Potential Regulatory Networks and Heterosis for Flavonoid and Terpenoid Contents in Pak Choi: Metabolomic and Transcriptome Analyses.

Pak choi exhibits a diverse color range and serves as a rich source of flavonoids and terpenoids. However, the mechanisms underlying the heterosis and coordinated regulation of these compounds-particularly isorhamnetin-remain unclear. This study involved three hybrid combinations and the detection of 528 metabolites from all combinations, including 26 flavonoids and 88 terpenoids, through untargeted metabolomics. Analysis of differential metabolites indicated that the heterosis for the flavonoid and terpenoid contents was parent-dependent, and positive heterosis was observed for isorhamnetin in the two hybrid combinations (SZQ, 002 and HMG, ZMG). Moreover, there was a high transcription level of flavone 3'-O-methyltransferase, which is involved in isorhamnetin biosynthesis. The third group was considered the ideal hybrid combination for investigating the heterosis of flavonoid and terpenoid contents. Transcriptome analysis identified a total of 12,652 DEGs (TPM > 1) in various groups that were used for comparison, and DEGs encoding enzymes involved in various categories, including "carotenoid bio-synthesis" and "anthocyanin biosynthesis", were enriched in the hybrid combination (SZQ, 002). Moreover, the category of anthocyanin biosynthesis also was enriched in the hybrid combination (HMG, ZMG). The flavonoid pathway demonstrated more differential metabolites than the terpenoid pathway did. The WGCNA demonstrated notable positive correlations between the dark-green modules and many flavonoids and terpenoids. Moreover, there were 23 ERF genes in the co-expression network (r ≥ 0.90 and p < 0.05). Thus, ERF genes may play a significant role in regulating flavonoid and terpenoid biosynthesis. These findings enhance our understanding of the heterosis and coordinated regulation of flavonoid and terpenoid biosynthesis in pak choi, offering insights for genomics-based breeding improvements.

BrABF3 promotes flowering through the direct activation of CONSTANS transcription in pak choi.

Drought stress triggers the accumulation of the phytohormone abscisic acid (ABA), which in turn activates the expression of the floral integrator gene CONSTANS (CO), accelerating flowering. However, the molecular mechanism of ABA-induced CO activation remains elusive. Here, we conducted a yeast one-hybrid assay using the CO promoter from Brassica campestris (syn. Brassica rapa) ssp. chinensis (pak choi) to screen the ABA-induced pak choi library and identified the transcription activator ABF3 (BrABF3). BrABF3, the expression of which was induced by ABA in pak choi, directly bound to the CO promoter from both pak choi and Arabidopsis. The BrABF3 promoter is specifically active in the Arabidopsis leaf vascular tissue, where CO is mainly expressed. Impaired BrABF3 expression in pak choi decreased BrCO expression levels and delayed flowering, whereas ectopic expression of BrABF3 in Arabidopsis increased CO expression and induced earlier flowering under the long-day conditions. Electrophoretic mobility shift assay analysis showed that BrABF3 was enriched at the canonical ABA-responsive element-ABRE binding factor (ABRE-ABF) binding motifs of the BrCO promoter. The direct binding of BrABF3 to the ABRE elements of CO was further confirmed by chromatin immunoprecipitation quantitative PCR. In addition, the induction of BrCO transcription by BrABF3 could be repressed by BrCDF1 in the morning. Thus, our results suggest that ABA could accelerate the floral transition by directly activating BrCO transcription through BrABF3 in pak choi.

BcNAC056 Interacts with BcWRKY1 to Regulate Leaf Senescence in Pak Choi.

Senescence is the final stage of leaf development. For leafy vegetables such as pak choi, leaf senescence is adverse to yield due to the harvest period shortening. However, the regulatory mechanisms of leaf senescence are largely unknown in leafy vegetables. Here, we isolated and characterized a NAC gene, BcNAC056, in pak choi [Brassica campestris (syn. Brassica rapa) ssp. chinensis cv. 49caixin]. BcNAC056-GFP was located in the nucleus at the subcellular level, and BcNAC056 was responsive to leaf senescence and different hormones at the transcriptional level. Heterologous overexpression of BcNAC056 in Arabidopsis promoted leaf senescence, accompanied by the increased expression of senescence-associated genes (SAGs), whereas virus-induced gene silencing-based silencing in pak choi delayed leaf senescence. The following transcriptome analysis showed that heterologous overexpression of BcNAC056 enhanced some AtSAG transcripts in Arabidopsis. Electrophoretic mobility shift assay (EMSA) and dual-luciferase (LUC) reporter assay revealed that BcNAC056 activated SAG12 by directly binding to the promoter. In addition, with the LUC reporter and transient overexpression assays, we proposed that BcNAC056-BcWRKY1 interaction promoted the activation of BcSAG12. Taken together, our findings revealed a new regulatory mechanism of leaf senescence in pak choi.

Diversity, Biosynthesis and Bioactivity of Aeruginosins, a Family of Cyanobacteria-Derived Nonribosomal Linear Tetrapeptides.

Aeruginosins, a family of nonribosomal linear tetrapeptides discovered from cyanobacteria and sponges, exhibit in vitro inhibitory activity on various types of serine proteases. This family is characterized by the existence of the 2-carboxy-6-hydroxy-octahydroindole (Choi) moiety occupied at the central position of the tetrapeptide. Aeruginosins have attracted much attention due to their special structures and unique bioactivities. Although many studies on aeruginosins have been published, there has not yet been a comprehensive review that summarizes the diverse research ranging from biogenesis, structural characterization and biosynthesis to bioactivity. In this review, we provide an overview of the source, chemical structure as well as spectrum of bioactivities of aeruginosins. Furthermore, possible opportunities for future research and development of aeruginosins were discussed.

Metabolic and Transcriptomic Analyses Reveal Different Metabolite Biosynthesis Profiles between Purple and Green Pak Choi.

Pak choi is one of the most important leafy vegetables planted in East Asia and provides essential nutrients for the human body. Purple pak choi differs mainly in leaf colour but exhibits distinct nutritional profiles from green pak choi. In this study, we performed metabolic and transcriptomic analyses to uncover the mechanisms underlying the differences in metabolite biosynthesis profiles between the two pak choi varieties. Metabolite profiling revealed significant differences in the levels of metabolites, mainly amino acids and their derivatives and flavonoids. Furthermore, 34 flavonoids significantly differed between green and purple pak choi leaves, and cyanidin and its derivative anthocyanins were abundant in purple pak choi. In addition, we found that the structural genes CHS, DFR, ANS, and UGT75C1, as well as the transcription factor MYB2, play a major role in anthocyanin synthesis. These results provide insight into the molecular mechanisms underlying leaf pigmentation in pak choi and offer a platform for assessing related varieties.

Effect of Plasma-Treated Water with Magnesium and Zinc on Growth of Chinese Cabbage.

Nonthermal biocompatible plasma (NBP) is an emerging technology in the field of agriculture to boost plant growth. Plasma is a source of various gaseous reactive oxygen and nitrogen species (RONS) and has a promising role in agricultural applications, as the long-lived RONS (H2O2, NO2-, NO3-) in liquid activate signaling molecules in plant metabolism. Plasma-treated water (PTW) has an acidic pH of around 3 to 4, which may be detrimental to pH-sensitive plants. Innovative techniques for producing PTW with a pH value of 6 to 7 under neutral circumstances are desperately required to broaden the application range of NBP in agriculture. Furthermore, Pak Choi (Brassica campestris L.) is a Brassicaceae family green vegetable that has yet to be investigated for its response to NBP. In this work, we proposed an alternate method for neutralizing the pH of PTW by immersing metal ions (Mg2+ and Zn2+) in the PTW and observing its effect on Pak Choi. After synthesizing PTW with MECDBD, we analyzed germination rate and growth parameters, then seedlings for 42 days to show physiological, biochemical, and molecular levels. The germination rate was observed to be higher with PTW and more efficient when metal ions were present. Seedling length and germination rates were dramatically boosted when compared to DI water irrigation. Because of the increased chlorophyll and protein content, the plants responded to the availability of nitrogen by generating highly green leaves. Furthermore, we observed that PTW increases the expression of NR genes and GLR1 genes, which are further increased when metals are submerged in the PTW. Furthermore, PTW and PTW with metals reduced ABI5 and CHO1 which is associated with a growth inhibitor. According to this study, nonthermal plasma might be utilized to significantly improve seed germination and seedlings' development.

BcWRKY1 confers salt sensitivity via inhibiting Reactive oxygen species scavenging.

WRKY transcription factors play important roles in abiotic stress by directly regulating stress-related genes. However, the molecular mechanism of its involvement in salt stress in pak-choi is still poorly understood. In this study, we elucidated the function of BcWRKY1 from pak-choi (Brassica rapa ssp. chinensis) in salt stress. The expression level of BcWRKY1 showed the highest in rosette leaves among different tissues and was induced by salt and ABA treatment in pak-choi. Subcellular localization showed that BcWRKY1 was located in nucleus. The transgenic Arabidopsis overexpressing BcWRKY1 exhibited enhanced salt sensitivity and higher H2O2 contents, which were further confirmed by silencing BcWRKY1 in pak-choi. In addition, the expression of ZAT12 was negatively regulated with BcWRKY1 under salt stress both in pak-choi and Arabidopsis. Yeast one-hybrid and dual luciferase reporter assay showed that BcWRKY1 could bind to the promoter of BcZAT12, and BcsAPX expression was activated by BcZAT12. To sum up, we propose a BcWRKY1-BcZAT12-BcsAPX regulatory model that involves in pak-choi salt stress response.

Comparison of Complications after Coronavirus Disease and Seasonal Influenza, South Korea.

We conducted a retrospective cohort study using claims data to determine the number and types of complications from coronavirus disease (COVID-19) that patients experience and which patients are more vulnerable to those complications compared with complications in patients with influenza. Among the cohort, 19.6% of COVID-19 patients and 28.5% of influenza patients had >1 new complication. In most complications, COVID-19 patients had lower or similar relative risk compared with influenza patients; exceptions were hair loss, heart failure, mood disorder, and dementia. Young to middle-aged adult COVID-19 patients and patients in COVID-19 hotspots had a higher risk for complications. Overall, COVID-19 patients had fewer complications than influenza patients, but caution is necessary in high-risk groups. If the fatality rate for COVID-19 is reduced through vaccination, management strategies for this disease could be adapted, similar to those for influenza management, such as easing restrictions on economic activity or requirements for close-contact isolation.

Exploratory analysis of tumor imaging in a Phase 2 trial with cabozantinib in gastrointestinal stromal tumor: lessons learned from study EORTC STBSG 1317 'CaboGIST'.

BACKGROUND: Gastrointestinal stromal tumors (GISTs) are malignant mesenchymal tumors arising in the gastrointestinal tract. Their systemic treatment is based on the use of tyrosine kinase inhibitors (TKIs) with imatinib, sunitinib, and regorafenib being the preferred agents. Assessment of tumor response to TKI treatment in GISTs is traditionally done according the Response Evaluation Criteria in Solid Tumors (RECIST), while Choi criteria have also been proposed as alternative tool assessing both volumetric and density changes on computer tomography (CT) scans. EORTC STBSG 1317 'CaboGIST' was a single-arm prospective Phase 2 trial which met its primary endpoint, as 60% of patients previously treated with imatinib and sunitinib were progression-free at 12 weeks (95% CI 45-74%) based on local RECIST assessment. MATERIALS AND METHODS: We report here an exploratory analysis of local versus central RECIST version 1.1 assessment and a comparison of RECIST version 1.1 versus Choi criteria. RESULTS: Comparisons between local and central RECIST version 1.1 at week 12 revealed discrepancies in 17/43 evaluable cases (39.5%). When comparing Choi with local and central RECIST version 1.1, discrepancies were observed in 27/43 (62.8%) and 21/43 (48.8%) cases, respectively. A total of 68% of evaluable patients were progression-free and alive at week 12 based on local RECIST, 84% according to central RECIST analysis and 81% when applying Choi criteria. Central assessment upgraded the treatment response both with RECIST version 1.1 and Choi. CONCLUSIONS: The results of this exploratory analysis support the conclusion that cabozantinib is active in patients with metastatic or recurrent GIST after treatment with imatinib and sunitinib and confirm once again the limitations of RECIST to capture response to TKI in GIST, and the importance to include density changes in the response evaluation in this setting. Clinical trial number: EORTC 1317, NCT02216578.

Foliar Application of Reaction Products Derived from Selenite Removal by Iron Monosulfide for Brassica rapa ssp. Chinensis L.

The extensive application of FeS in environmental remediation requires the recovery and reuse of reaction products between FeS and pollutants. Therefore, foliar application of reaction products derived from selenite [Se(IV)] removal by FeS for pak choi was performed. The removal rate of Se(IV) by 100 mg/L FeS was 0.047 h-1. 93.2% of Se(IV) was reduced to Se(0), and FeS was correspondingly oxidized to goethite (78.9%), lepidocrocite (21.1%), and S(0) (91.5%) based on the analysis of X-ray absorption fine structure. The reaction products promoted the growth of pak choi in terms of fresh biomass, vitamin C, and protein, ascribed to the key roles of Fe and S in enhancing the electron transfer rate and light conversion rate. Furthermore, the application of reaction products decreased by 64% of disease incidence as compared with the pathogen Pseudomonas syringae pv. maculicola-infected control. The total Se content in plants increased to 576 µg/kg and was composed of 11.9% of SeMeCys, 29.8% of SeMet, and 58.3% of SeCys after exposure to reaction products, which is beneficial to the human dietary intake from pak choi. This study demonstrated that the reaction products between FeS and Se(IV) could be recovered and applied as a nano-enabled strategy to prevent crop insecurity.

Effects of 35% hydrogen peroxide solution containing hydrated calcium silicate on enamel surface.

OBJECTIVES: The objectives of this study were to develop a novel bleaching material containing hydrated calcium silicate (hCS) particles and investigate the effects of hCS on the bleaching efficacy, microhardness, and surface morphology of bovine enamel. MATERIALS AND METHODS: To prepare the hCS particles, white Portland cement was mixed with distilled water and ground into a fine powder. The particles in various proportions were then mixed with 35% hydrogen peroxide solution (HP), while HP without hCS was used as a control (HP), and teeth whitening gel was used as a commercial control (CC). Following the thrice application of experimental and control solutions on the discolored bovine enamel surface for 15 min, color change (n = 10), microhardness (n = 10), and micromorphology (n = 2) of the enamel surface were analyzed. RESULTS: The Δ E* of the enamel surface treated with the experimental solution containing hCS was significantly higher than that of the CC, but there were no significant differences between the different hCS contents. The experimental solution containing hCS reduced the percentage of microhardness loss on the enamel surface, and the percentage of microhardness loss significantly decreased as the content of hCS increased (p < 0.05). The erosion pattern was only observed on enamel surfaces treated with HP and CC. CONCLUSIONS: This study suggests that HP containing hCS is effective in bleaching efficacy. In addition, hCS could also minimize the microhardness loss of tooth structure caused by HP and maintain enamel surface morphology. CLINICAL RELEVANCE: This novel bleaching material is promising for inhibiting demineralization and promoting the remineralization of teeth during bleaching treatment in dental clinics.

Enhanced photosynthetic activity in pak choi hybrids is associated with increased grana thylakoids in chloroplasts.

Increased photosynthetic activity is closely linked to heterosis in plants, but the underlying molecular mechanisms remain elusive. Pak choi (Brassica rapa ssp. chinensis) is a widely grown vegetable in Asia, and the most commercial cultivars are F1 hybrids. Here, the inbred pak choi lines WTC and 2Q, and their reciprocal F1 hybrids WQ and QW, were used to characterize the increased photosynthetic activity in these hybrids at the physiological, cellular and molecular levels. We found that the hybrids had larger leaves, with more grana thylakoids. Additionally, these hybrids had significantly increased net photosynthetic rates (Pn ) under both saturating and low irradiance conditions. These data indicate that the increased photosynthetic activity in pak choi hybrids was associated with an improved photosynthetic mechanism and larger leaves. Next, we obtained genome-wide data using transcriptome and bisulfite sequencing. Gene ontology (GO) analysis showed that the differentially expressed genes among the parents and hybrids were mostly enriched in the 'photosynthesis', 'thylakoid', and 'chloroplast' categories, indicating that the increased number of grana thylakoids contributes to the enhanced photosynthetic capacity in hybrids. Furthermore, we found that the increased number of grana thylakoids was associated with the upregulation of light-harvesting complex of photosystem II 1 (BrLhcb1). Yeast one-hybrid and transient assay showed that the BrLhcb1 promoter was directly bound by CIRCADIAN CLOCK ASSOCIATED 1 (BrCCA1), resulting in increased BrLhcb1 expression and enhanced carbon fixation in hybrids. Finally, our findings provide new insight into molecular mechanisms underlying enhanced photosynthesis in pak choi hybrids.

Efficacy of axitinib in metastatic head and neck cancer with novel radiographic response criteria.

BACKGROUND: There are limited treatment options for unresectable recurrent or metastatic (R/M) head and neck squamous cell carcinoma (HNSCC). Vascular endothelial growth factor is of significant interest for targeted therapy in R/M HNSCC because of its central role in tumorigenesis and immunosuppression. Axitinib is a potent inhibitor of vascular endothelial growth factor receptor (VEGFR) 1 , VEGFR2, VEGFR3, platelet-derived growth factor receptor, as well as c-kit and offers such an approach. METHODS: This article reports the results of a phase 2 trial evaluating axitinib in R/M HNSCC according to the Choi criteria for radiographic response assessment. The primary endpoint of this trial was 6-month overall survival. RESULTS: Twenty-nine patients were enrolled, and 28 were evaluable for a response. Patients were heavily pretreated with 61% having had at least 1 previous systemic treatment in the metastatic setting (range, 0-5). The median overall survival of 9.8 months and the 6-month overall survival rate of 70% met the protocol-defined criteria for clinical efficacy. The best overall response rate was 42%. Correlative analyses demonstrated that PI3K signaling pathway alterations were associated with an increased response to therapy (75% vs 17%). A marked response to therapy was seen in a subgroup of patients who were treated with an immune checkpoint inhibitor after progression on axitinib. CONCLUSIONS: Treatment with axitinib is associated with improved survival in patients with heavily pretreated head and neck cancer, and PI3K pathway alterations may serve as a biomarker for response. Further investigation is warranted to evaluate axitinib in biomarker-selected populations, especially in combination with immune checkpoint inhibitor therapy. LAY SUMMARY: Metastatic head and neck squamous cancer is an incurable disease with limited treatment options and a poor prognosis. This study is the first to demonstrate that the targeted oral drug axitinib improves survival in patients with heavily pretreated metastatic head and neck cancer. Furthermore, patients whose tumors have specific mutations derive the greatest benefit from therapy. The investigation of axitinib alone or in combination with immunotherapy in a genomic biomarker-selected population is warranted.

Assembly of the non-heading pak choi genome and comparison with the genomes of heading Chinese cabbage and the oilseed yellow sarson.

Brassica rapa displays a wide range of morphological diversity which is exploited for a variety of food crops. Here we present a high-quality genome assembly for pak choi (Brassica rapa L. subsp. chinensis), an important non-heading leafy vegetable, and comparison with the genomes of heading type Chinese cabbage and the oilseed form, yellow sarson. Gene presence-absence variation (PAV) and genomic structural variations (SV) were identified, together with single nucleotide polymorphisms (SNPs). The structure and expression of genes for leaf morphology and flowering were compared between the three morphotypes revealing candidate genes for these traits in B. rapa. The pak choi genome assembly and its comparison with other B. rapa genome assemblies provides a valuable resource for the genetic improvement of this important vegetable crop and as a model to understand the diversity of morphological variation across Brassica species.

Quantitative imaging decision support (QIDSTM) tool consistency evaluation and radiomic analysis by means of 594 metrics in lung carcinoma on chest CT scan.

OBJECTIVE: To evaluate the consistency of the quantitative imaging decision support (QIDSTM) tool and radiomic analysis using 594 metrics in lung carcinoma on chest CT scan. MATERIALS AND METHODS: We included, retrospectively, 150 patients with histologically confirmed lung cancer who underwent chemotherapy and baseline and follow-ups CT scans. Using the QIDSTM platform, 3 radiologists segmented each lesion and automatically collected the longest diameter and the density mean value. Inter-observer variability, Bland Altman analysis and Spearman's correlation coefficient were performed. QIDSTM tool consistency was assessed in terms of agreement rate in the treatment response classification. Kruskal Wallis test and the least absolute shrinkage and selection operator (LASSO) method with 10-fold cross validation were used to identify radiomic metrics correlated with lesion size change. RESULTS: Good and significant correlation was obtained between the measurements of largest diameter and of density among the QIDSTM tool and the radiologists measurements. Inter-observer variability values were over 0.85. HealthMyne QIDSTM tool quantitative volumetric delineation was consistent and matched with each radiologist measurement considering the RECIST classification (80-84%) while a lower concordance among QIDSTM and the radiologists CHOI classification was observed (58-63%). Among 594 extracted metrics, significant and robust predictors of RECIST response were energy, histogram entropy and uniformity, Kurtosis, coronal long axis, longest planar diameter, surface, Neighborhood Grey-Level Different Matrix (NGLDM) dependence nonuniformity and low dependence emphasis as Volume, entropy of Log(2.5 mm), wavelet energy, deviation and root man squared. CONCLUSION: In conclusion, we demonstrated that HealthMyne quantitative volumetric delineation was consistent and that several radiomic metrics extracted by QIDSTM were significant and robust predictors of RECIST response.

Plasma membrane-localized protein BcHIPP16 promotes the uptake of copper and cadmium in planta.

Cadmium (Cd) is one of the toxic heavy metals in soil, which not only suppresses crop production but also threatens human health. In this study, we aim to clarify the biological function of Cd-related gene BcHIPP16, so as to provide potential genetic solutions to decrease the Cd levels of pak choi. Tissue expression analysis showed that BcHIPP16 expressed in almost all the plant bodies. The transcriptional level of BcHIPP16 in roots was higher than that in shoots, which was significantly induced by copper (Cu) deficiency and Cd exposure conditions. Subcellular localization revealed that BcHIPP16 localized in plasma membrane. Expressing BcHIPP16 in yeast cells improved the sensitivity to Cu and Cd and improved their accumulation in yeast. Furthermore, the Cu and Cd content of Arabidopsis seedlings were increased and complemented, respectively when expressing BcHIPP16 in wild type (WT) and hip16 mutants. Non-invasive Micro-test Technology (NMT) was used to measure the real-time Cd2+ influx from the root surface of BcHIPP16 transgenic Arabidopsis lines, and the result demonstrated that BcHIPP16 promoted Cd2+ influx into Arabidopsis root cells. Taken together, our study showed that BcHIPP16 contributed to absorbing nutrient metal Cu and heavy metal Cd in planta.

Residue behavior and dietary risk assessment of six pesticides in pak choi using QuEChERS method coupled with UPLC-MS/MS.

A reliable and simple modified QuEChERS method with UPLC-MS/MS was developed for the simultaneous determination of six pesticides (dimethomorph, imidaclothiz, lufenuron, methoxyfenozide, pyridaben, spinetoram) and their metabolites in pak choi. Method validation indicated good linearity (R2 ≥ 0.99), accuracy (recoveries of 75%-112%), sensitivity (limits of quantification, 0.002-0.01 mg kg-1), and precision (relative standard deviations ≤ 21%), and matrix effects were -36-28%. The half-lives of the six pesticides in pak choi were 2.2-12 d under open field and greenhouse conditions. Considering the short growth cycle of pak choi, the terminal residue levels (0.046-7.8 mg kg-1) and the relevant maximum residue limits (MRLs) of some countries, 5 d was recommended as the pre-harvest interval for the six pesticides on pak choi. Dietary risk assessment revealed that the risk quotients were 3.1%-58% for different gender and age groups in China, indicating none unacceptable public health risk for general population. The results showed that all the six pesticides degraded faster and the terminal residues were much lower under open field conditions than those under greenhouse conditions, which was mainly due to the influence of rainfall, sunlight and other environmental factors. This work was thus significant in assessing the dissipation fate and food safety risks of the six pesticides on pak choi and facilitated the establishment of maximum residue limits.

First Report of Bacterial Soft Rot Disease on Pak Choi (Brassica rapa subsp. chinensis) caused by Pectobacterium brasiliense in the United States.

Pak choi (Brassica rapa subsp. chinensis) is an important vegetable crop native to China, known for high water content and low caloric value, containing high quality of protein, carbohydrates, fiber, vitamins, minerals, and secondary plant metabolites (Acikgoz, 2016). A pak choi field (8,000 sq. ft.) on Oahu, Hawaii, was visited in May 2020. About 10% plants were infected and showed characteristic symptoms of soft rot, wet lesions, macerated infected stem and necrotic leaves (Figure1A-D); leading to the suspect of one of the most devastating bacterial pathogens within genus Pectobacterium (Boluk et al. 2020; Li et al. 2019; Arizala et al. 2020; Arizala and Arif, 2019). Four infected plants were collected from the field, and stems were surface sterilized with 0.6% sodium hypochlorite solution for 30 sec, followed by three consecutive rinses in distilled water. The stems were aseptically macerated, streaked on Crystal violet pectate medium (CVP) (Hélias et al. 2011), and incubated for 48 h at 26 ± 2°C. The peculiar morphological characteristic of pectolytic bacterial pathogen, forming pits on CVP, were observed (Meng et al. 2016) (Figure 1E). Purification of bacterial colonies were done by re-streaking of a single colony on dextrose peptone agar (DPA-without tetrazolium chloride; Norman and Alvarez 1989). DNA was isolated from bacterial cultures using the DNeasy Blood and Tissue Kit (Qiagen, Germantown, MA), respectively. Molecular identification of four strains (PL243-246) were performed by the sequencing region of the housekeeping gene dnaA (chromosomal replication initiation protein) using Pec. dnaA-F1/R1 primer set (Dobhal et al. 2020). The amplified PCR product was enzymatically cleaned using ExoSAP-ITTM (Affymetrix Inc, Santa Clara, CA), and sent for sequencing at the GENEWIZ facility (Genewiz, La Jolla, CA) using both forward and reverse primers. The dnaA gene sequences were aligned using Geneious, and manually edited to remove the errors. The consensus sequences were analyzed with the NCBI BLASTn tool and were deposited in the NCBI GenBank under the accession numbers MT899920-MT899923. The NCBI BLASTn report indicated that all the sequences shared 99-100% identity and query cover with Pectobacterium brasiliense accession numbers MN544627-29. A phylogenetic analysis, using Geneious, was performed with the dnaA sequences representing different Pectobacterium spp., all strains grouped within the clade of P. brasiliense (Figure 2; Arizala et al, 2020). A pathogenicity assay was carried out in three replications on pak choi grown in pots containing commercial pot mixture, and maintained in the controlled-greenhouse (temperature 26-30°C; relative humidity 50-58%). Three-weeks old plant stems were artificially inoculated with 100 µl bacterial suspensions of PL243 (1.3x 108 CFU/ml), PL244 (1.2x 108 CFU/ml), PL 245 (1.2x 108 CFU/ml) and PL246 (1.1x 108CFU/ml); control plants were inoculated with 100 µl of distilled water (Figure 1F). Two days after inoculation, the soft rot and wilting symptoms (Figure 1G-H), similar to the ones observed on the field, were developed for all four strains tested. Bacteria was successfully re-isolated from the inoculated plants; DNA was isolated, amplified, sequenced for dnaA region and analyzed for 100% homology with original strains, to fulfill Koch's postulates. Based on the molecular characteristics re-isolates were identical to the original strains. To the best of our knowledge, this is the first report of P. brasiliense on pak choi in the USA. Recent reports indicated that the pathogen could potentially pose a threat to cruciferous crops, therefore, highlighting a need to conduct a state-wide survey for pectinolytic bacteria, and implement better management strategies to combat the vegetable crop losses.

An Automated High-Accuracy Detection Scheme for Myocardial Ischemia Based on Multi-Lead Long-Interval ECG and Choi-Williams Time-Frequency Analysis Incorporating a Multi-Class SVM Classifier.

Cardiovascular Disease (CVD) is a primary cause of heart problems such as angina and myocardial ischemia. The detection of the stage of CVD is vital for the prevention of medical complications related to the heart, as they can lead to heart muscle death (known as myocardial infarction). The electrocardiogram (ECG) reflects these cardiac condition changes as electrical signals. However, an accurate interpretation of these waveforms still calls for the expertise of an experienced cardiologist. Several algorithms have been developed to overcome issues in this area. In this study, a new scheme for myocardial ischemia detection with multi-lead long-interval ECG is proposed. This scheme involves an observation of the changes in ischemic-related ECG components (ST segment and PR segment) by way of the Choi-Williams time-frequency distribution to extract ST and PR features. These extracted features are mapped to a multi-class SVM classifier for training in the detection of unknown conditions to determine if they are normal or ischemic. The use of multi-lead ECG for classification and 1 min intervals instead of beats or frames contributes to improved detection performance. The classification process uses the data of 92 normal and 266 patients from four different databases. The proposed scheme delivered an overall result with 99.09% accuracy, 99.49% sensitivity, and 98.44% specificity. The high degree of classification accuracy for the different and unknown data sources used in this study reflects the flexibility, validity, and reliability of this proposed scheme. Additionally, this scheme can assist cardiologists in detecting signal abnormality with robustness and precision, and can even be used for home screening systems to provide rapid evaluation in emergency cases.

Blue and UV-A light wavelengths positively affected accumulation profiles of healthy compounds in pak-choi.

BACKGROUND: Recently, it become an important strategy using light to regulate plant growth and quality, especially on daily edible leafy vegetable. Pak-choi is rich in healthy functional compounds, e.g. flavonoid and glucosinolate. Many studies have focused on the plant response to increased radiation and transformed visible light quality, however, we know less about different blue and UV-A light wavelengths. Therefore, the goal of this study was to identify whether different blue and UV-A light wavelengths could improve quality in two cultivars of pak-choi and further cultivate potentially healthy functional plants. RESULTS: The different blue and UV-A light wavelength treatments significantly increased the fresh and dry weight in two cultivars of pak-choi. Compared with control, the content of soluble protein was higher after the different blue and UV-A light treatments. Similarly, the contents of total phenolics and total flavonoids increased significantly under the light treatments, and the highest content presented under T430 (supplemental blue light at 430 nm) in red-leaf pak-choi and under T400 (supplemental UV-A light at 400 nm) in green-leaf pak-choi. The total anthocyanins content and 2,2-diphenyl-1-picrylhydrazyl (DPPH) of two pak-choi cultivars improved positively with decreasing treatment wavelength, and other healthy compounds were affected to varying degrees under supplemental light treatments. CONCLUSION: The growth and healthy compound contents of pak-choi were significantly improved by supplemental blue and UV-A light, and there were wavelength- and cultivar-dependent effects. Compared with control, T430 presented the higher biomass and the contents of total phenolics, flavonoids and pigment in two pak-choi cultivars, and T380 was an efficient strategy to increase antioxidants and health-promoting compounds of red-leaf pak-choi. © 2020 Society of Chemical Industry.