A rare case of symptomatic congenital lobar emphysema in an adolescent female who underwent expectant management of a congenital pulmonary malformation: a case report.

Background: Asymptomatic congenital pulmonary airway malformations (CPAMs) and congenital lobar emphysema (CLE) may safely be observed, though little is known about how many patients later require surgery. The management of these lesions remains controversial, as limited research exists on which patients later require surgery. Notably, there are few reported cases where patients become symptomatic and require definitive treatment beyond childhood. Case Description: We present the rare case of a 17-year-old female who developed dyspnea on exertion associated with pre-syncope, following diagnosis of a CPAM at birth. She had undergone surveillance until 2 years of age. The decision to proceed with a left upper lobectomy via thoracotomy was made, for treatment of her symptoms. During the surgery, one lung ventilation suggested that oxygenation had likely limited to the patient's healthy lung, prior to intervention. The surgery was uncomplicated, and her recovery was uneventful. Pathology revealed CLE. Conclusions: This case highlights that congenital lung malformations should remain in the differential diagnosis of patients presenting in adolescence or adulthood with new onset respiratory symptoms. For patients with a history of untreated congenital lung pathology, it is important to consider the presence of adhesions and loss of domain in perioperative planning. This case also highlights the ongoing need for research on the prognostication of these lesions to better inform surveillance.

Endoscopic ultrasound classification for prediction of endoscopic submucosal dissection resectability: PREDICT classification.

Background and study aims The safety of endoscopic submucosal dissection (ESD) has been reported, and the risk of lymph node metastasis is low for colorectal cancer if depth of invasion is the only non-curative factor on histological evaluation. ESD is increasingly performed even if submucosal (SM) invasion is suspected. However, reports about endoscopic findings for the criteria to predict ESD resectability remain limited. Endoscopic ultrasound (EUS) can directly visualize the tomographic image of the gastrointestinal wall and may help predict ESD resectability. Therefore, we investigated the possibility of predicting ESD resectability using EUS. Patients and methods We compared the association between EUS findings and pathological results for gastric or colorectal lesions with suspected SM invasion using white light endoscopy between June 2020 and January 2023. EUS findings were grouped based on the status of the underlying the tumor, as follows: Type I, submucosal layer was observed with reproducibility; Type II, submucosal layer not fully visible; and Type III, submucosal layer disrupted and muscularis propria (MP) layer thickened. Results Forty-one gastric cancer and 22 colorectal cancer cases were analyzed. The proportions of pathological VM0 (no tumor exposed on any vertical margin) for ESD-resected specimens were 89% and 33% for Type I and II, respectively, ( P ≤ 0.01). The proportions of cancer involving MP or deeper were significantly higher for Type II/III than for Type I (41% vs 0%, P ≤ 0.01). Conclusions EUS may have an important role in predicting ESD resectability of gastric and colorectal cancers suspected of having SM invasion.

Epiglottic retraction is useful in diagnosis of exercise-induced laryngeal obstruction.

BACKGROUND: Exercise-induced laryngeal obstruction (EILO) is distinguished as upper airway obstruction accompanied by inspiratory dyspnoea and stridor during highly intensive exercises. Epiglottic retraction in the diagnosis of EILO has not been sufficiently explored. AIMS/OBJECTIVES: We highlight the importance of epiglottic retraction in patients with EILO by evaluation by several diagnostic methods for EILO. Consideration of epiglottic retraction may be important for accurate diagnosis of EILO. RESULTS: Epiglottic retraction could be observed in three patients by laryngoscopy during heavy breathing and in two patients by continuous laryngoscopy during exercise. Stridor occurred by the forward prolapse of the arytenoids, but not by epiglottic retraction. In comparison with three athletes from other sports, lung volume was significantly greater in four swimmers although it was not different related to depending on the existence of epiglottic retraction. CONCLUSIONS AND SIGNIFICANCE: Laryngoscopy during heavy breathing is suggested to be useful to detect the epiglottic retraction, which might be overlooked if only continuous laryngoscopy during exercise is used. Epiglottic retraction was not rare in Japanese swimmers' exercise-induced laryngeal obstruction in our cohort and it might be related to the greater lung volume.

The parallel narrative of RGF/GLV/CLEL peptide signalling.

Plant peptide families share distinct characteristics, and many members are in homologous signalling pathways controlling development and responses to external signals. The root meristem growth factor (RGF) peptides/GOLVEN (GLV)/CLAVATA3-ESR-related like (CLEL) are a family of short signalling peptides that are derived from a precursor protein and undergo post-translational modifications. Their role in root meristem development is well established and recent efforts have identified subtilase processing pathways and several downstream signalling components. This discovery has enabled the convergence of previously distinct pathways and enhanced our understanding of plant developmental processes. Here, we review the structure-function relationship of RGF peptides, the post-translational modification pathways, and the downstream signalling mechanisms and highlight components of these pathways that are known in non-RGF-mediated pathways.

An overview of S100 proteins and their functions in skin homeostasis, interface dermatitis conditions and other skin pathologies.

S100 proteins comprise a family of structurally related proteins that are calcium-sensitive. S100 proteins have been found to play various roles in regulation of cell apoptosis, cell proliferation and differentiation, cell migration and invasion, energy metabolism, calcium homeostasis, protein phosphorylation, anti-microbial activity and inflammation in a variety of cell types. While the specific function of many S100 proteins remains unknown, some of the S100 proteins serve as disease biomarkers as well as possible therapeutic targets in skin diseases. Interface dermatitis (ID) is a histopathological term that covers many different skin conditions including cutaneous lupus erythematosus, lichen planus, and dermatomyositis. These pathologies share similar histological features, which include basal cell vacuolization and lymphocytic infiltration at the dermal-epidermal junction. In this review, we summarize how the S100 protein family contributes to both homeostatic and inflammatory processes in the skin. We also highlight the role of S100 proteins in neuronal signalling, describing how this might contribute to neuroimmune interactions in ID and other skin pathologies. Last, we discuss what is known about the S100 family proteins as both biomarkers and potential treatment targets in specific pathologies.

Prognostic impact of deprivation on esophagogastroduodenoscopy outcome.

Background and study aims Socioeconomic deprivation has long been associated with many gastrointestinal diseases, yet its influence on esophagogastroduodenoscopy (EGD) diagnosis has not been evaluated. The aim of this study was to investigate the influence of deprivation on outcomes of EGD irrespective of referral reason. Patients and methods Two thousand consecutive patients presenting to four Health Boards in Wales beginning in June 2019 were studied retrospectively with deprivation scores calculated using the Wales Indices of Multiple Deprivation (WIMD). Patients were subclassified into quintiles for analysis (Q1 most, Q5 least deprived). Results Inhabitants of the most deprived areas were more likely to be diagnosed with peptic ulcer (Q1 7.9%, Q5 4.7%; odds ratio [OR] 0.498, P =0.018), severe esophagitis (LA4, Q1 2.7% v Q5 0%, OR 0.089, P 0.002), Helicobacter pylori infection (Q1 5.4%, Q5 1.7%; OR 0.284, P =0.002), but less likely to be diagnosed with Barrett's esophagus (Q1 6.3% v Q5 12.3%, OR 2.146, P =0.004) than those from the least deprived areas. New cancer diagnoses numbered 53 and were proportionately higher after presentation for urgent suspected cancer (USC, n=35, 4.6%) than for routine referrals (n=3, 0.6%, P < 0.001). Deprivation was associated with more advanced stage cancer (stage III Q1 16.7% v Q5 5.6%, OR 0.997, P =0.006: stage IV Q1 16.7% v Q2 38.9% v Q5 22.2%, OR 0.998, P =0.049). Conclusions Deprivation was associated with two-fold more peptic ulcer disease, three-fold more H. pylori infection, and 12-fold more severe esophagitis, and more advanced cancer stage.

Value of green sign and chicken skin aspects for detecting malignancy of colorectal neoplasia in a prospective characterization study.

Background and study aims Accurate endoscopic characterization of colorectal lesions is essential for predicting histology but is difficult even for experts. Simple criteria could help endoscopists to detect and predict malignancy. The aim of this study was to evaluate the value of the green sign and chicken skin aspects in detection of malignant colorectal neoplasia. Patients and methods We prospectively characterized and evaluated the histology of all consecutive colorectal lesions detected during screening or referred for endoscopic resection (Pro-CONECCT study). We evaluated the diagnostic accuracy of the green sign and chicken skin aspects for detection of superficial and deep invasive lesions. Results 461 patients with 803 colorectal lesions were included. The green sign had a negative predictive value of 89.6% (95% confidence interval [CI] 87.1%-91.8%) and 98.1% (95% CI 96.7%-99.0%) for superficial and deep invasive lesions, respectively. In contrast to chicken skin, the green sign showed additional value for detection of both lesion types compared with the CONECCT classification and chicken skin (adjusted odds ratio [OR] for superficial lesions 5.9; 95% CI 3.4-10.2; P <0.001), adjusted OR for deep lesions 9.0; 95% CI 3.9-21.1; P <0.001). Conclusions The green sign may be associated with malignant colorectal neoplasia. Targeting these areas before precise analysis of the lesion could be a way of improving detection of focal malignancies and prediction of the most severe histology.

Recurrent pneumonia in a 2-year-old: A rare case of congenital lobar emphysema.

INTRODUCTION AND SIGNIFICANCE: Emphysema is an uncommon but important condition that often appears in the neonatal period. Diagnosis is based on CT, which identifies the affected lung lobe, which is treated with complete surgical resection. CASE PRESENTATION: We present a case of a child who had been suffering for about a year from recurrent respiratory infections without arriving at a clear and correct diagnosis. He was evaluated by us and diagnosed correctly despite the difficulty of distinguishing it from pneumothorax. The final treatment was surgical removal. CLINICAL DISCUSSION: Emphysema is considered one of the important conditions that should be considered as a differential diagnosis if there is clear hyperinflated in the pulmonary lobe. The evaluation is mainly done through CT to reach the correct diagnosis and treatment. CONCLUSION: Congenital lobar emphysema is a rare condition that primarily affects children. The majority of children with CLE experience symptoms and necessitate surgery.

Usefulness of magnifying endoscopy for diagnosis of sessile serrated lesion with dysplasia or carcinoma: Large retrospective study.

Background and study aims Sessile serrated lesions (SSLs) are precursor lesions in the serrated neoplasia pathway that lead to invasive carcinoma from dysplasia arising from SSLs. This study aimed to elucidate the clinicopathological and endoscopic features of SSLs with and without dysplasia or carcinoma. Patients and methods We reviewed the clinicopathological and endoscopic data from all colorectal lesions pathologically diagnosed as SSLs at Juntendo University Hospital, Tokyo, Japan, between 2011 and 2022. In addition to conventional endoscopic findings, we retrospectively evaluated magnifying endoscopic findings with narrow-band imaging (NBI) or blue laser imaging (BLI) using the Japan NBI Expert Team system and analyzed pit patterns using magnified chromoendoscopic images. Results Of the 2,132 SSLs, 92.5%, 4.7%, 1.8%, and 0.9% had no dysplasia, low-grade dysplasia, high-grade dysplasia, and submucosal invasive carcinoma, respectively. Older age, the proximal colon, and larger lesions were more frequently associated with SSLs with dysplasia or carcinoma. However, 41.3% of the SSLs with dysplasia or carcinoma were ≤ 10 mm in size. Endoscopic findings, such as (semi)pedunculated morphology, double elevation, central depression, and reddishness, were frequently found in SSLs with dysplasia or carcinoma. Furthermore, magnifying endoscopy using NBI or BLI and magnifying chromoendoscopy showed high sensitivity, specificity, and accuracy for diagnosing dysplasia or carcinoma within SSLs. Conclusions SSLs with and without dysplasia or carcinoma exhibit distinct clinicopathological and endoscopic features. In an SSL series, conventional endoscopic characteristics in addition to use of magnifying endoscopy may be useful for accurately diagnosing advanced histology within an SSL.

Uncovering PheCLE1 and PheCLE10 Promoting Root Development Based on Genome-Wide Analysis.

Moso bamboo (Phyllostachys edulis), renowned for its rapid growth, is attributed to the dynamic changes in its apical meristem. The CLAVATA3/EMBRYO SURROUNDING REGION-RELATED (CLE) family genes are known to play crucial roles in regulating meristem and organ formation in model plants, but their functions in Moso bamboo remain unclear. Here, we conducted a genome-wide identification of the CLE gene family of Moso bamboo and investigated their gene structure, chromosomal localization, evolutionary relationships, and expression patterns. A total of 11 PheCLE genes were identified, all of which contained a conserved CLE peptide core functional motif (Motif 1) at their C-termini. Based on Arabidopsis classification criteria, these genes were predominantly distributed in Groups A-C. Collinearity analysis unveiled significant synteny among CLE genes in Moso bamboo, rice, and maize, implying potential functional conservation during monocot evolution. Transcriptomic analysis showed significant expression of these genes in the apical tissues of Moso bamboo, including root tips, shoot tips, rhizome buds, and flower buds. Particularly, single-cell transcriptomic data and in situ hybridization further corroborated the heightened expression of PheCLE1 and PheCLE10 in the apical tissue of basal roots. Additionally, the overexpression of PheCLE1 and PheCLE10 in rice markedly promoted root growth. PheCLE1 and PheCLE10 were both located on the cell membrane. Furthermore, the upstream transcription factors NAC9 and NAC6 exhibited binding affinity toward the promoters of PheCLE1 and PheCLE10, thereby facilitating their transcriptional activation. In summary, this study not only systematically identified the CLE gene family in Moso bamboo for the first time but also emphasized their central roles in apical tissue development. This provides a valuable theoretical foundation for the further exploration of functional peptides and their signaling regulatory networks in bamboo species.

High comorbidity rates in congenital lobar emphysema and the effect on clinical presentation.

The presence of comorbidities (CM) in congenital lobar emphysema (CLE) has been previously described with varying rates. However, the clinical implication of CM on the clinical presentation and patient outcome in CLE is unclear. This was a retrospective cohort study between 2000 and 2022 in a single institution. The study included pediatric patients diagnosed with CLE. During the study period, 36 patients were identified with CLE. The presence of respiratory symptoms at diagnosis was documented in 69% (24/35) of the patients with 48% admitted to intensive care units. The presence of CM was documented in 14/36 (39%) of the patients, with cardiovascular anomalies as the most common (22%), followed by genitourinary anomalies (11%) and large airway anomalies (8%). When comparing the CM group (n = 14) vs the non-CM group (N = 22), there was no significant difference regarding gender, term birth, age of diagnosis, and the location of the lesion. The CM group had significantly higher rates of intensive care unit (ICU) admissions (75% vs 33%, p = 0.02), longer duration of admission (median 46 days vs 9, p = 0.02), and need for pre-operative non-invasive respiratory support (50% vs 5%, p = 0.002).Conclusion: The presence of CM in CLE was associated with a more severe presentation as reflected with significantly higher rates of ICU admissions, prolonged admissions, and higher need for non-invasive respiratory support. Screening for CM, mostly for cardiac anomalies, is recommended at time of CLE diagnosis. What is Known: • Congenital lobar emphysema (CLE) is a developmental lung anomaly generally associated with other comorbidities; however, their clinical implication on presentation is unclear • The prevalence of comorbidities in CLE was previously reported around 15-20%, with cardiovascular anomalies as the most common What is New: • Comorbidities are very common (39%) in CLE, and their presence is related to a more severe clinical presentation and need for respiratory support • Screening for comorbidities is recommended in CLE, specifically for cardiovascular and genitourinary anomalies.

Novel physiological analysis using blood flow velocity for colonic polyps: Pilot study.

Real-time visualization of red blood cell flow inside subepithelial microvessels is performed with magnifying endoscopy. However, microvascular blood flow velocity in the colorectum has not been investigated. Here, we aimed to evaluate the blood flow velocity of microvessels of colonic polyps and to compare it with that of surrounding mucosa. We examined 50 lesions, including 30 adenomas (ADs) and 20 hyperplastic polyps (HPs). Blood flow velocities of lesions and their surrounding mucosa were evaluated using magnifying blue laser imaging (BLI) prior to endoscopic resection. Calculation of mean blood flow velocities was based on mean movement distance of one tagged red blood cell using split video images of magnifying BLI. Mean microvascular blood flow velocity was significantly lower in ADs (1.65±0.66 mm/sec; range 0.46-2.90) than in HPs (2.83±1.10 mm/sec; 1.07-4.50) or the surrounding mucosa (3.73±1.11 mm/sec; 1.80-6.20; P <0.001). The blood flow velocity rate compared with the surrounding mucosa was significantly lower in ADs (0.41±0.16; 0.10-0.82) than in HPs (0.89±0.25; 0.46-1.51; P <0.001). We found that mean microvascular blood flow velocity was significantly lower in ADs than in HPs and the surrounding non-neoplastic mucosa. These findings indicate that a novel dynamic approach with microvascular blood flow velocity using magnifying endoscopy may be useful in assessing physiological differences between ADs and HPs.

A rare case of congenital lobar emphysema presenting as recurrent pneumonia in early adulthood: A case report and literature review.

INTRODUCTION AND SIGNIFICANCE: Congenital lobar emphysema (CLE) is a rare but important lung malformation typically discovered in the newborn period. Some rare cases are reported in adults. It can present with various respiratory symptoms. Diagnosis relies primarily on chest CT scans, and the main treatment is surgery. CASE PRESENTATION: We present a case of a young girl with recurrent respiratory infections who was misdiagnosed with a pneumothorax. A chest tube was inserted. Later, CLE was identified, and the affected lung lobe was surgically removed. CLINICAL DISCUSSION: This case highlights the importance of including CLE in the differential diagnosis for hyperinflation of a lung lobe. A CT scan is crucial for confirmation. CONCLUSION: Congenital lobar emphysema is a rare disease that primarily affects children. Most children with CLE experience symptoms and require surgery. In adults, CLE is uncommon, and surgery is based on the severity of symptoms and radiological findings.

Whole-Genome Sequencing and Analysis of Tumour-Forming Radish (Raphanus sativus L.) Line.

Spontaneous tumour formation in higher plants can occur in the absence of pathogen invasion, depending on the plant genotype. Spontaneous tumour formation on the taproots is consistently observed in certain inbred lines of radish (Raphanus sativus var. radicula Pers.). In this paper, using Oxford Nanopore and Illumina technologies, we have sequenced the genomes of two closely related radish inbred lines that differ in their ability to spontaneously form tumours. We identified a large number of single nucleotide variants (amino acid substitutions, insertions or deletions, SNVs) that are likely to be associated with the spontaneous tumour formation. Among the genes involved in the trait, we have identified those that regulate the cell cycle, meristem activity, gene expression, and metabolism and signalling of phytohormones. After identifying the SNVs, we performed Sanger sequencing of amplicons corresponding to SNV-containing regions to validate our results. We then checked for the presence of SNVs in other tumour lines of the radish genetic collection and found the ERF118 gene, which had the SNVs in the majority of tumour lines. Furthermore, we performed the identification of the CLAVATA3/ESR (CLE) and WUSCHEL (WOX) genes and, as a result, identified two unique radish CLE genes which probably encode proteins with multiple CLE domains. The results obtained provide a basis for investigating the mechanisms of plant tumour formation and also for future genetic and genomic studies of radish.

The white ring sign is useful for differentiating between fundic gland polyps and gastric adenocarcinoma of the fundic gland type.

Background and study aims Gastric adenocarcinoma of the fundic gland type (GA-FG) is characterized by an elevated lesion with vessel dilation exhibiting branching architecture (DVBA). However, this feature is also found in fundic gland polyps (FGPs), posing a challenge in their differentiation. In this study, we aimed to investigate the clinicopathological features of gastric elevated lesions with DVBA and assess the efficacy of the white ring sign (WRS) as a novel marker for distinguishing between FGPs and GA-FGs. Methods We analyzed 159 gastric elevated lesions without DVBA and 51 gastric elevated lesions with DVBA, further dividing the latter into 39 in the WRS-positive group and 12 in the WRS-negative group. The clinicopathological features, diagnostic accuracy, and inter-rater reliability were analyzed. Results Univariate and multivariate analyses for gastric elevated lesions with DVBA identified the histological type consistent with FGPs and GA-FGs, along with the presence of round pits in the background gastric mucosa, as independent predictors. FGPs were present in 92.3% (36/39) of the WRS-positive group and GA-FGs were observed in 50.0% (6/12) of the WRS-negative group. WRS positivity and negativity exhibited high diagnostic accuracy, with 100% sensitivity, 80.0% specificity, and 94.1% accuracy for FGPs, and 100% sensitivity, 86.7% specificity, and 88.2% accuracy for GA-FGs. Kappa values for WRS between experts and nonexperts were 0.891 and 0.841, respectively, indicating excellent agreement. Conclusions WRS positivity and negativity demonstrate high diagnostic accuracy and inter-rater reliability for FGPs and GA-FGs, respectively, suggesting that WRS is a useful novel marker for distinguishing between FGPs and GA-FGs.

Outcomes in colorectal endoscopic submucosal dissection for large protruded lesions: A retrospective multicenter study.

Background and study aims Colorectal endoscopic submucosal dissection (ESD) is increasingly used for treating early-stage colorectal cancer, including large, protruded lesions (LPL). However, the challenges posed by LPLs, especially those accompanied by severe fibrosis or muscle-retracting sign (MRS), remain unclear. This study aims to investigate ESD outcomes for LPL, focusing on factors such as tumor size and, submucosal fibrosis. Patients and methods In a multicenter retrospective study (June 2012 to May 2023), data from 526 patients with 542 LPL lesions (≥ 2 cm) were analyzed. Parameters included lesion size, procedure time, dissection speed, physician experience, submucosal fibrosis, and adverse events. The tunnel method, including the double tunnel method, was used for cases with severe fibrosis or MRS. Multivariate analysis assessed factors affecting procedure difficulty, particularly LPLs ≥ 4 cm. Results The study revealed an impressive en bloc resection rate of 97.8% and a curative resection rate of 78.6% for LPLs. Notably, fibrosis and MRS were present in 25% and 18% of 4-cm LPLs, respectively, and their frequency tended to increase as the tumor diameter increased. One treatment strategy for LPLs was the tunneling method, which was used most frequently (41 cases, 7.6%). Factors affecting dissection speed included larger tumor size, submucosal fibrosis, MRS, and physician experience. Conclusions Treating LPLs through colorectal ESD presents significant challenges, especially in patients with fibrosis and MRS. This study highlights the importance of recognizing these complexities, and that more reliable resection strategy must be established for accurate pathological evaluation.

Identification and Application of CLE Peptides for Drought Resistance in Solanaceae Crops.

The CLE (CLAVATA3/Embryo Surrounding Region-related) family, a group of peptides with hormone-like features, plays a pivotal role in plant growth, development, and adaptation to stress. Through homology-based blast analysis of 32 Arabidopsis thaliana CLE peptide sequences, we have identified 5, 14, and 10 CLE family members in Nicotiana tabacum, Capsicum annuum, and Solanum melongena, respectively. Chemical synthesis and functional assays of the peptides led to the discovery that NtCLE3 substantially enhances the drought resistance of these three Solanaceae crops. Our transcriptome, RT-qPCR, and antioxidant enzyme activity data showed that NtCLE3 increased antioxidant capacity and ABA synthesis in tobacco. Moreover, the recombinant protein RPNtCLE3, composed of 6*NtCLE3, preserved the capacity to foster drought resilience and proved to be a promising drought resistance regulator, which presents a more favorable alternative for field applications compared to ABA which degrades rapidly under sunlight exposure. This research unveils the prospective utility of NtCLE3 in enhancing drought tolerance in Solanaceae crops and provides new ideas for the development of novel bioregulators aimed at mitigating drought stress.

The Compact Root Architecture 2 systemic pathway is required for the repression of cytokinins and miR399 accumulation in Medicago truncatula N-limited plants.

Legume plants can acquire mineral nitrogen (N) either through their roots or via a symbiotic interaction with N-fixing rhizobia bacteria housed in root nodules. To identify shoot-to-root systemic signals acting in Medicago truncatula plants at N deficit or N satiety, plants were grown in a split-root experimental design in which either high or low N was provided to half of the root system, allowing the analysis of systemic pathways independently of any local N response. Among the plant hormone families analyzed, the cytokinin trans-zeatin accumulated in plants at N satiety. Cytokinin application by petiole feeding led to inhibition of both root growth and nodulation. In addition, an exhaustive analysis of miRNAs revealed that miR2111 accumulates systemically under N deficit in both shoots and non-treated distant roots, whereas a miRNA related to inorganic phosphate (Pi) acquisition, miR399, accumulates in plants grown under N satiety. These two accumulation patterns are dependent on Compact Root Architecture 2 (CRA2), a receptor required for C-terminally Encoded Peptide (CEP) signaling. Constitutive ectopic expression of miR399 reduced nodule numbers and root biomass depending on Pi availability, suggesting that the miR399-dependent Pi-acquisition regulatory module controlled by N availability affects the development of the whole legume plant root system.