Tomato NADPH oxidase SlWfi1 interacts with the effector protein RipBJ of Ralstonia solanacearum to mediate host defence.

Reactive oxygen species (ROS) play a crucial role in regulating numerous functions in organisms. Among the key regulators of ROS production are NADPH oxidases, primarily referred to as respiratory burst oxidase homologues (RBOHs). However, our understanding of whether and how pathogens directly target RBOHs has been limited. In this study, we revealed that the effector protein RipBJ, originating from the phytopathogenic bacterium Ralstonia solanacearum, was present in low- to medium-virulence strains but absent in high-virulence strains. Functional genetic assays demonstrated that the expression of ripBJ led to a reduction in bacterial infection. In the plant, RipBJ expression triggered plant cell death and the accumulation of H2O2, while also enhancing host defence against R. solanacearum by modulating multiple defence signalling pathways. Through protein interaction and functional studies, we demonstrated that RipBJ was associated with the plant's plasma membrane and interacted with the tomato RBOH known as SlWfi1, which contributed positively to RipBJ's effects on plants. Importantly, SlWfi1 expression was induced during the early stages following R. solanacearum infection and played a key role in defence against this bacterium. This research uncovers the plant RBOH as an interacting target of a pathogen's effector, providing valuable insights into the mechanisms of plant defence.

Synthesis of 2-(pyridylvinyl)chromen-4-ones and their N-oxide analogs for assessment of their biological activities as anticancer agents.

2-Styrylchromones have been shown to possess a broad spectrum of biological activities. Replacing the carbon atom in 2-styrylchromones with a nitrogen atom in the benzene rings forms 2-(pyridylvinyl)chromen-4-ones (aza-2-styrylchromones). We have synthesized a series of novel 2-(pyridylvinyl)chromen-4-ones and their pyridine N-oxides to evaluate them as potential anticancer agents against human non-small-cell lung cancer cells (A549). Among the 18 synthesized molecules, compounds 18 and 8a exhibited comparable inhibitory effects to 5-fluorouracil and showed no toxicity against normal cells.

Synthesis of selenophene-containing flavonols and 2-styrylchromones: Evaluation of their activities compared with selenophene-containing chalcones as potential anticancer agents.

Previously, we documented the synthesis and assessed the biological effects of chalcones containing selenium against HT-29 human colorectal adenocarcinoma cells, demonstrating their significant potential. As research on selenium-containing flavonoids remains limited, this article outlines our design and synthesis of three selenium-based flavonols and three 2-styrylchromones. We conducted evaluations of these compounds to determine their impact on human lung cancer cells (A549, H1975, CL1-0, and CL1-5) and their influence on normal lung fibroblast MRC5 cells. Additionally, we included selenium-based chalcones in our testing for comparative purposes. Our findings highlight that the simplest compound, designated as compound 1, exhibited the most promising performance among the tested molecules.

Treatment Efficacy of Internet Gaming Disorder With Attention Deficit Hyperactivity Disorder and Emotional Dysregulaton.

BACKGROUND: Recent youth with Attention Deficit Hyperactivity Disorder (ADHD) noticed emotional dysregulation if they had Internet Gaming Disorder (IGD). This study aims to understand the treatment efficacy of IGD with ADHD and emotional dysregulaton. METHOD: A total of 101 ADHD youths were recruited. We used the Chen Internet Addiction Scale and IGD criteria of the diagnotsic statistical manual (DSM)-5 to confirm IGD. The Swanson, Nolan, and Pelham questionnaire Version IV was used for symptoms of ADHD and oppositional defiant disorder. Disruptive mood dysregulation disorder was assessed by psychiatrist. RESULTS: There is a new phenomenon that emotional dysregulation has been frequently noticed in severely gaming-addicted ADHD youth. Treatment efficacy of IGD is good when the underlying symptom of ADHD is controlled. Symptom scores of disruptive mood dysregulation (DMDD) were significantly reduced by 71.9%, 74.8%, and 84.4% at week 2, 3, and 4, respectively (P ≤ .001) after adjusting baseline symptom severity. CONCLUSION: IGD may strongly arouse emotional dysregulation. Future DSM criteria could consider these gaming-addicted youth as a specific subclass of ADHD.

Pharmacologically upregulated carcinoembryonic antigen-expression enhances the cytolytic activity of genetically-modified chimeric antigen receptor NK-92MI against colorectal cancer cells.

BACKGROUND: The natural killer cell line, NK-92MI, is cytotoxic against various types of cancer. The aim of this study was to develop chimeric antigen receptor-modified (CAR) NK-92MI cells targeting carcinoembryonic antigen-expressing (CEA) tumours and increase killing efficacy by pharmacologically modifying CEA-expression. RESULT: We generated anti-CEA-CAR NK-92MI cells by retroviral vector transduction. This genetically-modified cell line recognised and lysed high CEA-expressing tumour cell lines (LS174T) at 47.54 ± 12.60% and moderate CEA-expressing tumour cell lines (WiDr) at 31.14 ± 16.92% at a 5:1 effector: target (E/T) ratio. The cell line did not lyse low CEA-expressing tumour cells (HCT116) as they did their parental cells (NK-92MI cells). The histone deacetylase-inhibitor (HDAC) sodium butyrate (NaB) and the methylation-inhibitor 5-azacytidine (5-AZA), as epigenetic modifiers, induced CEA-expression in HCT116 and WiDr cells. Although the IC50 of 5 fluorouracil (5-FU) increased, both cell lines showed collateral sensitivity to anti-CEA-CAR NK-92MI cells. The cytolytic function of anti-CEA-CAR NK-92MI cells was increased from 22.99 ± 2.04% of lysis background to 69.20 ± 11.92% after NaB treatment, and 69.70 ± 9.93% after 5-AZA treatment, at a 10:1 E/T ratio in HCT116 cells. The WiDr cells showed similar trend, from 22.99 ± 4.01% of lysis background to 70.69 ± 10.19% after NaB treatment, and 59.44 ± 10.92% after 5-AZA treatment, at a 10:1 E/T ratio. CONCLUSIONS: This data indicates that the effector-ability of anti-CEA-CAR NK-92MI increased in a CEA-dependent manner. The combination of epigenetic-modifiers like HDAC-inhibitors, methylation-inhibitors, and adoptive-transfer of ex vivo-expanded allogeneic-NK cells may be clinically applicable to patients with in 5-FU resistant condition.

19-substituted benzoquinone ansamycin heat shock protein-90 inhibitors: biological activity and decreased off-target toxicity.

The benzoquinone ansamycins (BQAs) are a valuable class of antitumor agents that serve as inhibitors of heat shock protein (Hsp)-90. However, clinical use of BQAs has resulted in off-target toxicities, including concerns of hepatotoxicity. Mechanisms underlying the toxicity of quinones include their ability to redox cycle and/or arylate cellular nucleophiles. We have therefore designed 19-substituted BQAs to prevent glutathione conjugation and nonspecific interactions with protein thiols to minimize off-target effects and reduce hepatotoxicity. 19-Phenyl- and 19-methyl-substituted versions of geldanamycin and its derivatives, 17-allylamino-17-demethoxygeldanamycin and 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (17-DMAG), did not react with glutathione, whereas marked reactivity was observed using parent BQAs. Importantly, although 17-DMAG induced cell death in primary and cultured mouse hepatocytes, 19-phenyl and 19-methyl DMAG showed reduced toxicity, validating the overall approach. Furthermore, our data suggest that arylation reactions, rather than redox cycling, are a major mechanism contributing to BQA hepatotoxicity. 19-Phenyl BQAs inhibited purified Hsp90 in a NAD(P)H: quinone oxidoreductase 1 (NQO1)-dependent manner, demonstrating increased efficacy of the hydroquinone ansamycin relative to its parent quinone. Molecular modeling supported increased stability of the hydroquinone form of 19-phenyl-DMAG in the active site of human Hsp90. In human breast cancer cells, 19-phenyl BQAs induced growth inhibition also dependent upon metabolism via NQO1 with decreased expression of client proteins and compensatory induction of Hsp70. These data demonstrate that 19-substituted BQAs are unreactive with thiols, display reduced hepatotoxicity, and retain Hsp90 and growth-inhibitory activity in human breast cancer cells, although with diminished potency relative to parent BQAs.

A novel lectin receptor kinase gene, AtG-LecRK-I.2, enhances bacterial pathogen resistance through regulation of stomatal immunity in Arabidopsis.

The S-locus lectin receptor kinases (G-LecRKs) have been suggested as receptors for microbe/damage-associated molecular patterns (MAMPs/DAMPs) and to be involved in the pathogen defense responses, but the functions of most G-LecRKs in biotic stress response have not been characterized. Here, we identified a member of this family, G-LecRK-I.2, that positively regulates flg22- and Pseudomonas syringae pv. tomato (Pst) DC3000-induced stomatal closure. G-LecRK-I.2 was rapidly phosphorylated under flg22 treatment and could interact with the FLS2/BAK1 complex. Two T-DNA insertion lines, glecrk-i.2-1 and glecrk-i.2-2, had lower levels of reactive oxygen species (ROS) and nitric oxide (NO) production in guard cells, as compared with the wild-type Col-0, under Pst DC3000 infection. Also, the immunity marker genes CBP60g and PR1 were induced at lower levels under Pst DC3000 hrcC- infection in glecrk-i.2-1 and glecrk-i.2-2. The GUS reporter system also revealed that G-LecRK-I.2 was expressed only in guard cells. We also found that G-LecRK-I.2 could interact H+-ATPase AHA1 to regulate H+-ATPase activity in the guard cells. Taken together, our results show that G-LecRK-I.2 plays an important role in regulating stomatal closure under flg22 and Pst DC3000 treatments and in ROS and NO signaling specifically in guard cells.

Flavonol-ruthenium complexes as antioxidant and anticancer agents.

Flavonol-metal complexes can enhance the biological activity of flavonols. Inspired by the potential of ruthenium-based drugs in pharmaceutical applications, seven flavonol-Ru (II) complexes were synthesized to evaluate their biological activities. Among these compounds, compounds 8, 11, and 12 showed potent antioxidant activities. Compound 12 exhibited superior anti-inflammatory activity to natural quercetin, which served as a positive control. This study is the first to report the free radical scavenging abilities and antioxidant activity of flavonol-Ru (II) complexes. Furthermore, compound 12 demonstrated comparable efficacy to 5-FU against human non-small-cell lung cancer cells (A549). These results strongly support the potential of flavonol-Ru (II) agents.

A Brain-Protective Sterol from Soft Coral Inhibits Lipopolysaccharide-Induced Matrix Metalloproteinase-9-Mediated Astrocytic Migration.

Matrix metalloproteinases (MMPs), which are proteolytic enzymes, promote blood-brain barrier (BBB) disruption, leading to neuronal damage and neuroinflammation. Among them, MMP-9 upregulation serves as an inflammatory biomarker in the central nervous system (CNS). Currently, the development of marine organism-derived bioactive compounds or metabolites as anti-inflammatory drugs has received considerable attention. The 9,11-secosteroid, 3ß,11-dihydroxy-9,11-secogorgost-5-en-9-one (4p3f), is a novel sterol compound extracted from the soft coral Sinularia leptoclado with potential anti-inflammatory activity. However, the effect of and potential for brain protection of 4p3f on brain astrocytes remain unclear. Herein, we used rat brain astrocytes (RBAs) to investigate the effects and signaling mechanisms of 4p3f on lipopolysaccharide (LPS)-induced MMP-9 expression via zymographic, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blot, immunofluorescence staining, promoter-reporter, and cell migration analyses. We first found that 4p3f blocked LPS-induced MMP-9 expression in RBAs. Next, we demonstrated that LPS induced MMP-9 expression via the activation of ERK1/2, p38 MAPK, and JNK1/2, which is linked to the STAT3-mediated NF-κB signaling pathway. Finally, 4p3f effectively inhibited LPS-induced upregulation of MMP-9-triggered RBA cell migration. These data suggest that a novel sterol from soft coral, 4p3f, may have anti-inflammatory and brain-protective effects by attenuating these signaling pathways of MMP-9-mediated events in brain astrocytes. Accordingly, the soft coral-derived sterol 4p3f may emerge as a potential candidate for drug development or as a natural compound with neuroprotective properties.

Oridonin enhances cytotoxic activity of natural killer cells against lung cancer.

BACKGROUND: Oridonin is a Chinese herbal medicine exhibiting anti-tumor properties; however, its immune modulation capacity has yet to be elucidated. Our objective in this study was to determine whether oridonin enhances the anti-tumor activity of natural killer (NK) cells against lung cancer cells. METHODS: LDH-releasing assays were used to investigate the effects of oridonin on NK-92MI cell activity against lung cancer cells. Flow cytometry and real-time PCR were used to examine the effects of oridonin on degranulation markers, cytotoxic factors, activating receptors on NK-92MI cells, and ligands in lung cancer cells. Western blot analysis provided insight into the mechanisms underlying the observed effects. RESULTS: Oridonin enhanced the cytotoxic effects of NK-92MI cells against A549 lung cancer cells. This effect involved upregulating the expression of the degranulation marker CD107a and IFN-γ as well as activating receptors on NK cells and their ligand MICA/B. Oridonin also inhibited STAT3 phosphorylation in A549 cells and NK-92MI cells. A lung cancer mouse model confirmed the anti-tumor effects of oridonin and NK-92MI cells, wherein both treatments alone suppressed tumor growth. Oridonin was also shown to have a synergistic effect on the anti-tumor activity of NK-92MI cells. CONCLUSIONS: The ability of oridonin to enhance the cytotoxic effects of NK cells indicates its potential as a novel therapeutic agent for the treatment of lung cancer.

Comparing the Therapeutic Efficacies of Lung Cancer: Network Meta-Analysis Approaches.

BACKGROUND: In recent years, reduction of nuclear power generation and the use of coal-fired power for filling the power supply gap might have increased the risk of lung cancer. This study aims to explore the most effective treatment for different stages of lung cancer patients. METHODS: We searched databases to investigate the treatment efficacy of lung cancer. The network meta-analysis was used to explore the top three effective therapeutic strategies among all collected treatment methodologies. RESULTS: A total of 124 studies were collected from 115 articles with 171,757 participants in total. The results of network meta-analyses showed that the best top three treatments: (1) in response rate, for advanced lung cancer were Targeted + Targeted, Chemo + Immuno, and Targeted + Other Therapy with cumulative probabilities 82.9, 80.8, and 69.3%, respectively; for non-advanced lung cancer were Chemoradio + Targeted, Chemoradi + Immuno, and Chemoradio + Other Therapy with cumulative probabilities 69.0, 67.8, and 60.7%, respectively; (2) in disease-free control rate, for advanced lung cancer were Targeted + Others, Chemo + Immuno, and Targeted + Targeted Therapy with cumulative probabilities 93.4, 91.5, and 59.4%, respectively; for non-advanced lung cancer were Chemo + Surgery, Chemoradio + Targeted, and Surgery Therapy with cumulative probabilities 80.1, 71.5, and 43.1%, respectively. CONCLUSION: The therapeutic strategies with the best effectiveness will be different depending on the stage of lung cancer patients.

Structural Equation Modeling (SEM): Gaming Disorder Leading Untreated Attention-Deficit/Hyperactivity Disorder to Disruptive Mood Dysregulation.

(1) Background: Internet gaming disorder (IGD) in youths likely leads to disruptive mood dysregulation, especially among those with attention-deficit/hyperactivity disorder (ADHD). Whether IGD mediates the pathways leading ADHD to disruptive emotional dysfunction remains unclear. This study aims to elucidate the direct or indirect influence of IGD on ADHD; (2) Method: The Swanson, Nolan, and Pelham Version IV questionnaire was used to evaluate symptoms of ADHD and oppositional defiant disorder, and the Chen gaming disorder scale was used to measure IGD. A psychiatrist diagnosed ADHD, IGD, and disruptive mood dysregulation disorder (DMDD)-like symptoms. Structural equation modeling was applied to evaluate the role of IGD in mediating ADHD progression to disruptive mood dysregulation; (3) Results: Among a total of 102 ADHD youths, 53 (52%) of them with IGD were significantly more likely to have poor interpersonal relationships (p < 0.01) and DMDD-like symptoms (p < 0.01) than ADHD youths without IGD. IGD played a mediating role in increasing the risk of disruptive mood dysregulation in ADHD youths; (4) Conclusions: The findings suggest that IGD mediates ADHD's progression to disruptive mood dysregulation. Intensive biopsychosocial interventions are warranted for ADHD youths with IGD. More children and adolescents became mood-dysregulated after excessive gaming during the COVID-19 pandemic; this study's results suggest that child mental health experts develop earlier detection and prevention strategies for children and adolescents hidden behind internet addiction.

The Comparative Efficacy of Treatments for Children and Young Adults with Internet Addiction/Internet Gaming Disorder: An Updated Meta-Analysis.

Internet gaming disorder (IGD) is a formal mental disorder leading to bad outcomes for children and adolescents. This study comprehensively compared the estimated effect of various pharmacotherapy and psychosocial interventions for IGD from randomized controlled trials (RCT) through updated meta-analysis, using meta-regression. A search of PubMed/MEDLINE, Cochrane Library, and Airiti Library between 2000 and 2017 was conducted for various IA/IGD intervention modalities. A total of 124 studies from 29 selected papers involving 5601 children and young adults with IA/IGD were found. Meta-analyzing the pooled standardized mean difference (SMD) revealed a preliminary random effect of 1.399 with a 95% confidence interval of 1.272-1.527, suggesting highly effective treatment of IA/IGD. After adjusting for the confounding risks of age, publication year, type of subjects, and type of study, this study revealed that combining pharmacotherapy with cognitive behavioral therapy (CBT) or multi-level counseling (MLC) was the most effective treatment option. Using a scale of time spent online or a severity of IA symptoms scale was a more effective measurement, with p-values = 0.006 and 0.002, respectively. IA/IGD patients with comorbid depression showed worse outcomes than youth with another comorbidity. The corresponding model goodness-of-fit indices were τ2 = 1.188; I2-Residual = 89.74%; and Adjusted-R2 = 16.10%. This systematic review indicates that pharmacotherapy combined with CBT or MLC might be an effective therapeutic strategy for youth with gaming disorder.

Structural Equation Modeling (SEM): Childhood Aggression and Irritable ADHD Associated with Parental Psychiatric Symptoms.

BACKGROUND: Children with attention deficit hyperactivity disorder (ADHD) co-occurring with Oppositional Defiant Disorder (ODD) further present aggressive behavior and may have a depressive parent. A child with co-occurring ADHD and ODD has differentially higher levels of behavioral and emotional difficulties. Little is known about how the irritable subtype of ADHD in children mediates the development of parental symptomatology. This study aims to elucidate the direct or indirect influence of childhood disruptive ADHD with aggressive behavior on their parental symptom using Structural Equation Modeling (SEM). METHODS: A total of 231 ADHD children and their parents completed the Swanson, Nolan, and Pelham Version IV questionnaire for symptoms of ADHD, Oppositional Defiant Disorder (ODD) scale for irritable symptoms, Child Behavior Check List (CBCL) for aggression, and Symptom Checklist (SCL) for parental symptom. RESULTS: The three-factor confirmatory factor analysis (CFA) model found symptoms of inattention, hyperactivity/impulsivity, irritable ODD, and aggression were inter-related. Mediational analyses demonstrated ODD mediates symptoms directly predicting the risk of increasing ADHD severity. Disruptive child symptoms (ADHD + ODD + aggression) may increase the risk of depression-related symptoms in the parent. When the child's aggression increases by one standard deviation (SD), parental psychiatric symptoms increase by 0.235 SD (p < 0.001). CONCLUSIONS: By this SEM pathway analysis, there is the correlation between the disruptive, more aggressive subtype of ADHD in children/adolescents and the existence of psychopathological symptomatology of their parents. ADHD + ODD + aggression in children should be classified as an irritable subtype of ADHD, warranting early diagnosis and intensive treatment.

TNF-alpha mediates eosinophil cationic protein-induced apoptosis in BEAS-2B cells.

BACKGROUND: Eosinophilic granulocytes are important for the human immune system. Many cationic proteins with cytotoxic activities, such as eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN), are released from activated eosinophils. ECP, with low RNase activity, is widely used as a biomarker for asthma. ECP inhibits cell viability and induces apoptosis to cells. However, the specific pathway underlying the mechanisms of ECP-induced cytotoxicity remains unclear. This study investigated ECP-induced apoptosis in bronchial epithelial BEAS-2B cells and elucidated the specific pathway during apoptosis. RESULTS: To address the mechanisms involved in ECP-induced apoptosis in human BEAS-2B cells, investigation was carried out using chromatin condensation, cleavage of poly (ADP-ribose) polymerase (PARP), sub-G1 distribution in cell cycle, annexin V labeling, and general or specific caspase inhibitors. Caspase-8-dependent apoptosis was demonstrated by cleavage of caspase-8 after recombinant ECP treatment, accompanied with elevated level of tumor necrosis factor alpha (TNF-alpha). Moreover, ECP-induced apoptosis was effectively inhibited in the presence of neutralizing anti-TNF-alpha antibody. CONCLUSION: In conclusion, our results have demonstrated that ECP increased TNF-alpha production in BEAS-2B cells and triggered apoptosis by caspase-8 activation through mitochondria-independent pathway.

Thioredoxin mediates remodeling factors of human bronchial epithelial cells upon interaction with house dust mite-stimulated eosinophils.

Bronchial epithelial cells exposed to allergens typically secrete chemokines to recruit eosinophils. Persistent inflammation and repair responses result in airway remodeling and irreversible airflow limitation. House dust mite (HDM) is a common allergen causing allergic disorders. Thioredoxin (TRX) is a redox protein that scavenges reactive oxygen species (ROS). This study was to elucidate how TRX mediates gene expression of remodeling factors of human bronchial epithelial cells in response to HDM stimuli interacting with eosinophils. This study cultured normal human bronchial epithelial (BEAS-2B) cells with eosinophils exposed to 0.5 microg/ml recombinant Dermatophagoides pteronyssinus 1 (rDer p1) protease to mimic the allergen-immune reaction. Eosinophils were induced by rDer p1 protease to secrete tumor necrosis factor (TNF)-alpha and generate ROS. When cultured with rDer p1-stimulated eosinophils, BEAS-2B cells released interleukin-6 and underwent apoptosis. The HDM-stimulated eosinophils applied oxidative stress and apoptosis to BEAS-2B cells through the release of mediators. Damaged BEAS-2B cells interfered with gene expression of remodeling factors, such as transforming growth factor (TGF)-beta 1, epidermal growth factor receptor (EGFR), cyclin dependent kinase inhibitor (p21(waf)) and matrix metalloproteinase (MMP) 9, relevant to inflammatory response and epithelial repair in airway remodeling. Notably, BEAS-2B cells over-expressing TRX reduced eosinophil-derived apoptosis and suppressed underlying airway remodeling via attenuation of TGF-beta1, EGFR and p21(waf) and up-regulation of MMP9 expression. Results of this study indicated TRX-over-expressing bronchial epithelial cells attenuated TGF-beta1 and activated MMP9 expression to prevent airway remodeling from HDM-induced inflammation. The finding can be as a reference for further therapeutic studies of TRX.

Autism Associated With Anti-NMDAR Encephalitis: Glutamate-Related Therapy.

The purpose of this review is to correlate autism with autoimmune dysfunction in the absence of an explanation for the etiology of autism spectrum disorder. The anti-N-methyl-D-aspartate receptor (anti-NMDAR) autoantibody is a typical synaptic protein that can bind to synaptic NMDA glutamate receptors, leading to dysfunctional glutamate neurotransmission in the brain that manifests as psychiatric symptoms (psychosis, hallucinations, and personality changes). Detection of autoantibodies, cytokines, decreased lymphocytes, serum immunoglobulin level imbalance, T-cell mediated immune profile, maternal infection history, and children's infection history can all be vital biological markers of autoimmune autism. Diagnosing autoimmune encephalitis sooner can increase the effectiveness of curative treatments-such as immune therapy or immune modulatory therapy-that may prevent the long-term consequence of being misdiagnosed with autism spectrum disorder. Glutamate therapy primarily normalizes glutamate neurotransmission and can be a new add-on intervention alongside antipsychotics for treating autoimmune autism.