Significant Inhibition Research Articles

Synthesis of β-Amino Carbonyl 6-(Aminomethyl)- and 6-(Hydroxymethyl)pyrazolopyrimidines for DPP-4 Inhibition Study.

Type-2 diabetes is a chronic progressive metabolic disease resulting in severe vascular complications and mortality risk. Recently, DPP-4 inhibitors had been conceived as a favorable class of agents for the treatment of type 2 diabetes due to the minimal side effects. Sitagliptin is the first medicine approved for the DPP-4 inhibitor. Its structure involved three fragments: 2,4,5-triflorophenyl fragment pharmacophore, enantiomerically β-amino carbonyl linker, and tetrahydrotriazolopyridine. Herein, we are drawn to the possibility of substituting tetrahydrotriazolopyridine motif present in Sitagliptin with a series of new fused pyrazolopyrimidine bicyclic fragment to investigate potency and safety. Two series of fused 6-(aminomethyl)pyrazolopyrimidine and 6-(hydroxymethyl) pyrazolopyrimidine derivatives containing β-amino ester or amide as linkers were successfully designed for the new DPP-4 inhibitors. Most fused 6-methylpyrazolopyrimidines were evaluated against DPP-4 inhibition and selectivity capacity. Based on research study, β-amino carbonyl fused 6-(hydroxymethyl)pyrazolopyrimidine possesses the significant DPP-4 inhibition (IC50 ≤ 59.8 nM) and presents similar with Sitagliptin (IC50 = 28 nM). Particularly, they had satisfactory selectivity over DPP-8 and DPP-9, except for QPP. β-Amino esters and amides fused 6-(hydroxymethyl)pyrazolopyrimidine were developed as the new DPP-4 inhibitors. Those compounds with a methyl group or hydrogen in N-1 position and methyl substituted group in C-3 of pyrazolopyrimidine moiety showed better potent DPP-4 inhibition (IC50 = 21.4-59.8 nM). Furthermore, they had satisfactory selectivity over DPP-8 and DPP-9 Finally, the docking results revealed that compound 9n was stabilized at DPP-4 active site and would be a potential lead drug.

The Power of Molecular Dynamics Simulations and Their Applications to Discover Cysteine Protease Inhibitors.

A large family of enzymes with the function of hydrolyzing peptide bonds, called peptidases or cysteine proteases (CPs), are divided into three categories according to the peptide chain involved. CPs catalyze the hydrolysis of amide, ester, thiol ester, and thioester peptide bonds. They can be divided into several groups, such as papain-like (CA), viral chymotrypsin-like CPs (CB), papainlike endopeptidases of RNA viruses (CC), legumain-type caspases (CD), and showing active residues of His, Glu/Asp, Gln, Cys (CE). The catalytic mechanism of CPs is the essential cysteine residue present in the active site. These mechanisms are often studied through computational methods that provide new information about the catalytic mechanism and identify inhibitors. The role of computational methods during drug design and development stages is increasing. Methods in Computer-Aided Drug Design (CADD) accelerate the discovery process, increase the chances of selecting more promising molecules for experimental studies, and can identify critical mechanisms involved in the pathophysiology and molecular pathways of action. Molecular dynamics (MD) simulations are essential in any drug discovery program due to their high capacity for simulating a physiological environment capable of unveiling significant inhibition mechanisms of new compounds against target proteins, especially CPs. Here, a brief approach will be shown on MD simulations and how the studies were applied to identify inhibitors or critical information against cysteine protease from several microorganisms, such as Trypanosoma cruzi (cruzain), Trypanosoma brucei (rhodesain), Plasmodium spp. (falcipain), and SARS-CoV-2 (Mpro). We hope the readers will gain new insights and use our study as a guide for potential compound identifications using MD simulations.

Phytol and α-Bisabolol Synergy Induces Autophagy and Apoptosis in A549 Cells and Additional Molecular Insights through Comprehensive Proteome Analysis via Nano LC-MS/MS.

Non-Small Cell Lung Cancer (NSCLC) is a malignancy with a significant prevalence and aggressive nature, posing a considerable challenge in terms of therapeutic interventions. Autophagy and apoptosis, two intricate cellular processes, are integral to NSCLC pathophysiology, each affecting the other through shared signaling pathways. Phytol (Phy) and α-bisabolol (Bis) have shown promise as potential anticancer agents individually, but their combined effects in NSCLC have not been extensively investigated. The present study was to examine the synergistic impact of Phy and Bis on NSCLC cells, particularly in the context of autophagy modulation, and to elucidate the resulting differential protein expression using LCMS/ MS analysis. The A549 cell lines were subjected to the patented effective concentration of Phy and Bis, and subsequently, the viability of the cells was evaluated utilizing the MTT assay. The present study utilized real-time PCR analysis to assess the expression levels of crucial apoptotic genes, specifically Bcl-2, Bax, and Caspase-9, as well as autophagy-related genes, including Beclin-1, SQSTM1, Ulk1, and LC3B. The confirmation of autophagy marker expression (Beclin-1, LC3B) and the autophagy-regulating protein SQSTM1 was achieved through the utilization of Western blot analysis. Differentially expressed proteins were found using LC-MS/MS analysis. The combination of Phy and Bis demonstrated significant inhibition of NSCLC cell growth, indicating their synergistic effect. Real-time PCR analysis revealed a shift towards apoptosis, with downregulation of Bcl-2 and upregulation of Bax and Caspase-9, suggesting a shift towards apoptosis. Genes associated with autophagy regulation, including Beclin-1, SQSTM1 (p62), Ulk1, and LC3B, showed significant upregulation, indicating potential induction of autophagy. Western blot analysis confirmed increased expression of autophagy markers, such as Beclin-1 and LC3B, while the autophagy-regulating protein SQSTM1 exhibited a significant decrease. LC-MS/MS analysis revealed differential expression of 861 proteins, reflecting the modulation of cellular processes. Protein-protein interaction network analysis highlighted key proteins involved in apoptotic and autophagic pathways, including STOML2, YWHAB, POX2, B2M, CDA, CAPN2, TXN, ECHS1, PEBP1, PFN1, CDC42, TUBB1, HSPB1, PXN, FGF2, and BAG3, emphasizing their crucial roles. Additionally, PANTHER pathway analysis uncovered enriched pathways associated with the differentially expressed proteins, revealing their involvement in a diverse range of biological processes, encompassing cell signaling, metabolism, and cellular stress responses. The combined treatment of Phy and Bis exerts a synergistic inhibitory effect on NSCLC cell growth, mediated through the interplay of apoptosis and autophagy. The differential protein expression observed, along with the identified proteins and enriched pathways, provides valuable insights into the underlying molecular mechanisms. These findings offer a foundation for further exploration of the therapeutic potential of Phy and Bis in the management of NSCLC.

Preventive Effect of Astaxanthin on Diabetic Cataracts and Its Protective Effect on Lens Epithelial Cells

To better combat the development of diabetic cataracts (DC), we conducted a study on the use of astaxanthin (ATX) as an innovative antioxidant. Our research found that ATX exhibited good biocompatibility with lens epithelial cells (LECs) and increasing its concentration did not reduce cell activity. In the presence of high glucose, LECs experienced elevated oxidative stress levels, leading to a significant inhibition of their activity. However, the addition of ATX increased the antioxidant levels in LECs, resulting in a concentration-dependent increase in cell activity. Furthermore, when tested on diabetic rat models, higher concentrations of ATX application effectively prevented the development of DC and suppressed the level of oxidative stress in the lens. Our findings highlight the strong antioxidant effect of ATX, which can safeguard LEC activity in the presence of high glucose and inhibit the development of DC.

Abstract 4503: Preclinical characterization of PRT7732: A highly potent, selective, and orally bioavailable targeted protein degrader of SMARCA2

Abstract SWI/SNF (BAF) complexes play an important role in controlling gene expression by remodeling chromatin. SMARCA2 (BRM) and SMARCA4 (BRG1) are the core catalytic subunits of the SWI/SNF complexes, containing an ATPase domain and a bromodomain. SMARCA4 protein expression is lost in some cancers due to loss-of-function (LOF) mutations and homozygous deletions, and SMARCA4-deleted cancer cells are highly dependent on its paralog gene SMARCA2 for their survival. Therefore, targeting SMARCA2 in SMARCA4-deficient cancers using selective SMARCA2 degraders induces synthetic lethality while sparing SMARCA4 wild type (WT) normal cells. We have recently identified a series of orally bioavailable SMARCA2 selective degraders that demonstrate robust efficacy in pre-clinical animal models with favorable pharmacokinetic properties and safety profiles. Our nomination candidate PRT7732 exhibits >1000x selectivity for SMARCA2 over SMARCA4 in cell-based assays, with DC50 values in cancer cell lines in the low nanomolar range. The PRT7732-induced SMARCA2 degradation was rescued by a proteasome inhibitor and neddylation inhibitor, indicating ubiquitin-proteasome dependent degradation. Furthermore, PRT7732 does not change levels of known CRBN ligand neo-substrates such as IKZF2/3, GSPT1 and SALL4. PRT7732 inhibits only SMARCA4-deficient cancer cell proliferation with IC50 values ranging from 5.0-50 nM, but not SMARCA4 WT cells in vitro. Oral administration of PRT7732 resulted in near-total degradation of SMARCA2 protein with complete selectivity over SMARCA4 protein in a SMARCA4 WT lung cancer model in mice, consistent with the SMARCA2 degradation kinetics-based pharmacodynamic prediction model. PRT7732 oral daily administration showed significant tumor growth inhibition of SMARCA4-deficient lung cancer xenograft models at well tolerated doses. The treated tumor tissues show robust SMARCA2 protein reduction for 24h post dosing. In summary, our orally bioavailable SMARCA2 degraders induce synthetic lethality in SMARCA4-deficient cancers in vitro and in vivo. Efforts to further evaluate these compounds in additional models and in combination with other agents are ongoing. Citation Format: Artem Shvartsbart, Koichi Ito, Joseph Rager, Michael Hulse, Anjana Agarwal, Komali Vykuntam, Jessica Burtell, Min Wang, Justin Kurian, Miles Cowart, Joy Cote, Nick Stahl, Monisha Sivakumar, Anthony Reichelderfer, Jack Carter, Alexander Grego, Andrew Moore, Neha Bhagwat, Ross Kuskovsky, Shanthi Ganesan, Stefan Ruepp, Tom Emm, Philip Pitis, Corey Basch, Klare Bersch, Yongchun Pan, Song Mei, Raul Leal, John Rose, Dani Roth, Chaoyi Xu, Ganfeng Cao, Kris Vaddi, Sang Hyun Lee, Sandy Geeganage, Andrew Combs, Peggy Scherle. Preclinical characterization of PRT7732: A highly potent, selective, and orally bioavailable targeted protein degrader of SMARCA2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4503.

Abstract 2780: Induced pluripotent stem cell (iPSC) based vaccines potentializes anti-CTLA4 immune response in a murine lung carcinoma model

Abstract Despite the development of new immunotherapy strategies, lung cancer remains the leading cause of cancer-related deaths. One of the reasons is attributed to the persistence of cancer stem cells (CSCs) within the tumor, contributing to drug resistance, dormancy and metastatic spread. To eradicate CSCs, iPSC-based vaccination represents a novel approach based on the paradigm that CSCs and iPSCs share common tumor associated antigens (TAAs) with the ability to elicit a strong anti-tumor immune response.We previously showed that induced allogeneic pluripotent stem cells (miPSCs), combined with a histone deacetylase inhibitor (HDACi), induced a major reduction of tumor growth and metastatic spread to lungs in an aggressive triple negative breast cancer model (4T1) in syngeneic mice. We report here the use of the same protocol in a lung carcinoma model using LLC1 cell line in combination with an anti-CTLA4 antibody. We compared the effectiveness of allogenic miPSCs treatment alone or with an anti-CTLA4 mouse antibody on mice-bearing LLC1-Luc tumors (mean+se: 17,5+0,6 mm3). Five mice received 5 subcutaneous (SC) injection of 2 × 106 miPSCs, 5 mice received 3 intraperitoneal injections of anti-CTLA4 antibody at 5mg/kg and 5 mice were treated with the combinatory regimen (miPSCs and anti- CTLA4) with the same doses. In addition, all treated mice received a HDACi in the drink water throughout the experience.After 24 days, all tumor volumes from the 3 groups were significantly reduced compared to the control mice receiving only PBS. We obtained a reduction of 72% (p=0.007) and 74% (p=0.009) respectively for the mice treated solely with miPSCs or anti-CTLA4. An important synergic effect was obtained for the mice treated with miPSCs and anti- CTLA4 with a reduction of the tumor size by 92% (p=0.005) compared to the control mice. We also found a significant inhibition of metastatic spread to lungs for the 3 groups that was more pronounced in mice receiving the combination treatment with a significant reduction of 91 % compared to the control group. All these therapeutic effects were correlated with a significant increase of CD4+ and CD8+ cells that was more pronounced in mice receiving the combinatory regimen.We also have performed single-cell RNA sequencing analysis (10x Genomics) of LLC1 tumors treated with miPSCs and anti- CTLA4 compared to mice treated with monotherapy and untreated mice. T-distributed stochastic neighbor embedding plot reveals distinct clusters of stromal and immune cells across tumors with a significant upregulation of CD8+ and CD4+ T cells in mice receiving the combinatory regimen.Overall, these results demonstrate for the first time the clinical relevance of using an off-the-shelf allogeneic iPSC-based vaccine combined with an anti-CTLA4 antibody as a new anti-cancer immunotherapy strategy against aggressive non-small cell lung cancer (NSCLC) with metastatic potential. Citation Format: Frank Griscelli, Diana Chaker, Noufissa Oudrhiri, Christophe Desterke, Ali G. Turhan, Annelise Bennaceur Griscelli. Induced pluripotent stem cell (iPSC) based vaccines potentializes anti-CTLA4 immune response in a murine lung carcinoma model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2780.

Extracellular vesicles secreted from mesenchymal stem cells ameliorate renal ischemia reperfusion injury by delivering miR-100-5p targeting FKBP5/AKT axis

The incidence of acute kidney injury (AKI) due to ischemia–reperfusion (IR) injury is increasing. There is no effective treatment for AKI, and because of this clinical challenge, AKI often progresses to chronic kidney disease, which is closely associated with poor patient outcomes and high mortality rates. Small extracellular vesicles from human umbilical cord mesenchymal stem cells (hUCMSC-sEVs) play increasingly vital roles in protecting tissue function from the effects of various harmful stimuli owing to their specific biological features. In this study, we found that miR-100-5p was enriched in hUCMSC-sEVs, and miR-100-5p targeted FKBP5 and inhibited HK-2 cell apoptosis by activating the AKT pathway. HK-2 cells that were exposed to IR injury were cocultured with hUCMSC-sEVs, leading to an increase in miR-100-5p levels, a decrease in FKBP5 levels, and an increase in AKT phosphorylation at Ser 473 (AKT-473 phosphorylation). Notably, these effects were significantly reversed by transfecting hUCMSCs with an miR-100-5p inhibitor. Moreover, miR-100-5p targeted FKBP5, as confirmed by a dual luciferase reporter assay. In vivo, intravenous infusion of hUCMSC-sEVs into mice suffering from IR injury resulted in significant apoptosis inhibition, functional maintenance and renal histological protection, which in turn decreased FKBP5 expression levels. Overall, this study revealed an effect of hUCMSC-sEVs on inhibiting apoptosis; hUCMSC-sEVs reduced renal IR injury by delivering miR-100-5p to HK-2 cells, targeting FKBP5 and thereby promoting AKT-473 phosphorylation to activate the AKT pathway. This study provides novel insights into the role of hUCMSC-sEVs in the treatment of AKI.

Phytochemical analysis and evaluation of its antioxidant, antimicrobial, and cytotoxic activities for different extracts of Casuarina equisetifolia.

Casuarina equisetifolia belongs to the Casuarina species with the most extensive natural distribution, which contain various phytochemicals with potential health benefits. This study aimed to investigate the chemical composition and biological activities of different extracts of Casuarina equisetifolia. The n-hexane extract was analyzed for its unsaponifiable and fatty acid methyl esters fractions, while chloroform, ethyl acetate, and butanol extracts were studied for their phenolic components. Six different extracts of C. equisetifolia needles were evaluated for their total phenolic content, total flavonoid content, and their antioxidant, antimicrobial, and cytotoxic activities. The n-hexane extract contained mainly hydrocarbons and fatty acid methyl esters, while ten phenolic compounds were isolated and identified in the chloroform, ethyl acetate, and butanol extracts. The methanolic extract exhibited the highest total phenolic and flavonoid content, highest antioxidant activity, and most potent cytotoxic activity against HepG-2 and HCT-116 cancer cell lines. The ethyl acetate extract showed the most significant inhibition zone against Staphylococcus aureus and Bacillus subtilis. Casuarina equisetifolia extracts showed promising antioxidant, antimicrobial, and cytotoxic activities. Overall, Casuarina equisetifolia is a versatile tree with a variety of uses, and its plant material can be used for many different purposes.

Natural compound plumbagin based inhibition of hIAPP revealed by Markov state models based on MD data along with experimental validations.

Human islet amyloid polypeptide (amylin or hIAPP) is a 37 residue hormone co-secreted with insulin from β cells of the pancreas. In patients suffering from type-2 diabetes, amylin self-assembles into amyloid fibrils, ultimately leading to the death of the pancreatic cells. However, a research gap exists in preventing and treating such amyloidosis. Plumbagin, a natural compound, has previously been demonstrated to have inhibitory potential against insulin amyloidosis. Our investigation unveils collapsible regions within hIAPP that, upon collapse, facilitates hydrophobic and pi-pi interactions, ultimately leading to aggregation. Intriguingly plumbagin exhibits the ability to bind these specific collapsible regions, thereby impeding the aforementioned interactions that would otherwise drive hIAPP aggregation. We have used atomistic molecular dynamics approach to determine secondary structural changes. MSM shows metastable states forming native like hIAPP structure in presence of PGN. Our in silico results concur with in vitro results. The ThT assay revealed a striking 50% decrease in fluorescence intensity at a 1:1 ratio of hIAPP to Plumbagin. This finding suggests a significant inhibition of amyloid fibril formation by plumbagin, as ThT fluorescence directly correlates with the presence of these fibrils. Further TEM images revealed disappearance of hIAPP fibrils in plumbagin pre-treated hIAPP samples. Also, we have shown that plumbagin disrupts the intermolecular hydrogen bonding in hIAPP fibrils leading to an increase in the average beta strand spacing, thereby causing disaggregation of pre-formed fibrils demonstrating overall disruption of the aggregation machinery of hIAPP. Our work is the first to report a detailed atomistic simulation of 22 μs for hIAPP. Overall, our studies put plumbagin as a potential candidate for both preventive and therapeutic candidate for hIAPP amyloidosis.

Determination of the proteolytic profile in Balaustium murorum (Hermann, 1804) (Acari: Prostigmata: Erythraeidae)

ABSTRACT Balaustium murorum (Acariformes: Erythraeidae) is a pollen feeder and predatory mite that occasionally causes dermatitis. We sought to determine the protease profile in the whole-mite extract of B. murorum using general and specific substrates as well as inhibitors. The pH profiles using two general substrates, azacasein and haemoglobin, revealed an optimal pH of 6 for both substrates. Specific trypsin inhibitor TLCK showed no influence activity towards BApNA; however, TPCK and AEBSF inhibited chymotrypsin and elastase, respectively. Specific cysteine protease inhibitors Cystatin and E-64 influenced both types of cathepsins B and L, while DTT as a specific activator increased the enzymatic activities. Only aminopeptidase showed significant activity and inhibition using negative control and phenanthroline. Chymotrypsin and elastase had the highest activity at pH of 9–10 and 10, respectively, by using both substrates alone and substrates along with specific inhibitors. Cathepsins B and L had pH optima of 6. Aminopeptidase revealed a broad pH optimum of 5–7 on both substrates HA and HPA alone and substrate along with inhibitor phenanthroline. Finally, different concentrations of each inhibitor were used to find IC50 values of TPCK, AEBSF, cystatin, E-64, DTT and phenanthroline required to inhibit half of enzyme activity.

Green Ostrich fat waste extracts as a novel potential inhibitor to sustainable corrosion of steel in acidic environments: Electrochemical and DFT evaluation

The shift from traditional chemical corrosion inhibitors to green alternatives, focusing on plant extracts (GCIs), addresses environmental concerns in line with sustainable development goals. An innovative approach explores animal-derived extracts for eco-friendly corrosion inhibition, addressing environmental issues associated with conventional chemical inhibitors. Ostrich Oil, renowned for its nutritional, cosmetic, and pharmaceutical properties, is examined as a potential green corrosion inhibitor for C38 steel in 1 M hydrochloric. Extracted and characterized through GC-FID, Ostrich Oil exhibits corrosion protection analyzed via impedance spectroscopy, potentiodynamic polarization, and gravimetric methods. Complementary SEM and metallurgical microscopy with EDS offer surface insights. DFT and MD simulations explore the relationship between electronic properties and inhibition efficiency. Results show Ostrich Oil's significant corrosion inhibition (up to 94%), attributed to chemical adsorption of fatty compounds. The corrosion mechanism is influenced by Oil extract adsorption, increasing double electrochemical layer resistance. Gas chromatography identifies main compounds, and quantum chemical calculations highlight its reactivity. This research underscores the potential of animal-derived extracts, exemplified by Ostrich Oil, as effective and environmentally friendly corrosion inhibitors, contributing to sustainable corrosion protection practices.

Morus alba L. (Sangzhi) alkaloids mitigate atherosclerosis by regulating M1/M2 macrophage polarization

BackgroundAtherosclerosis (AS) is an important cause of cardiovascular disease, posing a substantial health risk. Recognized as a chronic inflammatory disorder, AS hinges on the pivotal involvement of macrophages in arterial inflammation, participating in its formation and progression. Sangzhi alkaloid (SZ-A) is a novel natural alkaloid extracted from the mulberry branches, has extensive pharmacological effects and stable pharmacokinetic characteristics. However, the effects and mechanisms of SZ-A on AS remain unclear. PurposeTo explore the effect and underlying mechanisms of SZ-A on inflammation mediated by macrophages and its role in AS development. MethodsAtherosclerosis was induced in vivo in apolipoprotein E-deficient mice through a high-fat and high-choline diet. We utilized macrophages and vascular endothelial cells to investigate the effects of SZ-A on macrophage polarization and its anti-inflammatory properties on endothelial cells in vitro. The transcriptomic analyses were used to investigate the major molecule that mediates cell-cell interactions and the antiatherogenic mechanisms of SZ-A based on AS, subsequently validated in vivo and in vitro. ResultsSZ-A demonstrated a significant inhibition in vascular inflammation and alleviation of AS severity by mitigating macrophage infiltration and modulating M1/M2 macrophage polarization in vitro and in vivo. Moreover, SZ-A effectively reduced the release of the proinflammatory mediator C-X-C motif chemokine ligand (CXCL)-10, predominantly secreted by M1 macrophages. This reduction in CXCL-10 contributed to improved endothelial cell function, reduced recruitment of additional macrophages, and inhibited the inflammatory amplification effect. This ultimately led to the suppression of atherogenesis. ConclusionSZ-A exhibited potent anti-inflammatory effects by inhibiting macrophage-mediated inflammation, providing a new therapeutic avenue against AS. This is the first study demonstrating the efficacy of SZ-A in alleviating AS severity and offers novel insights into its anti-inflammatory mechanism.

Antibacterial potentials of extracts from Gryllotalpa gryllotalpa, Pentodon algerinum grubs, and Gypsonoma euphraticana larva frass

Objectives: The overuse and abuse of antibiotics have accelerated antibiotic resistance, and to solve this problem, it has been found that many insect species have potential antimicrobial properties against a wide range of resistant pathogens. Our study tests the antibacterial activity of microbial defensive compounds included in body extract of insects inhabiting contaminated environments and frass of phytophagous insects. Materials and Methods: Through sequential extraction method by acidic methanol, chloroform, and hexane solvents, insect body extract of Gryllotalpa gryllotalpa, grubs of Pentodon algerinum besides feces of Gypsonoma euphraticana larvae were tested against Gram-positives Bacillus cereus, Bacillus coagulans, Staphylococcus aureus, and Salmonella typhi, Escherichia coli, and Klebsiella pneumoniae. The antibiotics ceftriaxone (CRO) and ampicillin (AM) were used as standard drugs. The antibacterial growth inhibition was estimated by well diffusion methods. Results: High significant antibacterial activity against the tested bacteria by acidic methanol then chloroform extracts, while hexane extract of all the three insect species only produced significant growth inhibition of S. aureus. In addition, growth inhibition 20.0 mm or more was induced by: MeOH extracts of G. gryllotalpa and P. algerinum for S. typhi and E. coli, besides chloroform G. gryllotalpa extract for S. typhi. The tested bacteria S. aureus, S. typhi, and K. pneumoniae were AM-resistant, while E. coli was both AM and CRO-resistant. Conclusion: Acidic meOH and chloroform body extract of G. gryllotalpa and P. algerinum and larvae G. euphraticana feces extract possess bioactive compounds with promising antibacterial properties, for overcoming antibiotic resistance.

Establishment of an immunogenic cell death-related model for prognostic prediction and identification of therapeutic targets in endometrial carcinoma.

Studies have firmly established the pivotal role of the immunogenic cell death (ICD) in the development of tumors. This study seeks to develop a risk model related to ICD to predict the prognosis of patients with endometrial carcinoma (EC). RNA-seq data of EC retrieved from TCGA database were analyzed using R software. We determined clusters based on ICD-related genes (ICDRGs) expression levels. Cox and LASSO analyses were further used to build the prediction model, and its accuracy was evaluated in the train and validation sets. Finally, in vitro and in vivo experiments were conducted to confirm the impact of the high-risk gene IFNA2 on EC. Patients were sorted into two ICD clusters, with survival analysis revealing divergent prognoses between the clusters. The Cox regression analysis identified prognostic risk genes, and the LASSO analysis constructed a model based on 9 of these genes. Notably, this model displayed excellent predictive accuracy when validated. Finally, increased IFNA2 levels led to decreased vitality, proliferation, and invasiveness in vitro. IFNA2 also has significant tumor inhibiting effect in vivo. The ICD-related model can accurately predict the prognosis of patients with EC, and IFNA2 may be a potential treatment target.

The potential of Paeonia lactiflora pall seeds oil as a pure natural cosmetics raw material: InVitro findings.

As a traditional Chinese herbal medicine, Paeonia lactiflora Pall is rich in various active ingredients such as polysaccharides and total flavonoids while having ornamental value. It has potential application value in the development of food and cosmetics. To study the invitro efficacy of Paeonia lactiflora Pall seeds oil. Firstly, the levels of linolenic acid and linoleic acid in Paeonia lactiflora Pall seeds oil were quantified using gas chromatography. The impact of Paeonia lactiflora Pall seeds oil on the proliferation rate of B16F10 cells was assessed through the CCK-8 method, while the melanin content of B16F10 cells was determined using the sodium hydroxide lysis method. The inhibitory effects of Paeonia lactiflora Pall seeds oil on elastase, collagenase and hyaluronidase were evaluated by biochemical techniques invitro. Lastly, the hen's egg chorioallantoic membrane test (HET-CAM) was conducted to confirm the absence of eye irritation caused by Paeonia lactiflora Pall seeds oil. Paeonia lactiflora Pall seeds oil within a certain volume concentration range (0.5%-4%) had no effect on the proliferation of B16F10 cells. Paeonia lactiflora Pall seeds oil showed significant inhibition of elastase, collagenase and hyaluronidase. Notably, the highest concentration tested, 4% Paeonia lactiflora Pall seed oil, yielded the most pronounced outcomes without causing any irritation. A certain concentration of Paeonia lactiflora Pall seeds oil has a significant effect on decreasing the melanin content in B16F10 cells and inhibiting the activities of elastase, collagenase, and hyaluronidase, which can provide a reference for the development of pure natural cosmetics raw materials.

Inhibitory Effects of the Polyphenols from the Root of Rhizophora apiculata Blume on Fatty Acid Synthase Activity and Human Colon Cancer Cells.

Marine mangrove vegetation has been traditionally employed in folk medicine to address various ailments. Notably, Rhizophora apiculata Blume has exhibited noteworthy properties, demonstrating efficacy against cancer, viruses, and bacteria. The enzyme fatty acid synthase (FAS) plays a pivotal role in de novo fatty acid synthesis, making it a promising target for combating colon cancer. Our study focused on evaluating the FAS inhibitory effects of both the crude extract and three isolated compounds from R. apiculata. The n-butanol fraction of R. apiculata extract (BFR) demonstrated a significant inhibition of FAS, with an IC50 value of 93.0 µg/mL. For inhibition via lyoniresinol-3α-O-β-rhamnopyranoside (LR), the corresponding IC50 value was 20.1 µg/mL (35.5 µM). LR competitively inhibited the FAS reaction with acetyl-CoA, noncompetitively with malonyl-CoA, and in a mixed manner with NADPH. Our results also suggest that both BFR and LR reversibly bind to the KR domain of FAS, hindering the reduction of saturated acyl groups in fatty acid synthesis. Furthermore, BFR and LR displayed time-dependent inhibition for FAS, with kobs values of 0.0045 min-1 and 0.026 min-1, respectively. LR also exhibited time-dependent inhibition on the KR domain, with a kobs value of 0.019 min-1. In human colon cancer cells, LR demonstrated the ability to reduce viability and inhibit intracellular FAS activity. Notably, the effects of LR on human colon cancer cells could be reversed with the end product of FAS-catalyzed chemical reactions, affirming the specificity of LR on FAS. These findings underscore the potential of BFR and LR as potent FAS inhibitors, presenting novel avenues for the treatment of human colon cancer.

Spatial-temporal characteristics and influencing factors of carbon emission in Chengdu-Chongqing area: an urban transportation perspective.

The Chengdu-Chongqing twin-city economic circle is a vital growth pole and a new power source for Chinese high-quality development. Studying the spatial-temporal characteristics of carbon emissions and the role of factors affecting them under the transportation perspective is of great significance for this region to realize the carbon peak and carbon neutrality and to formulate carbon emission reduction policies. We use the exploring spatial data analysis (ESDA) and spatial regression model combined with the STIRPAT model, and research finding: (1) The total carbon emissions in the research area gradually increased from 2014 to 2020, but the growth rate showed a significant decline in 2019. (2) There is significant spatial heterogeneity of carbon emissions in the study area; the hotspot areas of total carbon emissions are in Chongqing and Chengdu, forming a high-low aggregation of carbon emissions. Per capita carbon emissions show a high trend in the southwest and a low in the northeast. (3) From the factors of transportation perspective, highway density and private vehicles have a positive impact on carbon emissions, and urban road areas and public transportation have a very significant inhibition of carbon emissions and a spatial spillover effect. (4) Other factors, such as population size, national economic development, urbanization level, and industrial structure, all have a positive effect on carbon emissions, and disposable income has a negative effect on carbon emissions.

Relationship among runoff, soil erosion, and rill morphology on slopes of overburdened stockpiles under simulated rainfall

Overburdened stockpiles are difficult to achieve natural stability in a short period time because of their complex and diverse composition of materials and significant differences in physical and chemical properties, causing numerous cases of water loss and soil loss events. We conducted a series of experiments at different rainfall intensities (30, 60, 90, and 120 mm h−1) and different gravel contents (0 %, 10 %, 20 %, 30 %, and 40 %) to study the runoff, soil erosion, and rill morphology characteristics and relationship among three of overburdened stockpiles in production and construction projects with simulated rainfall. The runoff rate and soil loss rate basically showed a trend of increasing and then decreasing with gravel content and lower gravel content had a certain degree of inhibition on the flow velocity. 20 %, 0 %, 0 %, and 20 % gravel contents had the most significant inhibition on the flow velocity at 30, 60, 90, and 120 mm h−1 rainfall intensities, respectively. Flow shear stress, stream power, unit energy of water cross-section, maximum rill length, width and depth all increased with increasing rainfall intensity. The stream power, unit stream power, and unit energy of water cross-section were larger at higher gravel content. The maximum rill length and depth were positively correlated with gravel content, which were the minimum at 0 % gravel content and all rainfall intensities. Gravel content played a more important role in the runoff, soil loss, and rill morphology. The relationship between runoff rate and soil loss rate, flow velocity and rill morphology parameters were all expressed as linear functions, and flow velocity and soil loss rate was quadratic function, with a correlation coefficient of 0.339. In contrast, runoff rate and rill morphology parameters, soil loss rate and rill morphology parameters, erosion dynamics parameters and rill morphology parameters were all expressed as power functions. In addition, the runoff rate can better describe the process of soil loss (R2 = 0.798, P < 0.05). The unit energy of water cross-section was more suitable to describe the development of maximum rill length and depth (with rill length was R2 = 0.781, and with rill depth was R2 = 0.723, P < 0.05). At the same time, the stream power was more suitable to describe the development of maximum rill width on the slope of overburdened stockpiles (R2 = 0.841, P < 0.05). These results contributed to preventing soil loss on overburdened stockpiles and helping the development of estimation models for soil erosion.

Abstract B002: Exploiting JAK/STAT signaling to inhibit highly advanced and resistant forms of ovarian cancer

Abstract Ovarian cancer is the most fatal of all female reproductive cancers due to its late detection, aggressive nature, and propensity to develop resistance to current standards of care. Despite initial responses to first line platinum- and paclitaxel-based chemotherapies, &amp;gt;70% of ovarian cancers recur with increasingly resistant disease. Targeted therapies, including PARP-inhibitors (PARPi) and VEGF-inhibitors, are being included in the front-line setting; and while they have improved progression-free survival in a subset of patients, they have thus far failed to improve overall survival in patients without specific genetic signatures. With the broad uptake of PARPi, PARPi-resistance is becoming common. Once resistance develops, nearly all patients die of their disease and there is an obvious need to elucidate the basis for chemotherapy- and PARPi-resistance and devise novel approaches to overcome resistance. Using a small molecule drug screen, we identified lestaurtinib, a known tyrosine kinase inhibitor as a potent inhibitor of treatment naive and highly advanced models of ovarian cancer. Lestaurtinib was originally developed to target FLT3, a protein not expressed in any ovarian cancer models we’ve tested. To identify lestaurtinib targets, we preformed phospho-proteomics in parallel with RNAseq and discovered the JAK/STAT signally pathway as a top hit. We confirmed that essential components of JAK/STAT signaling are constitutively activated in chemotherapy- and PARPi-resistant cells, suggesting that induction of this pathway may drive resistance. Importantly, we found that lestaurtinib-sensitive patient-derived-xenografts (PDXs) exhibited higher expression of multiple JAK/STAT signaling pathway members compared to PDXs that did not respond to lestaurtinib. Genetic knockdown of STAT1 and STAT3 via siRNA, or knockout via CRISPR/Cas9, resulted in significant growth inhibition of sensitive and resistant models, confirming their importance in maintaining cell viability and progression. Combining lestuartinib with standard-of-care cisplatin or olaparib, a PARPi, was shown to be synergistic in multiple models, indicating that pharmacological inhibition of JAK/STAT signaling has the potential to alleviate drug resistance. Finally, we assessed a panel of JAK/STAT inhibitors that interfere at various levels of the pathway and found profound differences in their ability to block STAT-phosphorylation and inhibit cell proliferation. Thus, it is critical to further define the mechanisms by which these drugs function to selectively suppress JAK/STAT signaling in order to elucidate specific nodes of the pathway that drive ovarian cancer progression. Ongoing studies are aimed at identifying the unique roles of STAT1 and STAT3, the specific contributions of their tyrosine and serine phosphorylation, and the relevance of up-/down-stream mediators of the JAK/STAT pathway on ovarian cancer cell growth with the goal of informing future clinical trials. Citation Format: Esther Rodman, Michael Emch, Archit Bajaj, Xiaojia Tang, Xiaonan Hou, Scott Kaufmann, Krishna Kalari, John Weroha, John Hawse. Exploiting JAK/STAT signaling to inhibit highly advanced and resistant forms of ovarian cancer [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr B002.

Volatile compounds and bioactivities properties of Opuntia microdasys (Lehm.) and Opuntia macrorhiza (Engelm.) seeds

The seeds of Opuntia sp. require additional investigations because they could contain several active molecules. Therefore, the present study analyzed the chemical composition and biological activities of Opuntia microdasys and that of Opuntia macrorhiza using in vitro and in vivo models. The headspace of the powders of the seeds of Opuntia have been analyzed by gas chromatography–mass spectrometry (GC–MS). The analgesic activity was assessed using the acetic acid-induced abdominal constriction response on mouse. Chemical analyses showed that the powders of the seeds of O. microdasys and O. macrorhiza were rich in nonanal and camphor. Also, the O. macrorhiza seeds has a high antioxidant activity (EC50 =0.12 mg/ml). The results showed that the ethanolic extracts, at a dose of 200 mg/kg of body weight, caused a significant (P&lt; 0.05) inhibition of the abdominal constriction response (75.17% and 56.22% for O. microdasys and O. microdasys). The microdilution assay revealed that ethanolic extract of O. macrorhiza seeds is more active and could inhibit the growth of almost all the evaluated bacterial and fungal strains, with the minimal bactericidal concentration values (MIC) ranging from 0.039 to 2.50 mg/mL. The overall results highlighted the potential of the tested Opuntia seeds that can be used as a source of bioactive compounds that confer various important biological activities to it, which could imply other pharmacological applications.