Enrichment of grape berries and tomato fruit with health-promoting tartaric acid by expression of the Vitis vinifera transketolase VvTK2 gene.

Tartaric acid (TA) is a major non-fermentable plant soluble acid that abundantly occur in grapes and wines, imparting low pH and tart flavour to berries thereby regulating numerous quality attributes of wine, such as flavour, microbial stability, and aging potential. Evaluation of acidity in mature fruits of 21 wine grape (Vitis vinifera) varieties revealed significant variation between 'Beichun' and 'Gewürztraminer', which was correlated with TA content. RNA-seq analysis of fruits from the two cultivars at different developmental stages revealed that a transketolase gene, VvTK2, was significantly dominantly expressed in the high TA phenotype 'Beichun' variety. Subcellular localization assay showed that VvTK2 protein was located in the chloroplast. Virus-induced VvTK2 gene silencing significantly decreased the expression of 2-keto-L-gulonic acid reductase (Vv2-KGR) as well as L-idonate dehydrogenase (VvL-IdnDH3) and inhibited TA accumulation, while its transient over-expression in grape showed the opposite results. Heterologous VvTK2 over-expression in tomato demonstrated its obvious capacity to induce TA synthesis. Overall, these results highlights a novel role of VvTK2 in modulating TA biosynthesis, which could be an excellent strategy for future genetic improvement of grape flavour.

Hypofractionated concurrent chemoradiotherapy related lymphopenia and its association with survival in locally advanced non-small cell lung cancer patients.

Background: To evaluate longitudinal changes of concurrent chemoradiotherapy (CCRT) related lymphopenia and its association with survival in locally advanced non-small cell lung cancer (LA-NSCLC) patients. Methods: Total lymphocyte count (TLC) at baseline, weekly intervals during CCRT and monthly intervals up to 12 months after CCRT were documented. The Common Terminology Criteria for Adverse Events version 5.0 was used to grade the severity of lymphopenia. Cox regression analysis was performed to evaluate the association between overall survival (OS) and CCRT related lymphopenia at different timepoints. Logistic regression model was used to determine the clinical factors associated with TLC level. Results: 381 LA-NSCLC patients treated with definitive CCRT without consolidation therapy (NCT02573506/NCT02577341) between 2011 to 2020 were analyzed. With a median follow-up of 45.8 months, the median OS was 41.0 months for all patients. Univariable analysis demonstrated that the 3 weeks during CCRT Grade (G) 4 lymphopenia (P=0.018), 2 months after CCRT G1-4 lymphopenia (P=0.004), 6 months after CCRT (6m-post-CCRT) G1-4 lymphopenia (P=0.001), and TLC nadir (P=0.020) were significantly associated with poorer OS. Multivariable analysis suggested that 6m-post-CCRT G1-4 lymphopenia (HR 2.614; P=0.041) were one of the independent predictors of OS. Further analysis inferred that radiation dose (OR: 1.328; P=0.005), GTV volume (OR: 1.004; P=0.036), and baseline TLC (OR: 0.288; P=0.001) were associated with 6m-post-CCRT lymphopenia. Conclusion: The persistent lymphopenia at 6 months after CCRT was an independent prognostic factor of OS in LA-NSCLC patients. Higher radiation dose, larger gross tumor volume and lower baseline TLC were significantly related to 6m-post-CCRT lymphopenia.

PRMT5-activated c-Myc promote bladder cancer proliferation and invasion through up-regulating NF-κB pathway.

AIM: PRMT5 and c-Myc were considered as oncogene of bladder cancer. Nevertheless, whether the interaction between of PRMT5 and c-Myc affect bladder cancer progress is unknown. Herein, we explore the above points and discuss deeply its' potential mechanism. METHOD: 5637 and T24 cells were study subjects in vitro. Western blot was used to examined the protein expression. CCK8 and transwell assay were used to analyze proliferation and invasion ability. Additionally, xenograft tumor model was established. Mice imaging experiment, Immunochemistry assay and western blot were carried out. RESULT: Western blot result showed successful transfection of PRMT5-siRNA and c-Myc-siRNA. PRMT5-siRNA could inhibit c-Myc expression, and decrease the proliferation and invasion of bladder cells. And c-Myc overexpression could reverse inhibitory action caused by PRMT5 silence. And in vitro studies found low-expression of c-Myc reduced proliferation and invasion of tumor cells and make the NF-κB pathway inactivation. In vivo studies also demonstrated that inhibiting PRMT5 could downregulate c-Myc expression and inhibit the bladder cancer progress, and the potential mechanism was likely to be related to NF-κB signaling pathway. CONCLUSION: In a word, low-expression of PRMT5 suppressed c-Myc, and thus inhibited proliferation and invasion ability of 5637 and T24 cells through NF-κB pathway.

Sevoflurane induces inflammation of microglia in hippocampus of neonatal rats by inhibiting Wnt/ß-Catenin/CaMKIV pathway.

OBJECTIVE: To investigate the effect of sevoflurane on inflammation of microglia in hippocampus of neonatal rats, and to investigate whether the related mechanism is related to Wnt/ß-Catenin/CaMKIV pathway. METHODS: Neonatal rats were anesthetized with 2% or 3% sevoflurane for 4 h a day for 3 consecutive days. Water maze test was used to detect the effect of sevoflurane anesthesia on memory function of neonatal rats. H&E and Nissl staining were used to observe the pathological damage of hippocampal area of neonatal rats induced by sevoflurane anesthesia. The expression of microglial marker Iba-1 was detected by Immunofluorescence. Immunofluorescence and WB were used to detect the expression CD32b, CD86, TNF-α, IL-6, Wnt3a, ß-Catenin and CaMKIV in hippocampus. To further explore the related mechanism, Wnt-3α inhibitor and activator was treated to study the effect of sevoflurane on microglial inflammation in hippocampus of neonatal rats. RESULTS: Sevoflurane anesthesia significantly increased escape latency time, reduced platform crossing times, and damaged the learning and memory ability of neonatal rats. H&E and Nissl staining results showed that sevoflurane anesthesia caused obvious damage to the hippocampus of neonatal rats. Sevoflurane anesthesia promoted the expression of Iba-1 and activated microglia. Sevoflurane anesthesia not only significantly increased the positive expression of CD32b, CD86, TNF-α and IL-6, but also decreased the expression of Wnt3a, ß-Catenin and CaMKIV. These results suggested that sevoflurane inhibited Wnt/ß-Catenin/CaMKIV pathway. CONCLUSION: Sevoflurane induces inflammation of microglia in hippocampus of neonatal rats by inhibiting Wnt/ß-Catenin/CaMKIV pathway.

Quantifying the Interfractional motion of Esophagus Using Daily Cone Beam Computed Tomography with Oral Contrast During Radiation Therapy for Locally Advanced Non-Small Cell Lung Cancer.

PURPOSE: To quantify the interfractional motion of the esophagus during fractionated radiation therapy for locally advanced non-small cell lung cancer. METHODS AND MATERIALS: We registered simulation 4-dimensional computed tomography (CT) and daily cone beam CT (CBCT) and documented the motion of the esophagus centroid at 5-mm interval slices in right-left (RL) and anterior-posterior (AP) directions. Oral barium sulfate was administrated during CBCT to help localize the esophagus. Thirty-five patients were enrolled. Thirty-five 4-dimensional CT scans, 595 CBCT scans, and 25,970 slices were analyzed. The slice-derived motion values for all patients were presented as 2.5 to 97.5 percentiles and ranges stratified by segments. The magnitude of motion for each individual patient was defined as the standard deviation (SD) of daily motion values stratified by segments. Correlations between the magnitude of motion and clinical variables were explored. RESULTS: The 2.5 to 97.5 percentiles of RL and AP motion were -4.2 to 7.1 and -4.4 to 5.1; -10.3 to 6.0 and -4.3 to 3.8; -8.7 to 5.5 and -6.4 to 2.8; and -9.1 to 4.7 and -5.8 to 3.3 mm for cervical, proximal, middle, and distal thoracic esophagus, respectively. The interfractional motion was direction- and location-dependent. The magnitude of RL motion was greater than that of AP motion for the 4 segments (P < .05). In the RL direction, the magnitude of motion was greater for the middle thoracic esophagus than for the cervical (median SD 2.7 vs 2.0 mm, P = .001) and proximal thoracic esophagus (median SD 2.7 vs 2.1 mm, P = .002). Patients with right lung tumor and bulky lymph nodes tended to display greater RL esophageal motion. CONCLUSIONS: The interfractional motion of the esophagus can be considerable during radiation therapy in locally advanced non-small cell lung cancer, especially for middle thoracic esophagus in RL direction. Strategies to minimize the effect of interfractional esophageal motion on dosimetry should be considered.

Author Correction: Whole-genome resequencing of 472 Vitis accessions for grapevine diversity and demographic history analyses.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Whole-genome resequencing of 472 Vitis accessions for grapevine diversity and demographic history analyses.

Understanding the Vitis species at the genomic level is important for cultivar improvement of grapevine. Here we report whole-genome genetic variation at single-base resolution of 472 Vitis accessions, which cover 48 out of 60 extant Vitis species from a wide geographic distribution. The variation helps to identify a recent dramatic expansion and contraction of effective population size in the domesticated grapevines and that cultivars from the pan-Black Sea region have a unique demographic history in comparison to the other domesticated cultivars. We also find selective sweeps for berry edibility and stress resistance improvement. Furthermore, we find associations between candidate genes and important agronomic traits, such as berry shape and aromatic compounds. These results demonstrate resource value of the resequencing data for illuminating the evolutionary biology of Vitis species and providing targets for grapevine genetic improvement.

Analyses of the factors influencing the accuracy of three-dimensional ultrasound in comparison with cone-beam CT in image-guided radiotherapy for prostate cancer with or without pelvic lymph node irradiation.

BACKGROUND: Three-dimensional ultrasound (3DUS) is an attractive option in image-guided radiotherapy (IGRT) for prostate cancer (PCa) patients. However, the potential factors influencing the accuracy of 3DUS in comparison with cone-beam CT (CBCT) in IGRT for PCa patients haven't been clearly identified. METHODS: The differences between US/US and CBCT/CT registrations were analyzed over 586 and 580 sessions for 24 and 25 PCa patients treated with or without pelvic lymph node irradiation, respectively. The clinical factors that may influence registration differences were also evaluated. RESULTS: The average discrepancies between US/US and CBCT/CT registrations were - 0.28 ± 5.28 mm, - 0.16 ± 3.48 mm, and - 0.47 ± 4.31 mm in the superior-inferior (SI), left-right (LR), and anterior-posterior (AP) directions, respectively. The discrepancies were respectively less than 5 mm longitudinally, laterally, and vertically in 64.4 and 70.1%, 84.9 and 89.2%, and 75.9 and 79.1% of the patients treated with or without pelvic lymph node irradiation, respectively. The registration differences were significantly smaller at least in one direction in patients younger than 70 years, without pelvic lymph node irradiation, guided by transperineal ultrasonography and had a bladder volume smaller than 300 mL. CONCLUSIONS: Age, irradiated regions, 3DUS modality, and bladder volume are important factors that may influence the differences between US/US and CBCT/CT registrations. 3DUS guidance is more feasible for younger PCa patients with a better control of bladder volume during the treatment and those who did not undergo pelvic lymph node irradiation.

Candidate Genes Involved in the Biosynthesis of Triterpenoid Saponins in Platycodon grandiflorum Identified by Transcriptome Analysis.

BACKGROUND: Platycodon grandiflorum is the only species in the genus Platycodon of the family Campanulaceae, which has been traditionally used as a medicinal plant for its lung-heat-clearing, antitussive, and expectorant properties in China, Japanese, and Korean. Oleanane-type triterpenoid saponins were the main chemical components of P. grandiflorum and platycodin D was the abundant and main bioactive component, but little is known about their biosynthesis in plants. Hence, P. grandiflorum is an ideal medicinal plant for studying the biosynthesis of Oleanane-type saponins. In addition, the genomic information of this important herbal plant is unavailable. PRINCIPAL FINDINGS: A total of 58,580,566 clean reads were obtained, which were assembled into 34,053 unigenes, with an average length of 936 bp and N50 of 1,661 bp by analyzing the transcriptome data of P. grandiflorum. Among these 34,053 unigenes, 22,409 unigenes (65.80%) were annotated based on the information available from public databases, including Nr, NCBI, Swiss-Prot, KOG, and KEGG. Furthermore, 21 candidate cytochrome P450 genes and 17 candidate UDP-glycosyltransferase genes most likely involved in triterpenoid saponins biosynthesis pathway were discovered from the transcriptome sequencing of P. grandiflorum. In addition, 10,626 SSRs were identified based on the transcriptome data, which would provide abundant candidates of molecular markers for genetic diversity and genetic map for this medicinal plant. CONCLUSION: The genomic data obtained from P. grandiflorum, especially the identification of putative genes involved in triterpenoid saponins biosynthesis pathway, will facilitate our understanding of the biosynthesis of triterpenoid saponins at molecular level.