Inhibition mechanism of Rhizoctonia solani by pectin-coated iron metal-organic framework nanoparticles and evidence of an induced defense response in rice.

Nanocrop protectants have attracted much attention as sustainable platforms for controlling pests and diseases and improving crop nutrition. Here, we reported the fungicidal activity and disease inhibition potential of pectin-coated metal-iron organic framework nanoparticles (Fe-MOF-PT NPs) against rice stripe blight (RSB). An in vitro bacterial inhibition assay showed that Fe-MOF-PT NPs (80 mg/L) significantly inhibited mycelial growth and nucleus formation. The Fe-MOF-PT NPs adsorbed to the surface of mycelia and induced toxicity by disrupting cell membranes, mitochondria, and DNA. The results of a nontargeted metabolomics analysis showed that the metabolites of amino acids and their metabolites, heterocyclic compounds, fatty acids, and nucleotides and their metabolites were significantly downregulated after treatment with 80 mg/L NPs. The difference in metabolite abundance between the CK and Fe-MOF-PT NPs (80 mg/L) treatment groups was mainly related to nucleotide metabolism, pyrimidine metabolism, purine metabolism, fatty acid metabolism, and amino acid metabolism. The results of the greenhouse experiment showed that Fe-MOF-PT NPs improved rice resistance to R. solani by inhibiting mycelial invasion, enhancing antioxidant enzyme activities, activating the jasmonic acid signaling pathway, and enhancing photosynthesis. These findings indicate the great potential of Fe-MOF-PT NPs as a new RSB disease management strategy and provide new insights into plant fungal disease management.

Pectin-Coated Iron-Based Metal-Organic Framework Nanoparticles for Enhanced Foliar Adhesion and Targeted Delivery of Fungicides.

Conventional agrochemicals are underutilized due to their large particle sizes, poor foliar retention rates, and difficult translocation in plants, and the development of functional nanodelivery carriers with high adhesion to the plant body surface and efficient uptake and translocation in plants remains challenging. In this study, a nanodelivery system based on a pectin-encapsulated iron-based MOF (TF@Fe-MOF-PT NPs) was constructed to enhance the utilization of thifluzamide (TF) in rice plants by taking advantage of the pectin affinity for plant cell walls. The prepared TF@Fe-MOF-PT NPs exhibited an average particle size of 126.55 nm, a loading capacity of 27.41%, and excellent dual-stimulus responses to reactive oxygen species and pectinase. Foliar washing experiments showed that the TF@Fe-MOF-PT NPs were efficiently adhered to the surfaces of rice leaves and stems. Confocal laser scanning microscopy showed that fluorescently labeled TF@Fe-MOF-PT NPs were bidirectionally delivered through vascular bundles in rice plants. The in vitro bactericidal activity of the TF@Fe-MOF-PT NPs showed better inhibitory activity than that of a TF suspension (TF SC), with an EC50 of 0.021 mg/L. A greenhouse test showed that the TF@Fe-MOF-PT NPs were more effective than TF SC at 7 and 14 d, with control effects of 85.88 and 78.59%, respectively. It also reduced the inhibition of seed stem length and root length by TF SC and promoted seedling growth. These results demonstrated that TF@Fe-MOF-PT NPs can be used as a pesticide nanodelivery system for efficient delivery and intelligent release in plants and applied for sustainable control of pests and diseases.

Long-term toxic effects of iron-based metal-organic framework nanopesticides on earthworm-soil microorganism interactions in the soil environment.

With the widespread use of controlled-release nanopesticides in field conditions, the interactions between these nanopesticides and biological systems are complex and highly uncertain. The toxicity of iron-based metal organic frameworks (CF@MIL-101-SL) loaded with chlorfenapyr (CF) to terrestrial invertebrate earthworms in filter paper and soil environments and the potential mechanisms of interactions in the nanopesticide-earthworm-cornfield soil microorganism system were investigated for the first time. The results showed that CF@MIL-101-SL was more poisonous to earthworms in the contact filter paper test than suspension concentrate of CF (CF-SC), and conversely, CF@MIL-101-SL was less poisonous to earthworms in the soil test. In the soil environment, the CF@MIL-101-SL treatment reduced oxidative stress and the inhibition of detoxifying enzymes, and reduced tissue and cellular substructural damage in earthworms compared to the CF-SC treatment. Long-term treatment with CF@MIL-101-SL altered the composition and abundance of microbial communities with degradative functions in the earthworm intestine and soil and affected the soil nitrogen cycle by modulating the composition and abundance of nitrifying and denitrifying bacterial communities in the earthworm intestine and soil, confirming that soil microorganisms play an important role in reducing the toxicity of CF@MIL-101-SL to earthworms. In conclusion, this study provides new insights into the ecological risks of nanopesticides to soil organisms.

Enhanced Insecticidal Activity of Chlorfenapyr against Spodoptera frugiperda by Reshaping the Intestinal Microbial Community and Interfering with the Metabolism of Iron-Based Metal-Organic Frameworks.

Spodoptera frugiperda (S. frugiperda) is an invasive pest that threatens global crop production and food security and poses a serious threat to maize production worldwide. Metal-organic framework (MOF) nanocarriers have great potential for agricultural pest control applications. The present study successfully prepared the chemical cross-linking of iron-based metal-organic framework nanoparticles (MIL-101(Fe)-NH2 NPs) with sodium lignosulfonate (SL) as a pH/laccase double stimuli-responsive pesticide release system. The average particle size of the prepared chlorfenapyr (CF)-loaded nanoparticles (CF@MIL-101-SL NPs) was 161.54 nm, and the loading efficiency was 44.52%. Bioactivity assays showed that CF@MIL-101-SL NPs increased the toxicity of CF to S. frugiperda and caused the rupture of the peritrophic membrane and enlargement of the midgut. Data from 16S rRNA gene sequencing showed that CF@MIL-101-SL treatment reduced the resistance of S. frugiperda to pesticides and pathogens and affected nutrient and energy availability by remodeling the intestinal microbiota of S. frugiperda. The dysregulated microbial community interacted with the broken peritrophic membrane, which exacerbated damage to the host. Nontargeted metabolomic results showed that ABC transporters may be a potential mechanism for the enhanced toxicity of CF@MIL-101-SL to S. frugiperda. In summary, the present study provides effective strategies for toxicological studies of nanopesticides against insects.

Mechanism of Rotenone Toxicity against Plutella xylostella: New Perspective from a Spatial Metabolomics and Lipidomics Study.

The botanical pesticide rotenone can effectively control target pest Plutella xylostella, yet insights into in situ metabolic regulation of P. xylostella toward rotenone remain limited. Herein, we demonstrated metabolic expression levels and spatial distribution of rotenone-treated P. xylostella using spatial metabolomics and lipidomics. Specifically, rotenone significantly affected purine and amino acid metabolisms, indicating that adenosine monophosphate and inosine were distributed in the whole body of P. xylostella with elevated levels, while guanosine 5'-monophosphate and tryptophan were significantly downregulated. Spatial lipidomics results indicated that rotenone may significantly destroy glycerophospholipids in cell membranes of P. xylostella, inhibit fatty acid biosynthesis, and consume diacylglycerol to enhance fat oxidation. These findings revealed that high toxicity of rotenone toward P. xylostella may be ascribed to negative effects on energy production and amino acid synthesis and damage to cell membranes, providing guidelines for the toxicity mechanism of rotenone on target pests and rational development of botanical pesticide candidates.

Identification of Host Molecules Involved in the Proliferation of Nucleopolyhedrovirus in Bombyx mori.

The Bombyx mori nucleopolyhedrovirus (BmNPV), a foodborne infectious virus, is the pathogen causing nuclear polyhedrosis and high lethality in the silkworm. In this study, we characterized the molecules involved in BmNPV-silkworm interaction by RNA sequencing of the fat body isolated from the virus-susceptible strain P50. Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation showed that the upregulated differentially expressed genes (DEGs) were mainly involved in translation, signal transduction, folding, sorting, and degradation, as well as transport and catabolism, while the downregulated DEGs were predominantly enriched in the metabolism of carbohydrates, amino acids, and lipids at 72 h post BmNPV infection. Knockout of the upregulated somatomedin-B and thrombospondin type-1 domain-containing protein, probable allantoicase, trifunctional purine biosynthetic protein adenosine-3, and Psl and pyoverdine operon regulator inhibited the proliferation of BmNPV, while knockout of the downregulated clip domain serine protease 3 and carboxylesterase clade H, member 1 promoted it. The molecules herein identified provide a foundation for developing strategies and designing drugs against BmNPV.

Residue and distribution of drip irrigation and spray application of two diamide pesticides in corn and dietary risk assessment for different consumer groups.

BACKGROUND: As the use of diamide insecticides on corn continues to increase, there is growing concern about their residue levels on corn and dietary risks to populations. In this study, the distribution, dispersion and transfer efficiency of two diamide insecticides (tetrachlorantraniliprole (TCAP) and cyantraniliprole (CNAP)) in different parts of corn and soil were investigated in a 1-year field trial in Guangzhou and Lanzhou using two different application methods - spray and drip irrigation, respectively - and the dietary risk of the insecticides to different consumer populations was assessed under the two application methods. RESULTS: The results showed that drip irrigation had a longer persistence period than spraying, and there was a hysteresis in the absorption distribution of the agent in different parts of corn, which was gradually transferred to the leaves after absorption from the roots. The average TE1 (transfer efficiency) and TE2 were 0.230-0.261 and 1.749-1.851 for TCAP and 0.168-0.187 and 2.363-2.815 for CNAP, respectively. At corn harvest, both TCAP and CNAP were below detectable levels in soil and corn. For different consumer populations, hazard quotients ranged from 0.001 to 0.066 for TCAP and from 0.003 to 0.568 for CNAP - both well below 100%. CONCLUSION: This study indicates that TCAP and CNAP applied by spray or drip irrigation are safe for long-term risk of human intake and also provides guidance for the use of both insecticides in agricultural production to control corn pests, especially in arid and semi-arid areas. © 2022 Society of Chemical Industry.

Effects of sublethal azadirachtin on the immune response and midgut microbiome of Apis cerana cerana (Hymenoptera: Apidae).

As a wildly used plant-derived insecticide, azadirachtin (AZA) is commonly reported as harmless to a range of beneficial insects. However, with the research on the effect of AZA against pollinators in recent years, various negative physiological effects on other Apidae species have been demonstrated. Thus to explore the safety of azadirachtin to Apis cerana cerana, the different physiological effects of sublethal concentration of azadirachtin on worker bees A.c.cerana has been studied. With the exposure of 5 mg·L-1 and 10 mg·L-1 azadirachtin for 5 d, the relative expression of Apidaecin, Abaecin and Lysosome genes in workers has decreased significantly at 1, 2,3 and 5 d, and the mRNA levels of Defensin 2 and Hymenoptaecin were also significantly inhibited by 10 mg·L-1 azadirachtin at each check point. Besides, the activity of midgut antioxidant enzymes Superoxide Dismutase (SOD) and Catalase (CAT) which are the first line of defence in antioxidant systems was not affected by AZA, the activity of Peroxidase (POD) showed a fluctuating pattern at 24 h and 48 h, while the activity of polyphenol oxidase (PPO) has significantly inhibited by AZA. However, through 16sRNA analysis it was observed that 5 mg·L-1 AZA did not affect the midgut microbiome colony composition and relative abundance, as well as its main function. Therefore, to a certain extent, azadirachtin is safe for workers, but we should pay more attention to the sublethal effect of AZA that also detrimental to the healthy development of the honeybee colony.

Antibacterial and Cytotoxic Phenyltetracenoid Polyketides from Streptomyces morookaense.

Formicapyridine-type racemates, streptovertidines A (1) and B (2), a 7,24-seco-fasamycin, streptovertidione (3), and the fasamycin-type streptovertimycins I-T (4-15), together with 13 known fasamycin congeners (16-28), were isolated from soil-derived Streptomyces morookaense SC1169. Their structures were elucidated by extensive spectroscopic analysis and theoretical computations of ECD spectra. The fasamycin-type compounds 5, 8-12, 14, and 15 exhibited activity against the drug-resistant bacteria MRSA and VRE (MIC: 1.25-10.0 µg/mL). All isolates, except 3, 4, 10, and 24, displayed cytotoxicity against at least one of the human carcinoma A549, HeLa, HepG2, and MCF-7 cells (IC50 < 10.0 µM), of which some were also cytotoxic to the noncancerous Vero cells. Taken together, the activity data demonstrated that the fasamycin-type compounds were more selective to the tested bacteria over the mammalian cells. Structure-activity relationship analysis suggested that chlorination at C-2 in antibacterial fasamycin-type compounds improves the activity and selectivity to the bacteria. Theoretical simulations of reaction paths and chemical reactions for conversion of 3 to 1 were carried out and supported that the pyridine ring formation in formicapyridines proceeds nonenzymatically via 1,5-dicarbonyl condensation with ammonia.

Harnessing a Transient Gene Expression System in Nicotiana benthamiana to Explore Plant Agrochemical Transporters.

Functional characterization of plant agrichemical transporters provided an opportunity to discover molecules that have a high mobility in plants and have the potential to increase the amount of pesticides reaching damage sites. Agrobacterium-mediated transient expression in tobacco is simple and fast, and its protein expression efficiency is high; this system is generally used to mediate heterologous gene expression. In this article, transient expression of tobacco nicotine uptake permease (NtNUP1) and rice polyamine uptake transporter 1 (OsPUT1) in Nicotiana benthamiana was performed to investigate whether this system is useful as a platform for studying the interactions between plant transporters and pesticides. The results showed that NtNUP1 increases nicotine uptake in N. benthamiana foliar discs and protoplasts, indicating that this transient gene expression system is feasible for studying gene function. Moreover, yeast expression of OsPUT1 apparently increases methomyl uptake. Overall, this method of constructing a transient gene expression system is useful for improving the efficiency of analyzing the functions of plant heterologous transporter-encoding genes and revealed that this system can be further used to study the functions of transporters and pesticides, especially their interactions.

Insights into the degradation and toxicity difference mechanism of neonicotinoid pesticides in honeybees by mass spectrometry imaging.

Honeybees are essential for the pollination of a wide variety of crops and flowering plants, whereas they are confronting decline around the world due to the overuse of pesticides, especially neonicotinoids. The mechanism behind the negative impacts of neonicotinoids on honeybees has attracted considerable interest, yet it remains unknown due to the limited insights into the spatiotemporal distribution of pesticides in honeybees. Herein, we demonstrated the use of matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) for the spatiotemporal visualization of neonicotinoids, such as N-nitroguanidine (dinotefuran) and N-cyanoamidine (acetamiprid) compounds, administered by oral application or direct contact, in the whole-body section of honeybees. The MSI results revealed that both dinotefuran and acetamiprid can quickly penetrate various biological barriers and distribute within the whole-body section of honeybees, but acetamiprid can be degraded much faster than dinotefuran. The degradation rate of acetamiprid is significantly decreased when piperonyl butoxide (PBO) is applied, whereas that of dinotefuran remains almost unchanged. These two factors might contribute to the fact that dinotefuran affords a higher toxicity to honeybees than acetamiprid. Moreover, the toxicity and degradation rate of acetamiprid can be affected by co-application with tebuconazole. Taken together, the results presented here indicate that the discrepant toxicity between dinotefuran and acetamiprid does not lie in the difference in their penetration of various biological barriers of honeybees, but in the degradation rate of neonicotinoid pesticides within honeybee tissues. Moreover, new perspectives are given to better understand the causes of the current decline in honeybee populations posed by insecticides, providing guidelines for the precise use of conventional agrochemicals and the rational design of novel pesticide candidates.

Ionic Liquids Enhanced Alkynyl Schiff Bases Derivatives of Fipronil Synthesis and Their Cytotoxicity Studies.

To obtain highly selective toxic derivatives of fipronil, a series of Schiff bases with an alkynyl group (3a-3k) were designed and synthesized from 4-ethynylbenzaldehyde (2) and 4-substituted 5-amino-N-arylpyrazole (1a-1k) via a nucleophilic addition elimination reaction in ionic liquids. Utilization of ionic liquids was demonstrated to endow the yield of each compound beyond 50%, which was enhanced over 1.5 times of the synthetic productive rates comparing the conventional method by which longer reactive time was consumed. The derivatives were characterized via nuclear magnetic resonance hydrogen spectroscopy (1H-NMR), carbon-13 nuclear magnetic resonance spectroscopy (13C-NMR), and electrospray ionization high resolution mass spectrometry (ESI-HRMS). The cytotoxicity of these derivatives on Trichoplusia ni (Hi-5) cell and Spodoptera litura cell (SL cell) was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) bioassays. The results indicated that several compounds had potential cytotoxicity on Hi-5 cell, especially a 4-ethyl substituted alkynyl Schiff base derivative (3f) that was demonstrated to possess high selective toxicity to the Hi-5 cell than the SL cell. In addition, 3f exhibited comparable toxic activity to commercial fipronil on a Hi-5 cell while a little toxic effect on the SL cell, which satisfied the expectation for selective toxicity screening.

Development of Multifunctional Avermectin Poly(succinimide) Nanoparticles to Improve Bioactivity and Transportation in Rice.

Avermectin (AVM) as a nonsystemic pesticide possesses a low effective utilization rate. Studies of the multifunctional pesticide delivery system for improving biological activity are developing prosperously. In this study, multifunctional avermectin/polysuccinimide with glycine methyl ester nanoparticles (AVM-PGA) were prepared by the self-assembly process. The AVM loading capacity was up to 23.7%. After 24 h of UV irradiation, there was still about 70% of AVM remaining in PGA42 nanocarriers, as opposed to less than 5% of the free-form AVM. The rising ambient pH promoted the release of AVM using an in vitro releasing test, revealing a favorable pH-responsively controlled-release property. The mortality rate of Plutella xylostella with 2.5 µg/mL of AVM content of AVM-PGA42 was 96.3% after 48 h, while that of free AVM was only 51.5%. In addition, the AVM could be detected in stems and all leaves treated with AVM-PGA42 nanoparticles, whereas rare AVM was detected only in treated leaves for the free-form AVM, which achieved the transportation of nanocarriers carrying AVM in rice for the first time. Furthermore, the PGA nanoparticles performed a good growth promoting effect on rice. These results show that the AVM-PGA42 nanopesticides have a great potential application prospect to control the pest and improve the drug utilization efficiency on agriculture.

Limonoids from seeds of Azadirachta indica A. Juss. and their cytotoxic activity.

Four new limonoid-type nortriterpenoids, 1-detigloyl-1-O-methacryloylsalannin (1), 28-deoxo-2,3-dihydronimbolide (2), 12-acetoxy-3-O-acetyl-7-O-tigloylvilasinin (3) and 12-acetoxy-3-O-acetyl-7-O-methacryloylvilasinin (4), along with five known ones, were isolated from seeds of Azadirachta indica A. Juss. Their structures were elucidated by various spectroscopic methods, including UV, IR, MS, NMR, X-ray crystallography, quantum chemical calculation, as well as by comparison of their spectroscopic data with those reported. In the in vitro cytotoxic assay, 2 showed inhibitory activity against human breast cancer MDA-MB-231 cell line with IC50 value of 7.68±1.74 µmol/L, and 5 inhibited growth of human cervical cancer Hela cell line, melanoma A375 cell line and promyelocytic leukemia HL-60 cell line, with IC50 12.00±2.08, 17.44±2.11, and 13.95±5.74 µmol/L, respectively.

Induction of Autophagy and Apoptosis via PI3K/AKT/TOR Pathways by Azadirachtin A in Spodoptera litura Cells.

Azadirachtin is one of the most effective botanical insecticides and has been widely used in pest control. Toxicological reports show that azadirachtin can induce apoptosis in various insect cell lines. However, studies of azadirachtin-induced autophagy in cultured insect cells are lacking. This study reports that azadirachtin A significantly inhibits cell proliferation by inducing autophagic and apoptotic cell death in Spodoptera litura cultured cell line (SL-1 cell). Characteristic autophagolysosome and Atg8-PE (phosphatidylethanolamine) accumulation were observed by electron microscopy and western blotting, indicating that azadirachtin triggered autophagy in SL-1 cell. Furthermore, azadirachtin inhibited survival signaling by blocking the activation of PI3K, AKT and the down-stream target of rapamycin. Similar to the positive control of starvation, azadirachtin induced the activation of insulin receptor (InR) via a cellular feedback mechanism. In addition, the autophagy-related 5 (Atg5), a molecular switch of autophagy and apoptosis, was truncated (tAtg5) to trigger cytochrome c release into the cytoplasm under azadirachtin stress, which indicated that azadirachtin induced apoptosis through autophagy. Our findings suggest that azadirachtin primarily induced autophagy in SL-1 cell by dysregulating InR- and PI3K/AKT/TOR pathways, then stimulated apoptosis by activating tAtg5.

Preparation, Characterization and Intracellular Imaging of 2,4-Dichlorophenoxyacetic Acid Conjugated Gold Nanorods.

Visualizing the biodistribution of pesticides inside living cells is great importance for enhancing targeting of pesticides. Here we reported for the first time that gold nanorods (Au NRs) with size of 39.4 nm x 11.3 nm could be used as a fluorescent tracer to examine the distribution of a typical herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D), in tobacco bright yellow 2 (BY-2) cells. The nanostructures of hybrid materials were analyzed by using Raman spectra and X-ray photoelectron spectroscopy (XPS), including spectra assignments and electronic property. These data revealed 2,4-D has successfully conjugated MP-Au NRs according to Raman and XPS. The biodistribution of the conjugates inside BY-2 cells was directly examined at 12 and 24 h by the two-photon microscopy. The intensity of two-photon luminescence (TPL) inside cells demonstrated that the conjugates could be localized and excluded by BY-2 cells. Thus, this labeling approach opens up new avenues to the facile and efficient labeling of pesticides.

Biosynthesis of Gold Nanoparticles Using Novel Bamboo (Bambusa chungii) Leaf Extracts.

Gold nanoparticles (Au NPs) have drawn significant interest because of their antisotropic physical properties in biomedical applications. In this paper, we reported the application of bamboo (Bambusa chung) leaf extracts, previously not exploited, in the synthesis of Au NPs at ambient temperature. The average dimension of quasi-spherical Au NPs was 28.8±4.5 nm by transmission electron microscopy (TEM). The UV-vis spectroscopy gave an optimal reaction time of 180 min in the process of bioreduction. The organic shell of Au NPs was characterized by Fourier transform infrared (FTIR) spectra and thermogravimetric analysis (TGA), suggesting that the main compositions of the organic shell were hydroxyflavones. The X-ray diffraction (XRD) studies indicated the Au NPs were (111) oriented. This eco-friendly method for the synthesis of Au NPs was simple, amenable for large scale commercial production and biological applications to future in vivo imaging and cancer therapy.

Phloem mobility and translocation of fluorescent conjugate containing glucose and NBD in castor bean (Ricinus communis).

Phloem mobility is an important factor for long-distance transport of systemic pesticides in plants. Our previous study revealed that a fluorescent glucose-insecticide conjugate, N-{3-cyano-1-[2,6-dichloro-4-(trifluoromethyl)phenyl]-4-iodo-1H-pyrazol-5-yl}-N-{[1-(ß-D-glucopyranosyl)-1H-1,2,3-triazole-4-yl]methyl}-N-{[1-((N-(7-nitrobenz-2-oxa-1,3-diazole-4-amine))-propyl)-1H-1,2,3-triazole-4-yl]methyl}amine (IPGN), can be transported in tobacco cells. Several studies have also indicated that glucose moieties can guide the conjugates into plant cells. In this study, we investigated the phloem mobility of IPGN within castor bean seedlings. Cotyledon uptake experiment results show that IPGN could enter the phloem of the mid-veins of cotyledons. The results of further quantitative analysis show that IPGN was present in small amounts in the phloem sap despite the inconsistencies of physicochemical properties with diffusion through the plasma membrane. Its concentration in the phloem sap (about 370nM at 5h) was much lower than that in the incubation medium (100µM), which suggests that IPGN exhibited weak phloem mobility. After the leaves of Ricinus plantlets were treated with IPGN, green fluorescence could be observed in the phloem of the petioles, bud apical nodes, bud mid-veins, and mid-veins of the untreated leaves. The localization of the fluorescent conjugate at various levels of Ricinus plantlets indicates that it was translocated at a distance to sink organs via sieve tubes. The results proved that introducing a glucose group is a feasible approach to modify non-phloem-mobile pesticides and produce phloem-mobile pesticides.

Sasanquasaponin from Camellia oleifera Abel. induces cell cycle arrest and apoptosis in human breast cancer MCF-7 cells.

Sasanquasaponin (SQS) is a triterpenoid extracted from the Chinese medicinal plant Camellia oleifera Abel. Little knowledge about the role of SQS in the prevention and treatment of cancer is available. Recent studies have shown that SQS possesses potent anti-tumor activities in human leukemia Jurkat cells and human hepatoma HepG2 cells. However, research on the effects and mechanisms of SQS on the treatment of breast cancer, a major cause of mortality in women worldwide, is not available. In this work, the effects of SQS on cell growth characteristics, including proliferation, cell cycle progression, and apoptosis, in MCF-7 cells were investigated. SQS reduced the viability of MCF-7 cells, induced G1 phase arrest in the cell cycle, and triggered the apoptosis of MCF-7 cells. Stimulation of MCF-7 with SQS induced upregulation of p21 and downregulation of cyclin D1, which can cause G1 cell cycle arrest. Stimulation also induced activation of E2F1 and downregulation of p53, indicating that SQS induces cell cycle arrest and apoptosis via a p53-independent pathway; p53-independent apoptosis may be mediated by E2F1 activation. Our results demonstrate the potential application of SQS as an anti-breast cancer agent.

Novel fluorescent conjugate containing glucose and NBD and its carrier-mediated uptake by tobacco cells.

Some compounds that contain glucose groups can be transported across the plasma membrane into the cells through hexose transporters. To test the hypothesis that glucose-conjugated insecticides also have similar uptake and translocation properties, a novel fluorescent conjugate (12) was prepared by conjugating glucose and 7-nitrobenz-2-oxa-1,3-diazole with 4-iodo-1-phenylpyrazoles. Its fluorescence spectra and uptake by suspension-cultured tobacco (Nicotiana tabacum L.cv.) cells were studied. The fluorescence spectra showed long wavelengths with maximum emission at 530nm. After incubating tobacco cell suspensions in 10µM conjugate for 0.5h, green fluorescence of 12 was clearly visible in the cells under fluorescence microscopy. After 2h of incubation, more than 70% of 12 was absorbed. Carbonyl cyanide m-chlorophenylhydrazone, phloridzin and glucose drastically inhibited uptake. In concentration-dependent uptakes, the uptake rate of 12 showed a saturable component and was in accordance with Michaelis-Menten kinetics. The results proved that the glucose moiety can guide 12 into tobacco cells and that hexose transporters mediated the uptake.