Bis(4-carb­oxy­pyridine-2-carboxyl­ato-κ2 N,O 2)copper(II) dimethyl sulfoxide disolvate

In the title complex, [Cu(C7H4NO4)2]·2C2H6OS, the CuII atom is situated on an inversion centre and is N,O-chelated by two monoanionic 4-carb­oxy­pyridine-2-carboxyl­ate ligands in a slightly distorted square-planar coordination geometry. The dimethyl sulfoxide solvent mol­ecules and CuII complex mol­ecules are linked by O—H⋯O hydrogen bonding. In addition, C—H⋯O contacts and π–π inter­actions [centroid–centroid distance = 3.590 (1) Å] occur

The Role Of Indigenous Knowledge In Increasing Rural People Knowledge About Agriculture 1

ABSTRACT Our today's world is the contradictions and collision's world. Contradiction between cultures, religions, different societies and countries. In recently years, from Renaissance till now, as much as human had developed, they also had contradictions and collisions in their world . One of these contradictions is the contrast between tradition and modernism. Maybe we can find these contrast roots in colonial era, the time when colonists promote their innovation in their colonies. Mostly these techniques and innovations show their native knowledge and the way of their living is foolish and inefficient and tried to enter industrial ways in to their life to increase production efficiency through this way. Thus the way of their living which was been formed during thousands of years has gone to be forgotten little by little. We can say, agriculture part is bearing the most damage in this rapid industrialization process. Absolving old and compatible ways in agriculture part and replacing and using of implant, harvest patterns without any proportions with environment has caused decrease of production efficiency, soil erosion and hard destruction of environment during a long time. Finally, at the end of the 20 th century decades, some solutions were suggested to solve these inconsistencies and problems. So the importance of native knowledge and effort in compilation of that with modern knowledge were considered and it was tried to make general and stable view in relation with environment and the way of living through this way

Acridine–benzene-1,3,5-tricarb­oxy­lic acid (3/1)

In the title adduct, 3C13H9N·C9H6O6 or (acr)3(btc), associ­ations of one btc and three acr molecules linked by O—H⋯N hydrogen bonds occur. C—H⋯O interactions also occur, resulting in a cyclic hydrogen-bonded synthon R 2 1(6). The acr mol­ecules and the btc mol­ecules also form slipped or offset π–π stacking inter­actions [centroid–centroid distances of 3.5212 (17) Å for btc rings and 3.703 (2) and 3.731 (2) Å for acr rings]. Together these inter­actions lead to a three-dimensional network

DNA multiplex hybridization on microarrays and thermodynamic stability in solution: a direct comparison

Hybridization intensities of 30 distinct short duplex DNAs measured on spotted microarrays, were directly compared with thermodynamic stabilities measured in solution. DNA sequences were designed to promote formation of perfect match, or hybrid duplexes containing tandem mismatches. Thermodynamic parameters ΔH°, ΔS° and ΔG° of melting transitions in solution were evaluated directly using differential scanning calorimetry. Quantitative comparison with results from 63 multiplex microarray hybridization experiments provided a linear relationship for perfect match and most mismatch duplexes. Examination of outliers suggests that both duplex length and relative position of tandem mismatches could be important factors contributing to observed deviations from linearity. A detailed comparison of measured thermodynamic parameters with those calculated using the nearest-neighbor model was performed. Analysis revealed the nearest-neighbor model generally predicts mismatch duplexes to be less stable than experimentally observed. Results also show the relative stability of a tandem mismatch is highly dependent on the identity of the flanking Watson–Crick (w/c) base pairs. Thus, specifying the stability contribution of a tandem mismatch requires consideration of the sequence identity of at least four base pair units (tandem mismatch and flanking w/c base pairs). These observations underscore the need for rigorous evaluation of thermodynamic parameters describing tandem mismatch stability

Regulation of the DNA Damage Response and Gene Expression by the Dot1L Histone Methyltransferase and the 53Bp1 Tumour Suppressor

Dot1L, a histone methyltransferase that targets histone H3 lysine 79 (H3K79), has been implicated in gene regulation and the DNA damage response although its functions in these processes remain poorly defined.Using the chicken DT40 model system, we generated cells in which the Dot1L gene is disrupted to examine the function and focal recruitment of the 53Bp1 DNA damage response protein. Detailed kinetic and dose response assays demonstrate that, despite the absence of H3K79 methylation demonstrated by mass spectrometry, 53Bp1 focal recruitment is not compromised in these cells. We also describe, for the first time, the phenotypes of a cell line lacking both Dot1L and 53Bp1. Dot1L⁻/⁻ and wild type cells are equally resistant to ionising radiation, whereas 53Bp1⁻/⁻/Dot1L⁻/⁻ cells display a striking DNA damage resistance phenotype. Dot1L and 53Bp1 also affect the expression of many genes. Loss of Dot1L activity dramatically alters the mRNA levels of over 1200 genes involved in diverse biological functions. These results, combined with the previously reported list of differentially expressed genes in mouse ES cells knocked down for Dot1L, demonstrates surprising cell type and species conservation of Dot1L-dependent gene expression. In 53Bp1⁻/⁻ cells, over 300 genes, many with functions in immune responses and apoptosis, were differentially expressed. To date, this is the first global analysis of gene expression in a 53Bp1-deficient cell line.Taken together, our results uncover a negative role for Dot1L and H3K79 methylation in the DNA damage response in the absence of 53Bp1. They also enlighten the roles of Dot1L and 53Bp1 in gene expression and the control of DNA double-strand repair pathways in the context of chromatin

Associations of autozygosity with a broad range of human phenotypes

In many species, the offspring of related parents suffer reduced reproductive success, a phenomenon known as inbreeding depression. In humans, the importance of this effect has remained unclear, partly because reproduction between close relatives is both rare and frequently associated with confounding social factors. Here, using genomic inbreeding coefficients (F-ROH) for >1.4 million individuals, we show that F-ROH is significantly associated (p <0.0005) with apparently deleterious changes in 32 out of 100 traits analysed. These changes are associated with runs of homozygosity (ROH), but not with common variant homozygosity, suggesting that genetic variants associated with inbreeding depression are predominantly rare. The effect on fertility is striking: F-ROH equivalent to the offspring of first cousins is associated with a 55% decrease [95% CI 44-66%] in the odds of having children. Finally, the effects of F-ROH are confirmed within full-sibling pairs, where the variation in F-ROH is independent of all environmental confounding.Peer reviewe

Modèle de tumeur sphéroïde multicellulaire pour l'évaluation de l'efficacité de l'administration de médicaments à base de nanoparticules

Le développement d'un modèle 3D in vitro standard pour le criblage des nanothérapies reste un défi. Plusieurs modèles de culture de tumeurs en 3D ont été développés, dont les sphéroïdes multicellulaires de tumeurs qui gagnent en popularité en raison de leur capacité à imiter certaines des caractéristiques des tumeurs naturelles. Les sphéroïdes peuvent être fabriqués de différentes manières, mais les méthodes traditionnelles présentent plusieurs inconvénients. Dans cette étude, des micropuits à base d'agarose ont été utilisés pour la génération de sphéroïdes tumoraux multicellulaires uniformes d'une lignée de cancer colorectal (HCT-116). Selon les résultats de ce projet, la pénétration des nanoparticules AGuIX®- Cy5.5 dans les sphéroïdes dépend du temps et de la concentration. Alors que les nanoparticules ont été observées dans l'espace extracellulaire et intracellulaire des sphéroïdes, la localisation intracellulaire dominante des nanoparticules AGuIX®- Cy5.5 était dans les lysosomes ; les endosomes précoces et les mitochondries, par contre, ont montré une certaine colocalisation avec AGuIX®. Lorsque la localisation des nanoparticules AGuIX®- Cy5.5 dans les sphéroïdes et la culture cellulaire 2D a été comparée, la différence dans la localisation des nanoparticules en 2D et en 3D a mis en évidence les avantages de l'utilisation de ce modèle in vitro 3D pour le criblage des nanothérapies en raison de sa capacité à récapituler les caractéristiques tumorales influençant l'internalisation des nanoparticules et leur destin intracellulaire. Le modèle in vitro 3D mis au point s'est avéré bénéfique pour l'évaluation de la thérapie associée à la radiothérapie, car les cellules en sphéroïdes (3D) ont montré une radiorésistance plus élevée que les cellules monocouches (2D), ce qui indique que ce modèle in vitro 3D peut être utilisé pour évaluer l'efficacité thérapeutique des nanothérapies dans une configuration plus réaliste. Le suivi de la croissance des sphéroïdes HCT-116 irradiés a révélé leur capacité à croître pour toutes les doses 6 jours après l'irradiation, contrairement au test classique de survie clonogénique, qui a révélé qu'ils étaient incapables de former des colonies aux doses plus élevées. Le meilleur effet radiosensibilisant d'AGuIX® a été observé à une irradiation de 2 Gy, selon l'analyse de la prolifération de cellules uniques dans les sphéroïdes.Developing a standard 3D in vitro model for screening nanotherapeutics remains challenging. Several 3D tumor culture models have been developed, multicellular tumor spheroids among them are gaining popularity due to their ability to mimic some of the characteristics of natural tumors. Spheroids can be made in a variety of ways, but traditional methods have several drawbacks. In this study, agarose-based microwells were used for generation of uniform multicellular tumor spheroids of a colorectal cancer line (HCT-116). According to the findings of this project, the penetration of AGuIX®- Cy5.5 nanoparticles within spheroids is time and concentration dependent. While nanoparticles were observed in both extracellular and intracellular space of spheroids, the dominant intracellular localization of AGuIX®- Cy5.5 nanoparticles was in lysosomes; early endosomes and mitochondria, on the other hand, showed some colocalization with AGuIX®. When the localization of AGuIX®- Cy5.5 nanoparticles in spheroids and 2D cell culture was compared, the difference in nanoparticle localization in 2D and 3D highlighted the advantages of using this 3D in vitro model for nanotherapeutics screening because of its ability to recapitulate tumor features influencing nanoparticle internalization and intracellular fate. The developed 3D in vitro model has been shown to be beneficial in therapy assessment associated with radiotherapy, as cells in spheroids (3D) showed higher radioresistance than monolayer cells(2D), indicating that this 3D in vitro model can be used to assess therapeutic efficacy of nanotherapeutics in a more realistic set up. Growth monitoring of irradiated HCT-116 spheroids revealed their ability to grow for all doses, 6 days after irradiation, in contrast to the classical clonogenic survival assay, which revealed they were unable to form colonies at higher doses greater. The best radiosensitizing effect of AGuIX® was observed at 2 Gy irradiation, according to single cell proliferation analysis in spheroids

Modèle de tumeur sphéroïde multicellulaire pour l'évaluation de l'efficacité de l'administration de médicaments à base de nanoparticules

Developing a standard 3D in vitro model for screening nanotherapeutics remains challenging. Several 3D tumor culture models have been developed, multicellular tumor spheroids among them are gaining popularity due to their ability to mimic some of the characteristics of natural tumors. Spheroids can be made in a variety of ways, but traditional methods have several drawbacks. In this study, agarose-based microwells were used for generation of uniform multicellular tumor spheroids of a colorectal cancer line (HCT-116). According to the findings of this project, the penetration of AGuIX®- Cy5.5 nanoparticles within spheroids is time and concentration dependent. While nanoparticles were observed in both extracellular and intracellular space of spheroids, the dominant intracellular localization of AGuIX®- Cy5.5 nanoparticles was in lysosomes; early endosomes and mitochondria, on the other hand, showed some colocalization with AGuIX®. When the localization of AGuIX®- Cy5.5 nanoparticles in spheroids and 2D cell culture was compared, the difference in nanoparticle localization in 2D and 3D highlighted the advantages of using this 3D in vitro model for nanotherapeutics screening because of its ability to recapitulate tumor features influencing nanoparticle internalization and intracellular fate. The developed 3D in vitro model has been shown to be beneficial in therapy assessment associated with radiotherapy, as cells in spheroids (3D) showed higher radioresistance than monolayer cells(2D), indicating that this 3D in vitro model can be used to assess therapeutic efficacy of nanotherapeutics in a more realistic set up. Growth monitoring of irradiated HCT-116 spheroids revealed their ability to grow for all doses, 6 days after irradiation, in contrast to the classical clonogenic survival assay, which revealed they were unable to form colonies at higher doses greater. The best radiosensitizing effect of AGuIX® was observed at 2 Gy irradiation, according to single cell proliferation analysis in spheroids.Le développement d'un modèle 3D in vitro standard pour le criblage des nanothérapies reste un défi. Plusieurs modèles de culture de tumeurs en 3D ont été développés, dont les sphéroïdes multicellulaires de tumeurs qui gagnent en popularité en raison de leur capacité à imiter certaines des caractéristiques des tumeurs naturelles. Les sphéroïdes peuvent être fabriqués de différentes manières, mais les méthodes traditionnelles présentent plusieurs inconvénients. Dans cette étude, des micropuits à base d'agarose ont été utilisés pour la génération de sphéroïdes tumoraux multicellulaires uniformes d'une lignée de cancer colorectal (HCT-116). Selon les résultats de ce projet, la pénétration des nanoparticules AGuIX®- Cy5.5 dans les sphéroïdes dépend du temps et de la concentration. Alors que les nanoparticules ont été observées dans l'espace extracellulaire et intracellulaire des sphéroïdes, la localisation intracellulaire dominante des nanoparticules AGuIX®- Cy5.5 était dans les lysosomes ; les endosomes précoces et les mitochondries, par contre, ont montré une certaine colocalisation avec AGuIX®. Lorsque la localisation des nanoparticules AGuIX®- Cy5.5 dans les sphéroïdes et la culture cellulaire 2D a été comparée, la différence dans la localisation des nanoparticules en 2D et en 3D a mis en évidence les avantages de l'utilisation de ce modèle in vitro 3D pour le criblage des nanothérapies en raison de sa capacité à récapituler les caractéristiques tumorales influençant l'internalisation des nanoparticules et leur destin intracellulaire. Le modèle in vitro 3D mis au point s'est avéré bénéfique pour l'évaluation de la thérapie associée à la radiothérapie, car les cellules en sphéroïdes (3D) ont montré une radiorésistance plus élevée que les cellules monocouches (2D), ce qui indique que ce modèle in vitro 3D peut être utilisé pour évaluer l'efficacité thérapeutique des nanothérapies dans une configuration plus réaliste. Le suivi de la croissance des sphéroïdes HCT-116 irradiés a révélé leur capacité à croître pour toutes les doses 6 jours après l'irradiation, contrairement au test classique de survie clonogénique, qui a révélé qu'ils étaient incapables de former des colonies aux doses plus élevées. Le meilleur effet radiosensibilisant d'AGuIX® a été observé à une irradiation de 2 Gy, selon l'analyse de la prolifération de cellules uniques dans les sphéroïdes

Clearing spheroids for 3D fluorescent microscopy: combining safe and soft chemicals with deep convolutional neural network

In life sciences, there are increasing interest in 3D culture models to better reproduce the 3D environment encountered in-vivo. Imaging of such 3D culture models is instrumental for drug discovery, but face several issues before its use becomes widespread. Extensive microscopic investigation of these 3D cell models faces the challenge of light penetration in depth in opaque biological tissues. To overcome this limit, diverse clearing techniques have emerged over the past decades. However, it is not straightforward to choose the best clearing protocols, and assess quantitatively their clearing efficiency. Focusing on spheroids, we propose a combination of fast and cost-effective clearing procedure for such medium-sized samples. A generic method with local contrast metrics and deep convolutional neural network-based segmentation of nuclei is proposed to quantify the efficiency of clearing. We challenged this method by testing the possibility to transfer segmentation knowledge from a clearing protocol to another. The later results support the pertinence of training deep learning algorithms on cleared samples to further use the segmentation pipeline on non-cleared ones. This second step of the protocol gives access to digital clearing possibilities applicable to live and high-throughput optical imaging