Robust Activity Research Articles

Emergency medicine : what's new in 2024

Emergency medicine plays a crucial vital role as the gateway to the Swiss healthcare system. Although it has not yet been officially recognized with a specialist title, unlike most European countries - emergency medicine in Switzerland is characterized by robust research activity. This scientific article demonstrates a dynamic and rigorous evolution. In this edition, we present six articles authored by Swiss emergency physicians, each examining different aspects of our field. Topics include gender stereotypes, prehospital cardiac arrest, the unique needs of the older patients in emergency department, the administration of antifibrinolytics in trauma cases, and a predictive model for infectious endocarditis.

Anti-furfurative comparison of Kesh Kanti-Herbal Shampoos and synthetic shampoos against Malassezia furfur for dandruff management

Malassezia furfur is the primary etiological agent of dandruff (Pityriasis capitis). Although herbal shampoos are preferred for their natural, mild ingredients over synthetic counterparts, they are often perceived as less effective in managing flaky scalp conditions or furfuration causing dandruff. The study compares the antifungal efficacy of herbal and synthetic shampoos against M. furfur. Seven shampoos including herbal (HS_Adv, HS_M&P, HS_Aloe), synthetic (SYN_01, SYN_02, SYN_03) and an antifungal shampoo containing ketoconazole (KETO) were employed in the study. Experiments were designed to stimulate real-world conditions, utilizing disc-diffusion assay, 3-minute shampoo contact at mild dilutions (1% and 5%), recurrent 3-minute shampoo contact every 24 h with intermittent recovery. Both disc diffusion and 3-minute shampoo contact demonstrated that all shampoos were effectively inhibiting the viability of M. furfur. However, a single 3-minute shampoo contact followed by a prolonged recovery of 72 h revealed SYN_01 and KETO with maximal antifungal action. In contrast, herbal shampoos were as effective as synthetic options when M. furfur was subjected to 3-minute shampoo contact every 24 h with intermittent recovery. Comprehensive ingredient analysis revealed the robust antifungal activity in SYN_01 was probably because of the presence of various surfactants, allergens and a potent synthetic antifungal agent, Piroctone olamine. This study experimentally demonstrates that herbal shampoos are as effective as synthetic options in managing M. furfur-induced dandruff when applied consistently. The findings highlight the importance of regular scalp cleansing for dandruff management and provide valuable insights into the antifungal potential of both herbal and synthetic formulations.

Engineering an optimized hypercompact CRISPR/Cas12j-8 system for efficient genome editing in plants.

The Cas12j-8 nuclease, derived from the type V CRISPR system, is approximately half the size of Cas9 and recognizes a 5'-TTN-3' protospacer adjacent motif sequence, thus potentially having broad application in genome editing for crop improvement. However, its editing efficiency remains low in plants. In this study, we rationally engineered both the crRNA and the Cas12j-8 nuclease. The engineered crRNA and Cas12j-8 markedly improved genome editing efficiency in plants. When combined, they exhibited robust editing activity in soybean and rice, enabling the editing of target sites that were previously uneditable. Notably, for certain target sequences, the editing activity was comparable to that of SpCas9 when targeting identical sequences, and it outperformed the Cas12j-2 variant, nCas12j-2, across all tested targets. Additionally, we developed cytosine base editors based on the engineered crRNA and Cas12j-8, demonstrating an average increase of 5.36- to 6.85-fold in base-editing efficiency (C to T) compared with the unengineered system in plants, with no insertions or deletions (indels) observed. Collectively, these findings indicate that the engineered hypercompact CRISPR/Cas12j-8 system serves as an efficient tool for genome editing mediated by both nuclease cleavage and base editing in plants.

The high efficacy of claudin18.2-targeted CAR-T cell therapy in advanced pancreatic cancer with a strategy to ensure the safety of patients.

Pancreatic cancer (PC) is one of the most lethal digestive system tumors. Claudin18.2 is highly expressed in PC tissue and could serve as a suitable target for CAR-T therapy. In the present study, we reported the utilization of tEGFR-expressing claudin18.2-targeted CAR-T cells to treat 3 patients with advanced PC. Intriguingly, all 3 patients achieved disease remission after CAR-T cell infusion, with 1 complete remission (CR) and 2 partial remissions (PR). However, gastric mucosal injury was observed, which was recognized as on-target off-tumor toxicity (OTOT) and may be due to the expression of claudin18.2 on normal gastric tissues. To control the severe OTOT in patient 3, cyclophosphamide and cetuximab were administered to deplete CAR-T cells, and they successfully controlled OTOT. Single cell transcriptome and TCR sequencing revealed the objective alterations of CAR-T cell clones after cetuximab treatment. Collectively, the present study showed the robust anti-tumor activity of claudin18.2-targeted CAR-T cells against PC, and reported the feasibility of antibody-dependent safety switch strategy to control the OTOT caused by CAR-T cells in patients. Our study may pave the way for the development of a novel strategy to treat patients with advanced PC in the future.

Characterizing Microglial Signaling Dynamics During Inflammation Using Single-Cell Mass Cytometry.

Microglia play a critical role in maintaining central nervous system (CNS) homeostasis and display remarkable plasticity in their response to inflammatory stimuli. However, the specific signaling profiles that microglia adopt during such challenges remain incompletely understood. Traditional transcriptomic approaches provide valuable insights, but fail to capture dynamic post-translational changes. In this study, we utilized time-resolved single-cell mass cytometry (CyTOF) to measure distinct signaling pathways activated in microglia upon exposure to bacterial and viral mimetics-lipopolysaccharide (LPS) and polyinosinic-polycytidylic acid (Poly(I:C)), respectively. Furthermore, we evaluated the immunomodulatory role of astrocytes on microglial signaling in mixed cultures. Microglia or mixed cultures derived from neonatal mice were treated with LPS or Poly(I:C) for 48 h. Cultures were stained with a panel of 33 metal-conjugated antibodies targeting signaling and identity markers. High-dimensional clustering analysis was used to identify emergent signaling modules. We found that LPS treatment led to more robust early activation of pp38, pERK, pRSK, and pCREB compared to Poly(I:C). Despite these differences, both LPS and Poly(I:C) upregulated the classical reactivity markers CD40 and CD86 at later time points. Strikingly, the presence of astrocytes significantly blunted microglial responses to both stimuli, particularly dampening CD40 upregulation. Our studies demonstrate that single-cell mass cytometry effectively captures the dynamic signaling landscape of microglia under pro-inflammatory conditions. This approach may pave the way for targeted therapeutic investigations of various neuroinflammatory disorders. Moreover, our findings underscore the necessity of considering cellular context, such as astrocyte presence, in interpreting microglial behavior during inflammation.

A Breakthrough Straightforward Multifunctional Hydrogel Platform Releasing Therapeutic Gas for Advanced Wound Healing

ABSTRACTThe advancement of therapeutic gas treatment has significantly impacted on the biomaterial field, with nitric oxide (NO) gaining attention for its safety, multifunctionality, and role in regulating biological processes. Thus, this study introduces a novel biocatalytic NO‐generating in situ forming hydrogel (GTA/Cu) to address wound‐related issues, fabricated through a simple, one‐step process by incorporating copper ions (Cu2+) into tannic acid‐conjugated gelatin (GTA). Herein, Cu2+ functions simultaneously as a crosslinking agent, NO‐generating catalyst, and antibacterial agent, while the galloyl groups in GTA enable effective tissue adhesion and diverse crosslinking interactions. The hydrogels' mechanical properties are controlled by varying Cu2+ concentrations (25, 50, and 100 mg/mL), with higher concentration accelerating gelation and enhancing mechanical strength. At 100 mg/mL Cu2+, the hydrogel releases NO for up to 12 days, reaching a cumulative concentration of around 200 μM. It also demonstrated robust antioxidant activity, high tissue adhesion (~20 kPa), and comparable antibacterial effects to Cu‐only samples. Interestingly, the released NO facilitates endothelial cell proliferation, accelerates scratch closure within 36 h, and stimulates new tube formation on Matrigel, showing comparable effects to VEGF. Additionally, it clearly promotes new blood vessel formation in vivo following subcutaneous injection, further highlighting its potential for practical wound healing applications.

Novel design of potent anti-tumour activity of IL-2 prodrug by FAPα-mediated activation

Interleukin-2 (IL-2) is a T cell growth factor that is essential for the proliferation of T cells and the generation of effector and memory cells. The antitumor activity of high-dose IL-2 therapy requires maintaining the affinity between IL-2 and IL2-Rα, which can also bring serious toxic side effects. To address this issue, we designed ZGP-Cysteamine-IL-2-K64C and (ZGP-Cysteamine)2-IL-2-(K43C, K64C) based on the strategy of FAPα enzyme-activated prodrugs, and investigated their anti-tumour activity and side effects. In vitro FAPα enzyme cleavage results indicated that the side-chain modified ZGP-Cysteamine moiety could be precisely recognized and cleaved by FAPα, thereby restoring the activity of native IL-2 capable of binding to IL-2Rα in the tumour microenvironment, where it promotes the expansion of CD8+ T cells. Meanwhile, surface plasmon resonance analysis revealed that, compared to wt-IL-2, both ZGP-Cysteamine-IL-2-K64C and (ZGP-Cysteamine)2-IL-2-(K43C, K64C) exhibited significantly reduced affinity for IL-2Rα, while their affinity for IL-2Rβγ remained unchanged. Remarkably, ZGP-Cysteamine-IL-2-K64C and (ZGP-Cysteamine)2-IL-2-(K43C, K64C) almost completely eliminated the pulmonary edema and vascular permeability. Furthermore, the combination of ZGP-Cysteamine-IL-2-K64C and PD-1 blockade showed robust anti-tumour activity in mice tumour models. Our study provides new insights into the structural design of IL-2 prodrug with low side effect and robust anti-tumour efficacy.

Citronellol Induces Apoptosis via Differential Regulation of Caspase-3, NF-κB, and JAK2 Signaling Pathways in Glioblastoma Cell Line.

Citronellol (CT) is a naturally occurring lipophilic monoterpenoid which has shown anticancer effects in numerous cancerous cell lines. This study was, therefore, designed to examine CT's potential as an anticancer agent against glioblastoma (GBM). Network pharmacology analysis was employed to identify potential anticancer targets of CT. A comprehensive data mining was carried out to assess CT and GBM-associated target genes. Protein-protein interaction network was constructed to identify hub genes and later GO and KEGG enrichment analysis was performed to elucidate the possible mechanism. Human glioblastoma cell line "SF767" was used to confirm in silico findings. MTT, crystal violet, and trypan blue assays were performed to assess the cytotoxic effects of various concentrations of CT. Subsequently, ELISA and qPCR were performed to analyze the effects of CT on proapoptotic and inflammatory mediators. In silico findings indicated that CT differentially regulated proapoptotic and inflammatory pathways by activating caspase-3 and 8 and inhibiting nuclear factor-kappa B (NF-κB), tumor necrosis factor-α, Janus kinase 2 (JAK2). Molecular docking also demonstrated strong binding affinities of CT with the above-mentioned mediators when compared to 5-fluorouracil or temozolomide. In SF767 cell line, CT displayed dose-dependent cytotoxic and antioxidant effects, and upregulation of annexin-V, caspase-3, and 8 along with downregulation of inflammatory modulators. In a nutshell, it can be concluded from these findings that CT possesses robust anticancer activity which is mediated via differential regulation of caspase-3, JAK2, and NF-κB pathways.

Anti-tumor activity of an αPD-L1-PE38 immunotoxin delivered by engineered Nissle 1917.

Although immune checkpoint inhibitors specifically targeting the PD-1/PD-L1 axis have exhibited remarkable clinical success, they are not uniformly effective across all patient cohorts. Immunotoxins, a novel class of cancer therapeutics, offering a promising alternative. PD-L1, which is also present in certain normal tissues, limits its suitability as an ideal target for immunotoxins. The probiotic strain of E. coli Nissle 1917 (EcN) could target and colonize to solid tumors, which positions it as a promising candidate for tumor tissue-specific delivery of anti-tumor proteins. In this study, we constructed a PD-L1-targeted immunotoxin, designated as αPD-L1-PE38, by fusing an anti-PD-L1 nanobody and a clinically validated PE38 toxin. This immunotoxin exhibited potent cytotoxic activity against tumor cells while showed slightly cytotoxic activity against normal cells. To effectively deliver the αPD-L1-PE38 to tumor tissues, we engineered the EcN strain to release the immunotoxin induced by L-arabinose. Upon induction, the immunotoxin was efficiently secreted, and exhibited robust anti-tumor activity mainly by inducing cell apoptosis both in vitro and in vivo. Furthermore, we enhanced the immunotoxin's affinity for PD-L1 by optimizing the linker between the nanobody and PE38 toxin. The engineered EcN expressing the optimized immunotoxin, achieved superior anti-tumor activity. Collectively, our study suggests that the delivery of immunotoxins through live bacteria to improve safety and efficacy is a promising option in cancer therapeutics.

Adaptive radiation strategy with V20 limitation associates with survival benefit and lower incidence of symptomatic radiation pneumonitis in stage III NSCLC patients receiving concurrent immunotherapy and thoracic radiation.

To evaluate the efficacy and the incidence of symptomatic radiation pneumonitis (RP) of the adaptive radiation strategy with V20 limitation in stage III non-small cell lung cancer (NSCLC) patients receiving concurrent immunotherapy and radiotherapy Materials and Methods: We retrospectively reviewed stage III NSCLC patients received thoracic radiation with or without immunotherapy from January 2015 to September 2024 in the Third Xiangya Hospital. The overall survival (OS), progression free survival (PFS), objective response rate (ORR), and the incidence of symptomatic RP were compared among patients stratified by the sequential of immunotherapy and radiotherapy. 45 patients received concurrent immunotherapy and radiotherapy with application of the adaptive radiation strategy (the CIR group). 32 patients received simultaneous integrated boost intensity-modulated radiotherapy (SIB-IMRT), 26 patients received 2 to 4 cycles neoadjuvant immuno-chemotherapy before the concurrent immunotherapy and radiotherapy, 7 patients received thoracic radiotherapy with a prescribed dosage of 50 Gy, and 10 patients received radiotherapy with CTV omission. 86.67 % (39/45) patients had a V20 ≤ 20 %. The ORR was 86.67 %. The median PFS of these patients was significantly longer than those received concurrent chemo-radiotherapy followed by immunotherapy (the PACIFIC paradigm, HR, 2.40; 95 % CI, 1.15 to 5.02; log-rank p = 0.013; median, 28.6 vs. 16.1 months p = 0.013). The median OS was not reached. 13.3 % patients in the CIR group experienced grade ≥ 2 RP and the incidence was significantly lower than that of patients received radiotherapy without immunotherapy or concurrent chemo-radiotherapy with immunotherapy consolidation. The application of adaptive radiotherapy strategies with V20 limitation demonstrated robust antitumor activity and reduced pulmonary toxicity in stage III NSCLC patients receiving concurrent ICIs treatment and thoracic radiation. This treatment modality deserves further validation as a promising therapy in patients with treatment-naive, unresectable, stage III NSCLC.

Exploring perceptions and attitudes towards vaccines in a remote western DRC health zone

The Democratic Republic of the Congo (DRC) struggles with low full childhood vaccination coverage (around 50 %) and a high children-under-five mortality rate (79 deaths per 1000 live births). This situation is potentially exacerbated by vaccine hesitancy, which was identified by the World Health Organization (WHO) as one of the top 10 global health threats in 2019. To gain deeper insights into levels of vaccine confidence possibly influencing vaccination coverage, we explored perceptions and attitudes towards childhood and adult vaccines in Boende (Tshuapa province, western DRC), which experienced an Ebola outbreak in 2014 and hosted the EBL2007 Ebola vaccine trial (2019–2022). Using purposeful sampling, we conducted 29 individual interviews and 14 focus group discussions with diverse community members between July 2022 and March 2023. Our findings suggest the prevalence of a relatively low level of trust in COVID-19 vaccines compared to other vaccines. Additionally, the fear of Ebola disease seemed to lead to general acceptance of the EBL2007 trial vaccine, although some concerns were voiced about pre-testing and the decision to conduct the trial in the DRC. While trust in childhood vaccines and reported uptake appeared to be high, concerns existed regarding potential adverse effects and the possibility that vaccines targeted African children specifically. Our analysis further identified four recommendations to possibly enhance vaccine confidence in the region. This study highlights the multifaceted nature of vaccine confidence, influenced by the perceived risk of the targeted diseases, past experiences with medical interventions and staff, and sociopolitical contexts. However, we emphasise that increasing vaccine uptake requires a comprehensive approach, addressing not only vaccine confidence, but also crucial aspects like access to vaccines and robust disease surveillance activities. This would ultimately reduce the burden of vaccine-preventable diseases and lead to better public health outcomes in the region.

A curcumin-based HDACs inhibitor for targeted sonodynamic therapy of breast cancer.

Histone Deacetylases (HDACs) have emerged as key therapeutic targets in cancer treatment. In this study, we designed CURSAHA, a multifunctional anticancer agent, through the pharmacophore fusion of Vorinostat and curcumin. CURSAHA demonstrates broad-spectrum inhibitory activity against HDACs, effectively suppressing tumor cells with overexpressed HDACs. Notably, CURSAHA generates reactive oxygen species (ROS) under ultrasonic conditions, exhibiting sonodynamic therapeutic activity. Additionally, CURSAHA downregulates HDACs through redox reactions involving ROS. These properties enable CURSAHA to exhibit robust antitumor activity in both in vitro and in vivo models, highlighting its potential as a promising candidate for further development in cancer therapy.

Discovery of a non-nucleotide stimulator of interferon genes (STING) agonist with systemic antitumor effect.

Agonists of the stimulator of interferon genes (STING) pathway are increasingly being recognized as a promising new approach in the treatment of cancer. Although progress in clinical trials for STING agonists in antitumor applications has been slow, there is still an urgent need for developing new potent STING agonists with versatile potential applications. Herein, we developed and identified a non-nucleotide STING agonist called DW18343. DW18343 showed robust activation across different STING isoforms. Crystallography analysis revealed that DW18343 binds more deeply into the ligand binding domain (LBD) pocket of STING-H232 compared to other agonists such as MSA-2, SR-717, or cGAMP, which likely contributes to its high potency. DW18343 triggered downstream p-TBK1/p-IRF3 signaling, leading to the production of multiple cytokines. Additionally, DW18343 displayed broad and long-lasting antitumor effects in various syngeneic mouse tumor models, whether administered locally or systemically. Moreover, DW18343 induced immune memory to combat the growth of rechallenged tumors. Finally, DW18343 was shown to be an activator of both the innate and adaptive antitumor immunity in tumor tissue, potentially explaining its strong antitumor effects in vivo. In conclusion, DW18343 serves as a novel non-nucleotide STING agonist with systemic antitumor effect through the activation of antitumor immunity.

Exploring anticancer properties of new triazole-linked benzenesulfonamide derivatives against colorectal carcinoma: Synthesis, cytotoxicity, and in silico insights.

This study reports the design, synthesis, and characterization of a novel series of benzene sulfonamide-triazole hybrid derivatives, to evaluate their anticancer potential against colorectal cancer. The synthesized compounds were characterized using NMR and HRMS spectroscopic techniques. In vitro cytotoxicity assessments revealed that compounds 5g and 5j exhibited significant anticancer effects. 5g showed the highest potency in the DLD-1 cell line (IC50=11.84µM), while 5j demonstrated robust activity in the HT-29 cell line (IC50=9.35µM). Apoptotic analysis indicated that compound 5g effectively induced early and total apoptosis, surpassing the chemotherapeutic agent 5-fluorouracil (5-FU), highlighting its therapeutic potential. Molecular docking studies showed strong binding interactions with key proteins involved in colorectal cancer progression, such as TGFβ2 and VEGFR1. 5j displayed a high binding affinity for TGFβ2 (MM-GBSA ΔG = -92.52kcal/mol) and 5g showed promising interactions with VEGFR1 (ΔG = -70.63kcal/mol). Molecular dynamics simulations confirmed the stability of the ligand-protein complexes, indicating potential as targeted therapeutic agents. Compounds 5g and 5j demonstrate significant promise for further development in colorectal cancer treatment.

A generative framework for enhanced cell-type specificity in rationally designed mRNAs.

mRNA delivery offers new opportunities for disease treatment by directing cells to produce therapeutic proteins. However, designing highly stable mRNAs with programmable cell type-specificity remains a challenge. To address this, we measured the regulatory activity of 60,000 5' and 3' untranslated regions (UTRs) across six cell types and developed PARADE (Prediction And RAtional DEsign of mRNA UTRs), a generative AI framework to engineer untranslated RNA regions with tailored cell type-specific activity. We validated PARADE by testing 15,800 de novo-designed sequences across these cell lines and identified many sequences that demonstrated superior specificity and activity compared to existing RNA therapeutics. mRNAs with PARADE-engineered UTRs also exhibited robust tissue-specific activity in animal models, achieving selective expression in the liver and spleen. We also leveraged PARADE to enhance mRNA stability, significantly increasing protein output and therapeutic durability in vivo. These advancements translated to notable increases in therapeutic efficacy, as PARADE-designed UTRs in oncosuppressor mRNAs, namely PTEN and P16, effectively reduced tumor growth in patient-derived neuroglioma xenograft models and orthotopic mouse models. Collectively, these findings establish PARADE as a versatile platform for designing safer, more precise, and highly stable mRNA therapies.

Cloning of glucoamylase gene from Aspergillus niger and its expression in Pichia pastoris

Glucoamylase (1,4-α-glucosidase) is a crucial commercial enzyme responsible for the conversion of starch, glycogen, and oligosaccharides into D-glucose through the hydrolysis of their non-reducing terminal glycosidic bonds. While many microorganisms, including bacteria, yeasts, and fungi, can produce glucoamylase, fungal glucoamylase is the preferred choice for industrial applications. The goal of this study was to produce the glucoamylase enzyme recombinantly in Pichia pastoris. To achieve this, the glaA gene from Aspergillus niger, responsible for encoding the glucoamylase enzyme, was cloned into a plasmid (pGAPZα-A) under the control of the GAP promoter and subsequently transferred into P. pastoris. The gene was verified through sequence analysis, while the effectiveness of transfection was validated using colony PCR and enzyme activity assays. The results demonstrated that the recombinant P. pastoris strain successfully secreted a substantial amount of glucoamylase (307.05 mg/L). The activity of the recombinant enzyme was measured at 79 U/mL.min. The enzyme exhibited robust activity over a broad range of temperatures (50-80°C) and various pH levels (pH 5-10), retaining 92-60% of its maximum activity. In conclusion, this study highlights the potential for laboratory-scale production of the glucoamylase enzyme, crucial for various industries, from a cost-effective and easily cultivable recombinant yeast strain, P. pastoris.

Graphitic Carbon Nitride for Photocatalytic Hydrogen Production from Water Splitting: Nano-Morphological Control and Electronic Band Tailoring.

Semiconductor polymeric graphitic carbon nitride (g-C3N4) photocatalysts have garnered significant and rapidly increasing interest in the realm of visible light-driven hydrogen evolution reactions. This interest stems from their straightforward synthesis, ease of functionalization, appealing electronic band structure, high physicochemical and thermal stability, and robust photocatalytic activity. This review starts with the basic principle of photocatalysis and the development history, synthetic strategy, and structural properties of g-C3N4 materials, followed by the rational design and engineering of g-C3N4 from the perspectives of nano-morphological control and electronic band tailoring. Some representative results, including experimental and theoretical calculations, are listed to show the advantages of optimizing the above two characteristics for performance improvement in photocatalytic hydrogen evolution from water splitting. The existing opportunities and challenges of g-C3N4 photocatalysts are outlined to illuminate the developmental trajectory of this field. This paper provides guidance for the preparation of g-C3N4 and to better understand the current state of the art for future research directions.

The ATPase activity of yeast chromosome axis protein Hop1 affects the frequency of meiotic crossovers.

Saccharomyces cerevisiae meiosis-specific Hop1, a structural constituent of the synaptonemal complex, also facilitates the formation of programmed DNA double-strand breaks and the pairing of homologous chromosomes. Here, we reveal a serendipitous discovery that Hop1 possesses robust DNA-independent ATPase activity, although it lacks recognizable sequence motifs required for ATP binding and hydrolysis. By leveraging molecular docking combined with molecular dynamics simulations and biochemical assays, we identified an ensemble of five amino acid residues in Hop1 that could potentially participate in ATP-binding and hydrolysis. Consistent with this premise, we found that Hop1 binds to ATP and that substitution of amino acid residues in the putative ATP-binding site significantly impaired its ATPase activity, suggesting that this activity is intrinsic to Hop1. Notably, K65A and N67Q substitutions in the Hop1 N-terminal HORMA domain synergistically abolished its ATPase activity, noticeably impaired its DNA-binding affinity and reduced its association with meiotic chromosomes, while enhancing the frequency of meiotic crossovers (COs). Overall, our study establishes Hop1 as a DNA-independent ATPase and reveals a potential biological function for its ATPase activity in the regulation of meiotic CO frequency.

Phenolic and Flavonoids Contributions to the Antioxidant, Antidiabetic, and Anticholesterol Activities of Eriobotrya japonica Fruit Extract: An In Vitro Analysis

Eriobotrya japonica has been traditionally used for its medicinal properties. This study investigates its fruit ethanol extract's phenolic and flavonoids profiles and evaluates its antioxidant, antidiabetic, and anticholesterol activities. Eriobotrya japonica fruit ethanol extract was tested for total phenolic and flavonoid content using Folin-Ciocalteu and AlCl3 methods. The antioxidant capacity was assessed by DPPH and metal chelation assays. The antidiabetic action was assessed using α-glucosidase inhibition and the Nelson-Somogyi method, whereas the Liebermann-Burchard method was used to measure anticholesterol. The extract exhibited large quantities of phenolics (38.62 mg GAE/g) and flavonoids (4.23 mg QE/g), demonstrating robust antioxidant activity with IC50 of 55 μg/mL by DPPH method and substantial inhibition (71%) by metal ion chelation. In addition, it exhibited strong antidiabetic effects 2.42 μg/mL in the α-glucosidase inhibition test, and at 10 μg/mL using the Nelson-Somogyi method resulted in a 24.02% yield. The extract demonstrated a significant reduction in cholesterol levels during the process of cholesterol reduction, obtaining a maximum reduction of 47.31% at the highest concentration tested. Ethanol extract of Eriobotrya japonica can be used in treating oxidative stress and diabetes and introduce its potential to reduce cholesterol levels. The high content of phenolic and flavonoids total contributes to its bioactivities, indicating its potential as a functional food ingredient.

Immune complexes-mediated activation of neutrophils in systemic lupus erythematosus is dependent on RNA recognition by toll-like receptor 8

IntroductionNeutrophil activation is important in systemic lupus erythematosus (SLE). We previously demonstrated that ribonucleoprotein (RNP) immune complexes (ICs) promoted neutrophil activation in a TLR7/8-dependent manner. However, it remains unclear if this mechanism occurs in patients. Here, we investigated the role of RNA recognition by evaluating TLR7/8 in plasma-mediated neutrophil activation in SLE.MethodsPlasma levels of neutrophil activation markers and ICs were measured by ELISA and flow cytometry in SLE patients (n=151) and healthy controls (HCs, n=31). Neutrophils were incubated with plasma and assessed for CD66b and CD11b up-regulation by flow cytometry in the presence of select inhibitors to define the mechanisms of neutrophil activation by SLE plasma.ResultsSLE plasma induced higher levels of CD66b (p=0.0002) and CD11b (p=0.01) expression than plasma from HCs. Blocking FcγRIIA, targeting RNA sensing by adding RNase, or blocking TLR7/8, TLR8 only, or IRAK4, decreased plasma-mediated neutrophil activation (p<0.05). Consistent with the ability of selective TLR8 inhibitor to block plasma-mediated neutrophil activation, TLR8 agonists, but not TLR7 agonists induced robust neutrophil activation. Further, neutrophil mRNA expression of TLR8 was higher than TLR7. Finally, patients with plasma samples inducing neutrophil activation in RNA-dependent manner had increased levels of interferon alpha, IP-10 (p<0.05), ICs (p<0.05), and reduced complement C3 levels (p<0.01), indicative of IC-driven disease.ConclusionThe data support IC-driven RNA-sensing by TLR8 in neutrophils is a key mechanism of neutrophil activation in SLE. Patients with elevated neutrophil activation and presence of RNA-containing ICs, may benefit from TLR8 inhibition and other strategies targeting RNA removal.