Monitoring early responses to neoadjuvant chemotherapy and the factors affecting neoadjuvant chemotherapy responses in primary osteosarcoma.

Background: This study sought to predict the early responses to neoadjuvant chemotherapy (NACT) of patients with primary conventional osteosarcoma (COS) using the apparent diffusion coefficient (ADC) and to evaluate the factors affecting the tumor necrosis rate (TNR). Methods: The data of 41 patients who underwent magnetic resonance imaging (MRI) and diffusion-weighted imaging sequence scans before NACT, 5 days after the end of the first phase of NACT, after the end of the whole course of chemotherapy, were prospectively collected. ADC1 refers to the ADC before chemotherapy, ADC2 refers to the ADC after the first phase of chemotherapy, and ADC3 refers to the ADC before surgery. The change in values before and after the first phase of chemotherapy was calculated as follows: ADC2-1 = ADC2 - ADC1. The change in values before and after the last phase of chemotherapy was calculated as follows: ADC3-1 = ADC3 - ADC1. The change in values after the first phase and the last phase of chemotherapy was calculated as follows: ADC3-2 = ADC3 - ADC2. We recorded the patient characteristics, including age, gender, pulmonary metastasis, alkaline phosphatase (ALP), and lactate dehydrogenase (LDH). The patients were divided into the following 2 groups based on their histological TNR after postoperative: (I) the good-response group (≥90% necrosis, n=13) and (II) the poor-response group (<90% necrosis, n=28). Changes in the ADCs were compared between the good-response and poor-response groups. The different ADCs between the 2 groups were compared, and a receiver operating characteristic analysis was performed. A correlation analysis was performed to assess the correlations of the clinical features, laboratory features, and different ADCs with patients' histopathological responses to NACT. Results: The ADC2 (P<0.001), ADC3 (P=0.004), ADC3-1 (P=0.008), ADC3-2 (P=0.047), and ALP before NACT (P=0.019) were significantly higher in the good-response group than in the poor-response group. The ADC2 [area under the curve (AUC) =0.723; P=0.023], ADC3 (AUC =0.747; P=0.012), and ADC3-1 (AUC =0.761; P=0.008) showed good diagnostic performance. Based on the univariate binary logistic regression analysis, the ADC2 (P=0.022), ADC3 (P=0.009), ADC2-1 (P=0.041), and ADC3-1 (P=0.014) were correlated with the TNR. However, based on the multivariate analysis, these parameters were not significantly correlated with the TNR. Conclusions: In patients with COS who are undergoing neoadjuvant chemotherapy, the ADC2 is a promisingindicator for predicting tumor response to chemotherapy in early.

Chemical inhibition of Arabidopsis PIN-FORMED auxin transporters by the anti-inflammatory drug naproxen.

The phytohormone auxin plays central roles in many growth and developmental processes in plants. Development of chemical tools targeting the auxin pathway is useful for both plant biology and agriculture. Here we reveal that naproxen, a synthetic compound with anti-inflammatory activity in humans, acts as an auxin transport inhibitor targeting PIN-FORMED (PIN) transporters in plants. Physiological experiments indicate that exogenous naproxen treatment affects pleiotropic auxin-regulated developmental processes. Additional cellular and biochemical evidence indicates that naproxen suppresses auxin transport, specifically PIN-mediated auxin efflux. Moreover, biochemical and structural analyses confirm that naproxen binds directly to PIN1 protein via the same binding cavity as the indole-3-acetic acid substrate. Thus, by combining cellular, biochemical, and structural approaches, this study clearly establishes that naproxen is a PIN inhibitor and elucidates the underlying mechanisms. Further use of this compound may advance our understanding of the molecular mechanisms of PIN-mediated auxin transport and expand our toolkit in auxin biology and agriculture.

The functional evolution of architecturally different plant geranyl diphosphate synthases from geranylgeranyl diphosphate synthase.

Terpenoids constitute the largest class of plant primary and secondary metabolites with a broad range of biological and ecological functions. They are synthesized from isopentenyl diphosphate and dimethylallyl diphosphate, which in plastids are condensed by geranylgeranyl diphosphate synthases (GGPPSs) to produce GGPP (C20) for diterpene biosynthesis and by geranyl diphosphate synthases (GPPSs) to form GPP (C10) for monoterpene production. Depending on the plant species, unlike homomeric GGPPSs, GPPSs exist as homo- and heteromers, the latter of which contain catalytically inactive GGPPS-homologous small subunits (SSUs) that can interact with GGPPSs. By combining phylogenetic analysis with functional characterization of GGPPS homologs from a wide range of photosynthetic organisms, we investigated how different GPPS architectures have evolved within the GGPPS protein family. Our results reveal that GGPPS gene family expansion and functional divergence began early in nonvascular plants, and that independent parallel evolutionary processes gave rise to homomeric and heteromeric GPPSs. By site-directed mutagenesis and molecular dynamics simulations, we also discovered that Leu-Val/Val-Ala pairs of amino acid residues were pivotal in the functional divergence of homomeric GPPSs and GGPPSs. Overall, our study elucidated an evolutionary path for the formation of GPPSs with different architectures from GGPPSs and uncovered the molecular mechanisms involved in this differentiation.

The DnaJ-like Zinc Finger Protein ORANGE Promotes Proline Biosynthesis in Drought-Stressed Arabidopsis Seedlings.

Orange (OR) is a DnaJ-like zinc finger protein with both nuclear and plastidial localizations. OR, and its orthologs, are highly conserved in flowering plants, sharing a characteristic C-terminal tandem 4× repeats of the CxxCxxxG signature. It was reported to trigger chromoplast biogenesis, promote carotenoid accumulation in plastids of non-pigmented tissues, and repress chlorophyll biosynthesis and chloroplast biogenesis in the nucleus of de-etiolating cotyledons cells. Its ectopic overexpression was found to enhance plant resistance to abiotic stresses. Here, we report that the expression of OR in Arabidopsis thaliana was upregulated by drought treatment, and seedlings of the OR-overexpressing (OE) lines showed improved growth performance and survival rate under drought stress. Compared with the wild-type (WT) and OR-silencing (or) lines, drought-stressed OE seedlings possessed lower contents of reactive oxygen species (such as H2O2 and O2-), higher activities of both superoxide dismutase and catalase, and a higher level of proline content. Our enzymatic assay revealed a relatively higher activity of Δ1-pyrroline-5-carboxylate synthase (P5CS), a rate-limiting enzyme for proline biosynthesis, in drought-stressed OE seedlings, compared with the WT and or lines. We further demonstrated that the P5CS activity could be enhanced by supplementing exogenous OR in our in vitro assays. Taken together, our results indicated a novel contribution of OR to drought tolerance, through its impact on proline biosynthesis.

Gapless indica rice genome reveals synergistic contributions of active transposable elements and segmental duplications to rice genome evolution.

The ultimate goal of genome assembly is a high-accuracy gapless genome. Here, we report a new assembly pipeline that is used to produce a gapless genome for the indica rice cultivar Minghui 63. The resulting 397.71-Mb final assembly is composed of 12 contigs with a contig N50 size of 31.93 Mb. Each chromosome is represented by a single contig and the genomic sequences of all chromosomes are gapless. Quality evaluation of this gapless genome assembly showed that gene regions in our assembly have the highest completeness compared with the other 15 reported high-quality rice genomes. Further comparison with the japonica rice genome revealed that the gapless indica genome assembly contains more transposable elements (TEs) and segmental duplications (SDs), the latter of which produce many duplicated genes that can affect agronomic traits through dose effect or sub-/neo-functionalization. The insertion of TEs can also affect the expression of duplicated genes, which may drive the evolution of these genes. Furthermore, we found the expansion of nucleotide-binding site with leucine-rich repeat disease-resistance genes and cis-zeatin-O-glucosyltransferase growth-related genes in SDs in the gapless indica genome assembly, suggesting that SDs contribute to the adaptive evolution of rice disease resistance and developmental processes. Collectively, our findings suggest that active TEs and SDs synergistically contribute to rice genome evolution.

A novel metal organic gel with superior oxidase-like activity for efficient and sensitive chemiluminescence detection of uric acid.

It is found that MIL-100(Fe) gels, as a kind of metal-organic gels (MOGs), constitutting of iron (Fe3+) and trimesic acid (H3BTC), has been regarded as the efficient catalyst of luminol chemiluminescence (CL) system without the presence of extra oxidants in the present work. MIL-100(Fe) gels that have possessed mimicking oxidase-like activity can excellently enhanced luminol CL intensity by accelerating the generation of reactive oxygen species. Furthermore, with the addition of uric acid (UA), the CL signal has been dramatically inhibited under alkaline condition. Hence, the CL intensity inhibiting ratio (I0/IS) was proportional to the increasing concentration of UA in the rang from 10 nM to 4000 nM with the detection limit of 5.9 nM. This method has been successfully applied for analysis of UA with acceptable recoveries ranging from 97.0% to 107.9% in urine sample. These results indicates that this study open up a novel, sensitive and convenient method to detect UA in biological samples.

At3g53630 encodes a GUN1-interacting protein under norflurazon treatment.

Chloroplasts are semi-autonomous organelles, with more than 95% of their proteins encoded by the nuclear genome. The chloroplast-to-nucleus retrograde signals are critical for the nucleus to coordinate its gene expression for optimizing or repairing chloroplast functions in response to changing environments. In chloroplasts, the pentatricopeptide-repeat protein GENOMES UNCOUPLED 1 (GUN1) is a master switch that senses aberrant physiological states, such as the photooxidative stress induced by norflurazon (NF) treatment, and represses the expression of photosynthesis-associated nuclear genes (PhANGs). However, it is largely unknown how the retrograde signal is transmitted beyond GUN1. In this study, a protein GUN1-INTERACTING PROTEIN 1 (GIP1), encoded by At3g53630, was identified to interact with GUN1 by different approaches. We demonstrated that GIP1 has both cytosol and chloroplast localizations, and its abundance in chloroplasts is enhanced by NF treatment with the presence of GUN1. Our results suggest that GIP1 and GUN1 may function antagonistically in the retrograde signaling pathway.

The Nuclear Localization of the DnaJ-Like Zinc Finger Domain-Containing Protein EDA3 Affects Seed Development in Arabidopsis thaliana.

The DnaJ-like zinc finger domain-containing proteins are involved in different aspects of plastid function and development. Some of these proteins were recently reported to have dual subcellular localization in the nucleus and plastids. One member of this family, PSA2 (AT2G34860), was found to localize to the thylakoid lumen and regulate the assembly of photosystem I (PSI). However, PSA2 was also annotated as Embryo sac Development Arrest 3 (EDA3) from the observation that its embryo sac development was arrested at the two-nuclear stage. In this study, we characterized the eda3 mutant, and demonstrated that, as compared with the wild-type (WT) plants, the mutant has shorter siliques, fewer siliques per plant, and fewer seeds per silique. Both aborted and undeveloped ovules were observed in siliques of the mutant. By immunoblot analysis, we found that, different from the chloroplast localization in mature leaves, EDA3 localizes in the nucleus in seeds. A nuclear localization signal was identified from the deduced amino acid sequence of EDA3, and also proved to be sufficient for directing its fusion peptide into the nucleus.

ORANGE Represses Chloroplast Biogenesis in Etiolated Arabidopsis Cotyledons via Interaction with TCP14.

The conversion of etioplasts into chloroplasts in germinating cotyledons is a crucial transition for higher plants, enabling photoautotrophic growth upon illumination. Tight coordination of chlorophyll biosynthesis and photosynthetic complex assembly is critical for this process. ORANGE (OR), a DnaJ-like zinc finger domain-containing protein, was reported to trigger the biogenesis of carotenoid-accumulating plastids by promoting carotenoid biosynthesis and sequestration. Both nuclear and plastidic localizations of OR have been observed. Here, we show that Arabidopsis (Arabidopsis thaliana) OR physically interacts with the transcription factor TCP14 in the nucleus and represses its transactivation activity. Through this interaction, the nucleus-localized OR negatively regulates expression of EARLY LIGHT-INDUCIBLE PROTEINS (ELIPs), reduces chlorophyll biosynthesis, and delays development of thylakoid membranes in the plastids of germinating cotyledons. Nuclear abundance of OR decreased upon illumination. Together with an accumulation of TCP14 in the nucleus, this derepresses chloroplast biogenesis during de-etiolation. TCP14 is epistatic to OR and expression of ELIPs is directly regulated by the binding of TCP14 to Up1 elements in the ELIP promoter regions. Our results demonstrate that the interaction between OR and TCP14 in the nucleus leads to repression of chloroplast biogenesis in etiolated seedlings and provide new insights into the regulation of early chloroplast development.plantcell;31/12/2996/FX1F1fx1.

Ratiometric method based on silicon nanodots and Eu3+ system for highly-sensitive detection of tetracyclines.

A silicon nanodots (SiNDs)-based ratiometric fluorescent sensor was designed to establish highly sensitive and selective method for detection of tetracyclines (TCs). The sensor comprises the blue fluorescence SiNDs were prepared by hydrothermal method synthesis from (3-aminopropyl)triethoxysilane and low-cost diethylenetriaminepentaacetic acid as the precursor, which serve as co-ligands for covalently bond with europium ions (Eu3+), and also the internal reference signal. In the presence of TCs, the luminescence intensity of Eu3+ was significantly increased, while the luminescence intensity of SiNDs was reduced. The fluorescence intensity ratio of I616/I450 of the sensor exhibits a sensitive linear range from 0.2 to 20 µM with a detection limit of 3 nM for the detection of TCs. This method was successfully applied to determine the levels of TCs in nature river samples and milk samples with good recoveries ranges from 92.2% to 107.4% and relative standard deviations (RSD, n = 3) was below 5%, providing a promising application future in real samples.