Higher Growth Inhibition Rate Research Articles

Two Strains of Lentinula edodes Differ in Their Transcriptional and Metabolic Patterns and Respond Differently to Thermostress

Temperature type is one of the key traits determining the cultivation regime of Lentinula edodes. However, the molecular and metabolic basis underling temperature type remain unclear. Here, we investigated the phenotypic, transcriptomic, and metabolic features of L. edodes with different temperature types under both control (25 °C) and high (37 °C) temperature conditions. We found that under the control condition, the high- and low-temperature types of L. edodes harbored distinct transcriptional and metabolic profiles. The high-temperature (H-)-type strain had a higher expression level of genes involved in the toxin processes and carbohydrate binding, while the low-temperature (L-)-type strain had a high expression level of oxidoreductase activity. Heat stress significantly inhibited the growth of both H- and L-type strains, while the latter had a higher growth inhibition rate. Upon exposure to heat, the H-type strain significantly up-regulated genes associated with the components of the cellular membrane, whereas the L-type strain markedly up-regulated genes involved in the extracellular region and carbohydrate binding. Metabolome data showed that thermostress altered purine and pyrimidine metabolism in the H-type strain, whereas it altered cysteine, methionine, and glycerophospholipid metabolism in the L-type strain. Transcriptome and metabolome integrative analysis was able to identify three independent thermotolerance-related gene-metabolite regulatory networks. Our results deepen the current understanding of the molecular and metabolic basis underlying temperature type and suggest, for the first time, that thermotolerance mechanisms can be temperature-type-dependent for L. edodes.

Molecular diagnosis and possible control of the pathogenic Aspergillus flavus isolates associated with cultivated mushroom Agaricus bisporus in Iraq

Five isolates of Aspergillus flavus were isolated from casing soils, compost and fruiting bodies of Agaricus bisporus from several culture stations in Iraq. According to the pathogenicity test, the highest percentage of inhibition was recorded for isolate A.flavusB1 .It was molecularly diagnosed to the level of A.flavus on the basis of the nucleotide sequence analysis of the ITS genome region and I was recorded globally on the NCBI data base under the accession number ON738705.1. The effect of the hot aqueous extract of Cloves and Conocarpus, and the efficacy of the biocide VEROX on the production and phenotypic characteristics of the white edible fungus A. bisporus was studied in present of the pathogenic fungus. The results showed that the treatment of the combination of plant extracts + biocide + pathogenic fungus was recorded the highest rate of growth inhibition of the pathogenic fungus compared with other treatments ,and it was increased the number of fruiting bodies up to 505 compared with the control treatment of pathogenic fungus was 195. The total yield of the same treatment was 19.894 kg compared to 4.933 kg in the control treatment, and the biological efficiency was recorded by 33.7% compared to 12.6% in the control treatment. In general, this combination increased production and improved the qualitative characteristics of the fruiting bodies, as the average diameter of the cap was 43.1 mm and the stalk length was 39.1 mm compared to 28.5 mm and 23.5 mm in the control treatment, respectively.

Evaluation the pathogenicity and control of Penicillium citrinum isolates affecting the white fungus Agaricus bisporus

Five isolates of Penicillium citrinumwere isolated from casing soils, copmpost and fruiting bodies of Agaricusbisporus from several culture stations in Iraq. According to the pathogenicity test, the highest percentage of inhibition was recorded for isolate P.citrinum(B1). The causal agent was molecularly diagnosed to the level of P. citrinumaccording to the nucleotide sequence analysis of the ITS genome region and it was recorded globally on the NCBI data base under the accission number ON738710.1.The effect of the hot aqueous extract of cloves and conocarpus, and the efficacy of the biocide VEROX on the production and phenotypic characteristics of the white edable fungus Agaricusbisporus was studied with the presence of the pathogenic fungus. The results showed that the treatment of the combination of plant extracts + biocide + pathogenic fungus led to the highest rate of growth inhibition of the pathogenic fungus, increasing number of fruiting bodies up to 435 compared to the control treatment of pathogenic fungus 140. The total yield of the same treatment was 16.398 kg compared to 3.343 kg in the control treatment, and the biological efficiency was recorded by 37.2% compared to 10.2% in the control treatment. In general, this combination led to an increase in production and an improvement in the qualitative characteristics of the fruiting bodies, as the average diameter of the cap was 40.6 mm and the stalk length was 37.3 mm compared to 27.8 mm and 23.5 mm in the control treatment, respectively.

Chemical composition and bioactivities of thirteen non-host plant essential oils against Plutella xylostella L. (Lepidoptera: Plutellidae)

• Essential oils were isolated from 13 species of Chinese medicinal herbs and wild plants. • Five tested oils showed moderate to high bioactivities against DBM larvae. • The geranium and bishkatali oils exhibited remarkable toxicities to DBM larvae. • The basil oil revealed the highest antifeedant activity to DBM larvae. • The rue oil had strong inhibitory effects on the DBM larval growth. The diamondback moth (DBM), Plutella xylostella (L.), is a globally destructive pest of cruciferous vegetables. Excessive use of synthetic pesticides to control this species results in negative effects on the environment, human health, and nontargeted organisms. The essential oils (EOs) derived from plants may be developed as effective alternatives to conventional pesticides. In this study, thirteen EOs were extracted by hydrodistillation, respectively. Their chemical compositions were identified by gas chromatography-mass spectrometry (GC–MS). Furthermore, the biological activities of EOs such as toxicity, antifeedant activity, and growth inhibition effect, toward DBM larvae were investigated. Against DBM second-instar larvae, the most toxic EO was Pelargonium graveolens (LC 50 = 0.36 μg/μl) after 72 hr of exposure, followed by Polygonum hydropiper (LC 50 = 0.53 μg/μl). The Ocimum basilicum EO exhibited the highest antifeedant effect to third-instar larvae at at all set concentrations. At 15 μg/μl, the EOs of Acorus calamus , O. basilicum , and P . graveolens completely inhibited the feeding activities of larvae (100%). The Ruta graveolens EO showed the lowest relative growth rate and the highest growth inhibition rate towards third-instar larvae at diverse concentrations. And the EOs of P. hydropiper , A. calamus , and O. basilicum showed promising growth inhibition activities. Overall, the five EOs ( P . graveolens , O. basilicum , R. graveolens , P. hydropiper , and A. calamus ) showed moderate to high bioactivity, whereas eight EOs were found to be less active against DBM larvae. These results indicate that the five tested EOs are promising to be developed as novel botanical insecticides to control DBM population.

Cytotoxicity, apoptosis, and IL-8 gene expression induced by some foodborne pathogens in presence of Bacillus coagulans in HT-29 cells

Food poisoning caused by bacteria is one of the most important concerns in food hygiene. The use of probiotics in prevention, control, and treatment of these infections has been considerably increased in recent years. This study evaluated the effect of B. coagulans cell free supernatant (CFS) on growth of Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, non-pathogenic Escherichia coli, and Escherichia coli 0157:H7 by the broth dilution method. The cytotoxicity, and apoptosis induced by pathogens alone and in co-culture with B. coagulans or its CFS were measured by trypan blue, and fluorescence staining methods. The expression level of interleukin-8 (IL-8) cytokine-encoding genes was also investigated by a qRT-PCR assay in all pathogens and co-cultured groups in HT-29 cells. Our results showed that 4% B. coagulans CFS reduced pathogen growth. The highest rate of growth inhibition was observed in L. monocytogenes. We also found that B. coagulans, and its 4% CFS reduced the cytotoxic effects of pathogens, with the exception of S. aureus. Non-pathogenic E. coli also had no significant cytotoxic effect on the cells. Examination of the treated cells with acridine orange/ethidium bromide staining showed reductions in the rate of cell damage (including early apoptosis, late apoptosis, and necrosis) in pathogen-probiotic co-cultures. Furthermore, we showed that co-culture of pathogens with B. coagulans significantly down-regulated IL-8 gene expression (P < 0.05). The greatest down-regulation compared with pathogen alone was observed in S. aureus. Hence, B. coagulans can be considered as an appropriate probiotic to diminish cytotoxicity, and inflammatory response of enteropathogenic bacteria.

Effects of endoplasmic reticulum stress on the autophagy, apoptosis, and chemotherapy resistance of human breast cancer cells by regulating the PI3K/AKT/mTOR signaling pathway.

Nowadays, although chemotherapy is an established therapy for breast cancer, the molecular mechanisms of chemotherapy resistance in breast cancer remain poorly understood. This study aims to explore the effects of endoplasmic reticulum stress on autophagy, apoptosis, and chemotherapy resistance in human breast cancer cells by regulating PI3K/AKT/mTOR signaling pathway. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to detect the cell viability of six human breast cancer cell lines (MCF-7, ZR-75-30, T47D, MDA-MB-435s, MDA-MB-453, and MDA-MB-231) treated with tunicamycin (5 µM), after which MCF-7 cells were selected for further experiment. Then, MCF-7 cells were divided into the control (without any treatment), tunicamycin (8 µ), BEZ235 (5 µ), and tunicamycin + BEZ235 groups. Cell viability of each group was testified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Western blotting was applied to determine the expressions of endoplasmic reticulum stress and PI3K/AKT/mTOR pathway-related proteins and autophagy- and apoptosis-related proteins. Monodansylcadaverine and Annexin V-fluorescein isothiocyanate/propidium iodide staining were used for determination of cell autophagy and apoptosis. Furthermore, MCF-7 cells were divided into the control (without any treatment), tunicamycin (5 µM), cisplatin (16 µM), cisplatin (16 µM) + BEZ235 (5 µM), tunicamycin (5 µM) + cisplatin (16 µM), and tunicamycin (5 µM) + cisplatin (16 µM) + BEZ235 groups. Cell viability and apoptosis were also evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and Annexin V-fluorescein isothiocyanate/propidium iodide staining. In MCF-7 cells treated with tunicamycin, cell viability decreased significantly, but PEAK, eIF2, and CHOP were upregulated markedly and p-PI3K, p-AKT, and p-MTOR were downregulated in dose- and time-dependent manners. In the tunicamycin + BEZ235 group, the cell viability was lower and the apoptosis rate was higher than those of the control and monotherapy groups. Compared with the cisplatin group, the tunicamycin + cisplatin group showed a relatively higher growth inhibition rate; the growth inhibition rate substantially increased in the tunicamycin + cisplatin + BEZ235 group than the tunicamycin + cisplatin group. The apoptosis rate was highest in tunicamycin + cisplatin + BEZ235 group, followed by tunicamycin + cisplatin group and then cisplatin group. Our study provide evidence that endoplasmic reticulum stress activated by tunicamycin could promote breast cancer cell autophagy and apoptosis and enhance chemosensitivity of MCF-7 cells by inhibiting the PI3K/AKT/mTOR signaling pathway.

In vitro antitumor activity of patulin on cervical and colorectal cancer cell lines.

Patulin is a mycotoxin produced by some molds, especially Aspergillus and Penicilium, and is responsible for mycotoxicosis in animals and humans.There is still not very detailed data about the anti-cancer potency of patulin, but some reports demonstrated that it induces cellular apoptosis and toxicity. To determine the efficacy of patulin as a therapeutic strategy for cervical and colorectal cancers, we investigated its effects on HeLa,SW-48, and MRC-5 cell lines. Cell lines were exposed to various concentrations of patulin (i.e., 0.5, 1, 2, and 4 µM), then using methyl thiazolyl tetrazolium (MTT) and bromo-2'-deoxyuridine (BrdU) assays, the rates of apoptosis and cell viability were determined. The obtained results showed a significant reduction in cell viability and apoptosis induction in a dose-dependent manner. Among all the cell lines, the highest growth inhibition rate was obtained at the 4 μM concentration of patulin. Our results suggested that patulin could significantly decrease tumor growth in human cervical and colorectal cancer models.

Radioimmunotherapy of 131I-4E5 in nude mice bearing human B-cell lymphoma

Objective To explore the therapeutic efficacy of 131I labeled anti-CD80 monoclonal antibody (4E5) in nude mice bearing human B-cell lymphoma. Methods The 4E5 was radiolabeled with 131I using the Iodogen method at room temperature, and the immune activity and stability of 131I-4E5 were analyzed. The lethal effects of 131I-4E5 and 4E5 against Raji cells were evaluated by MTT assay. Three tumor-bearing nude mice were injected with 7.4 MBq of 131I-4E5 through tail vein, 72 h later the mice were sacrificed and the %ID/g was measured by calibrator and calculated. Fifteen tumor-bearing nude mice were randomly divided into 131I-4E5 group, 4E5 group and normal saline control group according to different injections. The tumor inhibition rate was calculated 15 d post-injection, and then the mice were sacrificed to get tumor tissues for histopathological examination. One-way analysis of variance and two-sample t test were used to analyze the data. Results The labeling rate of 131I-4E5 was (78.3±2.4)%, the radiochemical purity was (95.7±1.8)% and specific activity was 579.4 GBq/g. The specific binding efficiency of 131I-4E5 with Raji cells was (36.1±2.6)%. 131I-4E5 presented with a dose-dependent cytotoxicity against Raji cells. There were significant differences among the killing effects of different dosage groups (F=33.882, P<0.001). Furthermore, 131I-4E5 had higher growth inhibition rate on Raji cells than that of 4E5 (t: 3.986-5.515, all P<0.05). The biodistribution study showed that the tumor had high uptake of 131I-4E5, and the tumor/muscle ratio was 6.05±1.20. The radioimmunotherapy results showed that the tumor growth suppression effect was more obvious in 131I-4E5 group and the tumor inhibition rate was 71.69% on 15 d post-injection. The pathologic results showed necrosis in both 131I-4E5 group and 4E5 group, and the necrosis level was higher in 131I-4E5 group. Conclusion 131I-4E5 can target the tumor tissue and display an obvious antitumor effect in nude mice bearing human B-cell lymphoma. Key words: Lymphoma, B-cell; Radioimmunotherapy; Antibodies, monoclonal; Isotope labeling; Iodine radioisotopes; Mice, nude

Quality characteristics of Cheonggukjang containing Phellinus linteus extracts and antitumor effects in hep-2 and SK-MES cells

The quality characteristics of cheonggukjang containing Phellinus linteus extracts (PLE) and its cytotoxicity in Hep-2 and SK-MS-cells were investigated. Cheonggukjang containing 0.3% PLE showed the highest protease activity (7th aging day). The reducing sugar content and the amylase activity of cheonggukjang supplemented with 0.3% PLE increased immediately on the 1st aging day, and decreased slightly until the 15th aging day. All samples showed an increase in the amino-type nitrogen content. The dominant volatile compound detected using chromatography-mass spectrophotometry was 2,5-dimethyl pyrazine with the highest peak area value of 17.36% observed in cheonggukjang containing 0.3% PLE (3rd aging day). Cheonggukjang containing 0.3% PLE exhibited the best taste and acceptability with the least unpleasant odor. Cheonggukjang supplemented with PLE had a higher antitumor activity against SK-MES-1 and Hep-2 cells than a control. Hep-2 cells treated with a 10 mg/mL extract of cheonggukjang containing 0.3% PLE showed the highest growth inhibition rate of 84.74%.

Biocontrol Efficacy of Trichoderma Koningii Against some Plant Pathogenic fungi

In the present investigation, Trichoderma koningii was evaluated in vitro for its antagonistic activity against four fungal pathogen viz., Fusarium oxysporum, Fusarium solani, Alternari solani and Rhizoctonia solani causing vegetable diseases. T. koningii strongly antagonized with all pathogenic fungi in dual culture assay and gave maximum inhibition of mycelia growth (91.09%). Volatile metabolites produced by T. koningii exhibited highest growth inhibition rate (54.67%). The Metabolites released from T. koningii were tested in culture medium against all pathogens. Cell free metabolites of T. koningii inhibited the growth of all four pathogens and appeared to be fungicidal in its activity.

Matrine and anti-tumor drugs in inhibiting the growth of human lung cancer cell line

To study the activities of matrine and anti-tumor drugs on SPCA/I human lung adenocarcinoma cell line. Suppression effects of different concentrations of matrine and matrine combined with anti-tumor drugs on lung cancer cells were measured by methyl thiazolyl tetrazolium (MTT) colorimetric assay. Different concentrations of matrine could inhibit the growth of SPCA/I human lung adenocarcinoma cells and there was a positive correlation between the inhibition rate and the drug concentration. Different concentrations of matrine combined with anti-tumor drugs had higher growth inhibition rate than anti-tumor drugs alone. Matrine has direct growth suppression effect on SPCA/I human lung adenocarcinoma cells and matrine combined with anti-tumor drugs shows a significant synergistic effect on tumor cells.