neurogenesis in vivo. RESULTS: We identified in KIDINS220 a pathogenic nonsense variant (c.4177C > T, p.Q1393*) that associated with AD cerebral palsy. We demonstrated that the nonsense variants located in the terminal exon of KIDINS220 are gain-of-function (GoF) variants, which enable the mRNA to escape nonsense-mediated decay and produce a truncated yet functional KIDINS220 protein. The truncated protein exhibited significant resistance to calpain and consequently accumulated within cells, resulting in the hyperactivation of Rac1 and defects in neuronal development. CONCLUSIONS: Our findings demonstrate that the location of variants within KIDINS220 plays a crucial role in determining inheritance patterns and corresponding clinical outcomes. The proposed interaction between Rac1 and KIDINS220 provides new insights into the pathogenesis of cerebral palsy, implying potential therapeutic perspectives. © 2023 International Parkinson and Movement Disorder Society.

DHX33 mediates p53 to regulate mevalonate pathway gene transcription in human cancers.

Tumor suppressor p53 is frequently null or mutated in human cancers. Here in this study, DHX33 protein was found to be induced in p53 null cells in vitro, and in p53 mutant lung tumorigenesis in vivo. Cholesterol metabolism through mevalonate pathway is pivotal for cell proliferation and is frequently altered in human cancers. Mice carrying mutant p53 and KrasG12D alleles showed upregulation of mevalonate pathway gene expression. However upon DHX33 loss, their upregulation was significantly debilitated. Additionally, in many human cancer cells, DHX33 knockdown caused inhibition of mavelonate pathway gene transcription. We propose DHX33 locates downstream of mutant p53 and Ras to regulate mevalonate pathway gene transcription and thereby supports cancer development in vivo.

Preparation, characterization, antioxidant and antifungal activities of benzoic acid compounds grafted onto chitosan.

The current study created three novel chitosan derivatives named BACS, PIBACS, and MHBACS by grafting benzoic acid (BA), p-isopropyl benzoic acid (PIBA), and m-hydroxybenzoic acid (MHBA) onto chitosan (CS). The structures of the derivatives were investigated using infrared spectroscopy (FT-IR) and nuclear magnetic resonance (13C NMR). The derivatives were discovered to be 45.06 %-60.49 % substituted using elemental analysis (EA). Based on the findings of in vitro antioxidant experiments (hydroxyl radical scavenging activity, superoxide anion radical scavenging activity, and DPPH radical scavenging activity), all of the derivatives had a higher hydroxyl radical scavenging activity than the chitosan raw material. MHBACS scavenged (31.02 ± 0.90)% of hydroxyl radicals at 0.5 mg/mL, 28.69 % more than chitosan raw. The derivatives scavenged more superoxide anion radicals than the chitosan feedstock at a particular concentration. For instance, at a test dose of 0.2 mg/mL, the scavenging rate of MHBACS on superoxide anion radicals was 7.75 % greater than that of chitosan raw materials. DPPH radical scavenging activity, on the other hand, was not as competent as chitosan feedstock. The growth rate approach was used to assess the potential of the three derivatives to inhibit the development of four phytopathogenic fungi. Chitosan derivatives have better antifungal efficacy than chitosan raw materials. PIBACS, MHBACS, BACS, and Wuyiencin inhibited Phytophthora capsici by (98.03 ± 1.95)%, (81.73 ± 1.63)%, (66.38 ± 1.81)%, and (93.01 ± 2.69)%, respectively, at 1.0 mg/mL. PIBACS had a higher inhibitory impact on Phytophthora capsici than the positive control. Based on the evidence presented above, it is reasonable to conclude that the addition of benzoic acid molecules increased the antioxidant and antifungal capabilities of chitosan.

Association of dietary inflammatory index with all-cause and cardiovascular disease mortality in hyperuricemia population: A cohort study from NHANES 2001 to 2010.

Dietary management is a crucial component of non-pharmacological treatment for hyperuricemia, yet there is a paucity of research on the impact of dietary habits on the survival outcomes of individuals with hyperuricemia. The objective of this study is to examine the association between dietary inflammatory index (DII) and the all-cause and cardiovascular disease (CVD) mortality in individuals with hyperuricemia. This study included 3093 adult participants from National Health and Nutrition Examination Survey (NHANES) 2001 to 2010. Participants were categorized into 4 groups based on quartiles of DII to demonstrate data characteristics, with sample weights considered. The relationship between DII and the risk of hyperuricemia was examined using multivariable logistic regression models. Kaplan-Meier models and Cox proportional hazards models were employed to assess the relationship between DII levels and the all-cause mortality in individuals with hyperuricemia, with the non-linear relationship tested using restricted cubic splines (RCS). Competing risk models were employed to investigate the association between DII levels and the CVD mortality in individuals diagnosed with hyperuricemia. Subgroup and sensitivity analysis were performed to confirm the robustness and reliability of the findings. Among the participants, 47.95% were aged over 60 years. A positive association observed between the highest quartile of DII level and the incidence of hyperuricemia (OR: 1.34, CI [1.13, 1.57]). Elevated DII levels were correlated with increased all-cause mortality (P value < .001) and CVD mortality (P value < .001) in participants. In comparison to the lowest quartile, the highest quartile of DII exhibited a 31% rise in all-cause mortality (HR: 1.31, CI [1.01, 1.68]) and a 50% increase in CVD mortality (HR: 1.50, CI [1.00, 2.26]). No indication of a nonlinear association between DII levels and all-cause mortality (p-non-linear = .43). These findings indicate a positive correlation between the pro-inflammatory diet and the incidence of hyperuricemia. Additionally, a pro-inflammatory diet may elevate the all-cause and CVD mortality in individuals with hyperuricemia.

Development of small molecule inhibitors targeting RNA helicase DHX33 as anti-cancer agents.

RNA helicase DHX33 has been identified to be a critical factor in promoting cancer development. Genetic deletion of DHX33 significantly blocks tumorigenesis. Importantly, its helicase activity was found to be pivotal for exerting cellular functions. Herein we used a helicase-based high throughput screening (HTS) to discover DHX33 inhibitors from Chembridge chemical library containing 15,000 small molecules. We identified a hit compound containing benzimidazole ring that demonstrated activity against DHX33 with certain selectivity. Further structural optimization led to the design and synthesis of a series of analog inhibitors. Considering the potential role of DHX33 in cancer development, the compounds were evaluated based on the cytotoxicity activity in U251-MG cancer cells in vitro. Among them, compound IVa (KY386) was identified to be a selective inhibitor for DHX33 helicase with potent anti-cancer activity and moderate metabolic stability. These results support the promising role of DHX33 inhibitors for development of novel anti-cancer drugs.

ILC1-derived IFN-γ regulates macrophage activation in colon cancer.

BACKGROUND: Tumor-associated macrophages (TAMs) are an important subset of innate immune cells in the tumor microenvironment, and they are pivotal regulators of tumor-promoting inflammation and tumor progression. Evidence has proven that TAM numbers are substantially increased in cancers, and most of these TAMs are polarized toward the alternatively activated M2 phenotype; Thus, these TAMs strongly promote the progression of cancer diseases. Type 1 innate lymphocytes (ILC1s) are present in high numbers in intestinal tissues and are characterized by the expression of the transcription factor T-bet and the secretion of interferon (IFN)-γ, which can promote macrophages to polarize toward the classically activated antitumor M1 phenotype. However, the relationship between these two cell subsets in colon cancer remains unclear. METHODS: Flow cytometry was used to determine the percentages of M1-like macrophages, M2-like macrophages and ILC1s in colon cancer tissues and paracancerous healthy colon tissues in the AOM/DSS-induced mouse model of colon cancer. Furthermore, ILC1s were isolated and bone marrow-derived macrophages were generated to analyze the crosstalk that occurred between these cells when cocultured in vitro. Moreover, ILC1s were adoptively transferred or inhibited in vivo to explore the effects of ILC1s on tumor-infiltrating macrophages and tumor growth. RESULTS: We found that the percentages of M1-like macrophages and ILC1s were decreased in colon cancer tissues, and these populations were positively correlated. ILC1s promoted the polarization of macrophages toward the classically activated M1-like phenotype in vitro, and this effect could be blocked by an anti-IFN-γ antibody. The in vivo results showed that the administration of the Group 1 innate lymphocyte-blocking anti-NK1.1 antibody decreased the number of M1-like macrophages in the tumor tissues of MC38 tumor-bearing mice and promoted tumor growth, and adoptive transfer of ILC1s inhibited tumors and increased the percentage of M1-like macrophages in MC38 tumor-bearing mice. CONCLUSIONS: Our studies preliminarily prove for the first time that ILC1s promote the activation of M1-like macrophages by secreting IFN-γ and inhibit the progression of colon cancer, which may provide insight into immunotherapeutic approaches for colon cancer.

A Frequency-Dependent Dynamic Electric-Mechanical Network for Thin-Wafer Piezoelectric Transducers Polarized in the Thickness Direction: Physical Model and Experimental Confirmation.

This paper is concerned with electric-acoustic/acoustic-electric conversions of thin-wafer piezoelectric transducers polarized in the thickness direction. By introducing two mechanical components with frequency-dependent values, i.e., radiation resistance and radiation mass, into the equivalent circuit of the thin-wafer piezoelectric transducer, we established a frequency-dependent dynamic mechanic-electric equivalent network with four terminals for an arbitrary given frequency, an enhancement from the conventional circuit networks. We derived the analytic expressions of its electric-acoustic and acoustic-electric conversion impulse responses using the four-terminal equivalent circuit to replace the traditional six-terminal equivalent circuit for a thin-wafer transducer with harmonic vibrational motion. For multifrequency electrical/acoustic signals acting on the transducer, we established parallel electric-acoustic/acoustic-electric conversion transmission networks. These two transmission network models have simple structures and clear physical and mathematical descriptions of thin-wafer transducers for electric-acoustic/acoustic-electric conversion when excited by a multifrequency electric/acoustic signal wavelet. The calculated results showed that the transducer's center frequency shift relates to its mechanical load and vibration state. The method reported in this paper can be applied to conventional-sized and small-sized piezoelectric transducers with universal applicability.

RNA helicase DHX33 regulates HMGB family genes in human cancer cells.

RNA helicase DHX33 has been shown to be aberrantly expressed in various human cancers, however, its role in tumorigenesis remains incompletely understood. In this report, we uncovered that a family of DNA architecture proteins, HMGBs, can be regulated by DHX33 in cancer cells but not in normal cells. Specifically, DHX33 knockdown caused the downregulation of HMGBs at the levels of both gene transcription and protein expression. Notably, in RAS driven lung tumorigenesis, nuclear HMGBs proteins can be induced via DHX33. When DHX33 was knocked out, HMGBs overexpression was debilitated. Mechanistically, DHX33 was found to bind to the promoters of HMGB family genes and regulated their transcription through demethylation on gene promoters. Our study reveals a novel mechanism for DHX33 to promote tumorigenesis and highlights its therapeutic value in human cancers.

Divergent Total Syntheses of Illicium Sesquiterpenes through Late-Stage Skeletal Reorganization.

We disclose unified, protecting-group-free, bioinspired divergent total syntheses of eight allo-cedrane and seco-prezizaane Illicium sesquiterpenes and formal syntheses of five anislactone sesquiterpenes. The efficiency of our approach derives from rapid access to the 15-carbon tricyclic carboxylic acid through cationic epoxide-ene cyclization and HAT oxygenation, transformation of this intermediate into three distinct tricyclic precursors via Lewis acid-mediated skeletal reorganizations, subsequent programmed oxidation level enhancement, and a biomimetic oxidation-initiated skeletal rearrangement cascade. Consequently, we created a synthetic correlation map of the three most prevalent Illicium sesquiterpene families.

Identification of potential HDAC11 deacylase substrates by affinity pulldown MS.

Lysine fatty acylation is a protein posttranslational modification (PTM) that has been linked to various important biological processes. HDAC11, the sole member of class IV of histone deacetylases (HDACs), has been shown to have high lysine defatty-acylase activity. In order to better understand the functions of lysine fatty acylation and its regulation by HDAC11, it is important to identify the physiological substrates of HDAC11. This can be achieved through profiling the interactome of HDAC11 using a stable isotope labeling with amino acids in cell culture (SILAC) proteomics strategy. Here we describe a detailed method on using SILAC to identify the interactome of HDAC11. This method can be similarly used to identify the interactome, and thus potential substrates, of other PTM enzymes.