Full-length EFOP3 and EFOP4 proteins are essential for pollen intine development in Arabidopsis thaliana.

Pollen development is critical to plant reproduction, but the underlying regulatory molecular mechanisms have not been fully elucidated. The Arabidopsis (Arabidopsis thaliana) EFR3 OF PLANT 3 (EFOP3) and EFR3 OF PLANT 4 (EFOP4) genes encode members of the Armadillo (ARM) repeat superfamily that play key roles in pollen development. Herein, we demonstrate that EFOP3 and EFOP4 are co-expressed in pollen at anther stages 10-12, but loss-of-function of both EFOP3 and EFOP4 leads to male gametophyte sterility, irregular intine, and shriveled pollen grains at anther stage 12. We further established that full-length EFOP3 and EFOP4 specifically localize to the plasma membrane, and the integrity of these proteins is essential for pollen development. We observed uneven intine, less organized cellulose and reduced pectin content in mutant pollen compared with the wild-type. These, together with the misexpression of several genes related to cell wall metabolism in efop3-/- efop4+/- mutants, suggest that EFOP3 and EFOP4 may indirectly regulate the expression of these genes to affect intine formation, thus controlling Arabidopsis pollen fertility in a functionally redundant manner. Moreover, transcriptome analysis showed that the absence of EFOP3 and EFOP4 function affects multiple pollen development pathways. These results enhance our understanding of EFOPs proteins and their role in pollen development.

Sequencing and Genomic Analysis of Sorghum DNA Introgression Variant Line R21 and Recipient Rice Jin Hui 1 Revealed Repetitive Element Variation.

Transferring the genome of distant species to crops is an efficient way to create new germplasms. However, the molecular mechanisms involved are unclear. In this study, a new rice restorer line R21 with heat tolerance was created by introgressing the genomic DNA of sorghum into the recipient restorer line Jin Hui 1. Assembly of rice R21 and Jin Hui 1 genomes was performed using PacBio sequencing technology. Comparative genome analysis and coverage statistics showed that the repetitive sequence atr0026 was a candidate introgression fragment of sorghum DNA. Sequence similarity analysis revealed that atr0026 was distributed at different copy numbers on the telomeric position of chromosomes 9 or 10 in R21, Jin Hui 1, and several rice varieties, indicating that the repetitive sequence from sorghum was highly conserved in rice. The repeat annotation in Gramineae indicated that ribosomal DNA loci that existed in atr0026 may be cause a rearrangement of chromosomes 9 and 10 of the R21 genome, resulting in a copy number variation at the 5' end of it. Our study lays the foundation for further elucidation of the molecular mechanisms underlying the heat tolerance of sorghum DNA introgression variant line R21, which is of great significance for guiding crop genetic breeding.

VIRESCENT-ALBINO LEAF 1 regulates leaf colour development and cell division in rice.

The de novo synthesis of purine nucleotides is crucial to all living organisms, but limited information is available for plants. In this study, we isolated a virescent-albino leaf 1 (val1) mutant of rice (Oryza sativa) that produces dynamic green-revertible albino and narrow-leaf phenotypes. In albino leaves, chloroplast development was defective, pigment contents were reduced, and cell division was impaired compared with the wild-type. Map-based cloning revealed that VAL1 encodes a phosphoribosylamine-glycine ligase (PurD), the second enzyme in the de novo purine biosynthesis pathway. Subcellular localization analysis demonstrated that VAL1 was localized in the chloroplast. Our results demonstrate that VAL1 is a crucial enzyme in the de novo purine biosynthesis pathway and is involved in regulating chloroplast development, chlorophyll metabolism, and cell division during leaf development in rice.

Adaptor proteins GIR1 and GIR2. I. Interaction with the repressor GLABRA2 and regulation of root hair development.

Plants use specialized root outgrowths, termed root hairs, to enhance acquisition of nutrients and water, help secure anchorage, and facilitate interactions with soil microbiome. One of the major regulators of this process is GLABRA2 (GL2), a transcriptional repressor of root hair differentiation. However, regulation of the GL2-function is relatively well characterized, it remains completely unknown whether GL2 itself functions in complex with other transcriptional regulators. We identified GIR1 and GIR2, a plant-specific two-member family of closely related proteins that interact with GL2. Loss-of-function mutants of GIR1 and GIR2 enhanced development of root hair whereas gain-of-function mutants repressed it. Thus, GIR1 and GIR2 might function as adaptor proteins that associate with GL2 and participate in control of root hair formation.

Adaptor proteins GIR1 and GIR2. II. Interaction with the co-repressor TOPLESS and promotion of histone deacetylation of target chromatin.

Understanding how root hair development is controlled is important for understanding of many fundamental aspects of plant biology. Previously, we identified two plant-specific adaptor proteins GIR1 and GIR2 that interact with the major regulator of root hair development GL2 and suppress formation of root hair. Here, we show that GIR1 and GIR2 also interact with the co-repressor TOPLESS (TPL). This interaction required the GIR1 protein EAR motif, and was essential for the transcriptional repressor activity of GIR1. Both GIR1 and GIR2 promoted histone hypoacetylation of their target chromatin. Potentially, GIR1 and GIR2 might may link TPL to and participate in epigenetic regulation of root hair development.

CFL1, a WW domain protein, regulates cuticle development by modulating the function of HDG1, a class IV homeodomain transcription factor, in rice and Arabidopsis.

Plants have a chemically heterogeneous lipophilic layer, the cuticle, which protects them from biotic and abiotic stresses. The mechanisms that regulate cuticle development are poorly understood. We identified a rice (Oryza sativa) dominant curly leaf mutant, curly flag leaf1 (cfl1), and cloned CFL1, which encodes a WW domain protein. We overexpressed both rice and Arabidopsis CFL1 in Arabidopsis thaliana; these transgenic plants showed severely impaired cuticle development, similar to that in cfl1 rice. Reduced expression of At CFL1 resulted in reinforcement of cuticle structure. At CFL1 was predominantly expressed in specialized epidermal cells and in regions where dehiscence and abscission occur. Biochemical evidence showed that At CFL1 interacts with HDG1, a class IV homeodomain-leucine zipper transcription factor. Suppression of HDG1 function resulted in similar defective cuticle phenotypes in wild-type Arabidopsis but much alleviated phenotypes in At cfl1-1 mutants. The expression of two cuticle development-associated genes, BDG and FDH, was downregulated in At CFL1 overexpressor and HDG1 suppression plants. HDG1 binds to the cis-element L1 box, which exists in the regulatory regions of BDG and FDH. Our results suggest that rice and Arabidopsis CFL1 negatively regulate cuticle development by affecting the function of HDG1, which regulates the downstream genes BDG and FDH.

A respiratory compensating system: design and performance evaluation.

This study proposes a respiratory compensating system which is mounted on the top of the treatment couch for reverse motion, opposite from the direction of the targets (diaphragm and hemostatic clip), in order to offset organ displacement generated by respiratory motion. Traditionally, in the treatment of cancer patients, doctors must increase the field size for radiation therapy of tumors because organs move with respiratory motion, which causes radiation-induced inflammation on the normal tissues (organ at risk (OAR)) while killing cancer cells, and thereby reducing the patient's quality of life. This study uses a strain gauge as a respiratory signal capture device to obtain abdomen respiratory signals, a proposed respiratory simulation system (RSS) and respiratory compensating system to experiment how to offset the organ displacement caused by respiratory movement and compensation effect. This study verifies the effect of the respiratory compensating system in offsetting the target displacement using two methods. The first method uses linac (medical linear accelerator) to irradiate a 300 cGy dose on the EBT film (GAFCHROMIC EBT film). The second method uses a strain gauge to capture the patients' respiratory signals, while using fluoroscopy to observe in vivo targets, such as a diaphragm, to enable the respiratory compensating system to offset the displacements of targets in superior-inferior (SI) direction. Testing results show that the RSS position error is approximately 0.45 ~ 1.42 mm, while the respiratory compensating system position error is approximately 0.48 ~ 1.42 mm. From the EBT film profiles based on different input to the RSS, the results suggest that when the input respiratory signals of RSS are sine wave signals, the average dose (%) in the target area is improved by 1.4% ~ 24.4%, and improved in the 95% isodose area by 15.3% ~ 76.9% after compensation. If the respiratory signals input into the RSS respiratory signals are actual human respiratory signals, the average dose (%) in the target area is improved by 31.8% ~ 67.7%, and improved in the 95% isodose area by 15.3% ~ 86.4% (the above rates of improvements will increase with increasing respiratory motion displacement) after compensation. The experimental results from the second method suggested that about 67.3% ~ 82.5% displacement can be offset. In addition, gamma passing rate after compensation can be improved to 100% only when the displacement of the respiratory motion is within 10 ~ 30 mm. This study proves that the proposed system can contribute to the compensation of organ displacement caused by respiratory motion, enabling physicians to use lower doses and smaller field sizes in the treatment of tumors of cancer patients.

The Relationship between Corporate Social Responsibility and Firm Value of Chinese Firms: Exploring from Degree of Internationalization.

The purpose of this paper is to explore the relationship between corporate social responsibility (CSR) and firm value in the Chinese market and identify factors that may influence it. We discuss the relationship between CSR engagement in Chinese firms and firm value from a nonlinear perspective. In addition, we examine how the degree of internationalization in a firm may affect the relationship between CSR and firm value. We employ the Hausman test to compare a random-effects and a fixed-effects model, and after testing and comparison, the fixed-effects model was chosen in our paper. Using data from 314 firms listed in China's A-shares market from 2010 to 2017, we verify the U-shaped relationship between CSR and firm value. Meanwhile, the degree of internationalization will affect firm value but cannot positively regulate the relationship between corporate social responsibility and firm value.

Comprehensive MDCT evaluation of patients with suspected May-Thurner syndrome.

OBJECTIVE: The purpose of this essay is to introduce the MDCT protocol and interpretation techniques for optimal evaluation of patients with suspected May-Thurner syndrome. CONCLUSION: May-Thurner syndrome is always the working diagnosis when a patient presents with unilateral left lower limb swelling without signs of infection. MDCT is useful for fast, comprehensive evaluation of the vascular system to determine whether May-Thurner syndrome or an alternative condition is present.

A compensating system of respiratory motion for tumor tracking: design and verification.

Using the reverse motion of the treatment couch, this study offset the organ displacement generated by respiratory motion to solve the current clinical problem of increasing field sizes and safety margin expansions. This study used the self-designed simulated respiratory system (SRS) coupled with radiochromic EBT film to verify the self-developed respiratory compensation system. Pressure signals were generated from SRS to simulate abdomen movements during respiratory motion. The respiratory compensation system takes the phase of the pressure signals as the respiratory motion phase and adjusts the pressure signal gain to make the compensation signal amplitude close to the displacement of the target region. A linear accelerator is used to irradiate a 300 cGy dose on the EBT film. The experimental results suggested that the average dose percentage in the target region for the sine-wave amplitudes of 5, 10 and 15 mm with compensation improved by 6.9 ∼ 20.3% over the cases without compensation. The 80% isodose area with compensation improved by 22.8 ∼ 77.2% over the cases without compensation. The average dose percentage in the target region with compensation for respiratory motion distances of 5, 10 and 15 mm improved by 10.3 ∼ 18.7%. The 80% isodose area improved by 22.4 ∼ 55.1% after compensation. The average dose percentage of the compensated target region indicates that the proposed respiratory compensation system could improve the issue of the inability to constantly irradiate the target region caused by respiratory motion.

Over-expression of WOX1 leads to defects in meristem development and polyamine homeostasis in Arabidopsis.

In plants, the meristem has to maintain a separate population of pluripotent cells that serve two main tasks, i.e., self-maintenance and organ initiation, which are separated spatially in meristem. Prior to our study, WUS and WUS-like WOX genes had been reported as essential for the development of the SAM. In this study, the consequences of gain of WOX1 function are described. Here we report the identification of an Arabidopsis gain-of-function mutant wox1-D, in which the expression level of the WOX1 (WUSCHEL HOMEOBOX 1) was elevated and subtle defects in meristem development were observed. The wox1-D mutant phenotype is dwarfed and slightly bushy, with a smaller shoot apex. The wox1-D mutant also produced small and dark green leaves, and exhibited a failure in anther dehiscence and male sterility. Molecular evidences showed that the transcription of the stem cell marker gene CLV3 was down-regulated in the meristem of wox1-D but accumulated in the other regions, i.e., in the root-hypocotyl junction and at the sites for lateral root initiation. The fact that the organ size and cell size in leaves of wox1-D are smaller than those in wild type suggests that cell expansion is possibly affected in order to have partially retarded the development of lateral organs, possibly through alteration of CLV3 expression pattern in the meristem. An S-adenosylmethionine decarboxylase (SAMDC) protein, SAMDC1, was found able to interact with WOX1 by yeast two-hybrid and pull-down assays in vitro. HPLC analysis revealed a significant reduction of polyamine content in wox1-D. Our results suggest that WOX1 plays an important role in meristem development in Arabidopsis, possibly via regulation of SAMDC activity and polyamine homeostasis, and/or by regulating CLV3 expression.

Verification and compensation of respiratory motion using an ultrasound imaging system.

PURPOSE: The purpose of this study was to determine if it is feasible to use ultrasound imaging as an aid for moving the treatment couch during diagnosis and treatment procedures associated with radiation therapy, in order to offset organ displacement caused by respiratory motion. A noninvasive ultrasound system was used to replace the C-arm device during diagnosis and treatment with the aims of reducing the x-ray radiation dose on the human body while simultaneously being able to monitor organ displacements. METHODS: This study used a proposed respiratory compensating system combined with an ultrasound imaging system to monitor the compensation effect of respiratory motion. The accuracy of the compensation effect was verified by fluoroscopy, which means that fluoroscopy could be replaced so as to reduce unnecessary radiation dose on patients. A respiratory simulation system was used to simulate the respiratory motion of the human abdomen and a strain gauge (respiratory signal acquisition device) was used to capture the simulated respiratory signals. The target displacements could be detected by an ultrasound probe and used as a reference for adjusting the gain value of the respiratory signal used by the respiratory compensating system. This ensured that the amplitude of the respiratory compensation signal was a faithful representation of the target displacement. RESULTS: The results show that performing respiratory compensation with the assistance of the ultrasound images reduced the compensation error of the respiratory compensating system to 0.81-2.92 mm, both for sine-wave input signals with amplitudes of 5, 10, and 15 mm, and human respiratory signals; this represented compensation of the respiratory motion by up to 92.48%. In addition, the respiratory signals of 10 patients were captured in clinical trials, while their diaphragm displacements were observed simultaneously using ultrasound. Using the respiratory compensating system to offset, the diaphragm displacement resulted in compensation rates of 60%-84.4%. CONCLUSIONS: This study has shown that a respiratory compensating system combined with noninvasive ultrasound can provide real-time compensation of the respiratory motion of patients.

Hypofractionated radiotherapy for primary or secondary oligometastatic lung cancer using Tomotherapy.

BACKGROUND: To retrospectively review the outcome of patients with primary or secondary oligometastatic lung cancer, treated with hypofractionated Tomotherapy. METHODS: Between April 2007 and June 2011, a total of 33 patients with oligometastatic intrapulmonary lesions underwent hypofractionated radiotherapy by Tomotherapy along with appropriate systemic therapy. There were 24 primary, and 9 secondary lung cancer cases. The radiation doses ranged from 4.5 to 7.0 Gy per fraction, multiplied by 8-16 fractions. The median dose per fraction was 4.5 Gy (range, 4.5-7.0 Gy), and the median total dose was 49.5 Gy (range, 45-72 Gy). The median estimated biological effective dose at 10 Gy (BED10) was 71.8 Gy (range, 65.3-119.0 Gy), and that at 3 Gy (BED3) was 123.8 Gy (range, 112.5-233.3 Gy). The mean lung dose (MLD) was constrained mainly under 1200 cGy. The median gross tumor volume (GTV) was 27.9 cm3 (range: 2.5-178.1 cm3). RESULTS: The median follow-up period was 25.8 months (range, 3.0-60.7 months). The median overall survival (OS) time was 32.1 months for the 24 primary lung cancer patients, and >40 months for the 9 metastatic lung patients. The median survival time of the patients with extra-pulmonary disease (EPD) was 11.2 months versus >50 months (not reached) in the patients without EPD (p < 0.001). Those patients with smaller GTV (≦27.9 cm3) had a better survival than those with larger GTV (>27.9 cm3): >40 months versus 12.85 months (p = 0.047). The patients with ≦2 lesions had a median survival >40 months, whereas those with ≧3 lesions had 26 months (p = 0.065). The 2-year local control (LC) rate was 94.7%. Only 2 patients (6.1%) developed ≧grade 3 radiation pneumonitis. CONCLUSION: Using Tomotherapy in hypofractionation may be effective for selected primary or secondary lung oligometastatic diseases, without causing significant toxicities. Pulmonary oligometastasis patients without EPD had better survival outcomes than those with EPD. Moreover, GTV is more significant than lesion number in predicting survival.