ResearchPad - regular-paper https://www.researchpad.co Default RSS Feed en-us © 2020 Newgen KnowledgeWorks <![CDATA[Semi-automated prediction approach of target shifts using machine learning with anatomical features between planning and pretreatment CT images in prostate radiotherapy]]> https://www.researchpad.co/article/elastic_article_12628 The goal of this study was to develop a semi-automated prediction approach of target shifts using machine learning architecture (MLA) with anatomical features for prostate radiotherapy. Our hypothesis was that anatomical features between planning computed tomography (pCT) and pretreatment cone-beam computed tomography (CBCT) images could be used to predict the target, i.e. clinical target volume (CTV) shifts, with small errors. The pCT and daily CBCT images of 20 patients with prostate cancer were selected. The first 10 patients were employed for the development, and the second 10 patients for a validation test. The CTV position errors between the pCT and CBCT images were determined as reference CTV shifts (teacher data) after an automated bone-based registration. The anatomical features associated with rectum, bladder and prostate were calculated from the pCT and CBCT images. The features were fed as the input with the teacher data into five MLAs, i.e. three types of artificial neural networks, support vector regression (SVR) and random forests. Since the CTV shifts along the left–right direction were negligible, the MLAs were developed along the superior–inferior and anterior–posterior directions. The proposed framework was evaluated from the residual errors between the reference and predicted CTV shifts. In the validation test, the mean residual error with its standard deviation was 1.01 ± 1.09 mm in SVR using only one feature (one click), which was associated with positional difference of the upper rectal wall. The results suggested that MLAs with anatomical features could be useful in prediction of CTV shifts for prostate radiotherapy.

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<![CDATA[Analysis of cardiac toxicity after definitive chemoradiotherapy for esophageal cancer using a biological dose–volume histogram]]> https://www.researchpad.co/article/elastic_article_12625 This study aimed to evaluate the relationship between cardiac toxicity after definitive chemoradiotherapy (CRT) for esophageal cancer and the dose–volume histogram (DVH) of organs at risk (OARs) [using biological effective dose (BED)]. We analyzed the data of 83 patients with esophageal cancer treated using definitive CRT between 2001 and 2016. Furthermore, we evaluated pericardial effusion (PE) as a measure of cardiac toxicity. The median total irradiation dose was 60 (50.4–71) Gy. Symptomatic PE was observed in 12 (14%) patients. The heart and pericardium V5–V100-BED were significantly higher in patients with symptomatic PE than in those without symptomatic PE (heart: V5–V95-BED, P < 0.001; V100-BED, P = 0.0053, and pericardium: V5–V40-BED, V55–V95-BED, P < 0.001; V45–50-BED, V100-BED, P < 0.05, respectively). Receiver operating characteristic curve analysis showed that the dose–volume parameter of the pericardium and the heart that was most strongly associated with an adverse cardiac event was V80-BED, and the mean dose and the cut-off value were 27.38% and 61.7 Gy-BED, respectively. Multivariate analysis showed that the pericardium V80-BED and the mean heart dose-BED were risk factors for symptomatic PE (P < 0.001, respectively). We revealed the relationship between the irradiated dose of the OARs and symptomatic PE using a BED-based dose–volume histogram. Pericardium V80-BED and mean heart dose-BED were the most relevant risk factors for symptomatic PE.

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<![CDATA[Impact of RhoA overexpression on clinical outcomes in cervical squamous cell carcinoma treated with concurrent chemoradiotherapy]]> https://www.researchpad.co/article/elastic_article_12624 The Rho-associated coiled-coil-containing protein kinase (ROCK) pathway is known to influence metastasis in several cancers; however, the impact of the pathway on clinical outcomes in patients undergoing radiotherapy remains unknown. In the present study, the expression of RhoA, RhoC, ROCK-1, ROCK-2 and p53 was immunohistochemically evaluated using biopsy specimens obtained from 49 patients with stage II–III cervical squamous cell carcinoma treated with concurrent chemoradiotherapy (CCRT). The relationship between the expression of these proteins and patient outcomes was investigated. RhoA overexpression was associated with significantly impaired disease-free survival and distant metastasis-free survival (P = 0.045 and P = 0.041, respectively) in stage III cancer patients. No differences in survival were observed based on the expression of the other proteins among stage III cancer patients. In stage II cancer patients, no differences in survival were noted based on the expression of any of the proteins. The expression of RhoA was able to successfully differentiate cervical cancer patients with distant metastasis after CCRT. This information may help stratify patients according to the risk of metastasis, thereby leading to the potential to provide individualized treatment.

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<![CDATA[PARP inhibitor olaparib sensitizes esophageal carcinoma cells to fractionated proton irradiation]]> https://www.researchpad.co/article/elastic_article_12622 Proton beam therapy (PBT) combined with chemotherapy, such as cis-diamminedichloroplatinum (II) (CDDP) and 5-fluorouracil (5-FU), has been employed as an alternative approach to improve clinical outcomes. PBT has been reported to be effective against esophageal cancer. However, apart from 5-FU and CDDP, almost no other drug has been tested in combined chemotherapy with PBT. Therefore, we investigated the effects of a poly (ADP-ribose) polymerase inhibitor on enhancing proton beam effects using esophageal cancer cell lines that exhibit resistance to radiation and CDDP. Esophageal squamous cell carcinoma cell lines OE-21 and KYSE-450 were exposed to the drugs for 1 h prior to irradiation. The cell survival curve was obtained using a clonogenic assay and the sensitizing effect ratio (SER) was calculated. The clonogenic assay was used to compare the effect of multi-fractioned irradiation between 8 Gy/1 fraction (fr) and 8 Gy/4 fr. γH2AX, Rad51, BRCA1, BRCA2 and 53BP1 foci were detected via immunofluorescence. Olaparib exhibited an SER of 1.5–1.7 on PBT. The same sensitizing effect was exhibited in multi-fractioned irradiation, and the combined use increased the expression of double-strand breaks and homologous recombination-related genes in an additive manner. Such additive effects were not observed on non-homologous end joining-related genes. We demonstrated that olaparib has a high sensitizing effect on PBT in platinum- and radiation-resistant esophageal cancer cells. Our results suggest a potential clinical application of olaparib-proton irradiation (PT) against platinum- and radiation-resistant esophageal cancer.

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<![CDATA[Investigation of 4D dose in volumetric modulated arc therapy-based stereotactic body radiation therapy: does fractional dose or number of arcs matter?]]> https://www.researchpad.co/article/elastic_article_12621 The aim of this study was to assess the impact of fractional dose and the number of arcs on interplay effects when volumetric modulated arc therapy (VMAT) is used to treat lung tumors with large respiratory motions. A three (fractional dose of 4, 7.5 or 12.5 Gy) by two (number of arcs, one or two) VMAT plan was created for 10 lung cancer cases. The median 3D tumor motion was 17.9 mm (range: 8.2–27.2 mm). Ten phase-specific subplans were generated by calculating the dose on each respiratory phase computed tomography (CT) scan using temporally assigned VMAT arcs. We performed temporal assignment of VMAT arcs using respiratory information obtained from infrared markers placed on the abdomens of the patients during CT simulations. Each phase-specific dose distribution was deformed onto exhale phase CT scans using contour-based deformable image registration, and a 4D plan was created by dose accumulation. The gross tumor volume dose of each 4D plan (4D GTV dose) was compared with the internal target volume dose of the original plan (3D ITV dose). The near-minimum 4D GTV dose (D99%) was higher than the near-minimum 3D internal target volume (ITV) dose, whereas the near-maximum 4D GTV dose (D1%) was lower than the near-maximum 3D ITV dose. However, the difference was negligible, and thus the 4D GTV dose corresponded well with the 3D ITV dose, regardless of the fractional dose and number of arcs. Therefore, interplay effects were negligible in VMAT-based stereotactic body radiation therapy for lung tumors with large respiratory motions.

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<![CDATA[Treatment outcomes of patients with adenocarcinoma of the uterine cervix after definitive radiotherapy and the prognostic impact of tumor-infiltrating CD8<sup>+</sup> lymphocytes in pre-treatment biopsy specimens: a multi-institutional retrospective study]]> https://www.researchpad.co/article/elastic_article_12619 The current study aimed to evaluate the outcomes of patients with adenocarcinoma (AC) of the uterine cervix after definitive radiotherapy (RT) and to evaluate prognostic factors, including immunity-related molecules. A total of 71 patients with AC of the uterine cervix from multiple Japanese institutions were retrospectively analysed. Histological subtypes were diagnosed according to the 2014 World Health Organization classification. All patients underwent definitive RT comprising external beam RT and intracavitary brachytherapy with or without concurrent chemotherapy. Immunohistochemical studies were performed to detect the expression of programmed cell death-ligand 1(PD-L1) and CD8. The 5-year locoregional control (LC), overall survival (OS) and progression-free survival (PFS) rates for all patients were 61.8, 49.7 and 36.1%, respectively. The LC, OS and PFS rates were not significantly different among the histological subtypes. Membranous PD-L1 expression was not significantly associated with prognosis. Patients with CD8-positive tumor-infiltrating lymphocytes (CD8+TILs) in the tumor nests had significantly better OS than patients without CD8+TILs in the tumor nests (5-year OS: 53.8 vs 23.8%, P = 0.038). As expected, the International Federation of Gynecology and Obstetrics (FIGO) stage (2008) III–IVA and maximum tumor diameter > 40 mm were significantly associated with worse prognosis. In summary, the presence of CD8+TILs in the tumor nests has the potential to be an independent favorable prognostic factor for patients with AC of the uterine cervix after definitive RT.

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<![CDATA[Retrospective DVH analysis of point A based intracavitary brachytherapy for uterine cervical cancer]]> https://www.researchpad.co/article/elastic_article_12618 Combining external beam radiotherapy (EBRT) with intracavitary brachytherapy (ICBT) is important for definitive treatment of cervical cancer. In cervical cancer patients receiving radiotherapy, we evaluated treatment outcomes in relation to dose–volume histogram parameters, including the computed tomography (CT)-based high-risk clinical target volume (HR-CTV) for ICBT. Between 2010 and 2015, 89 consecutive cervical cancer patients were mostly treated with 40 Gy of EBRT in 20 fractions and 18 Gy of ICBT prescribed to point A in 3 fractions. CT scans were obtained during ICBT. The HR-CTV D90 was calculated and the total doses of ICBT and EBRT were converted to the equivalent dose in 2 Gy fractions (EQD2). When the patients were divided into four groups according to EQD2 of the HR-CTV D90, the 3-year local recurrence-free survival rates were 95.2, 78.4, 52.7 and 42.9% for patients receiving >80 , 70–80 , 60–70 and <60 Gy, respectively. There was a significant negative correlation between EQD2 of the HR-CTV D90 and the HR-CTV volume at first ICBT (r = −0.713). Local recurrence was more frequent when the HR-CTV volume was ≥22 cc and EQD2 of the HR-CTV D90 was <70 Gy. Multivariate analysis showed that EQD2 of the HR-CTV D90 and concurrent chemotherapy (≥4 cycles) were significant determinants of overall survival. HR-CTV D90 was an important prognostic indicator for local recurrence. HR-CTV D90 >70 Gy is required for the better local control, especially in patients with a larger HR-CTV (≥22 cc at initial ICBT).

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<![CDATA[Testing of behavioral and cognitive development in rats after prenatal exposure to 1800 and 2400 MHz radiofrequency fields]]> https://www.researchpad.co/article/elastic_article_12617 The objective of the study was to explore the effects of behavioral and cognitive development in rats after prenatal exposure to 1800 and 2400 MHz radiofrequency fields. Pregnant female rats were exposed to radiofrequency fields beginning on the 21st day of pregnancy. The indicators of physiological and behavioral development were observed and measured in the offspring rats: Y maze measured at 3-weeks postnatal, open field at 7-weeks postnatal, and the expression of N-methyl-D-aspartate receptors (NMDARs) measured by reverse transcription-PCR in the hippocampus at 9-weeks postnatal. The body weight of the 1800 MHz group and the 1800 MHz + WiFi group showed a downward trend. The eye opening time of newborn rats was much earlier in the WiFi group than in the control group. Compared to the control group, the overall path length of the 1800 MHz + WiFi group was shortened and the stationary time was delayed. The path length of the WiFi group was shortened and the average velocity was increased in the error arm. The 1800 MHz + WiFi group displayed an increased trend in path length, duration, entry times and stationary time in the central area. In both the 1800 MHz + WiFi and WiFi groups, NR2A and NR2B expression was down-regulated, while NR2D, NR3A and NR3B were up-regulated. Moreover, NR1 and NR2C in the WiFi group were also up-regulated. Prenatal exposure to 1800 MHz and WiFi radiofrequency may affect the behavioral and cognitive development of offspring rats, which may be associated with altered mRNA expression of NMDARs in the hippocampus.

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<![CDATA[Inter-unit variability of multi-leaf collimator parameters for IMRT and VMAT treatment planning: a multi-institutional survey]]> https://www.researchpad.co/article/elastic_article_12616 Modern treatment machines have shown small inter-unit variability regarding beam data. Recently, vendor-provided average beam data, such as the Representative Beam Data (RBD) of the TrueBeam (Varian Medical Systems, Palo Alto, CA, USA), has been used for modeling of the Eclipse (Varian Medical Systems) treatment planning system. However, RBD does not provide multi-leaf collimator (MLC) parameters, such as MLC leaf transmission factor (LTF) and dosimetric leaf gap (DLG). We performed a web-based multi-institutional survey to investigate these parameters as well as the measurement protocols and customization of the parameters for intensity-modulated radiotherapy (IMRT) and/or volumetric modulated radiotherapy (VMAT) commissioning. We collected 69 sets of linear accelerator (linac) data from 58 institutions. In order to measure MLC parameters, most institutions used farmer-type ionization chambers with a sensitive volume of 0.6 cm3, water phantoms, source surface distance of 90 cm with 10 cm depth, and a vendor-provided plan. The LTF showed small inter-unit variabilities, although the DLG showed large variations. For optimization of the parameters for IMRT/VMAT calculations, DLG values were upwardly adjusted at many institutions, whereas the LTF values were modestly changed. We clarified that MLC parameters were measured under the same conditions at more than half of the facilities. Most institutions customized parameters in a similar manner for IMRT/VMAT. The median measured and customized values obtained in our study will be valuable to verify MLC installation accuracy and to shorten the iterative processes of finding the optimal values.

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<![CDATA[Development of a hybrid magnetic resonance/computed tomography-compatible phantom for magnetic resonance guided radiotherapy]]> https://www.researchpad.co/article/elastic_article_12613 The purpose of the present study was to develop a hybrid magnetic resonance/computed tomography (MR/CT)-compatible phantom and tissue-equivalent materials for each MR and CT image. Therefore, the essential requirements necessary for the development of a hybrid MR/CT-compatible phantom were determined and the development process is described. A total of 12 different tissue-equivalent materials for each MR and CT image were developed from chemical components. The uniformity of each sample was calculated. The developed phantom was designed to use 14 plugs that contained various tissue-equivalent materials. Measurement using the developed phantom was performed using a 3.0-T scanner with 32 channels and a Somatom Sensation 64. The maximum percentage difference of the signal intensity (SI) value on MR images after adding K2CO3 was 3.31%. Additionally, the uniformity of each tissue was evaluated by calculating the percent image uniformity (%PIU) of the MR image, which was 82.18 ±1.87% with 83% acceptance, and the average circular-shaped regions of interest (ROIs) on CT images for all samples were within ±5 Hounsfield units (HU). Also, dosimetric evaluation was performed. The percentage differences of each tissue-equivalent sample for average dose ranged from −0.76 to 0.21%. A hybrid MR/CT-compatible phantom for MR and CT was investigated as the first trial in this field of radiation oncology and medical physics.

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<![CDATA[The effects of short-term calorie restriction on mutations in the spleen cells of infant-irradiated mice]]> https://www.researchpad.co/article/elastic_article_12611 The risk of cancer due to exposure to ionizing radiation is higher in infants than in adults. In a previous study, the effect of adult-onset calorie restriction (CR) on carcinogenesis in mice after early-life exposure to X-rays was examined (Shang, Y, Kakinuma, S, Yamauchi, K, et al. Cancer prevention by adult-onset calorie restriction after infant exposure to ionizing radiation in B6C3F1 male mice. Int J Cancer. 2014; 135: 1038-47). The results showed that the tumor frequency was reduced in the CR group. However, the mechanism of tumor suppression by CR is not yet clear. In this study, we examined the effects of CR on radiation-induced mutations using gpt delta mice, which are useful to analyze mutations in various tissues throughout the whole body. Infant male mice (1-week old) were exposed to 3.8 Gy X-rays and fed a control (95 kcal/week/mouse) or CR (65 kcal/week/mouse) diet from adult stage (7-weeks old). Mice were sacrificed at the age of 7 weeks, 8 weeks and 100 days, and organs (spleen, liver, lung, thymus) were harvested. Mutations at the gpt gene in the DNA from the spleen were analyzed by using a gpt assay protocol that detects primarily point mutations in the gpt gene. The results showed that mutation frequencies were decreased in CR groups compared with non-CR groups. Sequence analysis of the gpt gene in mutants revealed a reduction in the G:C to T:A transversion in CR groups. Since it is known that 8-oxoguanine could result in this base substitution and that CR has an effect of reducing oxidative stress, these results indicate that the suppression of oxidative stress by CR is the cause of the reduction of this transversion.

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<![CDATA[Microdosimetric quantities of an accelerator-based neutron source used for boron neutron capture therapy measured using a gas-filled proportional counter]]> https://www.researchpad.co/article/elastic_article_12610 Boron neutron capture therapy (BNCT) is an emerging radiation treatment modality, exhibiting the potential to selectively destroy cancer cells. Currently, BNCT is conducted using a nuclear reactor. However, the future trend is to move toward an accelerator-based system for use in hospital environments. A typical BNCT radiation field has several different types of radiation. The beam quality should be quantified to accurately determine the dose to be delivered to the target. This study utilized a tissue equivalent proportional counter (TEPC) to measure microdosimetric and macrodosimetric quantities of an accelerator-based neutron source. The micro- and macro-dosimetric quantities measured with the TEPC were compared with those obtained via the the particle and heavy ion transport code system (PHITS) Monte Carlo simulation. The absorbed dose from events >20 keV/μm measured free in air for a 1-h irradiation was calculated as 1.31 ± 0.02 Gy. The simulated result was 1.41 ± 0.07 Gy. The measured and calculated values exhibit good agreement. The relative biological effectiveness (RBE) that was evaluated from the measured microdosimetric spectrum was calculated as 3.7 ± 0.02, similar to the simulated value of 3.8 ± 0.1. These results showed the PHITS Monte Carlo simulation can simulate both micro- and macro-dosimetric quantities accurately. The RBE was calculated using a single-response function, and the results were compared with those of several other institutes that used a similar method. However, care must be taken when using such a single-response function for clinical application, as it is only valid for low doses. For clinical dose ranges (i.e., high doses), multievent distribution inside the target needs to be considered.

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<![CDATA[Efficacy evaluation of 2D, 3D U-Net semantic segmentation and atlas-based segmentation of normal lungs excluding the trachea and main bronchi]]> https://www.researchpad.co/article/elastic_article_12609 This study aimed to examine the efficacy of semantic segmentation implemented by deep learning and to confirm whether this method is more effective than a commercially dominant auto-segmentation tool with regards to delineating normal lung excluding the trachea and main bronchi. A total of 232 non-small-cell lung cancer cases were examined. The computed tomography (CT) images of these cases were converted from Digital Imaging and Communications in Medicine (DICOM) Radiation Therapy (RT) formats to arrays of 32 × 128 × 128 voxels and input into both 2D and 3D U-Net, which are deep learning networks for semantic segmentation. The number of training, validation and test sets were 160, 40 and 32, respectively. Dice similarity coefficients (DSCs) of the test set were evaluated employing Smart Segmentation Knowledge Based Contouring (Smart segmentation is an atlas-based segmentation tool), as well as the 2D and 3D U-Net. The mean DSCs of the test set were 0.964 [95% confidence interval (CI), 0.960–0.968], 0.990 (95% CI, 0.989–0.992) and 0.990 (95% CI, 0.989–0.991) with Smart segmentation, 2D and 3D U-Net, respectively. Compared with Smart segmentation, both U-Nets presented significantly higher DSCs by the Wilcoxon signed-rank test (P < 0.01). There was no difference in mean DSC between the 2D and 3D U-Net systems. The newly-devised 2D and 3D U-Net approaches were found to be more effective than a commercial auto-segmentation tool. Even the relatively shallow 2D U-Net which does not require high-performance computational resources was effective enough for the lung segmentation. Semantic segmentation using deep learning was useful in radiation treatment planning for lung cancers.

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<![CDATA[Clinical utilization of radiation therapy in Korea, 2016]]> https://www.researchpad.co/article/elastic_article_12608 Radiotherapy (RT) is one of the primary cancer treatment modalities. To estimate the actual utilization of RT and infrastructure in Korea, the current study was performed. Data from 2012 to 2016 were extracted from the Health Insurance Review and Assessment Service. In addition, a nationwide survey was conducted to collect the statistics of RT facilities, equipment and human resources in Korea. The total number of patients treated with RT was 72 563 in 2016. The five cancers that were most commonly treated with RT in 2016 were breast, lung, colorectal, liver and prostate cancer. According to analyses of specific treatment modalities, the number of patients treated with intensity-modulated radiotherapy (IMRT), stereotactic radiation therapy (SRT) and proton therapy increased from 6670, 6306 and 50 in 2012 to 21584, 9048 and 703 in 2016, respectively. Ninety radiation oncology centers were working in 2015 and there were a total of 213 megavoltage teletherapy machines. In 2015, 310 patients were treated per megavoltage RT machine, 246 patients per radiation oncologist, 501 patients per medical physicist and 111 patients per radiotherapy technologist. In conclusion, the number of patients who underwent RT in Korea has increased steadily from 2012 to 2016. The IMRT utilization rate remarkably increased in 2016, and the number of patients treated with advanced treatment modalities such as IMRT, SRT and proton therapy is expected to increase.

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<![CDATA[Wortmannin, a specific inhibitor of phosphatidylinositol-3-kinase, induces accumulation of DNA double-strand breaks]]> https://www.researchpad.co/article/elastic_article_12607 Wortmannin, a fungal metabolite, is a specific inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, which includes double-stranded DNA dependent protein kinase (DNA-PK) and ataxia telangiectasia mutated kinase (ATM). We investigated the effects of wortmannin on DNA damage in DNA-PK-deficient cells obtained from severe combined immunodeficient mice (SCID cells). Survival of wortmannin-treated cells decreased in a concentration-dependent manner. After treatment with 50 μM wortmannin, survival decreased to 60% of that of untreated cells. We observed that treatment with 20 and 50 μM wortmannin induced DNA damage equivalent to that by 0.37 and 0.69 Gy, respectively, of γ-ray radiation. The accumulation of DNA double-strand breaks (DSBs) in wortmannin-treated SCID cells was assessed using pulsed-field gel electrophoresis. The maximal accumulation was observed 4 h after treatment. Moreover, the presence of DSBs was confirmed by the ability of nuclear extracts from γ-ray-irradiated SCID cells to produce in vitro phosphorylation of histone H2AX. These results suggest that wortmannin induces cellular toxicity by accumulation of spontaneous DSBs through inhibition of ATM.

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<![CDATA[Transition from C<sub>3</sub> to proto-Kranz to C<sub>3</sub>–C<sub>4</sub> intermediate type in the genus <i>Chenopodium</i> (Chenopodiaceae)]]> https://www.researchpad.co/article/N9bfdbe47-1d4d-4f72-b062-f3ff5f5fe963 The Chenopodiaceae is one of the families including C4 species among eudicots. In this family, the genus Chenopodium is considered to include only C3 species. However, we report here a transition from C3 photosynthesis to proto-Kranz to C3–C4 intermediate type in Chenopodium. We investigated leaf anatomical and photosynthetic traits of 15 species, of which 8 species showed non-Kranz anatomy and a CO2 compensation point (Γ) typical of C3 plants. However, 5 species showed proto-Kranz anatomy and a C3-like Γ, whereas C. strictum showed leaf anatomy and a Γ typical of C3–C4 intermediates. Chenopodium album accessions examined included both proto-Kranz and C3–C4 intermediate types, depending on locality. Glycine decarboxylase, a key photorespiratory enzyme that is involved in the decarboxylation of glycine, was located predominantly in the mesophyll (M) cells of C3 species, in both M and bundle-sheath (BS) cells in proto-Kranz species, and exclusively in BS cells in C3–C4 intermediate species. The M/BS tissue area ratio, number of chloroplasts and mitochondria per BS cell, distribution of these organelles to the centripetal region of BS cells, the degree of inner positioning (vacuolar side of chloroplasts) of mitochondria in M cells, and the size of BS mitochondria also changed with the change in glycine decarboxylase localization. All Chenopodium species examined were C3-like regarding activities and amounts of C3 and C4 photosynthetic enzymes and δ13C values, suggesting that these species perform photosynthesis without contribution of the C4 cycle. This study demonstrates that Chenopodium is not a C3 genus and is valuable for studying evolution of C3–C4 intermediates.

Electronic supplementary materialThe online version of this article (10.1007/s10265-019-01135-5) contains supplementary material, which is available to authorized users. ]]>
<![CDATA[Cell cycle inhibitors improve seed storability after priming treatments]]> https://www.researchpad.co/article/Nf359965c-ee4c-41bb-a6c8-dd70c3d08025 Seed priming is a treatment that controls seed water content to partially activate germination processes such as metabolism but prevents full germination of the seeds. The treatment is well known to enhance seed performance, including germination, but sometimes reduces seed storability or longevity as a side effect. Toward developing a novel priming technique that can maintain seed longevity for a longer time period, chemicals that suppress the seed deterioration under a controlled condition were screened from 80 known biologically active compounds contained in the RIKEN NPDepo authentic library using Arabidopsis thaliana seeds. Seeds primed with mimosine, a cell cycle inhibitor, retained higher survival rate after a controlled deterioration treatment compared to seeds primed without the chemical. In addition, other cell cycle inhibitors such as aphidicolin, hydroxyurea and oryzalin had similar effects on the seed storability after priming. Our results suggest that progression of the cell cycle during priming is an important checkpoint that determines the storability of seeds after the treatment.

Electronic supplementary materialThe online version of this article (10.1007/s10265-018-01084-5) contains supplementary material, which is available to authorized users. ]]>
<![CDATA[Extrafloral nectary-bearing plant <i>Mallotus japonicus</i> uses different types of extrafloral nectaries to establish effective defense by ants]]> https://www.researchpad.co/article/Nba90c64d-0e49-4f6a-9325-94c8a68b62ce Extrafloral nectary (EFN)-bearing plants attract ants to gain protection against herbivores. Some EFN-bearing plants possess different types of EFNs, which might have different effects on ants on the plants. Mallotus japonicus (Thunb.) Muell. Arg. (Euphorbiaceae) bears two types of EFNs, including a pair of large EFNs at the leaf base and many small EFNs along the leaf edge. This study aimed to determine the different roles of the two types of EFNs in biotic defense by ants. A field experiment was conducted to investigate the effect of leaf damage on EFN production and on the distribution pattern of ants. After leaf damage, the number of leaf edge EFNs increased in the leaves first-produced. The number of ants on the leaves also increased, and the foraging area of ants extended from the leaf base to the leaf tip. An EFN-covering field experiment revealed that leaf edge EFNs had a greater effect than leaf base EFNs on ant dispersal on leaves. The extended foraging area of ants resulted in an increase of encounter or attack rate against an experimentally placed herbivore, Spodoptera litura. These results suggest that M. japonicus plants control the foraging area of ants on their leaves using different types of EFNs in response to leaf damage, thus achieving a very effective biotic defense against herbivores by ants.

Electronic supplementary materialThe online version of this article (10.1007/s10265-019-01119-5) contains supplementary material, which is available to authorized users. ]]>
<![CDATA[Salicylic acid-induced ROS production by mitochondrial electron transport chain depends on the activity of mitochondrial hexokinases in tomato (<i>Solanum lycopersicum</i> L.)]]> https://www.researchpad.co/article/Nc1fffbb4-7370-4289-943a-43651b3ea960 The growth regulator, salicylic acid (SA) plays an important role in the induction of cell death in plants. Production of reactive oxygen species (ROS) by mitochondrial electron transport chain (mtETC), cytochrome c (cyt c) release from mitochondria and loss of mitochondrial integrity can be observed during cell death execution in plant tissues. The aim of this work was to study the putative role of hexokinases (HXKs) in the initiation of cell death using tomato (Solanum lycopersicum L.) leaves and mitochondria isolated from plants exposed to a sublethal, 0.1 mM and a cell death-inducing, 1 mM concentrations of SA. Both treatments enhanced ROS and nitric oxide (NO) production in the leaves, which contributed to a concentration-dependent loss of membrane integrity. Images prepared by transmission electron microscopy showed swelling and disorganisation of mitochondrial cristae and vacuolization of mitochondria after SA exposure. Using post-embedding immunohistochemistry, cyt c release from mitochondria was also detected after 1 mM SA treatment. Both SA treatments decreased the activity and transcript levels of HXKs in the leaves and the total mtHXK activity in the mitochondrial fraction. The role of mitochondrial hexokinases (mtHXKs) in ROS and NO production of isolated mitochondria was investigated by the addition of HXK substrate, glucose (Glc) and a specific HXK inhibitor, N-acetylglucosamine (NAG) to the mitochondrial suspension. Both SA treatments enhanced ROS production by mtETC in the presence of succinate and ADP, which was slightly inhibited by Glc and increased significantly by NAG in control and in 0.1 mM SA-treated mitochondria. These changes were not significant at 1 mM SA, which caused disorganisation of mitochondrial membranes. Thus the inhibition of mtHXK activity can contribute to the mitochondrial ROS production, but it is not involved in NO generation in SA-treated leaf mitochondria suggesting that SA can promote cell death by suppressing mtHXK transcription and activity.

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<![CDATA[Heat and chilling stress induce nucleolus morphological changes]]> https://www.researchpad.co/article/N86759da0-1127-4bfc-949d-94e314902e0c The nucleolus, where components of the ribosome are constructed, is known to play an important role in various stress responses in animals. However, little is known about the role of the plant nucleolus under environmental stresses such as heat and chilling stress. In this study, we analyzed nucleolus morphology by determining the distribution of newly synthesized rRNAs with an analog of uridine, 5-ethynyl uridine (EU). When EU was incorporated into the root of the Arabidopsis thaliana, EU signals were strongly localized in the nucleolus. The results of the short-term incorporation of EU implied that there is no compartmentation among the processes of transcription, processing, and construction of rRNAs. Nevertheless, under heat and chilling stress, EU was not incorporated into the center of the nucleolus. Morphological analyses using whole rRNA staining and differential interference contrast observations revealed speckled and round structures in the center of the nucleolus under heat and chilling stress, respectively.

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