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Background: Cancer therapy is one of the most important challenges that human beings are facing. The abnormal activity of epidermal growth factor receptor tyrosine kinase (EGFR1) in tumors has been reported in many studies. Tyrosine kinase inhibitors are now commercially available for the treatment of a variety of cancers. Based on our previous studies, we assumed that a hybrid of aminopyrimidine derivatives as EGFR inhibitors and benzocheromen derivatives as cytotoxic agents can induce apoptosis in EGFR positive cancer cells. In the present study, the cytotoxic effect, ability of EGFR inhibition and apoptosis induction of some synthetic benzochromene pyrimidine derivatives were investigated on MDA-MB231, SKBR3 and PC3 cell lines. Methods: The EGFR inhibition activity was determined using cell-based EGFR ELISA kit. Cell viability was determined by MTT assay in 2D and 3D cultures. The apoptosis was confirmed through different methods such as fluorescent staining, annexin V– propidium iodide double staining, DNALadder assay, caspase-3 colorimetric assay, and nitric oxide assay. Results: The results of the MTT assay showed that derivatives with different substituents exhibited differential cytotoxicity in three cancer cell lines, although in MDA-MB231 the cytotoxicity effect of compounds is more obvious than the other cell lines. Production of nitric oxide, caspase-3 activity and DNA-fragmentation was significant in MDA-MB231 and PC3 cells. SKBR3 cells, despite having the lowest apoptosis among these three cell lines, showed a significant EGFR inhibition in the ELISA assay. Conclusion: In this research, we proved that hybrids of benzochromene and amino pyrimidine could be effective on growth inhibition of cancer cell lines and may be used as a drug candidate for cancer therapy in the future.

Ultrasound stimulated microbubbles (USMB) are being investigated for their potential to promote the uptake of anticancer agents into tumor tissue by exploiting their ability to enhance microvascular permeability. At sufficiently high ultrasound transmit amplitudes it has also recently been shown that USMB treatments can, on their own, induce vascular damage, shutdown blood flow, and inhibit tumor growth. The objective of this study is to examine the antitumor effects of ‘antivascular’ USMB treatments in conjunction with chemotherapy, which differs from previous work which has sought to enhance drug uptake with USMBs by increasing vascular permeability. Conceptually this is a strategy similar to combining vascular disrupting agents with a chemotherapy, and we have selected the taxane docetaxel (Taxotere) for evaluating this approach as it has previously been shown to have potent antitumor effects when combined with small molecule vascular disrupting agents. Experiments were conducted on PC3 tumors implanted in athymic mice. USMB treatments were performed at a frequency of 1 MHz employing sequences of 50 ms bursts (0.00024 duty cycle) at 1.65 MPa. USMB treatments were administered on a weekly basis for 4 weeks with docetaxel (DTX) being given intravenously at a dose level of 5 mg/kg. The USMB treatments, either alone or in combination with DTX, induced an acute reduction in tumor perfusion which was accompanied at the 24 hour point by significantly enhanced necrosis and apoptosis. Longitudinal experiments showed a modest prolongation in survival but no significant growth inhibition occurred in DTX–only and USMB-only treatment groups relative to control tumors. The combined USMB-DTX treatment group produced tumor shrinkage in weeks 4–6, and significant growth inhibition and survival prolongation relative to the control (p

The transcriptional repressor BCL11A is involved in hematological malignancies, B-cell development, and fetal-to-adult hemoglobin switching. However, the molecular mechanism by which it promotes the development of myeloid leukemia remains largely unknown. We find that Bcl11a cooperates with the pseudokinase Trib1 in the development of acute myeloid leukemia (AML). Bcl11a promotes the proliferation and engraftment of Trib1-expressing AML cells in vitro and in vivo. Chromatin immunoprecipitation sequencing analysis showed that, upon DNA binding, Bcl11a is significantly associated with PU.1, an inducer of myeloid differentiation, and that Bcl11a represses several PU.1 target genes, such as Asb2, Clec5a, and Fcgr3. Asb2, as a Bcl11a target gene that modulates cytoskeleton and cell-cell interaction, plays a key role in Bcl11a-induced malignant progression. The repression of PU.1 target genes by Bcl11a is achieved by sequence-specific DNA-binding activity and recruitment of corepressors by Bcl11a. Suppression of the corepressor components HDAC and LSD1 reverses the repressive activity. Moreover, treatment of AML cells with the HDAC inhibitor pracinostat and the LSD1 inhibitor GSK2879552 resulted in growth inhibition in vitro and in vivo. High BCL11A expression is associated with worse prognosis in humans with AML. Blocking of BCL11A expression upregulates the expression of PU.1 target genes and inhibits the growth of HL-60 cells and their engraftment to the bone marrow, suggesting that BCL11A is involved in human myeloid malignancies via the suppression of PU.1 transcriptional activity.

Recent studies show that triclosan (TCS) exposure causes reduction in pigments, suppression of photosynthesis, and induction of oxidative stress at the physiological level, resulting in morphological alteration and growth inhibition in algae including Raphidocelis subcapitata (R. subcapitata, a freshwater model green alga). However, the underlying molecular mechanisms remain to be elucidated, especially at environmentally relevant concentrations. The present study uncovered the transcriptional profiles and molecular mechanisms of TCS toxicity in R. subcapitata using next-generation sequencing. The algal growth was drastically inhibited following a 7-day exposure at both 75 and 100 μg/L TCS, but not at 5 μg/L (environmentally realistic level). The transcriptomic analysis shows that molecular signaling pathways including porphyrin and chlorophyll metabolism, photosynthesis – antenna proteins, and photosynthesis were suppressed in all three TCS treatments, and the perturbations of these signaling pathways were exacerbated with increased TCS exposure concentrations. Additionally, signaling of replication-coupled DNA repair was only activated in 100 μg/L TCS treatment. These results indicate that photosynthesis systems were sensitive targets of TCS toxicity in R. subcapitata, which is distinct from the inhibition of lipid synthesis by TCS in bacteria. This study provides novel knowledge on molecular mechanisms of TCS toxicity in R. subcapitata.

The current study was conducted to evaluate the effect of eight Palestinian indigenous plant essential oils (EOs) under in vitro and in vivo conditions against Botrytis cinerea Pers.: Fr., Penicillium italicum Wehmer, and Penicillium digitatum (Pers.) Sacc., three common post- harvest pathogens of tomato and strawberry fruits. In vivo tests showed that thyme, sesame and sage EOs exhibited high antifungal activity against B. cinerea on strawberry and toma- to fruits, compared with rosemary, mint and eucalyptus. In vitro agar, disk-diffusion tests showed that B. cinerea, P. digitatum and P. italicum mycelium growth was completely inhi- bited when treated with clove and sage EOs caused 50% inhibition of B. cinerea and P. itali- cum mycelium growth. Fruit decay and fruit quality index values measured in total soluble solids and fruit flesh firmness showed that EO coated strawberries had significantly less fruit decaying and ripping compared to control, while EO coated tomatoes showed no sig- nificant difference compared to control. EO constituents fall into different chemical classes, including sterols, caffeoylquinic acids, flavonoids, terpenoids, coumarins, and acetylenes. Chemical analysis of the EO preparations using gas chromatography-mass spectrometry determined that the main components in sesame oil were octadecenoic acid-(56%) and hexadecanoic acid (26%), while clove oil consisted of eugenol (53%). In the other EOs, the principal compounds were: menthol (44% in mint oil), eucalyptol (37% in sage oil), while bornanone (18% in rosemary oil) and γ-terpinene (21% in thyme oil) were present at lower concentrations. The EO of sage plants could potentially be a useful alternative to synthetic pesticides to control post-harvest diseases and prolong the shelf life of fruit products.

A Candida spp. possui elevada patogenicidade nos seres humanos imunocomprometidos, acometendo órgãos genitais, mucosa do trato respiratório e oral, entre outras partes do corpo, tornando-se cada vez mais resistente aos antifúngicos. Uma das opções para seu tratamento está relacionado ao uso de plantas medicinais as quais possuem ação antibacteriana e antifúngica. O presente estudo buscou avaliar a atividade antifúngica dos extratos in natura do Rosmarinus officinalis frente as cinco cepas de Candida spp. por meio de uma pesquisa experimental na qual foram testados dois extratos diferentes do alecrim in natura, sendo um do caule e outro das folhas da planta, frente as cepas de C. albicans, C. tropicalis, C. krusei, C. parapsilosis e C. glabrata. O alecrim apresentou atividade inibitória frente as cepas com variações de inibição relacionadas à diferentes concentrações, com inibição de crescimento em todas as concentrações dos extratos testados contra C. albicans e C. tropicalis. As cepas de C. krusei, C. parapsilosis e C. glabrata testadas demonstraram resultados distintos entre si relacionados a inibição do crescimento conforme as diluições propostas. Considerando o apresentado, é possível considerar que o R. officinalis in natura apresenta atividade inibitória frente a Candida albicans e Candidas não-albicans .

Previously undescribed species of Limosilactobacillus fermentum isolated from wholemeal buckwheat sourdough were identified by MALDI-TOF/MS and 16S rRNA analysis. The metabolism of 6 hydroxycinnamic acids by L. fermentum 6P1, 6P2 and 7P2 strains provided mostly o-, m-, p-dihydrocoumaric, dihydroferulic, dihydrocaffeic and dihydrosinapic acids. The ratio of hydroxyphenylpropionic acids to 4-vinylphenols was strain-to-strain specific. Decarboxylation of free cinnamic acid to potentially toxic styrene was low (1.8–23.9 mol. %) in all Limosilactobacillus strains tested. At a concentration of 13.6 mmol/L, reduced p-dihydrocoumaric and dihydroferulic acids were shown to cause less growth inhibition than their precursors, while specific growth rates μ increased significantly. L. fermentum POH with the highest sensitivity to phenolic acids exhibited the lowest ability to reduce hydroxycinnamic acids (11.8–44.5%) by phenolic acid reductase. Metabolomic experiments with chemically labelled D-glucose-d12 provided a deeper insight into electron transfer to polyphenols as the final acceptor. To the best of our knowledge, deuterium transfer to dideuteroferulic acid was observed for the first time.

The present study was aimed to examine the antibacterial potential of Brassica nigra essential oil (BNEO) against Ralstonia solanacearum, causal agent of bacterial wilt and Nitrosomonas sp., the nitrifying bacteria. In poisoned food assay, BNEO showed 100% growth inhibition of R. solancearum at ≥ 125 µg mL-1. Revalidation of findings by volatile assay employing inverted Petri plate technique exhibited 100% bacterial growth inhibition caused by vapors of BNEO, even at 50 µg mL-1 concentration. In the broth microdilution assay, the BNEO exhibited significant antibacterial activity only at higher concentrations (>500 µg mL-1). At 500 µg mL-1, BNEO showed 80% bacterial growth inhibition over control, which was at par with that of streptomycin (5 µg mL-1). In resazurin microtitre-plate assay, the maximum concentration of BNEO, at which color change occurred was 512 µg mL-1 (T9), and thus 512 µg mL-1 was concluded as the minimum inhibitory concentration (MIC). BNEO effectively inhibited the activity of Nitrosomonas spp. with 30-65% nitrification inhibition at the dose of 400 mkg-1 of Urea-N. Homology modeled protein targets assisted computational tool-based novel analysis helped to understand that the antibacterial potency of BNEO is due to preferable binding efficiency of allyl isothiocyanate (AITC), the major active ingredient of BNEO.

3-Deoxy-d-arabinoheptulosonate-7-phosphate (DAHP) synthase catalyzes the first step in the shikimate biosynthetic pathway and is an antimicrobial target. We used an inhibitor-in-pieces approach, based on the previously reported inhibitor DAHP oxime, to screen inhibitor fragments in the presence and absence of glycerol 3-phosphate to occupy the distal end of the active site. This led to DAHP hydrazone, the most potent inhibitor to date, Ki = 10 ± 1 nM. Three trifluoropyruvate (TFP)-based inhibitor fragments were efficient inhibitors with ligand efficiencies of up to 0.7 kcal mol-1/atom compared with 0.2 kcal mol-1/atom for a typical good inhibitor. The crystal structures showed the TFP-based inhibitors binding upside down in the active site relative to DAHP oxime, providing new avenues for inhibitor development. The ethyl esters of TFP oxime and TFP semicarbazone prevented E. coli growth in culture with IC50 = 0.21 ± 0.01 and 0.77 ± 0.08 mg mL-1, respectively. Overexpressing DAHP synthase relieved growth inhibition, demonstrating that DAHP synthase was the target. Growth inhibition occurred in media containing aromatic amino acids, suggesting that growth inhibition was due to depletion of some other product(s) of the shikimate pathway, possibly folate.

The objective of the present study was to elucidate the mechanism by which oxalic acid (OA) alleviates cadmium (Cd) stress. Chickpea (Cicer arietinum L.) seeds were treated with 200 µM CdCl2 for 6 days or 3 days followed by the combination 200 µM Cd + 100 µM OA for three additional days. Exogenous OA mitigated the growth inhibition by Cd of both roots and shoots. This positive effect can be attributed to the increased OA exudation from roots and the alleviation of the Cd-induced depletion of endogenous OA in shoots (80% increase versus Cd-stressed). The combination Cd + OA proved to be effective in protecting cell membrane integrity. This beneficial effect was reflected by 36 and 24% decrease in lipoxygenase (LOX) activity in roots and shoots, respectively, and 60% decrease in the accumulation of the harmful aldehyde, 4-hydroxy-2-nonenal (4-HNE) in roots when compared to Cd-treated samples. Besides, OA restored the Cd-imposed disruption of the plasma membrane function. This effect was depicted by an optimal H+-ATPase activity and proton exudation rate. Oxalic acid counteracted the Cd-induced overproduction of hydroxyl radical (OH•) and NADPH-oxidase activity increase, leading to restore a steady cellular redox state. Furthermore, OA modulated the impact of Cd on Cu/Zn-superoxide dismutase (Cu/Zn-SOD), ascorbate peroxidase (APX), and catalase (CAT) activities in chickpea seedlings. Present results indicate that OA is able to alleviate Cd toxicity by protecting plasma membrane integrity and modulating antioxidant activities. Moreover, our findings shed light on the interplay between plasma membrane H+-ATPase and the accumulation and root exudation of OA.

The in vitro effects on mycelial growth of the antagonistic fungi Trichoderma asperellum (ICC 012) and Trichoderma gamsii (ICC 080) were studied based on the active ingredients of 12 commonly used herbicides: amidosulfuron, dicamba, fluazifop-P-butyl, fluorochloridone, foramsulfuron, isoxaflutole, metribuzin, pendimethalin, prosulfocarb, pyridate, S-metolachlor and tembotrione. The fungal growth rates were evaluated at 20 °C and 25 °C using PDA plates that included 100%, 75%, 50%, 25%, 12.5%, 6.25% and 0% of the recommended herbicide application rates. These herbicides were classified into four categories based on inhibition of mycelial growth in the toxicity testing. Growth inhibition of Trichoderma spp. was seen for all of these herbicides at almost all doses and at both temperatures, although some major differences were seen. Lower herbicide concentrations in the agar also reduced the inhibitory effects, but generally not linearly. Temperature had no significant effects on the levels of inhibition of mycelial growth. Amidosulfuron, dicamba and isoxaflutole showed the least inhibition of mycelial growth of both Trichoderma spp., and the lowest doses with amidosulforone (6.25%, 12.5%) even significantly stimulated mycelial growth of both Trichoderma spp. Fluazifop-P-butyl, pyridate and tembotrione showed moderately harmful to harmful effects on mycelial growth, with 70% to 80% inhibition at the recommended herbicide application rates. Fluorochloridone, S-metolachlor, prosulfocarb, metribuzin and pendimethalin inhibited mycelial growth the most, in descending order of inhibition. At 50% to 100%, fluorochloridone, prosulfocarb and S-metolachlor had fungicidal effects. Fluorochloridone and prosulfocarb had significant inhibitory effects (> 75%) even at the lowest concentration of 6.25%. Of these herbicides tested, the least adverse effects were seen for amidosulfuron, dicamba and isoxaflutole, which are thus likely to be compatible with Trichoderma spp. in the field.

The infection caused by Entamoeba histolytica is still a serious public health problem, especially in developing countries. The goal of this study was to evaluate the effect of terfenadine against Entamoeba histolytica. The trophozoites were exposed to 1, 2, 3, and 4 μM of terfenadine, for 24 and 48 h. Consequently, the viability of cells was determined by trypan blue exclusion test. The effect of terfenadine on adhesion of Entamoeba histolytica was evaluated in Caco-2 cells. In addition, the effect of terfenadine on the erythrophagocytic capacity of the parasite was investigated. The results show that terfenadine affects the growth and cell viability in a time- and dose-dependent manner. The higher inhibitory effects were observed with 4 µM at 48 h; 91.6% of growth inhibition and only 22.5% of trophozoites were viable. Additionally, we demonstrate that terfenadine is highly selective for the parasite and has low toxicity on Caco-2 cells. Furthermore, adhesion to Caco-2 cells and erythrophagocytic capacity were significantly inhibited. These findings demonstrate that terfenadine exerts significant effects on the virulence of Entamoeba histolytica. This is the first study demonstrating the amoebicidal activity of terfenadine and the results suggest it may be effective in the treatment of amoebiasis.

Introduction Leishmania aethiopica (L. aethiopica) is responsible for different forms of cutaneous leishmaniasis (CL) in Ethiopia. Treatment heavily depends on limited drugs, together with drawbacks like toxicity and microbial resistance. The current research aimed to investigate in vitro growth inhibitory activity of Medicines for Malaria Ventures - Pathogen Box (MMV - PB) compounds against L. aethiopica clinical isolate. Methodology Four hundred MMV – PB compounds were screened against L. aethiopica using resazurin based colourimetric assay. Compounds with > 70% inhibition were further tested using macrophage based intracellular amastigote assay. Cytotoxic and hemolytic activity of candidate hits were assessed on THP1- cells and sheep red blood cells (RBCs), respectively. In vitro drug interaction study was also conducted for the most potent hit using the combination index method. Results At the test concentration of 1 μM, twenty-three compounds showed > 50% inhibition of promastigotes parasite growth, of which 11 compounds showed > 70% inhibition. The 50% growth inhibition (IC50) of the 11 compounds was ranged from 0.024 to 0.483 μM in anti-promastigote assay and from 0.064 to 0.899 μM in intracellular amastigote assay. Candidate compounds demonstrated good safety on sheep RBCs and THP-1 cell lines. MMV688415 demonstrated a slight hemolytic activity on sheep RBC (5.3% at 25 μM) and THP-1 cell line (CC20 = 25 μM) while MMV690102 inhibited half of THP-1 cells at 36.5 μM (selectivity index = 478). No synergistic activity was observed from the combinations of MMV690102 and amphotericin B (CI > 1), and MMV690102 and Pentamidine (CI > 1) at lower and higher combination points. Conclusion The present study identified a panel of compounds that can be used as a novel starting point for lead optimization. MMV690102 appears to be the most potent inhibitor against L. aethiopica promastigotes and amastigotes. Future works should investigate the antileishmanial mechanism of action and in vivo antileishmanial activities of identified hits.

Cancer cells develop several hallmark changes over the progress of the tumor process. Cell assistance in multicellular organisms is regulated by the division of cell coordination by aggressive growth modulation. In this perspective, the use of molecular indicators triggering cell division is a mystery, because a cancer cell can manipulate any molecule that induces and helps growth, disturbing cellular assistance. An effective alteration proceeding to tumors must develop to be competitive, allowing a cancer cell to pass a signal resulting in better selection chances. The subjective simulation of physiological systems has become increasingly valuable in recent years, and there is now a wide range of mathematical models of signalling pathways that have contributed to some groundbreaking discoveries and hypotheses as to how this system works. Here we discuss various modeling methods and their application to the physiology of medical systems, focusing on the identification of parameters in ordinary differential equation models and their significance for forecasting cellular decisions in network modeling. In situations of global and local cell-to-cell rivalry, we quantify how this mechanism impacts a mutated cell's fixing chance of producing such a signal, and consider that this process will play a vital role in reducing cancer.

The investigation on the n-hexane lipophilic extractives of Swietenia mahagoni, alongside the antifungal and antioxidant properties was conducted. The leaf sample was collected from 2- and 3-years-old trees in Perhutani enterprise of Temanggung, Central Java, Indonesia. In addition, the antifungal activity was tested using the white-rot of Phanerochaete chrysosporium, while the antioxidant property involved the DPPH (1,1-diphenyl-2-picrylhydrazyl) method. The results show antifungal of 28.6% and 37.5% growth inhibition in 2- and 3 -years-old leaf, respectively, while 21.7% and 25.4% antioxidant activity were also recorded. Furthermore, the GC-MS results showed the domination of S. mahagoni leaf by fatty acids and hydo-carbons, while the terpenoids and steroids were in minor quantities. These components possibly exhibit growth inhibitory and antioxidant effects against P. chrysosporium and DPPH radicals, respectively.

Microalgae, Botryococcus braunii in particular, have received increasing interest owing to their potential as biofuel sources. Although the fertilizer components present in wastewater are useful in reducing the cost of commercial production of microalgae, the ammonium nitrogen (NH4+-N) acidifies the medium and may inhibit the growth of B. braunii. In this study, we aimed to investigate the effectiveness of calcium carbonate in preventing the growth inhibition of B. braunii by suppressing pH decrease caused by NH4+-N. Four types of modified Chu13 media were prepared: a control, one with NH4+-N, one with calcium carbonate, and one with calcium carbonate and NH4+-N. Then, the pH, NH4+-N concentration, B. braunii growth, and hydrocarbon content were measured. We found that in the NH4+-N treatment without calcium carbonate, the pH decreased to approximately 4, and there was almost no algal growth even after 20 days. In contrast, there were no differences in the growth rate and medium pH (7–8) between the treatment with calcium carbonate and the control medium. In addition, calcium ions were leached into the medium with calcium carbonate as the culture progressed, confirming the effectiveness of calcium carbonate in maintaining a constant pH and thus preventing growth inhibition. As such, the addition of calcium carbonate effectively prevents B. braunii growth inhibition by suppressing pH decrease and is more cost-effective than aggressive pH adjustment with alkali or acid or by adding buffer solutions into the medium. This simple and inexpensive method can be used for the removal of ammonia from wastewater.

Plant fungal diseases cause major problems for the global economy. Antimicrobial peptides have aroused great interest in the control of phytopathogens, as they are natural molecules and have a broad spectrum of inhibitory activity. Herein, we have tried to identify and characterize antimicrobial peptides present in fruits of Capsicum chinense and to evaluate their enzymatic and antifungal activities. The retained fraction obtained in the anion exchange chromatography with strong antifungal activity was subjected to molecular exclusion chromatography and obtained four fractions named G1, G2, G3, and G4. The 6.0-kDa protein band of G2 showed similarity with protease inhibitors type II, and it was able to inhibit 100% of trypsin and α-amylase activities. The protein band with approximately 6.5 kDa of G3 showed similarity with sequences of protease inhibitors from genus Capsicum and showed growth inhibition of 48% for Colletotrichum lindemuthianum, 49% for Fusarium lateritium, and 51% for F. solani and F. oxysporum. Additionally, G3 causes morphological changes, membrane permeabilization, and ROS increase in F. oxysporum cells. The 9-kDa protein band of G4 fraction was similar to a nsLTP type 1, and a protein band of 6.5 kDa was similar to a nsLTP type 2. The G4 fraction was able to inhibit 100% of the activities of glycosidases tested and showed growth inhibition of 35 and 50% of F. oxysporum and C. lindemuthianum, respectively. C. chinense fruits have peptides with antifungal activity and enzyme inhibition with biotechnological potential.

Prymnesium parvum is a toxin-producing harmful alga that has caused ecological and economic damage worldwide. Effective methods to control blooms of this species in the field, however, are unavailable. This study examined five natural compounds present in the invasive plant Arundo donax and one synthetic derivative (5,6-dichlorogramine) for their effect on P. parvum growth. All compounds except one inhibited growth in the following order of potency: ellipticine > > 5,6-dichlorogramine > 1 H-indole = 2,4,6-trimethyl-benzonitrile > gramine. Ellipticine was by far the most potent inhibitor, with full algicidal activity at concentrations as low as 0.04 mg L−1 and 3- and 9-day IC50 values of 0.012 and 0.007 mg L−1, respectively. A reduction in chlorophyll content and swimming activity and an increase in length and volume (swelling) were documented in algal cells exposed to 0.01–0.02 mg ellipticine L−1. These results show that ellipticine is among the most potent natural algicides identified to date. The sixth compound tested, oleamide, unexpectedly stimulated algal growth above control levels. Overall, these observations confirm the existence of highly potent anti-P. parvum allelochemicals in giant reed and demonstrate potential for using products derived from this plant in the development of natural, environmentally friendly methods to control harmful algal blooms.

The development of novel drugs against Gram-negative bacteria represents an urgent medical need. To overcome their outer cell membrane, we synthesized conjugates of antibiotics and artificial siderophores based on the MECAM core, which are imported by bacterial iron uptake systems. Structures, spin states, and iron binding properties were predicted in silico using density functional theory. The capability of MECAM to function as an effective artificial siderophore in Escherichia coli was proven in microbiological growth recovery and bioanalytical assays. Following a linker optimization focused on transport efficiency, five β-lactam and one daptomycin conjugates were prepared. The most potent conjugate 27 showed growth inhibition of Gram-positive and Gram-negative multidrug-resistant pathogens at nanomolar concentrations. The uptake pathway of MECAMs was deciphered by knockout mutants and highlighted the relevance of FepA, CirA, and Fiu. Resistance against 27 was mediated by a mutation in the gene encoding ExbB, which is involved in siderophore transport.

Cyanobacterial mutants defective in acyl-acyl carrier protein synthetase (Aas) produce free fatty acids (FFAs) because the FFAs generated by deacylation of membrane lipids cannot be recycled. An engineered Aas-deficient mutant of Synechocystis sp. PCC 6803 grew normally under low-light (LL) conditions (50 µmol photons m−2 s−1) but was unable to sustain growth under high-light (HL) conditions (400 µmol photons m−2 s−1), revealing a crucial role of Aas in survival under the HL conditions. Several-times larger amounts of FFAs were produced by HL-exposed cultures than LL-grown cultures. Palmitic acid accounted for ∼85% of total FFAs in HL-exposed cultures, while C18 fatty acids (FAs) constituted ∼80% of the FFAs in LL-grown cultures. Since C16 FAs are esterified to the sn-2 position of lipids in the Synechocystis species, it was deduced that HL irradiation activated deacylation of lipids at the sn-2 position. Heterologous expression of FarB, the FFA exporter protein of Neisseria lactamica, prevented intracellular FFA accumulation and rescued the growth defect of the mutant under HL, indicating that intracellular FFA was the cause of growth inhibition. FarB expression also decreased the ‘per-cell’ yield of FFA under HL by 90% and decreased the proportion of palmitic acid to ∼15% of total FFA. These results indicated that the HL-induced lipid deacylation is triggered not by strong light per se but by HL-induced damage to the cells. It was deduced that there is a positive feedback loop between HL-induced damage and lipid deacylation, which is lethal unless FFA accumulation is prevented by Aas.

Salt elicitation in therapeutic plants is one of the most popular techniques to enrich targeted secondary metabolites at cellular levels. Therefore, the appropriate concentration of NaCl salt that may regulate secondary metabolites without growth retardation needs to be established for maximum productivity. The aim of this study was to investigate the optimum NaCl elicitation for improving centellosides’ yield and associated physio-morphological adaptations in Indian pennywort (Centella asiatica (L.) Urban). Sodium ions in the salt-treated leaf tissues were significantly increased, while potassium ions were declined, leading to an increase in Na:K ratio. Centellosides yield of Indian pennywort was maximized at 656.11 mg plant−1 (1.52 folds over control) when treated with 12.5 mM NaCl for 14 days. Similarly, madecassoside, asiaticoside and total centellosides were enriched under 12.5 and 25.0 mM NaCl, whereas those compounds were significantly declined upon 50 mM NaCl treatment. Under 50 mM NaCl, reduction in both non-stomatal functions [leaf greenness (SPAD), maximum quantum yield of PSII (Fv/Fm) and photon yield of PSII (ΦPSII)] and stomatal functions [net photosynthetic rate (Pn), stomatal conductance (gs) and transpiration rate (E)] were found, leading to increased leaf temperature (TLeaf) and crop stress index (CSI) as well as overall growth inhibition. Total soluble sugars, glucose, and fructose under 50 mM NaCl were enriched by 1.21, 1.28 and 1.21 folds over control, respectively to play a major role in osmotic adjustment. In summary, the production of centelloside in Indian pennywort can be practiced using 12.5–25.0 mM NaCl elicitation without any negative effects on physio-morphological performance.

Recent studies suggest that RRP15 (Ribosomal RNA Processing 15 Homolog) might be a potential target for cancer therapy. However, the role of RRP15 in hepatocarcinogenesis remains poorly delineated. In this study, we aimed to evaluate the expression and biological function of RRP15 in human hepatocellular carcinoma (HCC). We show that RRP15 was up regulated in HCC cell lines and tumours. Up-regulation of RRP15 in HCC tumours was also correlated with unfavorable prognosis. We further show that the frequent up-regulation of RRP15 in HCCs is at least partly driven by recurrent gene copy gain at chromosome 1q41. Functional studies indicated that RRP15 knockdown suppresses HCC proliferation and growth both in vitro and in vivo. Mechanistically, RRP15 depletion in p53-wild-type HepG2 cells induced senescence via activation of the p53-p21 signalling pathway through enhanced interaction of RPL11 with MDM2, as well as inhibition of SIRT1-mediated p53 deacetylation. Moreover, RRP15 depletion in p53-mutant PLC5 and p53-deleted Hep3B cells induced metabolic shift from the glycolytic pentose-phosphate to mitochondrial oxidative phosphorylation via regulating a series of key genes such as HK2 and TIGAR, and thus, promoted the generation of ROS and apoptosis. Taken together, our findings provide evidence for an important role of the RRP15 gene in hepatocarcinogenesis through regulation of HCC proliferation and growth, raising the possibility that targeting RRP15 may represent a potential therapeutic strategy for HCC treatment.

Prosopis strombulifera (Lam.) Benth. is a halophytic shrub abundant in high-salinity areas in central Argentina, with high tolerance against NaCl but strong growth inhibition by Na2SO4. In the present study, the modulation of the antioxidant systems (enzymatic and non-enzymatic components) was analyzed under different salt treatments (NaCl, Na2SO4 and the iso-osmotic mixture) in hydroponic cultivation. Na2SO4-treated plants showed strong indications of oxidative stress (H2O2 and O2-• increase). Modifications in antioxidant enzymes activities were observed mainly under Na2SO4 treatment, where CAT seems to play an important role in early detoxification of H2O2 in roots, whereas SOD and APX have a predominant role in leaves. As part of the non-enzymatic system, 21 compounds were identified in leaves, being polyphenols the most abundant. Control plants contained the major variety of detected phytochemicals (14). Na2SO4-treated plants contained 10 compounds and NaCl-treated plants nine compounds, but with a different profile. NaCl-treated plants showed the highest antioxidant capacity. Our findings confirm that different types of salt treatments provoke a differential modulation of the antioxidant systems. Polyphenols and other ROS-detoxifying compounds, in a joint action with the enzymatic antioxidant system, are proposed to have a fundamental role in the cellular protection of P. strombulifera plants under severe oxidative stress. Our findings also highlight the potential of this halophyte as a valuable source of bioactive compounds with high antioxidant activity and health benefits.

Lavender essential oil (LEO), a natural antimicrobial agent, is generally recognized as safe and effective in the inhibition of phytopathogenic fungi. Direct contact and fumigation (in vivo and in vitro) were used to study the fungistatic effect of LEO on Monilinia fructicola. Additionally, the effect on the ultrastructure of cells and the degree of destruction of the cell membrane of M. fructicola were revealed. In addition, the effects of LEO on the expression levels of apoptosis-related genes in M. fructicola cells were detected, and GC-MS was used to analyze the main components of LEO. LEO had a good inhibitory efficacy against M. fructicola in flat peaches, with almost complete growth inhibition at 800 μL/L. These effects were associated with the leakage of cytoplasmic contents, hyphal distortion, and spore disruption. Moreover, the expression of apoptosis RTG1 and RLM1 genes increased with LEO treatment. These results demonstrate that LEO can inhibit M. fructicola by inducing cytoplasmic membrane damage and cell apoptosis in fungi, and that the major ingredients of LEO are monoterpenes and sesquiterpenes, which are presumed to contribute to the inhibitory effects.

Inhibition of tight junction formation between two malaria parasite proteins, apical membrane antigen 1 and rhoptry neck protein 2, crucial for red blood cell invasion, prevents the disease progression. In this work, we have utilized a unique approach to design a chimeric peptide, prepared by fusion of the best features of two peptide inhibitors, that has displayed parasite growth inhibition, in-vitro, with nanomolar IC50, which is hundredfold better than any of its parent peptides. Further, to gain structural insights, we computationally modeled the hybrid peptide on its receptor.

Aberrant lipid metabolism has recently been recognized as a new hallmark of malignancy, but the characteristics of fatty acid metabolism in breast cancer stem cells (BCSC) and potential interventions targeting this pathway remain to be addressed. Here, by using the in vitro BCSC models, mammosphere-derived MCF-7 cells and HMLE-Twist-ER cells, we found that the cells with stem cell-like properties exhibited a very distinct profile of fatty acid metabolism compared with that of their parental cancer cells, characterized by increased lipogenesis, especially the activity of stearoyl-CoA desaturase 1 (SCD1) responsible for the production of monounsaturated fatty acids, and augmented synthesis and utilization of the omega-6 arachidonic acid (AA). Suppression of SCD1 activity by either enzyme inhibitors or small interfering RNA (siRNA) knockdown strikingly limited self-renewal and growth of the BCSC, suggesting a key role for SCD1 in BCSC proliferation. Furthermore, elevated levels of SCD1 and other lipogenic enzymes were observed in human breast cancer tissues relative to the noncancer tissues from the same patients and correlated with the pathological grades. Interestingly, treatment of BCSC with omega-3 fatty acids, eicosapentaenoic acid and docosahexaenoic acid, effectively downregulated the expression of the lipogenic enzymes and markedly suppressed BCSC self-renewal and growth. Dietary supplementation of nude mice bearing BCSC-derived tumors with omega-3 fatty acids also significantly reduced their tumor load. These findings have demonstrated that increased lipogenesis is critical for self-renewal and growth of BCSC, and that omega-3 fatty acids are effective in targeting this pathway to exert their anticancer effect.

Atractylodes is the dry root of atractylodes macrocephala koidz and has been commonly used as a traditional Chinese medicine (TCM). Atractylenolide III, a main component of atractylodes, has displayed significant effects on anti-inflammation and anticancer. However, the effects of atractylenolide III on growth inhibition and apoptosis induction in colon cancer remain unclear. The results showed that atractylenolide III significantly inhibited the cell growth and induce cellular apoptosis in HCT-116 cells in a concentration dependence manner in vitro. Mechanistic studies further showed that atractylenolide III could regulate the Bax/Bcl-2 apoptotic signaling pathway through promoting the expression of proapoptotic related gene/proteins Bax, caspase-9 and caspase-3 but inhibiting the expression of antiapoptotic related gene/protein Bcl-2 in HCT-116 cells. Furthermore, atractylenolide III also significantly inhibited the tumor growth of HCT-116 tumor xenografts bearing in nude mice through inducing apoptosis by upregulation of the expressions of Bax, cleaved caspase-3 and p53 but downregulation of the expressions of Bcl-2 in HCT-116 tumor tissues in vivo. The studies may provide the scientific rationale for the understanding of the anticancer effect of atractylenolide III. Therefore, atractylenolide III may have the potential to be developed as a promising novel anticancer agent for the treatment of colorectal cancer clinically.

The rise of antimicrobial resistance for infectious bacteria has become an alarming issue to human health. New antimicrobial drugs are in dire need and pivotal to overcome this issue. In this study, aspirinate azo ligands bearing different halogens L1-5 has been prepared via diazo-coupling reaction. The ligands L1-5 were coordinated with silver, Ag (I) metal to produce Ag (I) aspirin-azo complexes C1-5. The antibacterial properties of L1-5 and C1-5 were evaluated against Staphylococcus aureus and Escherichia coli using turbidimetric kinetic method. The complexes C1-5 showed comparable growth inhibition activity towards E. coli (MIC 82-105 ppm) and S. aureus (MIC 80-105 ppm) compared to ligands L1-5 with E. coli (MIC 83-200 ppm), S. aureus (80-131 ppm) and ampicillin (MIC 93 and 124 ppm, respectively). The excellent bacterial resistance of both L1-5 and C1-5 indicates the potential of aspirinate azo and their complexes as new antibacterial agents, which significantly benefit to the pharmaceutical industries.

Pneumocandin B0, the precursor of the antifungal drug caspofungin, is a lipohexapeptide produced by the fungus Glarea lozoyensis. Oxidative stress and the resulting production of reactive oxygen species (ROS) are known to be involved in the regulation of pneumocandin B0 biosynthesis. In this study, the Glyap1 gene of Glarea lozoyensis, a homologue of the yeast redox regulator YAP1, was knocked out. The intracellular ROS levels of the resulting ΔGlyap1 strain were higher than in the wild-type strain, which was caused by the downregulated expression of superoxide dismutase (SOD) and catalase (CAT). Compared with the wild-type strain, ΔGlyap1 exhibited an oxidative phenotype throughout its life cycle, which resulted in significantly higher pneumocandin B0 production per unit biomass. In addition, ΔGlyap1 showed growth inhibition and decreased pneumocandin B0 production in the presence of CCl4, which leads to strong oxidative stress. To overcome the strain’s sensitivity, a three-stage antioxidant addition strategy was developed. This approach significantly improved the growth of ΔGlyap1 while maintaining a high pneumocandin B0 production per unit biomass, which reached 38.78 mg/g DCW. Notably, this result represents a 50% increase over the wild-type strain. These findings provide new insights into the regulatory mechanisms that control pneumocandin B0 production under oxidative stress, which may be applied to improve the production of other secondary metabolites. • Glyap1 is involved in expression of redox and pneumocandin B0 synthesis-related genes. • Addition of a three-stage antioxidant alleviated the sensitivity of ΔGlyap1 strain. • The yield of pneumocandin B0 per unit biomass of ΔGlyap1 strain was 38.78 mg/g DCW.

Salinity is one of the major abiotic stresses that limits agricultural productivity worldwide. Many proteins with defined functions in salt stress adaptation are controlled through interactions with members of the 14-3-3 family. In the present study, we generated three 14-3-3 quadruple knockout mutants (qKOs: klpc, klun, and unpc) to study the role of six non-epsilon group 14-3-3 proteins for salt stress adaptation. The relative growth inhibition under 100 mM of NaCl stress was the same for wild-type (Wt) and qKOs, but the accumulation of Na+ in the shoots of klpc was significantly lower than that in Wt. This difference correlated with the higher expression of the HKT1 gene in klpc. Considering the regulatory role of 14-3-3 proteins in metabolism and the effect of salt stress on metabolite accumulation, we analyzed the effect of a 24-h salt treatment on the root metabolome of nutrient solution-grown genotypes. The results indicated that the klpc mutant had metabolome responses that were different from those of Wt. Notably, the reducing sugars, glucose and fructose, were lower in klpc under control and salt stress. On the other hand, their phosphorylated forms, glucose-6P and fructose-6P, were lower under salt stress as compared to Wt. This study provided insight into the functions of the 14-3-3 proteins from non-epsilon group members. In summary, it was found that these proteins control ion homeostasis and metabolite composition under salt stress conditions and non-stressed conditions. The analyses of single, double, and triple mutants that modify subsets from the most effective qKO mutant (klpc) may also reveal the potential redundancy for the observed phenotypes.

Toxin–antitoxin (TA) systems are genetic modules composed of a toxin interfering with cellular processes and its cognate antitoxin, which counteracts the activity of the toxin. TA modules are widespread in bacterial and archaeal genomes. It has been suggested that TA modules participate in the adaptation of prokaryotes to unfavorable conditions. The Bosea sp. PAMC 26642 used in this study was isolated from the Arctic lichen Stereocaulon sp. There are 12 putative type II TA loci in the genome of Bosea sp. PAMC 26642. Of these, nine functional TA systems have been shown to be toxic in Escherichia coli. The toxin inhibits growth, but this inhibition is reversed when the cognate antitoxin genes are coexpressed, indicating that these putative TA loci were bona fide TA modules. Only the BoVapC1 (AXW83_01405) toxin, a homolog of VapC, showed growth inhibition specific to low temperatures, which was recovered by the coexpression of BoVapB1 (AXW83_01400). Microscopic observation and growth monitoring revealed that the BoVapC1 toxin had bacteriostatic effects on the growth of E. coli and induced morphological changes. Quantitative real time polymerase chain reaction and northern blotting analyses showed that the BoVapC1 toxin had a ribonuclease activity on the initiator tRNAfMet, implying that degradation of tRNAfMet might trigger growth arrest in E. coli. Furthermore, the BoVapBC1 system was found to contribute to survival against prolonged exposure at 4°C. This is the first study to identify the function of TA systems in cold adaptation.

The extracts of decay-resistant tree species are important research objects for the future development of wood preservatives. To understand the antifungal mechanisms of Coriolus versicolor inhibition with methanol extracts of C. camphora xylem, the protein profiles of C. versicolor were analyzed using 2-DE followed by MALDI-TOF/MS and bioinformatic analyses. The results showed that 41 protein spots were obviously changed among the 366-385 protein spots of C. versicolor treated with methanol extracts of C. camphora xylem. Twenty-one protein spots were upregulated, and 20 protein spots were downregulated. Cellular localization was performed to identify these differential proteins, and biological process and functional analysis found that 9 of these proteins were in the cytoplasm, 6 were intracellular, and 5 were in the mitochondrion. A total of 18.8% were mapped to small-molecule metabolic processes, 12.5% to cellular amino acid metabolic processes, and 10.9% to cellular nitrogen compound metabolic processes. Twenty-five percent of the differential proteins were associated with ion bonding, 15% with oxidoreductase activity, and 15% with ATPase activity and transmembrane transport activity. Downregulated expression of aspartate aminotransferase, ATP synthase alpha chain, DEAD/DEAH-box helicase, and phosphoglycerate kinase showed that the methanol extracts of C. camphora xylem disrupted functional aspects such as nitrogen and carbon metabolism, energy metabolism, hormone signal response, and glucose metabolism, eventually leading to C. versicolor inhibition.

The unicellular alga Chlamydomonas reinhardtii and the bacterium Pseudomonas protegens serve as a model to study the interactions between photosynthetic and heterotrophic microorganisms. P. protegens secretes the cyclic lipopeptide orfamide A that interferes with cytosolic Ca2+ homeostasis in C. reinhardtii resulting in deflagellation of the algal cells. Here, we studied the roles of additional secondary metabolites secreted by P. protegens using individual compounds and co-cultivation of algae with bacterial mutants. Rhizoxin S2, pyrrolnitrin, pyoluteorin, 2,4-diacetylphloroglucinol (DAPG) and orfamide A all induce changes in cell morphology and inhibit the growth of C. reinhardtii. Rhizoxin S2 exerts the strongest growth inhibition, and its action depends on the spatial structure of the environment (agar versus liquid culture). Algal motility is unaffected by rhizoxin S2 and is most potently inhibited by orfamide A (IC50 = 4.1 μM). Pyrrolnitrin and pyoluteorin both interfere with algal cytosolic Ca2+ homeostasis and motility whereas high concentrations of DAPG immobilize C. reinhardtii without deflagellation or disturbance of Ca2+ homeostasis. Co-cultivation with a regulatory mutant of bacterial secondary metabolism (ΔgacA) promotes algal growth under spatially structured conditions. Our results reveal how a single soil bacterium uses an arsenal of secreted antialgal compounds with complementary and partially overlapping activities.

The hypoxia-induced transcription factor HIF1 inhibits cell growth in normoxia through poorly understood mechanisms. A constitutive upregulation of hypoxia response is associated with increased malignancy, indicating a loss of antiproliferative effects of HIF1 in cancer cells. To understand these differences, we examined the control of cell cycle in primary human cells with activated hypoxia response in normoxia. Activated HIF1 caused a global slowdown of cell cycle progression through G1, S and G2 phases leading to the loss of mitotic cells. Cell cycle inhibition required a prolonged HIF1 activation and was not associated with upregulation of p53 or the CDK inhibitors p16, p21 or p27. Growth inhibition by HIF1 was independent of its Asn803 hydroxylation or the presence of HIF2. Antiproliferative effects of hypoxia response were alleviated by inhibition of lactate dehydrogenase and, more effectively, by boosting cellular production of NAD+, which was decreased by HIF1 activation. In comparison to normal cells, various cancer lines showed several fold-higher expressions of NAMPT, which is a rate-limiting enzyme in the main biosynthetic pathway for NAD+. Inhibition of NAMPT activity in overexpressor cancer cells sensitized them to antigrowth effects of HIF1. Thus, metabolic changes in cancer cells, such as enhanced NAD+ production, create resistance to growth-inhibitory activity of HIF1 permitting manifestation of its tumor-promoting properties.Abbreviations: DMOG: dimethyloxalylglycine, DM-NOFD: dimethyl N-oxalyl-D-phenylalanine, NMN: β-nicotinamide mononucleotide.

Ralstonia solanacearum is a harmful pathogen that causes severe wilt disease in several vegetables. In the present study, we identified R. solanacearum from wilt of papaya by 16S rRNA PCR amplification. Virulence ability of R. solanacearum was determined by amplification of approximately 1500 bp clear band of hrpB gene. Further, in-vitro seed germination assay showed that R. solanacearum reduced the germination rate up to 26.21%, 34% and 33.63% of cucumber, bottle guard and pumpkin seeds, respectively whereas shoot and root growth were also significantly decreased. Moreover, growth inhibition of R. solanacearum was recorded using antibacterial compound from medicinal plant and antagonistic B. subtilis. Petroleum ether root extract of Rauvolfia serpentina showed highest 22 ± 0.04 mm diameter of zone of inhibition where methanolic extract of Cymbopogon citratus and ethanolic extract of Lantana camara exhibited 20 ± 0.06 mm and 20 ± 0.01 mm zone of inhibition against R. solanacearum, respectively. In addition, bioactive compounds of B. subtilis inhibited R. solanacearum growth by generating 17 ± 0.09 mm zone of inhibition. To unveil the inhibition mechanism, we adopted chemical-protein interaction network and molecular docking approaches where we found that, rutin from C. citratus interacts with citrate (Si)-synthase and dihydrolipoyl dehydrogenase of R. solanacearum with binding affinity of −9.7 kcal/mol and −9.5 kcal/mol while quercetin from B. subtillis interacts with the essential protein F0F1 ATP synthase subunit alpha of the R. solancearum with binding affinity of −6.9 kcal/mol and inhibit the growth of R. solanacearum. Our study will give shed light on the development of eco-friendly biological control of wilt disease of papaya.

Thymus daenensis is a native medicinal plant of Iran with high pharmacological effects. The present survey was performed to evaluate the antibacterial effects of T. daenensis extract toward Staphylococcus aureus, Listeria monocytogenes, Escherichia coli and Pseudomonas aeruginosa food-borne bacteria. Ethanolic extract of T. daenensis was prepared from plant’s aerial parts. The diameters of the growth inhibition zones of bacteria against this extract and amoxicillin antibiotic disk was assessed by the disk diffusion test. Minimum inhibitory concentration was assessed using the broth micro-dilution method. The diameter of the growth inhibition zones of different concentrations of T. daenensis extract had the ranges between 7.00±0.54 to 15.00±1.09 mm. These findings revealed that the increase in the concentration of T. daenensis extract have caused significant increase in the diameter of the growth inhibition zone (P ≤0.05). Antibacterial effects of different concentrations of T. daenensis extract were lower than that of amoxicillin disk. T. daenensis extract had the highest antibacterial effects against P. aeruginosa. Additionally, T. daenensis extract had the highest antibacterial effects at lower concentrations (25 mg/mL) toward S. aureus. T. daenensis extract had the lowest MIC against S. aureus (6.00±1.00 mg/mL). The MIC of T. daenensis extract toward P. aeruginosa was 10.00±1.00 mg/mL. Antibacterial effects of T. daenensis ethanolic extract was determined in this survey. According to antibacterial effect of T. daenensis extract against food-borne bacteria, its application as a food preservative should potentially assess.

Treatments for toxoplasmosis such as pyrimethamine have shown numerous side effects. It has been reported that the likelihood of relapse associated with pyrimethamine-based therapy in patients with HIV and toxoplasmic encephalitis (TE) can have significant implications, even for patients who often develop new lesions in areas of the brain previously free of infection. This led us to research for new agents against Toxoplasma gondii. Recent findings have shown the potent biological activity of 4-thiazolidinones. We proposed to design and synthesize a new series of 2-hydrazono-4-thiazolidinones derivatives to evaluate the in vitro growth inhibition effect on T. gondii. The growth rates of T. gondii tachyzoites in Human Foreskin Fibroblast (HFF) cell culture were identified by two in vitro methodologies. The first one was by fluorescence in which green fluorescent RH parasites and cherry-red fluorescent ME49 parasites were used. The second one was a colorimetric methodology using β-Gal parasites of the RH strain constitutively expressing the enzyme beta-galactosidase. The 4-thiazolidinone derivatives 1B, 2B and 3B showed growth inhibition at the same level of Pyrimethamine. These compounds showed IC50 values of 1B (0.468–0.952 μM), 2B (0.204–0.349 μM) and 3B (0.661–1.015 μM) against T. gondii. As a measure of cytotoxicity the compounds showed a TD50 values of: 1B (60 μM), 2B (206 μM) and 3B (125 μM). The in vitro assays and molecular modeling results suggest that these compounds could act as possible inhibitors of the Calcium-Dependent Protein Kinase 1 of T. gondii. Further, our results support the fact that of combining appropriate detection technologies, combinatorial chemistry and computational biology is a good strategy for efficient drug discovery. These compounds merit in vivo analysis for anti-parasitic drug detection., Graphical abstract Image 1, Highlights • The structures of 16 4-Thiazolidinones synthesized were characterized by spectral data. • The 2-pyridine that contains 5-substituted 4-thiazolidinone showed low cytotoxicity and high selectivity index. • The 2-hidroxyphenyl and 2-pyridinyl hydrazinyl derivatives showed the best anti-T. gondii activities.

Abnormal microRNA (miR) expression has frequently been reported to be implicated in cancer-related drug resistance. Herein, we planned to investigate whether miR-381-3p contributes to doxorubicin (DOX) resistance in anaplastic thyroid carcinoma (ATC). DOX-resistant ATC tissues and cell lines were prepared to detect miR-381-3p and homeobox A9 (HOXA9) expression. CCK8, transwell and TUNEL assays were performed to evaluate cell proliferation, migration and invasion, and apoptosis in in vitro experiments. HOXA9 expression is intensively expressed in ATC tissues compared with benign thyroid tissues. Compared with parental ATC cell lines, HOXA9 protein expression is significantly up-regulated in DOX-resistant SW1736 and CAL62 cells. The knockdown of HOXA9 leads to growth inhibition and apoptosis of DOX-resistant SW1736 and CAL62 cells. Our results also indicate a significant decrease in miR-381-3p expression levels in DOX-resistant ATC tissues and cell lines. miR-381-3p may function as a tumour suppressor to impede proliferation, migration and invasion and induce apoptosis of DOX-resistant SW1736 and CAL62 cells by inhibiting HOXA9 protein expression. Our results present a novel signalling axis miR-381-3p/HOXA9 that mediates DOX resistance in ATC. miR-381-3p and HOXA9 may be promising molecular targets for preventing ATC progression and drug resistance.

Citrus hystrix Peel Extract and Carica papaya Leaves Extract contain several active components that can be used as antimicrobial compounds. The aim was to test a mixture of extracts from Citrus hystrix peel and Carica papaya leaves as growth inhibiting compounds for Candida albicans and Escherichia coli. This study used 4 levels of treatment, namely a combination of papaya leaves extract and orange fruit peel at concentrations of 0%, 25%, 50%, 75% and 100%. The research data were in the form of the diameter of the growth inhibition of Escherichia coli and Candida fungi, which were analyzed using the Kruskall-Wallis test at 5% confidence level and 5% Duncan's test. showed a significant difference between treatments, but the concentration of 100% showed better results to inhibit Escherichia coli (1.58 ± 0.28 d) and the growth of Candida albicans (1.53 ± 0.57b) compared to controls and other concentrations.

Embryonal rhabdomyosarcoma (eRMS) is the most common type of rhabdomyosarcoma in children. eRMS is characterized by malignant skeletal muscle cells driven by hyperactivation of several oncogenic pathways including the MYC pathway. Targeting MYC in cancer has been extremely challenging. Recently, we have demonstrated that the heart failure drug, proscillaridin A, produced anticancer effects with specificity toward MYC expressing leukemia cells. We also reported that decitabine, a hypomethylating drug, synergizes with proscillaridin A in colon cancer cells. Here, we investigated whether proscillaridin A exhibits epigenetic and anticancer activity against eRMS RD cells, overexpressing MYC oncogene, and its combination with decitabine. We investigated the anticancer effects of proscillaridin A in eRMS RD cells in vitro. In response to drug treatment, we measured growth inhibition, cell cycle arrest, loss of clonogenicity and self-renewal capacity. We further evaluated the impact of proscillaridin A on MYC expression and its downstream transcriptomic effects by RNA sequencing. Then, we measured protein expression of epigenetic regulators and their associated chromatin post-translational modifications in response to drug treatment. Chromatin immunoprecipitation sequencing data sets were coupled with transcriptomic results to pinpoint the impact of proscillaridin A on gene pathways associated with specific chromatin modifications. Lastly, we evaluated the effect of the combination of proscillaridin A and the DNA demethylating drug decitabine on eRMS RD cell growth and clonogenic potential. Clinically relevant concentration of proscillaridin A (5 nM) produced growth inhibition, cell cycle arrest and loss of clonogenicity in eRMS RD cells. Proscillaridin A produced a significant downregulation of MYC protein expression and inhibition of oncogenic transcriptional programs controlled by MYC, involved in cell replication. Interestingly, significant reduction in total histone 3 acetylation and on specific lysine residues (lysine 9, 14, 18, and 27 on histone 3) was associated with significant protein downregulation of a series of lysine acetyltransferases (KAT3A, KAT3B, KAT2A, KAT2B, and KAT5). In addition, proscillaridin A produced synergistic growth inhibition and loss of clonogenicity when combined with the approved DNA demethylating drug decitabine. Proscillaridin A produces anticancer and epigenetic effects in the low nanomolar range and its combination with decitabine warrants further investigation for the treatment of eRMS.

OBJECTIVES To investigate the role of polymorphism rs11200638 of high-temperature requirement factor A-1 (HtrA1) gene in the pathogenesis of age-related macular degeneration (AMD). METHODS Cultured adult retinal pigment epithelial cells (ARPE-19) expressing HtrA1 gene were treated with H2O2 or lipopolysaccharides (LPS) and analysed using western blot and quantitative polymerase chain reaction to illustrate the effects of oxidative and inflammatory stress on HtrA1 gene expression. Luciferase reporter plasmid driven by HtrA1 promoter with either normal allele G or risk allele A at SNP rs11200638 was transfected to ARPE-19 cells to investigate the effect of the G/A variation on HtrA1 promoter activity. The effects of HtrA1 overexpression on ARPE-19 cells were analysed with respect to percentage of cell proliferation inhibition and cell apoptosis. RESULTS HtrA1 expression was significantly increased with LPS or H2O2 stimulations (p

This report highlights the synthesis and characterization of ten new bis(N‐picolinamido)cobalt(II) complexes of the type [(L)2CoX2]0/2+, whereby L=N‐picolinamide ligand and X=diisothiocyanato (−NCS), dichlorido (−Cl) or diaqua (−OH2) ligands. Single crystal X‐ray (SC‐XRD) analysis for nine of the structures are reported and confirm the picolinamide ligand is bound to the Co(II) center through a neutral N,O binding mode. With the addition of powder X‐ray diffraction (PXRD), we have confirmed the cis and trans ligand arrangements of each complex. All complexes were screened against several fungal species and show increased antifungal activity. Notably, these complexes had significant activity against strains of Candida albicans and Aspergillus fumigatus, with several compounds exhibiting growth inhibition of >80 %, and onecompound inhibiting Aspergillus fumigatus hyphal growth by >90 %. Conversely, no antifungal activity was exhibited toward Cryptococcus neoformans and no cytotoxicity towards mammalian cell lines., Bis(N‐picolinamide)cobalt(II) complexes containing trans isothiocyanate ligands are reported to have significantly enhanced activity toward fungal species, when compared the cis complexes. They display >80 % growth inhibition of fungal strains and no reported toxicity toward bacterial species or mammalian cells.

The methylase METTL3 is the writer enzyme of the N6‐methyladenosine (m6A) modification of RNA. Using a structure‐based drug discovery approach, we identified a METTL3 inhibitor with potency in a biochemical assay of 280 nM, while its enantiomer is 100 times less active. We observed a dose‐dependent reduction in the m6A methylation level of mRNA in several cell lines treated with the inhibitor already after 16 h of treatment, which lasted for at least 6 days. Importantly, the prolonged incubation (up to 6 days) with the METTL3 inhibitor did not alter levels of other RNA modifications (i. e., m1A, m6Am, m7G), suggesting selectivity of the developed compound towards other RNA methyltransferases., Writer's block: We developed and characterized a small‐molecule inhibitor of m6A writer METTL3 by protein crystallography, biochemical and cellular assays. It shows high‐nanomolar potency in the biochemical assay, good selectivity against a panel of protein methyltransferases and kinases, and it reduced m6A/A ratio in mRNAs of different cell lines. In addition, we confirmed the selectivity of our compound towards other RNA methyltransferases in living cells.

Environment-friendly sound measures with high algal growth inhibition efficiency are required to control and eliminate CyanoHABs. This study examined the effects of protopine on growth, gene expression, and antioxidant system of the M. aeruginosa TY001 and explored possible damage mechanism. The results revealed that higher concentrations of protopine seriously inhibited the growth of M. aeruginosa. Quantitative real-time PCR analysis showed downregulated expression of stress response genes (prx and fabZ), and DNA repair gene (recA) on days 3 and 5. The activities of antioxidant enzymes were also decreased markedly, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD). Additionally, protopine stress can significantly increase the malondialdehyde (MDA) level in cells. In conclusion, oxidative damage and DNA damage are the main mechanisms of protopine inhibition on M. aeruginosa TY001. Our studies provide evidence that alkaloid compounds such as protopine may have a potential use value as components of aquatic management strategies.

Using a panel of cancer cell lines, we characterized a novel degrader of AKT, MS21. In mutant PI3K–PTEN pathway cell lines, AKT degradation was superior to AKT kinase inhibition for reducing cell growth and sustaining lower signaling over many days. AKT degradation, but not kinase inhibition, profoundly lowered Aurora kinase B (AURKB) protein, which is known to be essential for cell division, and induced G2–M arrest and hyperploidy. PI3K activated AKT phosphorylation of AURKB on threonine 73, which protected it from proteasome degradation. A mutant of AURKB (T73E) that mimics phosphorylation and blocks degradation rescued cells from growth inhibition. Degrader-resistant lines were associated with low AKT phosphorylation, wild-type PI3K/PTEN status, and mutation of KRAS/BRAF. Pan-cancer analysis identified that 19% of cases have PI3K–PTEN pathway mutation without RAS pathway mutation, suggesting that these patients with cancer could benefit from AKT degrader therapy that leads to loss of AURKB. Significance: MS21 depletes cells of phosphorylated AKT (pAKT) and a newly identified AKT substrate, AURKB, to inhibit tumor growth in mice. MS21 is superior to prior agents that target PI3K and AKT due to its ability to selectively target active, pAKT and sustain repression of signaling to deplete AURKB. This article is highlighted in the In This Issue feature, p. 2945

Glioma is one of the most commonly observed tumours, representing approximately 75% of brain tumours in the adult population. Generally, glioma therapy includes surgical resection followed by radiotherapy and chemotherapy. The transcription factor STAT3 (signal transducer and activator of transcription 3) is a promising target for the treatment of cancer and several other diseases. At nanomolar concentrations, SD‐36 induces rapid cellular degradation of STAT3 but cannot degrade other STAT proteins. The current study demonstrates the therapeutic efficacies of the STAT3 degraders SD‐36 against glioma, as well as understanding the elucidating mechanisms and identifying molecular markers that determine cell sensitivity to STAT3 degraders. Glioma cell lines possessed similar response patterns to SD‐36 but different responses to the STAT3 inhibitor Stattic. SD‐36 potently induced apoptosis in glioma cells along with a reduction in Mcl‐1 levels, which are critical for mediating the induction of apoptosis and enhancing TMZ‐induced apoptosis. Accordingly, SD‐36 sensitizes the antitumour effect of TMZ in patient‐derived xenograft. In addition, the downregulation of Mcl‐1 expression‐mediated antitumour effect of SD‐36 was analysed in cell‐derived xenograft. These observations need to be validated clinically to confirm the efficacy of STAT3 degraders in glioma.