Essential contribution of CCL3 to alkali-induced corneal neovascularization by regulating vascular endothelial growth factor production by macrophages

金沢大学がん研究所がん病態制御Purpose: To evaluate the roles of CCL3 and its specific chemokine receptors, CCR1 and CCR5, in alkali-induced corneal neovascularization (CNV). Methods: Chemical denudation of corneal and limbal epithelium was performed on wild-type (WT) BALB/c mice and CCL3-, CCR1-, and CCR5-deficienct (knockout [KO]) counterparts. Two weeks after injury CNV was quantified by immunostaining with anti-CD31. Angiogenic factor expression and leukocyte accumulation in the early phase after injury were quantified by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical analysis, respectively. Results: Alkali injury augmented the intraocular mRNA expression of CCL3 and its receptors, CCR1 and CCR5, together with a transient infiltration of F4/80 positive macrophages and Gr-1 positive neutrophils. Compared with WT mice, CCL3-KO and CCR5-KO mice but not CCR1-KO mice exhibited reduced CNV two weeks after injury both macroscopically and microscopically as evidenced by CD31 positive areas. Concomitantly, the infiltration of F4/80 positive macrophages but not Gr-1 positive neutrophils was significantly attenuated in CCL3-KO mice compared with WT mice. Intracorneal infiltration of CCR5 expressing cells was significantly impaired in CCL3-KO mice compared with WT mice. Alkali injury induced a massive increase in the intraocular mRNA expression of a potent angiogenic factor, vascular endothelial growth factor (VEGF), in WT mice whereas these increments were severely retarded in CCL3-KO mice. Moreover, CCL3 enhanced VEGF expression by murine peritoneal macrophages at both the mRNA and the protein level. Furthermore, topical CCL3 application restored CNV, which was macroscopically and microscopically reduced in CCL3-KO mice after two weeks to levels similar to those found in WT mice. Conclusions: In alkali-induced CNV, CCL3 induced macrophages to infiltrate and produce VEGF by binding to CCR5 but not to CCR1 and eventually promoted angiogenesis. © 2008 Molecular Vision

BrLAS, a GRAS Transcription Factor From Brassica rapa, Is Involved in Drought Stress Tolerance in Transgenic Arabidopsis

GRAS proteins belong to a plant-specific transcription factor family and play roles in diverse physiological processes and environmental signals. In this study, we identified and characterized a GRAS transcription factor gene in Brassica rapa, BrLAS, an ortholog of Arabidopsis AtLAS. BrLAS was primarily expressed in the roots and axillary meristems, and localized exclusively in the nucleus of B. rapa protoplast cells. qRT-PCR analysis indicated that BrLAS was upregulated by exogenous abscisic acid (ABA) and abiotic stress treatment [polyethylene glycol (PEG), NaCl, and H2O2]. BrLAS-overexpressing Arabidopsis plants exhibited pleiotropic characteristics, including morphological changes, delayed bolting and flowering time, reduced fertility and delayed senescence. Transgenic plants also displayed significantly enhanced drought resistance with decreased accumulation of ROS and increased antioxidant enzyme activity under drought treatment compared with the wild-type. Increased sensitivity to exogenous ABA was also observed in the transgenic plants. qRT-PCR analysis further showed that expression of several genes involved in stress responses and associated with leaf senescence were also modified. These findings suggest that BrLAS encodes a stress-responsive GRASs transcription factor that positively regulates drought stress tolerance, suggesting a role in breeding programs aimed at improving drought tolerance in plants

BrRLP48, Encoding a Receptor-Like Protein, Involved in Downy Mildew Resistance in Brassica rapa

Downy mildew, caused by Hyaloperonospora parasitica, is a major disease of Brassica rapa that causes large economic losses in many B. rapa-growing regions of the world. The genotype used in this study was based on a double haploid population derived from a cross between the Chinese cabbage line BY and a European turnip line MM, susceptible and resistant to downy mildew, respectively. We initially located a locus Br-DM04 for downy mildew resistance in a region about 2.7 Mb on chromosome A04, which accounts for 22.3% of the phenotypic variation. Using a large F2 mapping population (1156 individuals) we further mapped Br-DM04 within a 160 kb region, containing 17 genes encoding proteins. Based on sequence annotations for these genes, four candidate genes related to disease resistance, BrLRR1, BrLRR2, BrRLP47, and BrRLP48 were identified. Overexpression of both BrRLP47 and BrRLP48 using a transient expression system significantly enhanced the downy mildew resistance of the susceptible line BY. But only the leaves infiltrated with RNAi construct of BrRLP48 could significantly reduce the disease resistance in resistant line MM. Furthermore, promoter sequence analysis showed that one salicylic acid (SA) and two jasmonic acid-responsive transcript elements were found in BrRLP48 from the resistant line, but not in the susceptible one. Real-time PCR analysis showed that the expression level of BrRLP48 was significantly induced by inoculation with downy mildew or SA treatment in the resistant line MM. Based on these findings, we concluded that BrRLP48 was involved in disease resistant response and the disease-inducible expression of BrRLP48 contributed to the downy mildew resistance. These findings led to a new understanding of the mechanisms of resistance and lay the foundation for marker-assisted selection to improve downy mildew resistance in Brassica rapa

Ubiquitin-Proteasome-Collagen (CUP) Pathway in Preterm Premature Rupture of Fetal Membranes

Spontaneous preterm birth (sPTB) occurs before 37 gestational weeks, with preterm premature rupture of the membranes (PPROM) and spontaneous preterm labor (sPTL) as the predominant adverse outcomes. Previously, we identified altered expression of long non-coding RNAs (lncRNAs) and message RNAs (mRNAs) related to the ubiquitin proteasome system (UPS) in human placentas following pregnancy loss and PTB. We therefore hypothesized that similar mechanisms might underlie PPROM. In the current study, nine pairs of ubiquitin-proteasome-collagen (CUP) pathway–related mRNAs and associated lncRNAs were found to be differentially expressed in PPROM and sPTL. Pathway analysis showed that the functions of their protein products were inter-connected by ring finger protein. Twenty variants including five mutations were identified in CUP-related genes in sPTL samples. Copy number variations were found in COL19A1, COL28A1, COL5A1, and UBAP2 of sPTL samples. The results reinforced our previous findings and indicated the association of the CUP pathway with the development of sPTL and PPROM. This association was due not only to the genetic variation, but also to the epigenetic regulatory function of lncRNAs. Furthermore, the findings suggested that the loss of collagen content in PPROM could result from degradation and/or suppressed expression of collagens

Locally advanced rectal cancer with dMMR/MSI-H may be excused from surgery after neoadjuvant anti-PD-1 monotherapy: a multiple-center, cohort study

ObjectiveExamine patients with locally advanced rectal cancer (LARC) with deficient mismatch repair (dMMR) or microsatellite instability-high (MSI-H) who received neoadjuvant immunotherapy (nIT), and compare the outcomes of those who chose a watch-and-wait (WW) approach after achieving clinical complete response (cCR) or near-cCR with those who underwent surgery and were confirmed as pathological complete response (pCR).MethodsLARC patients with dMMR/MSI-H who received nIT were retrospectively examined. The endpoints were 2-year overall survival (OS), 2-year disease-free survival (DFS), local recurrence (LR), and distant metastasis (DM). The efficacy of programmed cell death protein-1 (PD-1) inhibitor, immune-related adverse events (irAEs), surgery-related adverse events (srAEs), and enterostomy were also recorded.ResultsTwenty patients who received a PD-1 inhibitor as initial nIT were examined. Eighteen patients (90%) achieved complete response (CR) after a median of 7 nIT cycles, including 11 with pCR after surgery (pCR group), and 7 chose a WW strategy after evaluation as cCR or near-cCR (WW group). Both groups had median follow-up times of 25.0 months. Neither group had a case of LR or DM, and the 2-year DFS and OS in each group was 100%. The two groups had similar incidences of irAEs (P=0.627). In the pCR group, however, 2 patients (18.2%) had permanent colostomy, 3 (27.3%) had temporary ileostomy, and 2 (18.2%) had srAEs.ConclusionNeoadjuvant PD-1 blockade had high efficacy and led to a high rate of CR in LARC patients with dMMR/MSI-H. A WW strategy appears to be a safe and reliable option for these patients who achieve cCR or near-cCR after nIT

Image Object Extraction Based on Semantic Segmentation and Label Loss

Object extraction refers to the operation of obtaining an object area from an image based on a small amount of mark information given by users, which is a key step in image processing. In order to obtain a complete object profile, current methods usually require a large number of manual annotations, especially for objects with irregular contours. Since traditional algorithms rely on low-level pixel features without semantic information, and are based on obvious mathematical assumptions (ie, strong inductive bias), it is difficult to completely identify objects. At present, in order to improve the integrity of object extraction, semantic segmentation-based methods increase the complexity and latancy by adding more pre-processing and post-processing steps. In this paper, we propose a novel model named IOEBSS, which includes a fast binary plane pre-processing, an improved Deeplab v3+ semantic segmentation model, and an auxiliary loss function named Label Loss. Through the fast binary plane pre-processing, the model can accelerate the transformation of interactive inputs. The improved semantic segmentation model makes the extracted results more semantically complete, and Label Loss is more conducive to gradient flow and accelerates training convergence. For the above reasons, IOEBSS can accurately and quickly identify objects with complex contours and colors. On Pascal VOC and COCO datasets, compared to current methods, IOEBSS has a significant improvement in accuracy, inference speed, and convergence speed

Numerical Simulation of Soil Water–Salt Dynamics and Agricultural Production in Reclaiming Coastal Areas Using Subsurface Pipe Drainage

Soil salinization induced by shallow saline groundwater in coastal areas can be managed using subsurface pipe drainage (SPD) for agricultural land reclamation. However, a reasonable SPD system layout should comprehensively consider local hydrological conditions and crop physiological characteristics based on long-term model evaluations. The objectives of this study were to test the applicability of a crop growth model (AquaCrop) for simulating winter wheat growth in SPD-applied fields by employing the water table behaviors predicted by the soil hydrologic model HYDRUS. Model calibration and validation based on field observations suggested that HYDRUS accurately predicted the distributions of soil water–salt dynamics, and the seasonal variations of canopy cover and biomass production predicted by AquaCrop were close to the measured values. The simulation scenarios considering the long-term effect of groundwater salinity (10.53, 21.06, and 31.59 g L−1 for low, medium, and high levels), drain spacing (10, 20, 30, 40 m, and no-SPD), and precipitation category (dry, normal, and wet year) on soil solute transport, grain yield (GY), water productivity (WP), and groundwater supply (GS) were further explored using a combination of HYDRUS and AquaCrop. The simulation results indicated that narrowing the drain spacing could improve the desalination performance of SPD, but there was no continuous downward trend of soil solute concentration during the long-term application of SPD when groundwater salinity was constant. The SPD application could improve grain yield by 0.81–1.65 t ha−1, water productivity by 0.13–0.35 kg m−3, and groundwater supply by 6.06–31.03 mm compared to the no-SPD scenarios, but such increases would be less pronounced in dry years with groundwater salinity at the low level. This study demonstrated that the co-application of hydrologic and crop growth models is a feasible method for revealing the effects of SPD on agricultural land reclamation in coastal areas

Evaluating Soil Water–Salt Dynamics under Brackish Water Drip Irrigation in Greenhouses Subjected to Localized Topsoil Compaction

Localized soil compaction in greenhouses resulting from less frequent tillage operations and frequent trampling by farmers inevitably disturbs the continuity and homogeneity of soil’s hydraulic properties, which impacts the precision of greenhouse cultivation regarding water supply and salinity control. However, predicting water–salt dynamics under partly compacted topsoil is difficult because of the interactions between many factors related to soil properties, including irrigation method and water quality, which are especially subjected to varied compaction sizes and positions. Here, two field treatments were conducted in brackish water (3 g L−1) drip-irrigated plots, with the designed soil compaction region (40 cm width and 30 cm depth) adjacent to (T1) and below (T2) the drip lines. The calibrated and validated HYDRUS-2D model was applied to analyze salt exchanges across the vertical and horizontal interfaces between the compacted and non-compacted zones and the associated solute concentration variations within these two zones. The results indicated that the limited horizontal solute flux under T1 enhanced the subsequent downward flux below the drip lines, whereas, under T2, the restricted downward flux with relatively limited improved horizontal salt spreading resulted in more salt retention in the soil profile. Additional scenario simulations considering the vertical and horizontal extension of soil compaction sizes (ranging from 10 × 10 cm to 40 × 40 cm) were also conducted and revealed that, with the same increment in compaction size, the vertical extension of the compacted zone aggravated salt accumulation compared with that of horizontal extension, while the simulated cumulative water and salt downward fluxes were positive in relation to the compaction sizes in both vertical and horizontal directions under T1, but negative under T2. The findings of this study explore the effect of relative positions between drip lines and the soil compaction zone on salt transports under brackish water irrigation and reveal the potential soil salinization trend as extending compaction regions in the vertical or horizontal direction