Growth Inhibition Research Articles

ГЕТЕРОГЕННОСТЬ СИБИРСКИХ ПОПУЛЯЦИЙ ФУЗАРИОИДНЫХ ГРИБОВ ПО ФИТОПАТОГЕННЫМ СВОЙСТВАМ

The purpose of the work is to study the phytopathogenicity of Siberian strains of fusarium fungi (Fusarium, Neocosmospora) associated with grain crops, and their intraspecific heterogeneity in pathogenic properties. Highly toxic species have been identified F. sporotrichioides and F. oxysporum, the metabolites of which reduce the average laboratory germination of Triticumaestivum and Larixsibirica seeds by 50 % or more compared to the control. The species F. sporotrichioides and F. fujikuroi SC inhibit the development of the stem and root of wheat to an equal extent; N. solani and F. oxysporum suppress the root system to a greater extent. Intraspecific differences in the degree of phytotoxicity in relation to Zeamays seedlings and the dynamics of accumulation of metabolites for the species F. sporotrichioides, F. avenaceum, F. oxysporum, N. solani and F. fujikuroi SC are shown. Average root growth inhibition rates range from 52 to 89 %. For all strains, the phytotoxic effect increased (up to 98 %) with increasing cultivation time (14 days or more). The level of synthesis of T-2 fusariotoxin was studied for the most common species in the region, F. sporotrichioides. The maximum concentration (5.3–5.5 mg/kg) was observed in a strain isolated from barley and its monosporous crops; the toxin concentration in monoconidial clones varied from 0.94 to 5.4 mg/kg. The level of toxin synthesis in strains isolated from wheat was 0.05–0.84 mg/kg. The heterogeneity of F.sporotrichioides in terms of phytopathogenic properties was revealed: the studied strains inhibit the development of wheat within a wide range (by 36–79 % compared to the control) with the formation of necrosis with an intensity of up to 1.6 points. Monoconidial lines of F.sporotrichioides were identified, differing from the natural isolate in phytopathogenicity, mainly in the direction of its increase (growth inhibition up to 95%, necrosis up to 2.6 points).

Comparative Evaluation of the Antibacterial and Antitumor Activities of Marine Alkaloid 3,10-Dibromofascaplysin

Fascaplysins form a group of marine natural products with unique cationic five-ring coplanar backbone. Native fascaplysin exhibits a broad spectrum of bioactivities, among which the cytotoxic activity has been the most investigated. Several fascaplysin derivatives have more selective biological effects and are promising as lead compounds. Thus, the introduction of a substituent at C-9 of fascaplysin leads to a strong increase in its antimicrobial properties. Here, a comparative assessment of the antimicrobial activity of synthetic analogs of the marine alkaloids 3-bromofascaplysin, 10-bromofascaplysin, and 3,10-dibromofascaplysin, along with some of their isomers and analogs, was carried out against a panel of Gram-positive bacteria in vitro. For the first time, a significant increase in the antimicrobial activity of fascaplysin was observed when a substituent was introduced at C-3. The introduction of two bromine atoms at C-2 and C-9 enhances the antimicrobial properties by 4 to 16 times, depending on the tested strain. Evaluation of the antimicrobial potential in vivo showed that fascaplysin and 3,10-dibromofascaplysin had comparable efficacy in the mouse staphylococcal sepsis model. Additionally, 3,10-dibromofascaplysin demonstrated a strong and reliable antitumor effect in vivo on the Ehrlich carcinoma inoculated subcutaneously, with a value of tumor growth inhibition by 49.2% 20 days after treatment. However, further studies on alternative chemical modifications of fascaplysin are needed to improve its chemotherapeutic properties.

TAp63γ is the primary isoform of TP63 for tumor suppression but not development

Abstract TP63 is expressed as TAp63 and ΔNp63 from the P1 and P2 promoters, respectively. While TAp63 and ΔNp63 are expressed as three TAp63α/β/γ and ΔNp63α/β/γ due to alternative splicing, only p63α (TA and ΔN) and p63γ (TA and ΔN) proteins are found to be detectable and likely to be responsible for p63-dependent activity. Previous studies implied and/or demonstrated that TAp63α, which contains an N-terminal activation domain conserved in p53, functions as a tumor suppressor by regulating an array of genes for growth suppression. By contrast, ΔNp63α, which also contains an N-terminal activation domain but is different from that in TAp63, regulates a unique set of genes and functions as a master regulator for development of epidermis and other stratified epithelial tissues. However, the biological function of p63γ is largely unexplored. To explore this, we generated a mouse model in that exon 10’, a coding exon specific for p63γ, was deleted by CRISPR-cas9. We showed that mice deficient in p63γ are viable and futile, which is different from mice deficient in total TP63 or p63α. Like TAp63-deficient mice, p63γ-deficient mice have a short lifespan and are prone to spontanenous tumors. Additionally, loss of p63γ shortens the lifespan of tumor-free mice potentially via increased cellular senescence. Moreover, mice deficient in p63γ are prone to chronic inflammation in multiple organs and liver steatosis potentially via altered lipid metabolism. Single-cell RNA-seq revealed that loss of p63γ increases the expression of SCD1, a rate-limiting enzyme for synthesis of monounsaturated fatty acids, leading to altered lipid homeostasis. Together, our data indicate that TP63γ is the primary isoform of TP63 for tumor suppression but not development by maintaining normal inflammatory response and lipid homeostasis.

Antibiotics change the population growth rate heterogeneity and morphology of bacteria.

A better understanding of the system-level effects of antibiotics on bacterial cells is essential to address the growing challenge of antibiotic resistance. Utilising Multipad Agarose Plate (MAP) platforms, we monitor the growth rate and cell morphology of three clinically relevant species (E.coli, S.aureus and P.aeruginosa) following exposure to 14 antibiotics across 11 concentrations (31 microbe-antibiotic combinations in total). Our results reveal a consistent increase in population growth rate heterogeneity (PGRH) as drug concentrations approach the minimum inhibitory concentration (MIC). Strikingly, the magnitude of this heterogeneity correlates with the functional distance between the ribosome and the specific cellular processes targeted by the antibiotics. Among the seven antibiotic classes studied, protein synthesis inhibitors and disruptors cause the lowest PGRH, while heterogeneity progressively increases with RNA synthesis inhibitors, DNA replication inhibitors, cell membrane disruptors and cell wall synthesis inhibitors. Because the ribosome is central to growth rate control, we hypothesize that heterogeneity might arise at the system level as a result of the propagation of damage to protein synthesis. Low heterogeneity is desirable from a clinical perspective, as high heterogeneity is often associated with persistence and treatment survival. Additionally, we observed a strong correlation between morphological alterations and growth inhibition across all antibiotics and species tested. This led to the development of a novel morphological parameter, MOR50, which enables rapid estimation of MIC for antibiotic susceptibility testing (AST) with a single snapshot after just 2.5 hours of incubation. In addition to introducing a novel, resource-efficient and rapid AST method, our findings shed new light on the system-level effects of antibiotic perturbations on bacteria, which might inform treatment design.

Nanobody-Mediated Cellular Uptake Maximizes the Potency of Polylysine Dendrimers While Preserving Solid Tumor Penetration.

Dendrimers are branched macromolecular structures that are useful nanocarriers for small-molecule drugs, such as cancer therapeutics. Their small size permits penetration into solid tumors, coupled with functionalization with a low-fouling PEG coating that minimizes transient cellular interactions and enhances plasma circulation time. While PEGylated dendrimers show significant promise as anticancer therapeutics, there is potential to increase tumor cell specificity and drive uptake of drugs into cells by conjugating cell-targeting ligands onto the dendrimers. To achieve this, we used an expanded genetic code and bio-orthogonal click chemistry to functionalize monomethyl auristatin E (MMAE)-loaded PEGylated dendrimers with a single tumor cell-targeting nanobody per dendrimer. The uniform addition of a single nanobody ligand facilitated greater intracellular uptake of the drug payload into HER2-positive target cells, while preserving the desirable circulatory characteristics of dendrimers. While the nanobody-dendrimer conjugates show similar levels of tumor infiltration over 24 h compared to unmodified dendrimers, the targeted dendrimers had significantly greater inhibition of tumor growth and long-term retention in the tumors. Our results highlight that biodistribution studies alone are poor predictors of therapeutic performance. The controlled conjugation strategy presented here preserves the size advantage and tissue penetration of dendrimers while maximizing targeted cellular uptake and potency in difficult-to-access solid tumor tissue.

Single intraluminal delivery of a nitric oxide-donor results in inhibition of intimal thickening in the rabbit femoral artery.

Vascular smooth muscle cell proliferation and vascular homeostasis is thought to be regulated by nitric oxide and prostaglandins. We examined the effect of exogenous linsidomine, a NO releasing compound, in a model of balloon injury iliac arteries of rabbits. On the cellular levels NO can exert effects like cellular survival, growth and proliferation inhibition. Smooth muscle cell (SMC) proliferation was quantified as change of intima-media-ratio. Linsidomine injection or its vehicle was performed with an infusion-balloon catheter directly into the vessel wall. Balloon angioplasty resulted in a significant increase of intima-media-ratio during three weeks. Linsidomine treatment decreased the intima media ratio significantly (0.65±0.05 vs 1,2±0.2 intima-media-ratio, p<0.05). However, control vessels had an intima media ratio of 0.15±0.02. This effect was similar to NOS-2 transfected vessels following angioplasty. In in vitro experiments linsidomine inhibited significantly and dose-dependently rabbit smooth muscle cell proliferation measured by BrdU incorporation. Further linsidomine increased rate of apoptotic SMC's, however this effect was p53 independent. Simultaneous angioplasty of the rabbit iliac artery and direct injection of linsidomine into the vessel wall seem to be an effective strategy to reduce neointima formation. Despite single administration, local application of linsidomine resulted in potent inhibition of intima proliferation.

Antimicrobial, antibiofilm, and antiurease activities of green-synthesized Zn, Se, and ZnSe nanoparticles against Streptococcus salivarius and Proteus mirabilis.

This study assesses the antimicrobial, antibiofilm, and antiurease properties of selenium (Se), zinc (Zn), and zinc selenide (ZnSe) nanoparticles (NPs) against clinically pathogenic strains of Streptococcus salivarius and Proteus mirabilis. The Se, Zn, and ZnSe NPs, synthesized by Pseudomonas aeruginosa OG1, were characterized using transmission electron microscopy (TEM) revealing average sizes of approximately 30 ± 10nm, 30 ± 15nm, and 40 ± 10 nm, respectively. Atomic force microscopy (AFM) was used to examine the morphological and topological characteristics of the NPs. The structural and crystal characteristics were investigated using X-ray diffraction (XRD). Among the evaluated NPs, Zn NPs at a concentration of 200mg/mL exhibited the most substantial growth inhibition zone against S. salivarius. Conversely, the highest antibiofilm activity was observed against P. mirabilis, notably with 200µg/mL Zn NPs. In the context of antiurease activity, both 100μg Zn and ZnSe NPs caused complete urease inhibition (100%) in P. mirabilis within the initial 5h, with notable inhibition rates of 94% and 80%, respectively, observed against S. salivarius. Significantly, in the current landscape of NP research primarily focused on antimicrobial and antibiofilm properties, our study stands out due to its pioneering exploration of antiurease activities with these NPs. This distinctive emphasis on antiurease effects contributes original and unique value to our study, offering novel insights into the broader spectrum of NP applications, and paving the way for potential therapeutic advancements.

Anticancer Potential of Azatetracyclic Derivatives: In Vitro Screening and Selective Cytotoxicity of Azide and Monobrominated Compounds

This study investigated the antiproliferative activity of three classes of benzo[f]pyrrolo[1,2-a]quinoline azatetracyclic derivatives. All compounds were screened against 60 cancer cell lines at a single dose of 10 μM. When we compared the activity of the three classes of azatetracyclic derivatives (azide, monobrominated and dibrominated), we found that the dibrominated compounds were less active, while the azides were the most active molecules. Compounds 3b and 5a, showing the best growth inhibition profile of all the drugs evaluated, were selected for the second stage of a full five-dose testing. According to the results of the in vitro screening, compounds 3b and 5a exhibit good to moderate anticancer activity (in micromolar range) against all nine cancer sub-panels, with compound 5a being more selective than compound 3b. Both compounds presented better activity than phenstatin on T–47D breast cancer cells, with compound 3b also being more active on SK–MEL–28 melanoma cells, while compound 5a was more active than phenstatin on COLO 205 colon cancer cells. As for the probable mechanism of action, the benzoquinoline derivatives could act as PI5P4Kα and PI5P4Kβ inhibitors or topoisomerase II inhibitors.

Synthetic lethality of mRNA quality control complexes in cancer.

Synthetic lethality exploits the genetic vulnerabilities of cancer cells to enable a targeted, precision approach to treat cancer1. Over the past 15 years, synthetic lethal cancer target discovery approaches have led to clinical successes of PARP inhibitors2 and ushered several next-generation therapeutic targets such as WRN3, USP14, PKMYT15, POLQ6 and PRMT57 into the clinic. Here we identify, in human cancer, a novel synthetic lethal interactionbetween the PELO-HBS1L and SKI complexes of the mRNA quality control pathway. In distinct genetic contexts, including 9p21.3-deleted and high microsatellite instability(MSI-H)tumours, we found that phenotypically destabilized SKI complex leads to dependence on the PELO-HBS1L ribosomal rescue complex. PELO-HBS1L and SKI complex synthetic lethality alters the normal cell cycle and drives the unfolded protein response through the activation of IRE1, as well as robust tumour growth inhibition. Our results indicate that PELO and HBS1L represent novel therapeutic targets whose dependence converges upon SKI complex destabilization, a common phenotypic biomarker in diverse genetic contexts representing a significant population of patients with cancer.

Evaluation of Novel Broad-spectrum Fungicides for Management of Rice Blast and Sheath Blight under In-vitro and Field Conditions

The present study evaluated the efficacy of eight novel fungicidal treatments against Magnaporthe oryzae (Blast) and Rhizoctonia solani (Sheath blight) under both in-vitro and field conditions. In-vitro efficacy against M. oryzae revealed that, complete pathogen growth inhibition (100%) was exhibited by treatments T2 (Picoxystrobin 7% + Propiconazole 12% EC @ 2.0 ml/L), T3 (Pyraclostrobin 100 g/L @ 2.0 ml/L), T4 (Fluxapyroxad 62.5 g/L + Epoxiconazole 62.5 g/L EC @ 1.5 ml/L), T6 (Propiconazole 20% EC @ 1.0 ml/L) and T7 (Isoprothiolane 40% EC @ 1.5 ml/L) at recommended doses. Whereas, at half-recommended doses, T2 and T3 still maintained high efficacy with inhibition rate of 91.8% and 90.7%. Similarly, for R. solani, treatments T2, T3, T4, T5 and T6 also showed complete inhibition at their recommended doses, while T1 and T7 achieved 82.6% and 91.2% inhibition respectively. At half-recommended doses, T2, T3, T4 and T5 registered the cent per cent inhibition, while T1 and T7 showed 79.2% and 81.9%, respectively. In field trials conducted during kharif, 2020 to 2022 revealed that, the treatment T1 notably reduced sheath blight severity to 36.1%, compared to 61.8% in untreated control and achieved the lowest neck blast severity (6.3%) and realised highest yield (7097 kg/ha) and ICBR (1:7.9), and significantly far surpassing the control yield (5144 kg/ha). The triazole-strobilurins combination demonstrated broad-spectrum efficacy, achieving 62.7% and 18.3% reduction in neck blast and sheath blight respectively compared to standard fungicides. These findings underscore the potential of novel combination fungicides in enhancing disease management and yield while lowering the number of sprays and costs to the rice farmers.

Mycochemistry, antioxidant, anticancer activity, and molecular docking of compounds of F12 of ethyl acetate extract of Astraeus asiaticus with BcL2 and Caspase 3

Globally the current research is going for cancer disease management by different strategies as frequency and severity of this disease are increasing day by day. The fruit body of edible and tasty Astraeus asiaticus mushroom was extracted in ethyl acetate solvent (EA) and quantitative analysis revealed that it contained a significant amount of total phenols, flavonoids, and ascorbic acids. The FT-IR study revealed different functional groups in the extracts with different characteristic peak values. The GC/MS profile of AAEA (Astraeus asiaticus ethyl acetate) extract exhibited 61 compounds. The column chromatography of AAEA extract was performed and the F12 fraction demonstrated the greatest radical scavenging activity with an EC50 of 25.65 ± 4.82 µg. mLˉ¹. Mycochemistry (GC and mass spectrum) analysis showed that F12 was a mixture of six important compounds like Hexadecanoic acid, 3,4,5,6 Tetramethyloctane, 9,12-Octadecadienoic acid, 9,12-Octadecadienoic acid, methyl ester, 1-cyclododecyne, cis-9,10-Epoxyoctadecan-1-ol. The chemical properties of all six compounds were screened by SwissADME and pkCSM and AdmetSAR predictors. Out of them Hexadecanoic acid, 9,12-Octadecadienoic acid and 3,4,5,6 Tetramethyloctane, exhibited suitable properties for drug -preparation and they showed anticancer activity and antioxidant activity as per NIST data base and library search. We have tried to focus on anticancer compounds derived from the partial purification (F12) of mushroom extract (AAEA) from this edible mushroom against cancer (cervical, lung, and breast) cell lines. After 24 h of treatment, the percentages of cell growth inhibition of HeLa, MCF-7, and A549 cell lines by highest concentration (1500 µg. mL− 1) of F12 were 92.03 ± 6.21 a, 90.38 ± 4.53a, and 87.51 ± 5.36a % respectively and the IC50 values were 701 ± 11.54, 728.71 ± 10.53, and 806.88 ± 11.52 µg. mL− 1 respectively but the growth of normal cell HEK 293T was inhibited slightly (3.0%). The mechanism of anticancer effect of F12 (AAEAE) on cancer cell lines included induction of apoptosis, LDH leakage, and up regulation of gene expression levels of Caspase 3, Caspase 9, P53, and down regulation of BcL2 of all three cell lines. Molecular docking of the three important compounds (Hexadecanoic acid, 3,4,5,6 Tetramethyloctane and 9,12-Octadecadienoic acid), with apoptotic protein Caspase 3 and antiapoptotic protein BcL2 was done to find out the binding affinity, stability and drug- likeness properties of these chemicals. In conclusion, F12 fraction of AAEA extract of this mushroom containing six bioactive compounds was a promising antioxidant and anticancer agent and the use of this fraction in cancer treatment will be a novel study for future drug development.

Identifying CSNK1E as a therapeutic target in thyroid cancer among the core circadian clock genes.

Previous studies have shown that thyroid malignancies can alter the transcriptional oscillations of circadian clock genes. In this study, we screened the expression of core circadian clock genes in thyroid neoplasms and found that CSNK1E, NPAS2, and TIMELESS were upregulated, while ARNTL, CRY1, CRY2, PER2, and RORA were downregulated during the progression and dedifferentiation of thyroid cancer. Immunohistochemical analysis further confirmed an increase in CSNK1E expression parallel to the loss of tumor differentiation. To investigate the potential therapeutic implications, we treated thyroid cancer cell lines with two different CSNK1E inhibitors: PF670462 and IC261. Both inhibitors resulted in growth inhibition in monolayer and three-dimensional spheroid cultures. This growth inhibition was accompanied by G2/M cell cycle arrest and a decrease in CDK4 and cyclin D1 expression. Moreover, CSNK1E inhibitors suppressed cell migration and invasion and reduced the expression of epithelial-mesenchymal transition markers. In vivo experiments using xenograft models showed that the administration of IC261 significantly restrained tumor growth and decreased the Ki-67 index of the xenograft tumors. In conclusion, our study provides evidence of aberrant CSNK1E expression in thyroid cancer dedifferentiation and highlights the potential therapeutic value of targeting CSNK1E.

Efficacy of Botanicals as Biocontrol Agents Against Leaf Spot and Blossom Blight Pathogen of Marigold: A First Report from Longleng District of Nagaland, India

Aims: To study about etiology and organic disease management strategies of Fusarium oxysporum induced blossom blight in marigold (Tagetes spp.), cultivated in and around Longleng district of Nagaland. The experiment aimed to formulate an effective organic disease management module to combat the disease and assess the efficacy of plant extracts and biocontrol agents in suppressing fungal growth. Study Design: A completely randomized design (CRD) was employed for laboratory-based trials, and an exploratory survey was conducted during the winter season of 2024 to monitor disease incidence in marigold-growing areas around Longleng district of Nagaland. Place and Duration of Study: Field survey has been carried out in Longleng district of Nagaland. All the laboratory works has been carried out in laboratory of Assam Agricultural University(AAU), Jorhat, Assam. Duration of research is 6 months. Methodology: The tested phytoextracts at 10% and 20% concentrations included leaves of neem (Azadirachta indica), tulsi (Ocimum sanctum), garlic (Allium sativum), and other local plant species. Biocontrol agents such as Trichoderma viride, Trichoderma harzianum, and Pseudomonas fluorescens were collected from the rhizosperic soil of Marigold as well as from local fields and research institutions for evaluation. Disease incidence and severity were measured based on Percent Disease Intensity (PDI). Results: The study demonstrated that garlic extract, at both tested concentrations, achieved complete inhibition (100%) of fungal growth. Additionally, several other plant extracts exhibited notable antifungal properties. Among the biocontrol agents, Pseudomonas fluorescens showed a strong antagonistic effect against Fusarium oxysporum (82.65%). These findings underscore the potential for incorporating phytoextracts and biocontrol agents into effective organic disease management strategies for marigold cultivation. Conclusion: From the study, it is recommended to optimize the application of garlic extract, exploring its effectiveness in field conditions, and evaluating its potential in combination with other biocontrol agents or plant extracts.

Allicin Binds to Cysteine Residues of Fungal Dihydrolipoamide Dehydrogenase to Control Gray Mold (Botrytis cinerea) of Tomato.

Allicin exhibits strong inhibitory activity against phytopathogenic fungi; however, its antifungal mechanism remains unclear. This study assessed allicin's inhibitory effects on several phytopathogenic fungi, revealing a half-maximal effective concentration of 125.47 μg/ml against the hyphal growth of Botrytis cinerea. Micromorphological analysis showed that allicin caused abnormalities in the hyphae, including unclear organelle boundaries and organelle dissolution. Integrated transcriptomic, and proteomic, and metabolomic assays indicated that allicin induced differential gene and protein expression, particularly in the plasma membrane, oxidative stress processes, and energy metabolism pathways. Additionally, differentially expressed metabolites were involved in the inhibition of hyphal growth. Biochemical assays demonstrated that allicin inhibited ATP production and damaged hyphal cell membranes. Molecular docking revealed that allicin could bind to dihydrolipoamide dehydrogenase (DLDH), an enzyme rich in cysteine residues, with a binding free energy of -6.322 kcal/mol. Through antimicrobial activity testing of allicin analogues and molecular docking analysis, the active groups of allicin and its interaction with DLDH were identified. This study shows that allicin interferes with energy metabolism, impacts cell membrane and wall integrity by targeting cysteine-containing proteins, and inhibits the proliferation of plant pathogenic fungi. These insights into the antifungal mechanism will provide valuable data for the development and field application of allicin analogues.

Quantifying Baryonic Feedback on the Warm–Hot Circumgalactic Medium in CAMELS Simulations

Abstract The baryonic physics shaping galaxy formation and evolution are complex, spanning a vast range of scales and making them challenging to model. Cosmological simulations rely on subgrid models that produce significantly different predictions. Understanding how models of stellar and active galactic nucleus (AGN) feedback affect baryon behavior across different halo masses and redshifts is essential. Using the SIMBA and IllustrisTNG suites from the Cosmology and Astrophysics with MachinE Learning Simulations (CAMELS) project, we explore the effect of parameters governing the subgrid implementation of stellar and AGN feedback. We find that while IllustrisTNG shows higher cumulative feedback energy across all halos, SIMBA demonstrates a greater spread of baryons, quantified by the closure radius and circumgalactic medium (CGM) gas fraction. This suggests that feedback in SIMBA couples more effectively to baryons and drives them more efficiently within the host halo. There is evidence that the different feedback modes are highly interrelated in these subgrid models. The parameters controlling the stellar feedback efficiency significantly impact AGN feedback, as seen in the suppression of black hole mass growth and delayed activation of AGN feedback to higher-mass halos with increasing stellar feedback efficiency in both simulations. Additionally, the AGN feedback efficiency parameters affect the CGM gas fraction at low halo masses in SIMBA, hinting at complex, nonlinear interactions between the AGN and supernova feedback modes. Overall, we demonstrate that stellar and AGN feedback are intimately interwoven, especially at low redshift, due to subgrid implementation, resulting in halo property effects that might initially seem counterintuitive.

L-glutaminase synthesis by Klebsiella pneumoniae (AS KP 23) isolated from clinical strain, and its efficacy against human hepatocellular and breast cancer cell lines.

Cancer is a major cause of morbidity and mortality worldwide. The proliferation of cancer cells depends largely on glutamine for survival and proliferation. Glutamine serves as a carbon source for the synthesis of metabolites and lipids via the TCA cycle, as well as a source of nitrogen for the synthesis of amino acids and nucleotides. Recently, the role of glutamine metabolism in cancer has been explored in many studies. Therefore, it provides a scientific relationship for targeting glutamine metabolism for cancer treatment. L-glutaminase which is a powerful anticancer medication that is widely used around the world, works by removing L-glutamine from cancerous cells. L-glutaminase has been cited as the most potent molecule that inhibits the proliferation of cancer cells, which significantly raises the possible applicability of cancer therapy and the possibility of its application as an alternative drug to chemotherapy. The first investigation into the antitumor property of L-glutaminase revealed its inhibitory effect on the growth of Gardner lymphosarcoma (6C3HED) and L-1210 leukemia cells. In the same study, glutaminase from Pseudomonas spp., in combination with azaserine enhanced the degree of tumor growth inhibition. Subsequently, L-glutaminase was administered intravenously in patients with acute lymphoblastic leukemia and acute myeloid leukemia. Recently, a purified L-glutaminase from Streptomyces sp. D214 was shown to be the most effective, with an IC50 value of 10mg/ml against the MCF-7 tumor cell line. Also, various in vitro studies have revealed that the activity of glutaminase against the proliferation of tumor cell lines using the MTT (3-(4,5- dimethylthiazol-2-yl)- 2, 5-diphenyltetrazolium bromide) cell proliferation assay. Alcaligenes faecalis KLU102 glutaminase was able to reduce the viability of HeLa cells in a dose-dependent manner, with an IC50 value of 12.5mg/ml within 24h. In this study, a bacterium extracellular from human stool samples was extracted and identified using morphological, biochemical, and molecular methods. The 16S rRNA gene was 100% identical to the sequence from Klebsiella pneumoniae and was submitted to GenBank under accession number OQ703039. Thus, this strain was named Klebsiella pneumoniae AS KP 23. Further kinetic studies on the purified enzyme were performed. In addition, the pH stability of the L-glutaminase enzyme was slightly affected over the pH range of 7.0-9.0 after 2h of pre-incubation, and the rate of thermal inactivation of the L-glutaminase enzyme increased with higher temperatures and longer preheating periods. In addition, the stability of the tested enzyme decreased with an increasing storage period at -20°C. The SDS-PAGE revealed that the L-glutaminase subunits had a molecular weight of around 97kDa. L-glutaminase was purified 1.33-fold with a final specific activity of 799.9 U/mg protein using gel filtration chromatography. The enzyme´s cytotoxic activity showed severe toxicity against the HepG-2 human hepatocellular and breast cancer cell lines. Klebsiella pneumoniae glutaminase was able to reduce the viability of HeLa cells in a dose-dependent manner, with an IC50 value of 305.78µg/ml in human hepatocellular carcinoma and an IC50 value of 400.51µg/ml in breast cancer cell lines. Klebsiella pneumoniae AS KP 23 was a genetically determined microbial species isolated from human stool samples. The production of extracellular enzymes was examined. Additionally, purified L-glutaminase inhibited the growth of normal cells and showed potent anticancer activity against numerous cancer cell lines in the study. Its broad pH and temperature range, combined with its unique and highly stable catalytic activity, make it an excellent choice for use as an effective cancer inhibitor.

Discovery, Isolation, and Characterization of 23-Spirocholestane Derivatives as Novel Plant Root Growth Inhibitors from Ypsilandra thibetica.

In the present study, we first discovered that the 70% ethanol extract of Ypsilandra thibetica significantly inhibited Arabidopsis root growth. Through a bioactivity-guided strategy, 18 rare 23-spirocholestane derivatives (1-18), including 12 new ones (1-12), featuring a dioxane ring between C-24/C-23 of the aglycone moiety and C-1'/C-2' of the d-fucose ligand, were obtained from the strongest active elution fraction. Their structures were elucidated via a comprehensive method of MS and NMR spectroscopy, single-crystal X-ray diffraction, and chemical methods. Evaluation of their inhibitory effects of all isolates on Arabidopsis root growth revealed that 5/6, 3/4, 1, and 2 exhibited pronounced activity, with EC50 values of 6.89, 9.13, 18.01, and 26.83 μM, respectively. Then, a preliminary structure-activity relationship (SAR) analysis of these compounds was conducted. Furthermore, transcriptome data indicated that Arabidopsis responded to the stress caused by these bioactive compounds through the modulation of ethylene and auxin signaling pathways and the activation of various detoxification-related genes. Finally, the herbicidal evaluation showed that 3/4 and 5/6 significantly inhibited the root growth of Echinochloa crusgalli, while 3/4 remarkably suppressed the root growth of Lolium multiflorum. These findings highlight that 23-spirocholestane derivatives represent promising candidates for the development of new bioherbicides.

Pharmacological characterization of GB1908, a selective galectin-1 carbohydrate binding domain inhibitor for the treatment of cancer.

Galectin-1 (Gal-1) is a lectin that has been shown to be involved in a number of pro-tumorigenic mechanisms and has also been shown to be immune-suppressive. Therefore, pharmacological blockade of Gal-1 has the potential to be therapeutically beneficial in cancers that overexpress this lectin where it is hypothesized to be driving cancer progression. GB1908 is a novel, selective and high affinity inhibitor of the Gal-1 carbohydrate recognition domain and in this study we have pharmacologically characterized this small molecule in a range of in vitro and in vivo systems in the context of cancer therapy. In addition, we used a data-driven approach to identify the cancer types which may benefit from Gal-1 inhibitor therapy. The selectivity of GB1908 for Gal-1 compared with galectin-3 (Gal-3) was confirmed in biophysical and cellular assays. GB1908 attenuated Gal-1-induced T cell (Jurkat) apoptosis and reduced the production of immunosuppressive cytokines in a stromal non-small cell lung cancer (NSCLC) tumor microenvironment model. Breast carcinoma and metastatic skin cutaneous melanoma were identified as cancers in which high Gal-1 expression correlated with poorer survival outcomes in patients. Treatment with GB1908 slowed tumor growth in syngeneic mouse models of these cancers. The inhibition of both tumor growth and immune-suppressive cytokines, in cancers in which high Gal-1 is associated with poorer survival outcomes, suggests a potential therapeutic benefit for Gal-1 inhibitors such as GB1908.

Clioquinol inhibits angiogenesis by promoting VEGFR2 degradation and synergizes with AKT inhibition to suppress triple-negative breast cancer vascularization

Inhibition of angiogenesis, either as monotherapy or in conjunction with other treatments, holds significant promise in cancer treatment. However, the limited efficacy of clinically approved anti-angiogenic agents underscores the urgent need for the development of novel drugs and therapeutic strategies. In this study, we demonstrate the highly selective inhibitory effects of clioquinol, a topical antifungal and antibiotic agent, on the angiogenic activity of endothelial cells (ECs) in a series of in vitro angiogenesis assays. Moreover, clioquinol effectively suppressed blood vessel formation in ex vivo aortic ring and in vivo Matrigel plug assays. Mechanistic studies revealed that clioquinol directly binds to the ATP-binding site of vascular endothelial growth factor receptor 2 (VEGFR2), promoting its degradation through both proteasome and lysosome pathways. This led to the down-regulation of the downstream extracellular signal-regulated kinase (ERK) pathway. In addition, the combination with the AKT inhibitor MK-2206 synergistically boosted the anti-angiogenic efficacy of clioquinol in vitro and in an in vivo dorsal skinfold chamber model of triple-negative breast cancer (TNBC), leading to the suppression of TNBC growth. Accordingly, clioquinol, either alone or in combination with AKT inhibitors, represents a promising therapeutic agent for future anti-angiogenic cancer treatment.

Toxicological Effects of Metal-Doped Carbon Quantum Dots.

Multi-domain biological and environmental research highlights the efficacy of carbon quantum dots (CQDs) as a safer alternative to toxic metal-based quantum dots (QDs) and expensive conventional organic dyes, particularly in biomedical applications. CQDs are often functionalized by metal heteroatoms to improve their electron-donating properties and modify charge density, thereby enhancing their physicochemical characteristics. However, metal doping may re-introduce toxicity concerns similar to traditional QDs and further increase environmental risks. Thus, detailed ecotoxicology studies are necessary to understand the environmental impact of these CQDs in different organisms. To address this, we synthesized metal-doped CQDs (Mn, Fe, Cu and Ag) using microwave-assisted technique and conducted in-vitro experiments on diverse biological models belonging to different trophic levels, including bacteria (E. coli and B. subtilis), plants (Vigna radiata) and mammalian cells (mouse myoblast cells- C2C12). Results revealed that among all the CQDs explored, Ag-CQDs exhibited highest toxicity causing ~85% bacterial and 100% mammalian cell death even at 10 μg mL-1 and ~60% radicle growth inhibition after 5 days of exposure at 50 μg mL-1, whereas Mn-CQD showed the least toxicity. These findings contribute significantly to the critical need for determining optimal concentration ranges for metal-doped CQDs and enhance our understanding of their environmental implications.