RESUMO
The unsymmetrical biaryls (Ar1-Ar2) produced by the catalytic cross-couplings of aryl halides (Ar1-halo) with aryl metallics (Ar2-M) in the loading ratio of 1:1 are popular in chemical synthesis. In contrast, there has been less precedence on the same biaryls produced effectively from two normal aryl C-H bonds with equivalent loading. Here, we report that, in a palladium/oxidant/acid catalytic system at room temperature, one arene (Ar1-H, 1 equiv) can highly selectively couple with the other one (Ar2-H, 1 equiv) to afford the target Ar1-Ar2 just by controlling the directing groups and the substituted groups on their phenyl rings. The utility of this one-one cross-coupling is also demonstrated by synthesis of a few bioactive molecules.
RESUMO
The preparation of secondary 2,2'-bisanilides has been successfully achieved through an oxidative coupling of aryl ortho-sp2 C-H bonds of anilides in the presence of catalytic Pd(OAc)2 and K2S2O8 as an oxidant in MsOH/CF3CO2H (TFA) at room temperature (25 °C). The aromatic rings of anilides substituted by various electron-donating or electron-withdrawing groups are tolerant in these coupling reactions.
RESUMO
The acute cellular response to stress generates a subpopulation of reversibly stress-tolerant cells under conditions that are lethal to the majority of the population. Stress tolerance is attributed to heterogeneity of gene expression within the population to ensure survival of a minority. We performed whole transcriptome sequencing analyses of metastatic human breast cancer cells subjected to the chemotherapeutic agent paclitaxel at the single-cell and population levels. Here we show that specific transcriptional programs are enacted within untreated, stressed, and drug-tolerant cell groups while generating high heterogeneity between single cells within and between groups. We further demonstrate that drug-tolerant cells contain specific RNA variants residing in genes involved in microtubule organization and stabilization, as well as cell adhesion and cell surface signaling. In addition, the gene expression profile of drug-tolerant cells is similar to that of untreated cells within a few doublings. Thus, single-cell analyses reveal the dynamics of the stress response in terms of cell-specific RNA variants driving heterogeneity, the survival of a minority population through generation of specific RNA variants, and the efficient reconversion of stress-tolerant cells back to normalcy.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias da Mama , Resistencia a Medicamentos Antineoplásicos , Paclitaxel/farmacologia , RNA Neoplásico , Análise de Sequência de RNA , Transcrição Gênica , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Humanos , RNA Neoplásico/biossíntese , RNA Neoplásico/genética , Transcrição Gênica/efeitos dos fármacos , Transcrição Gênica/genéticaRESUMO
In the title compound, [Ni(C(20)H(13)O(5)P)(C(12)H(10)N(2))(H(2)O)](n), the Ni(II) cation is coordinated by three O atoms from two 5-(diphenyl-phosphino-yl)isophthalate anions, two N atoms from two 1,2-bis-(pyridin-4-yl)ethene ligands and one water mol-ecule in a distorted octa-hedral geometry. Both 1,2-bis-(pyridin-4-yl)ethene and 5-(diphenyl-phosphino-yl)iso-phthal-ate bridge the Ni(II) cations to form polymeric layers parallel to (001). In the crystal, O-Hâ¯O hydrogen bonding links layers into a three-dimensional supra-molecular structure.
RESUMO
An 11-mer peptide taken from a subsequence of the human protein ubiquitin was synthesized. The peptide has been fully characterized by NMR spectroscopy using chemical shift analysis and by NOE measurements. The conformation was calculated using state of the art MD methods of protein chemistry. A hairpin conformation was found which is to a large part identical with the structure of this peptide fragment within the human ubiquitin. The surprising result that already an 11-mer peptide adopts a hairpin conformation in aqueous solution is discussed in terms of initials sites for protein folding.
Assuntos
Peptídeos/química , Humanos , Sequências Repetidas Invertidas , Espectroscopia de Ressonância Magnética , Estrutura Secundária de Proteína , Ubiquitina/químicaRESUMO
In the title compound, [Zn(C(9)H(4)O(6))(C(12)H(12)N(2))(H(2)O)(2)], the Zn(II) atom is five-coordinated by two N atoms from a 4,4'-dimethyl-2,2'-bipyridine ligand, one O atom from a 5-carb-oxy-benzene-1,3-dicarboxyl-ate ligand and two water mol-ecules in a distorted trigonal-bipyramidal geometry. The complex mol-ecules are linked by inter-molecular O-Hâ¯O hydrogen bonds and partly overlapping π-π inter-actions [centroid-centroid distance = 4.017â (2)â Å] into a three-dimensional supra-molecular network.
RESUMO
In the title compound, {[Zn(3)(C(9)H(3)O(6))(2)(C(12)H(12)N(2))(2)(H(2)O)(6)]·6H(2)O}(n), one Zn(II) atom, lying on an inversion center, is six-coordinated by two O atoms from two benzene-1,3,5-tricarboxyl-ate (btc) ligands and four water mol-ecules in a distorted octa-hedral geometry. The other Zn(II) atom is five-coordinated by two N atoms from a 5,5'-dimethyl-2,2'-bipyridine (dmbpy) ligand, two O atoms from two btc ligands and one water mol-ecule in a distorted trigonal-bipyramidal geometry. The compound features a one-dimensional ladder structure, with windows of ca 10.245â (1) × 15.446â (2)â Å. The ladders are linked together by inter-molecular O-Hâ¯O hydrogen bonds and π-π inter-actions between the benzene rings and between the pyridine rings [centroid-to-centroid distances 3.858â (2) and 3.911â (3)â Å, respectively] to form a three-dimensional supra-molecular structure. One of the lattice water molecules is disordered over two positions in a 0.592:0.408 ratio.
RESUMO
In the title compound, [Ni(C(9)H(4)O(6))(C(12)H(12)N(2))(H(2)O)(2)]·7H(2)O, the Ni(II) atom is six-coordinated by two O atoms from a chelating carboxyl-ate group of a 5-carb-oxy-benzene-1,3-dicarboxyl-ate ligand, two O atoms of two water mol-ecules and two N atoms from a 6,6'-dimethyl-2,2'-bipyridine ligand in a distorted octa-hedral geometry. The compound exhibits a three-dimensional supra-molecular structure composed of the complex mol-ecules and lattice water mol-ecules, which are linked together by inter-molecular O-Hâ¯O hydrogen bonds and partly overlapping π-π inter-actions between the pyridine and benzene rings [centroid-centroid distances = 3.922â (2) and 3.921â (2)â Å]. One of the lattice water mol-ecules is disordered over two positions in an occupancy ratio of 0.521â (6):0.479â (6).
RESUMO
In the crystal structure of the title compound, [MnCl(2)(C(14)H(14)N(4))(2)](n), the Mn(II) atom, lying on an inversion center, is coordinated by four N atoms from four 1,4-bis-(imidazol-1-ylmeth-yl)benzene (bimb) ligands and two Cl(-) anions in a distorted octa-hedral geometry. The bimb ligands bridge the Mn(II) atoms, forming a two-dimensional polymeric complex, which is composed of a 52-membered [Mn(4)(bimb)(4)] ring with distances of 7.7812â (2) and 27.4731â (9)â Å between opposite metal atoms. Weak C-Hâ¯π inter-actions are present in the crystal structure.
RESUMO
In the title compound, [Cu(C(14)H(8)O(5))(C(12)H(12)N(2))(H(2)O)](n), the Cu(II) ion is penta-coordinated in a square-pyramidal geometry. Two N atoms of the chelating 5,5'-dimethyl-2,2'-bipyridine (dbp) ligand and two O atoms of two different 2,2'-oxydibenzoic (odb) ligands occupy the basal plane while the water O atom completes the square-pyramidal geometry at the apical site. The non-water N(2)O(2) donor atoms are nearly coplanar, with a mean deviation from the least-squares plane of 0.0518â (11)â Å and the Cu atom is displaced by 0.1507â (11)â Å from this plane towards the apical water O atom. Further coordination via the 2,2'-oxydibenzoate anions forms a one-dimensional coordination polymer extending parallel to [010]. In the crystal structure, O-Hâ¯O hydrogen bonds link the mol-ecules into a two-dimensional supra-molecular structure.
RESUMO
In the title salt, (C(13)H(10)N(3)O(2))[Ni(C(3)S(5))(2)], the Ni(III) cation is S,S'-chelated by two 2-thioxo-1,3-dithiole-4,5-dithiol-ate anions in a distorted square-planar geometry. The complex anion is approximately planar with a maximum deviation of 0.097â (1)â Å. In the 1-(4-nitro-benz-yl)-4-cyano-pyridinium cation, the pyridine ring is twisted at a dihedral angle of 73.84â (16)° with respect to the benzene ring. π-π stacking is observed between nearly parallel [dihedral angle = 4.71â (7)°] dithiole and benzene rings, the centroid-centroid distance being 3.791â (2)â Å.
RESUMO
Here we report whole genome sequencing of four individuals (H3, H4, H5, and H6) from a family of Pakistani descent. Whole genome sequencing yielded 1084.92, 894.73, 1068.62, and 1005.77 million mapped reads corresponding to 162.73, 134.21, 160.29, and 150.86 Gb sequence data and 52.49x, 43.29x, 51.70x, and 48.66x average coverage for H3, H4, H5, and H6, respectively. We identified 3,529,659, 3,478,495, 3,407,895, and 3,426,862 variants in the genomes of H3, H4, H5, and H6, respectively, including 1,668,024 variants common in the four genomes. Further, we identified 42,422, 39,824, 28,599, and 35,206 novel variants in the genomes of H3, H4, H5, and H6, respectively. A major fraction of the variants identified in the four genomes reside within the intergenic regions of the genome. Single nucleotide polymorphism (SNP) genotype based comparative analysis with ethnic populations of 1000 Genomes database linked the ancestry of all four genomes with the South Asian populations, which was further supported by mitochondria based haplogroup analysis. In conclusion, we report whole genome sequencing of four individuals of Pakistani descent.
Assuntos
Genoma Humano , Sequenciamento Completo do Genoma , Humanos , Paquistão , Polimorfismo de Nucleotídeo ÚnicoRESUMO
OBJECTIVE: To research the chemical constituents from Daphne tangutica. METHODS: Column chromatography with silica gel, ODS-C18, Sephadex LH-20 and re-crystallization were employed to isolate and purify the constituents. According to physical and chemical properties and spectral data to identify the structure of compounds. RESULTS: Eight compounds were isolated and identified as palmitic acid (1), laurostearic acid(2), beta-sitosterol(3), 7-methoxy-8-hydroxycumarin(4), daphnetin(5), genkwanin(6), hydroxygenkwanin(7), p-hydroxybenzoic acid(8). CONCLUSION: Compounds 1,2,7 and 8 are isolated from Daphne tangutica for the first time.
Assuntos
Daphne/química , Flavonoides/isolamento & purificação , Ácido Palmítico/isolamento & purificação , Parabenos/isolamento & purificação , Plantas Medicinais/química , Flavonas/química , Flavonas/isolamento & purificação , Flavonoides/química , Espectroscopia de Ressonância Magnética , Ácido Palmítico/química , Parabenos/química , Casca de Planta/química , Sitosteroides/química , Sitosteroides/isolamento & purificaçãoRESUMO
We previously investigated the transcriptome and proteome profiles of the murine ocular lens at six developmental time points including two embryonic (E15 and E18) and four postnatal time points (P0, P3, P6, and P9). Here, we extend our analyses to identify novel transcripts and peptides in developing mouse lens. We identified a total of 9,707 novel transcripts and 325 novel fusion genes in developing mouse lens. Additionally, we identified 13,281 novel alternative splicing (AS) events in mouse lens including 6,990 exon skipping (ES), 2,447 alternative 3' splice site (A3SS), 1,900 alternative 5' splice site (A5SS), 1,771 mutually exclusive exons (MXE), and 173 intron retention (IR). Finally, we integrated our OMIC (Transcriptome and Proteome) datasets identifying 20 novel peptides in mouse lens. All 20 peptides were validated through matching MS/MS spectra of synthetic peptides. To the best of our knowledge, this is the first report integrating OMIC datasets to identify novel peptides in developing murine lens.
Assuntos
Processamento Alternativo/genética , Cristalino/embriologia , Cristalino/crescimento & desenvolvimento , Organogênese/genética , Peptídeos/genética , Proteoma/genética , Transcriptoma/genética , Algoritmos , Animais , Cromatografia Líquida , Bases de Dados Genéticas , Éxons/genética , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Íntrons/genética , Camundongos , Gravidez , Sítios de Splice de RNA/genética , Análise de Sequência de RNA , Espectrometria de Massas em TandemRESUMO
UNLABELLED: The intracellular protozoan Toxoplasma gondii dramatically reprograms the transcriptome of host cells it infects, including substantially up-regulating the host oncogene c-myc. By applying a flow cytometry-based selection to infected mouse cells expressing green fluorescent protein fused to c-Myc (c-Myc-GFP), we isolated mutant tachyzoites defective in this host c-Myc up-regulation. Whole-genome sequencing of three such mutants led to the identification of MYR1 (Myc regulation 1; TGGT1_254470) as essential for c-Myc induction. MYR1 is a secreted protein that requires TgASP5 to be cleaved into two stable portions, both of which are ultimately found within the parasitophorous vacuole and at the parasitophorous vacuole membrane. Deletion of MYR1 revealed that in addition to its requirement for c-Myc up-regulation, the MYR1 protein is needed for the ability of Toxoplasma tachyzoites to modulate several other important host pathways, including those mediated by the dense granule effectors GRA16 and GRA24. This result, combined with its location at the parasitophorous vacuole membrane, suggested that MYR1 might be a component of the machinery that translocates Toxoplasma effectors from the parasitophorous vacuole into the host cytosol. Support for this possibility was obtained by showing that transit of GRA24 to the host nucleus is indeed MYR1-dependent. As predicted by this pleiotropic phenotype, parasites deficient in MYR1 were found to be severely attenuated in a mouse model of infection. We conclude, therefore, that MYR1 is a novel protein that plays a critical role in how Toxoplasma delivers effector proteins to the infected host cell and that this is crucial to virulence. IMPORTANCE: Toxoplasma gondii is an important human pathogen and a model for the study of intracellular parasitism. Infection of the host cell with Toxoplasma tachyzoites involves the introduction of protein effectors, including many that are initially secreted into the parasitophorous vacuole but must ultimately translocate to the host cell cytosol to function. The work reported here identified a novel protein that is required for this translocation. These results give new insight into a very unusual cell biology process as well as providing a potential handle on a pathway that is necessary for virulence and, therefore, a new potential target for chemotherapy.
Assuntos
Interações Hospedeiro-Patógeno , Proteínas de Protozoários/metabolismo , Toxoplasma/fisiologia , Fatores de Virulência/metabolismo , Animais , Deleção de Genes , Macrófagos/parasitologia , Camundongos , Proteínas de Protozoários/genética , Toxoplasmose Animal/parasitologia , Toxoplasmose Animal/patologia , Fatores de Virulência/genéticaRESUMO
FOXE3 is a lens-specific transcription factor that has been associated with anterior segment ocular dysgenesis. To determine the transcriptional target(s) of FOXE3 that are indispensable for the anterior segment development, we examined the transcriptome and the proteome of cells expressing truncated FOXE3 responsible for Peters anomaly identified through linkage-coupled next-generation whole-exome sequencing. We found that DNAJB1, an autophagy-associated protein, was the only candidate exhibiting differential expression in both screens. We confirmed the candidacy of DNAJB1 through chromatin immunoprecipitation and luciferase assays while knockdown of DNAJB1 in human lens epithelial cells resulted in a mitotic arrest. Subsequently, we targeted dnajb1a in zebrafish through injection of a splice-blocking morpholino. The dnajb1a morphants exhibited underdeveloped cataractous lenses with persistent apoptotic nuclei. In conclusion, here we report DNAJB1 is a transcriptional target of FOXE3 in a novel pathway that is crucial for the development of the anterior segment of the eye.
Assuntos
Autofagia/genética , Opacidade da Córnea/genética , Anormalidades do Olho/genética , Fatores de Transcrição Forkhead/genética , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP40/genética , Animais , Opacidade da Córnea/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Anormalidades do Olho/metabolismo , Saúde da Família , Feminino , Fatores de Transcrição Forkhead/metabolismo , Perfilação da Expressão Gênica/métodos , Técnicas de Silenciamento de Genes , Células HEK293 , Proteínas de Choque Térmico HSP40/metabolismo , Humanos , Cristalino/patologia , Masculino , Linhagem , Sequenciamento do Exoma/métodos , Peixe-Zebra/genética , Peixe-Zebra/metabolismoRESUMO
PURPOSE: Transcriptome is the entire repertoire of transcripts present in a cell at any particular time. We undertook a next-generation whole transcriptome sequencing approach to gain insight into the transcriptional landscape of the developing mouse lens. METHODS: We ascertained mouse lenses at six developmental time points including two embryonic (E15 and E18) and four postnatal stages (P0, P3, P6, and P9). The ocular tissue at each time point was maintained as two distinct pools serving as biological replicates for each developmental stage. The mRNA and small RNA libraries were paired-end sequenced on Illumina HiSeq 2000 and subsequently analyzed using bioinformatics tools. RESULTS: Mapping of mRNA and small RNA libraries generated 187.56 and 154.22 million paired-end reads, respectively. We detected a total of 14,465 genes in the mouse ocular lens at the above-mentioned six developmental stages. Of these, 46 genes exhibited a 40-fold differential (higher or lower) expression at one the five developmental stages (E18, P0, P3, P6, and P9) compared with their expression level at E15. Likewise, small RNA profiling identified 379 microRNAs (miRNAs) expressed in mouse lens at six developmental time points. Of these, 49 miRNAs manifested an 8-fold differential (higher or lower) expression at one the five developmental stages, as mentioned above compared with their expression level at E15. CONCLUSIONS: We report a comprehensive profile of developing murine lens transcriptome including both mRNA and miRNA through next-generation RNA sequencing. A complete repository of the lens transcriptome of six developmental time points will be monumental in elucidating processes essential for the development of the ocular lens and maintenance of its transparency.
Assuntos
Catarata/genética , Biologia Computacional/métodos , Perfilação da Expressão Gênica/métodos , Cristalino/metabolismo , Organogênese/genética , Prenhez , RNA Mensageiro/genética , Animais , Animais Recém-Nascidos , Catarata/metabolismo , Catarata/patologia , Modelos Animais de Doenças , Feminino , Biblioteca Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Cristalino/patologia , Camundongos , Gravidez , RNA Mensageiro/metabolismoRESUMO
A new class of organic photovoltaic materials, poly(rod-coil) polymers composed of alternatively definite conjugated and non-conjugated segments, have been proposed. The first five examples based on polyurethane chemistry showed photovoltaic performance surpassing the reference compound, but less dependent on their molecular weight.
RESUMO
The DinB (PolIV) protein of Escherichia coli participates in several cellular functions. We investigated a dinB mutation, Δ(dinB-yafN)883(::kan) [referred to as ΔdinB883], which strongly sensitized E. coli cells to both UV- and X-radiation killing. Earlier reports indicated dinB mutations had no obvious effect on UV radiation sensitivity which we confirmed by showing that normal UV radiation sensitivity is conferred by the ΔdinB749 allele. Compared to a wild-type strain, the ΔdinB883 mutant was most sensitive (160-fold) in early to mid-logarithmic growth phase and much less sensitive (twofold) in late log or stationary phases, thus showing a growth phase-dependence for UV radiation sensitivity. This sensitizing effect of ΔdinB883 is assumed to be completely dependent upon the presence of UmuDC protein; since the ΔdinB883 mutation did not sensitize the ΔumuDC strain to UV radiation killing throughout log phase and early stationary phase growth. The DNA damage checkpoint activity of UmuDC was clearly affected by ΔdinB883 as shown by testing a umuC104 ΔdinB883 double-mutant. The sensitivities of the ΔumuDC strain and the ΔdinB883 ΔumuDC double-mutant strain were significantly greater than for the ΔdinB883 strain, suggesting that the ΔdinB883 allele only partially suppresses UmuDC activity. The ΔdinB883 mutation partially sensitized (fivefold) uvrA and uvrB strains to UV radiation, but did not sensitize a ΔrecA strain. A comparison of the DNA sequences of the ΔdinB883 allele with the sequences of the Δ(dinB-yafN)882(::kan) and ΔdinB749 alleles, which do not sensitize cells to UV radiation, revealed ΔdinB883 is likely a "gain-of-function" mutation. The ΔdinB883 allele encodes the first 54 amino acids of wild-type DinB followed by 29 predicted residues resulting from the continuation of the dinB reading frame into an adjacent insertion fragment. The resulting polypeptide is proposed to interfere directly or indirectly with UmuDC function(s) involved in protecting cells against the lethal effects of radiation.
Assuntos
Proteínas de Escherichia coli/genética , Escherichia coli/metabolismo , Viabilidade Microbiana , Mutação , Raios Ultravioleta , Adenosina Trifosfatases/genética , Adenosina Trifosfatases/metabolismo , Alelos , DNA Helicases/genética , DNA Helicases/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , DNA Polimerase Dirigida por DNA/genética , DNA Polimerase Dirigida por DNA/metabolismo , Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Viabilidade Microbiana/genética , Viabilidade Microbiana/efeitos da radiação , Raios XRESUMO
To evaluate the relationship between atherosclerosis and hemodynamic of coronary artery in mice detecting by ultrasound bio‐microscopy flow imaging . Methods Double 14 20‐week‐old LDL‐R‐/‐and C57BL/6 male mice were selected ,and randomly divided into two groups in each genotype according to weight . Each two groups were fed to 28 weeks or 36 weeks age respectively with west diet . Coronary artery hemodynamics in these mice were assessed in vivo by Vevo ?2100 ultrasound imaging system ,then the intima‐media thickness( IM T ) of aorta in histopathology were analyzed . T he differences of coronary artery hemodynamic parameters such as maximum velocity ( Vmax ) ,mean velocity ( Vmean) and velocity time integral ( V T I) were compared between mice of different genotypes of the same week and mice of different weeks of the same genotype . And the relationship between coronary artery hemodynamic in ultrasound and aortic IM T in histopathology were analyzed . Results ① All coronary hemodynamic parameters in LDL‐R‐/‐ mice were significantly lower than those of wild‐type mice except the Vmax between two 28‐week‐old genotypes group at the same weeks of age of different genotypes ( all P <0 .05) . But there was no significant difference in coronary artery hemodynamic parameters between mice of the same genotype at different weeks of age( P >0 .05) . ②T he histopathological measurements of aortic IM T in LDL‐R‐/‐mice were significantly higher than those of wild type mice ( all P < 0 .05 ) ,and those of 36‐week‐old mice were significantly higher than those of 28‐week‐old mice ( all P < 0 .05 ) . ③ All coronary hemodynamic parameters such as Vmax ,Vmean and V TI were negatively correlated with pathological measurements of aortic IM T ( r = -0 .532 , -0 .423 , -0 .524 ; all P < 0 .05 ) . Conclusions The parameters of coronary artery hemodynamics obtained by ultrasound bio‐microscopy are well correlated with the pathological results of atherosclerosis . Ultrasound bio‐microscopic flow imaging can be used as a new method to evaluate the degree of atherosclerosis in mice by detecting the hemodynamic parameters of coronary artery .