Mediator complex subunit 19 regulates the proliferation, migration and invasion of human breast cancer cells

Purpose: To investigate the therapeutic implication of mediator complex subunit 19 (Med19) in breast cancer cells. Methods: The mRNA expression of Med19 was assayed using qRT-PCR. Cell viability was determined with 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay, while 4′,6-diamidino-2- phenylindole (DAPI) and annexin V/propidium iodide (PI) assays were used for determination of apoptosis. Wound healing and Transwell assays were used for the determination of cell migration and invasion. Western blotting analysis was used for assay of protein expression levels. Results: The results showed that Med19 was significantly (p < 0.05) upregulated in human breast cancer cell lines, relative to normal cells. The up-regulations ranged from 3.7-fold in UACC-2087 cells to 6.4-fold in BT-20 cells. Moreover, Med19 silencing caused significant decrease in the proliferation of BT-20 breast cancer cells (p < 0.05). The inhibition of cell proliferation was due to the induction of apoptosis, as was evident in increased Bax/Bcl-2 ratio. Annexin V/PI staining revealed 6 % apoptosis in si-NC-transfected, and about 13.30 % in si-Med19-transfected BT-20 cells. Wound healing and Transwell assays revealed that the invasion of BT-20 breast cancer cells significantly decreased upon Med19 silencing. Conclusion: Med19 regulates the proliferation, migration and invasion of human breast cancer cells. Thus, Med19 may be beneficial in the treatment of breast cancer

Perfluorocarbon restrains inflammation and cell apoptosis in rats with lung ischemia reperfusion injury via down regulation of TLR4/NF κB signaling pathway

Purpose: To exaluate the effect of perfluorocarbon on lung ischemia-reperfusion injury in rats, and to unravel the potential underlying mechanism.Methods: A total of 36 Sprague-Dawley (SD) rats were randomly assigned to sham group, model group, and perfluorocarbon group (12 rats per group). The levels of inflammatory factors (TNF-α and IL1β) were determined using enzyme-linked immunosorbent assay (ELISA). Cell apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, while Western blotting was conducted to determine the protein expressions of TLR4 and NF-κB.Results: The levels of inflammatory factors in the model and perfluorocarbon groups were significantly higher than those in operation group (p < 0.05), while their levels in perfluorocarbon group were significantly lower than in model group (p < 0.05). The mRNA expression levels of TNF-α and IL-1β in lung tissues rose significantly in both model and perfluorocarbon groups when compared with those in sham group (p < 0.05), but declined significantly in the perfluorocarbon group in comparison with those in model group (p < 0.05). Furthermore, the perfluorocarbon group exhibited a significantly lower cell apoptosis than model group (p < 0.05). The relative protein expression levels of TLR4 and NF-κBdeclined significantly in perfluorocarbon group than in model group.Conclusions: Perfluorocarbon down-regulates the TLR4/NF-κB signaling pathway, and inhibits inflammation a

Novel bi-allelic variants of CHMP1A contribute to pontocerebellar hypoplasia type 8: additional clinical and genetic evidence

Pontocerebellar hypoplasia type 8(PCH8) is a rare neurodegenerative disorder, reportedly caused by pathogenic variants of the CHMP1A in autosomal recessive inheritance, and CHMP1A variants have also been implicated in other diseases, and yet none of the prenatal fetal features were reported in PCH8. In this study, we investigated the phenotype and genotype in a human subject with global developmental delay, including clinical data from the prenatal stage through early childhood. Prenatally, the mother had polyhydramnios, and the bilateral ventricles of the fetus were slightly widened. Postnatally, the infant was observed to have severely delayed psychomotor development and was incapable of visual tracking before 2 years old and could not fix on small objects. The young child had hypotonia, increased knee tendon reflex, as well as skeletal malformations, and dental crowding; she also had severe and recurrent pulmonary infections. Magnetic resonance imaging of the brain revealed a severe reduction of the cerebellum (vermis and hemispheres) and a thin corpus callosum. Through whole exome sequencing and whole genomics sequencing, we identified two novel compound heterozygous variations in CHMP1A [c.53 T > C(p.Leu18Pro)(NM_002768.5) and exon 1 deletion region (NC_000016.10:g.89656392_89674382del)]. cDNA analysis showed that the exon1 deletion region led to the impaired expression, and functional verification with zebrafish embryos using base edition indicated variant c.53 T > C (p.Leu18Pro), causing dysplasia of the cerebellum and pons. These results provide further evidence that CHMP1A variants in a recessive inheritance pattern contribute to the clinical characteristics of PCH8 and further expand our knowledge of the phenotype and genotype spectrum of PCH8

TRIM37 interacts with PTEN to promote the growth of human T-cell acute lymphocytic leukemia cells through regulating PI3K/AKT pathway

BackgroundTRIM37 has been reported to be associated with the tumorigenesis of cancers. However, the role of TRIM37 in T-cell acute lymphoblastic leukemia (T-ALL) remains unclear. This study aimed to characterize the effect of TRIM37 on T-ALL.MethodsTRIM37 expression in T-ALL patients and T-ALL cell lines was determined by qRT-PCR and Western blot. Knockdown or overexpression of TRIM37 was conducted by transferring small-interfering TRIM37 or lentivirus-mediated transducing into T-ALL cells. CCK-8 assay and flow cytometry assay were conducted to analyze the proliferation and apoptosis of T-ALL cells. Co-immunoprecipitation experiments were conducted to investigate the relationship between TRIM37 and PTEN and the ubiquitination of PTEN.ResultsOur results suggested that TRIM37 expression was upregulated in the blood of T-ALL patients and T-ALL cell lines. Knockdown of TRIM37 noticeably inhibited the proliferation and promoted apoptosis of T-ALL cells. Ectopic expression of TRIM37 promoted the proliferation and suppressed the apoptosis rate of MOLT-4 cells and enhanced the phosphorylation of AKT. Moreover, TRIM37 interacted with PTEN and accelerated the degradation of PTEN via TRIM37-mediated ubiquitination in T-ALL cells. Moreover, TRIM37 reduced the sensitivity of T-ALL cells to bortezomib treatment. Additionally, PI3K/AKT signaling pathway was involved in the function of TRIM37 in T-ALL. TRIM37 contributed to the proliferation of T-ALL cells and reduced the susceptibility of T-ALL cells to bortezomib treatment through ubiquitination of PTEN and activating PI3K/AKT signaling pathway.ConclusionsOur study suggested that TRIM37 could be considered as a therapeutic target for T-ALL

Comparable Genital Tract Infection, Pathology, and Immunity in Rhesus Macaques Inoculated with Wild-Type or Plasmid-Deficient Chlamydia trachomatis Serovar D

Rhesus macaques were studied to directly address the potential for plasmid-deficient Chlamydia trachomatis to serve as a live attenuated vaccine in the genital tract. Five repeated cervical inoculations of rhesus macaques with wild-type serovar D strain D/UW-3/Cx or a plasmid-deficient derivative of this strain, CTD153, resulted in infections with similar kinetics and induced comparable levels of protective immunity. After all animals received five challenges with D/UW-3/Cx, levels of inflammation observed grossly and histologically were similar between the groups. Animals in both groups developed evidence of oviduct dilatation; however, reduced oviduct dilatation was observed for “controllers,” i.e., animals without detectable chlamydial DNA in the fimbriae at weeks 5 and 12. Grouping animals into “ascenders” and “controllers” revealed that elevated early T cell responses were associated with protection, whereas higher antibody responses were associated with ascension. Protected animals shared common major histocompatibility complex (MHC) alleles. Overall, genetic differences of individual animals, rather than the presence or absence of the chlamydial plasmid in the primary infecting strain, appeared to play a role in determining the outcome of infection

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Interactions indirectes médiées par la plante sous contraintes biotiques multiples

In the agroecosystem, plant crops are the primary trophic producer that can be consumed/parasitized by a plethora of organisms, including viruses, bacteria, fungi, nematodes and many herbivore insects. On one hand, plants support these organisms as food resources; on the other hand, plants defend themselves against those consumers by employing constitutive and induced defenses. Both features might be involved in plant-mediated indirect interactions when multiple pests share the same host plant. It means that the presence of one pest might have an indirect effect on another plant through the modification of some plant host features (nutritional quality or defense). During my Ph.D. study, I assessed the modulating factors (both abiotic and biotic factors) that might affect the plant-mediated indirect interactions, and also investigated the influences of such indirect interactions at long term and on a tri-trophic system, and finally analyzed the underlying phytochemical mechanisms by mechabolomics analyses. Firstly, sublethal effects of beta-cypermethrin, a pyrethroid pesticide as the abiotic factor, were assessed on the interspecific interactions between the specialist aphid Aphis glycines, and the generalist aphid Aulacorthum solani on soybeans. Sublethal concentration (LC5) of beta-cypermethrin accelerated the interspecific competition between the two species of aphids. Induced susceptibility of soybean plants to one species of aphids caused by the other species would be significantly impaired by the exposure to sublethal beta-cypermethrin.Secondly, various pest species abundance and feeding strategies, i.e. the piercing-sucking aphid, the chewing caterpillar, the plant fungus and the root-knot nematode, were introduced on tomato plants, Solanum lycopersicum to evaluate the plant-mediated indirect interactions among these pests. In the results, insect performances, i.e. the number of aphid individuals and development rate of caterpillar larva, were correlated to the abundance of indirect interactions whether pests attacked tomato plants sequentially or simultaneously. When pests attacked tomato plants at the same time, insect performances were positively correlated to the abundance of indirect interactions. Nevertheless, when pests infested tomato plants in sequence, the insect performances were negatively correlated to the abundance of indirect interactions. However, such a correlation was not detected with the pathogenic organisms (fungi and nematodes). Thirdly, the population dynamic of aphid M. euphorbiae was also monitored under a tri-trophic system. We found that the aphid population, involved in the above-belowground interactions, slowed down in both the short term and long term via reduced survival of nymph aphids and facilitating induction of winged aphids. Furthermore, parasitism efficiency of the natural enemy Aphidius ervi to aphids and tomato yields were also reduced by such above-belowground interactions. Lastly, following an untargeted metabolomic approach, we detected many significant chemical variations between non-infested and infested plants, disregarding the pest infestation. Most of them are probably induced by the presence of nematodes, the only belowground pest. On the opposite, we did not observe many chemical differences in roots metabolome when we compared the different types of infestation, meaning that the presence of nematodes probably lead the metabolomic signal in infested roots, disregarding the presence of other pests aboveground. All the studies may contribute to a comprehensive understanding of the diverse interactions among plants, pesticides, various pests, and natural enemies, and optimizing the integrated pest management in the agroecosystem.Les plantes, en tant que producteurs primaires, peuvent être consommées/attaquées par de nombreux organismes, notamment des virus, des pathogènes, des insectes herbivores ou encore des nématodes phytophages. D'une part, les plantes sont une ressource alimentaire principale pour ces organismes ; d'autre part, les plantes peuvent se défendre contre ces consommateurs en mobilisant des défenses constitutives et/ou induites. En cas de défenses induites suite à l’attaque d’un ravageur, des interactions indirectes médiées par la plante peuvent apparaitre entre ce ravageur et d’autres ravageurs partageant le même hôte i.e. la présence d’un ravageur va provoquer des modifications au niveau de la plante qui peuvent à leur tour impacter les autres ravageurs présents sur la même plante. Tout d’abord, les effets sublétaux de la bêta-cyperméthrine, un pesticide de type pyréthrinoïdes, ont été évalués sur les interactions interspécifiques entre le puceron spécialiste Aphis glycines et le puceron généraliste Aulacorthum solani sur soja. La concentration sublétale (LC5) de bêta-cyperméthrine accélère la compétition interspécifique entre les deux espèces de pucerons. La sensibilité des plants de soja aux pucerons induite par une pré-infestation d’une autre espèce de puceron pourrait ainsi être considérablement réduite par une exposition préalable à la bêta-cyperméthrine sublétaux. Dans un deuxième temps, nous nous sommes intéressés aux facteurs biotiques susceptibles de moduler les interactions indirectes. Dans ce cadre, divers ravageurs, à savoir le puceron, la chenille, le phytopathogène et le nématode, ont été introduits et suivis sur des plants de tomate, Solanum lycopersicum. La performance des ravageurs arthropodes, (e.g. le nombre d'individus de pucerons et le taux de développement des larves de lépidoptères) était corrélée au nombre d’interactions indirectes (i.e. au nombre d’espèces de ravageurs partageant le même hôte), que les ravageurs attaquent les plants de tomate séquentiellement ou simultanément. En effet, lorsque les ravageurs étaient mis en présence d’un plant de tomate de manière simultanée, la performance des insectes était positivement corrélée au nombre d’interactions indirectes. A l’inverse, lorsque les ravageurs étaient mis en présence de plants de tomate de façon séquentielle, la performance des insectes ravageurs était négativement corrélée à l'abondance des interactions indirectes. Enfin, aucune corrélation n'a pas été détectée pour les pathogènes, champignons et nématodes. D’autre part, nous avons pu mettre en évidence que la présence de nématodes et d’oïdium réduisait à court-terme i.e. à l’échelle d’une génération l’abondance de pucerons sur un plant de tomate attaqués par ces trois ravageurs. Nous avons voulu tester cette interaction indirecte sur du long-terme i.e. à l’échelle d’un suivi de la dynamique de populations des pucerons. De plus, nous avons ajouté à cette expérimentation des parasitoïdes du puceron afin de voir l’impact de ces interactions indirectes sur le troisième niveau trophique. La population de pucerons impliquée dans les interactions indirectes testées a été réduite à court et à long terme parce-que la survie des nymphes diminuait. De plus, le nombre de pucerons ailés a augmenté. Enfin, le taux de parasitisme de l’ennemi naturel Aphidius ervi. Ainsi, ce travail démontre la présence d’interactions indirectes fortes entre les différents ravageurs partageant une même plante hôte et ces interactions sont susceptibles d’être modulées par de nombreux facteurs comme le nombre ou encore la diversité des ravageurs. De plus, elles sont susceptibles de modifier le service de biocontrôle rendu par les auxiliaires des cultures ainsi que le rendement de la culture. Il est donc nécessaire de comprendre et prendre en compte ces interactions indirectes dans le cadre d’attaques multiples pour adopter les stratégies IPM les plus adéquates

무역 회사의 지속적 성장을 위한 경쟁 전략 수립에 대한 연구

Maste

How multiple-pest attack impacts plant-mediated indirect interactions on tomato crops?

Les plantes, en tant que producteurs primaires, peuvent être consommées/attaquées par de nombreux organismes, notamment des virus, des pathogènes, des insectes herbivores ou encore des nématodes phytophages. D'une part, les plantes sont une ressource alimentaire principale pour ces organismes ; d'autre part, les plantes peuvent se défendre contre ces consommateurs en mobilisant des défenses constitutives et/ou induites. En cas de défenses induites suite à l’attaque d’un ravageur, des interactions indirectes médiées par la plante peuvent apparaitre entre ce ravageur et d’autres ravageurs partageant le même hôte i.e. la présence d’un ravageur va provoquer des modifications au niveau de la plante qui peuvent à leur tour impacter les autres ravageurs présents sur la même plante. Tout d’abord, les effets sublétaux de la bêta-cyperméthrine, un pesticide de type pyréthrinoïdes, ont été évalués sur les interactions interspécifiques entre le puceron spécialiste Aphis glycines et le puceron généraliste Aulacorthum solani sur soja. La concentration sublétale (LC5) de bêta-cyperméthrine accélère la compétition interspécifique entre les deux espèces de pucerons. La sensibilité des plants de soja aux pucerons induite par une pré-infestation d’une autre espèce de puceron pourrait ainsi être considérablement réduite par une exposition préalable à la bêta-cyperméthrine sublétaux. Dans un deuxième temps, nous nous sommes intéressés aux facteurs biotiques susceptibles de moduler les interactions indirectes. Dans ce cadre, divers ravageurs, à savoir le puceron, la chenille, le phytopathogène et le nématode, ont été introduits et suivis sur des plants de tomate, Solanum lycopersicum. La performance des ravageurs arthropodes, (e.g. le nombre d'individus de pucerons et le taux de développement des larves de lépidoptères) était corrélée au nombre d’interactions indirectes (i.e. au nombre d’espèces de ravageurs partageant le même hôte), que les ravageurs attaquent les plants de tomate séquentiellement ou simultanément. En effet, lorsque les ravageurs étaient mis en présence d’un plant de tomate de manière simultanée, la performance des insectes était positivement corrélée au nombre d’interactions indirectes. A l’inverse, lorsque les ravageurs étaient mis en présence de plants de tomate de façon séquentielle, la performance des insectes ravageurs était négativement corrélée à l'abondance des interactions indirectes. Enfin, aucune corrélation n'a pas été détectée pour les pathogènes, champignons et nématodes. D’autre part, nous avons pu mettre en évidence que la présence de nématodes et d’oïdium réduisait à court-terme i.e. à l’échelle d’une génération l’abondance de pucerons sur un plant de tomate attaqués par ces trois ravageurs. Nous avons voulu tester cette interaction indirecte sur du long-terme i.e. à l’échelle d’un suivi de la dynamique de populations des pucerons. De plus, nous avons ajouté à cette expérimentation des parasitoïdes du puceron afin de voir l’impact de ces interactions indirectes sur le troisième niveau trophique. La population de pucerons impliquée dans les interactions indirectes testées a été réduite à court et à long terme parce-que la survie des nymphes diminuait. De plus, le nombre de pucerons ailés a augmenté. Enfin, le taux de parasitisme de l’ennemi naturel Aphidius ervi. Ainsi, ce travail démontre la présence d’interactions indirectes fortes entre les différents ravageurs partageant une même plante hôte et ces interactions sont susceptibles d’être modulées par de nombreux facteurs comme le nombre ou encore la diversité des ravageurs. De plus, elles sont susceptibles de modifier le service de biocontrôle rendu par les auxiliaires des cultures ainsi que le rendement de la culture. Il est donc nécessaire de comprendre et prendre en compte ces interactions indirectes dans le cadre d’attaques multiples pour adopter les stratégies IPM les plus adéquates.In the agroecosystem, plant crops are the primary trophic producer that can be consumed/parasitized by a plethora of organisms, including viruses, bacteria, fungi, nematodes and many herbivore insects. On one hand, plants support these organisms as food resources; on the other hand, plants defend themselves against those consumers by employing constitutive and induced defenses. Both features might be involved in plant-mediated indirect interactions when multiple pests share the same host plant. It means that the presence of one pest might have an indirect effect on another plant through the modification of some plant host features (nutritional quality or defense). During my Ph.D. study, I assessed the modulating factors (both abiotic and biotic factors) that might affect the plant-mediated indirect interactions, and also investigated the influences of such indirect interactions at long term and on a tri-trophic system, and finally analyzed the underlying phytochemical mechanisms by mechabolomics analyses. Firstly, sublethal effects of beta-cypermethrin, a pyrethroid pesticide as the abiotic factor, were assessed on the interspecific interactions between the specialist aphid Aphis glycines, and the generalist aphid Aulacorthum solani on soybeans. Sublethal concentration (LC5) of beta-cypermethrin accelerated the interspecific competition between the two species of aphids. Induced susceptibility of soybean plants to one species of aphids caused by the other species would be significantly impaired by the exposure to sublethal beta-cypermethrin.Secondly, various pest species abundance and feeding strategies, i.e. the piercing-sucking aphid, the chewing caterpillar, the plant fungus and the root-knot nematode, were introduced on tomato plants, Solanum lycopersicum to evaluate the plant-mediated indirect interactions among these pests. In the results, insect performances, i.e. the number of aphid individuals and development rate of caterpillar larva, were correlated to the abundance of indirect interactions whether pests attacked tomato plants sequentially or simultaneously. When pests attacked tomato plants at the same time, insect performances were positively correlated to the abundance of indirect interactions. Nevertheless, when pests infested tomato plants in sequence, the insect performances were negatively correlated to the abundance of indirect interactions. However, such a correlation was not detected with the pathogenic organisms (fungi and nematodes). Thirdly, the population dynamic of aphid M. euphorbiae was also monitored under a tri-trophic system. We found that the aphid population, involved in the above-belowground interactions, slowed down in both the short term and long term via reduced survival of nymph aphids and facilitating induction of winged aphids. Furthermore, parasitism efficiency of the natural enemy Aphidius ervi to aphids and tomato yields were also reduced by such above-belowground interactions. Lastly, following an untargeted metabolomic approach, we detected many significant chemical variations between non-infested and infested plants, disregarding the pest infestation. Most of them are probably induced by the presence of nematodes, the only belowground pest. On the opposite, we did not observe many chemical differences in roots metabolome when we compared the different types of infestation, meaning that the presence of nematodes probably lead the metabolomic signal in infested roots, disregarding the presence of other pests aboveground. All the studies may contribute to a comprehensive understanding of the diverse interactions among plants, pesticides, various pests, and natural enemies, and optimizing the integrated pest management in the agroecosystem