The young plants grown under LED tended to have increased photosynthetic capacity. Furthermore, the water stress indices CWSI and IG, which were obtained using thermal imaging, were positively correlated with gas exchange-derived sg and E, putting forward the use of thermal imaging for the phenotyping of transpiration. Under LED light, photosynthetic gas exchange was generally increased, which agreed with the water stress indices. The extent of this increase was genotype-dependent. All differences between LED and HPS were smaller in the experiments where the fraction of lamp light over natural light was smaller.Using light emitting diodes (LED) instead of conventionally used high pressure sodium (HPS) lamps as a supplemental light source in greenhouses results in a higher efficacy (µmol light per J electricity) and makes it possible to customize the light spectrum. To explore the effects of LED and HPS on gas exchange, thermal relations, photosynthesis, and water status of young tomato plants, seven genotypes were grown in a greenhouse under LED  or HPS lamps in four experiments differing in the fraction of lamp light over natural light. HPS lights emit a broader spectrum of red (40%), green–yellow (50%), blue (5%), and far-red (5%) and a substantial amount of infrared radiation (heat). Young tomato plants grown under LED showed lower leaf temperature and higher stomatal density, stomatal conductance ( LED alss lowest . Acclimas lowest . Acclimas lowest . For inss lowest  and Wangs lowest  found ths lowest  found ths lowest  found thr status . The ligr status ,18. Eisir status  found thr status ,21. It ir status . Leaf gar status . Therefor status . In thisr status . From thductance ,27.G.LED can improve the biomass production of young tomato plants due to the higher photosynthetic efficiency of the plants, compared to HPS lamps , but thesg  than plants grown under HPS lamps; on average, sg was 6% higher under LED , measured using the LI-6400, were again found in plants grown under LED , and 0.8 °C in experiment 2 (p < 0.001). No leaf temperature differences were found in the two consecutive experiments  lower, by 5–18%, in plants grown under HPS than under LED light  correlations between these variables are to be expected. Between sg and E, the correlation coefficient was 0.99, while between CWSI and Ig, this was 0.98 (p < 0.05) negative correlations were found between leaf temperature and sg and E , while no correlations were found between these and other physiological parameters . While leaf temperature was negatively correlated with sg and E, stomatal density was positively correlated (p ≤ 0.001) with both (sg can be used as an indicator of plant water status [E) is closely related to water movement through a plant and its transpiration from leaves, stems, and flowers. E is integral to the calculation of sg, as is leaf temperature. Leaves from plants grown under HPS showed lower sg and E than from plants grown under LEDs  and electron transport rate at 1500 μmol m−2 s−1 PPFD (J1500) were significantly higher under LED , we had 3 h per day with only SL as the light source . Therefogs and E ,35,36. Wgs and E . Accordigs and E ,22,35,37than HPS , except observed . MoreoveG. We measured sg with a porometer and a LI6400. Based on the studies of Jones [sg can be derived from IG and CWSI non-invasively and potentially as high-throughput scalable measurements. CWSI can vary between 0 (no transpiration/ bad water status) and 1 . Young tomato plants grown under supplemental LED compared to HPS showed better water status  to use Solanum lycopersicum L.: ‘Moneymaker’, ‘Momotaro’, ‘Rutgers’, ‘Ailsa Craig’, ‘Kentuky Beefsteak’ and ‘Nunhems–FM001’; and Solanum pimpinellifolium L.: ‘LA1578’) were grown in four greenhouse experiments during the 2018–2019 winter/spring season: experiment 1 (Oct.–Nov.), experiment 2 (Dec.–Jan.), experiment 3 (Jan.–Feb.) and experiment 4 (Mar.–Apr.). Seeds were sown on stonewool plugs  and germinated in the glasshouse facilities (Unifarm) of Wageningen University, the Netherlands , under supplemental HPS or LED light. Seven days after sowing, seedlings in plugs were transferred to 10 × 10 × 6 cm stonewool blocks (Grodan) and placed on ebb and flow benches, for four weeks in experiment 1 and for ca. three weeks in the other experiments (three plants per genotype were placed on each bench), under either HPS and LED red/blue supplemental lighting. The experimental layout consisted of three benches per supplemental lighting condition, with three plants of each genotype on each ebb and flow bench. Nine plants for each genotype were grown under HPS and nine plants under LED (−2 s−1 photosynthetic photon flux density (PPFD) at plant level, and lamps were switched on 16 h before sunset and switched off at sunset. HPS light contains a substantial fraction of yellow (Y) to G (501–600 nm), while LED did not  was used for fertigation. Plants were watered daily, without differences between the compartments. Average air temperature during the day from the first to the fourth experiment was, respectively: 22.8, 22.9, 23.2 and 24.8 °C; air temperature during the night was, respectively: 19.2, 19.2, 19.4 and 19.5 °C  was closed to prevent excessive sunlight from entering the greenhouse. To prevent light pollution from adjacent compartments, all side walls of the greenhouse compartments were closed off, using white horticultural plastic foil. The greenhouse climate was recorded by a horticultural sensor box . During each experiment, the environmental conditions between the two experimental compartments, apart from SL treatment spectra, were kept identical. For experiment 1, the daily light integral (DLI) received by the plants  was on average 20 mol m−2 d−1 and 75% of the total light was SL  outside the greenhouse using a solarimeter . The fraction of photosynthetic photon flux  in the total global radiation was assumed to be 47% (Britton and Dodd (1976)), and the conversion factor from energy flux to quantum flux in the PPF region of sunlight 4.57 µmol J−1 [To determine the greenhouse transmissivity, PPFD at plant level was measured on a cloudy day with a quantum sensor  while concomitantly measuring global radiation , the number of hours during which SL was switched on was multiplied by its intensity. Light quality of a HPS and LED lamp was measured at 1 m distance with a field spectroradiometer . Light intensity at plant level was measured with a quantum sensor .sg was measured with a porometer , which was calibrated daily with fresh desiccant to ensure accurate measurements. To confirm the results obtained with porometer and to measure sg and E at the same time, during experiments 3 and 4, the LI-6400 photosynthesis system (Li-Cor) was used. Using a transparent cuvette (enclosed leaf area: 6 cm2), leaflets were enclosed at 400 ± 1.3 µbar CO2, 22.4 ± 0.2 °C cuvette temperature, 70 ± 1% RH and a flow rate of 500 µmol s−1. After they had reached stability, sg and E were logged 10 times at 30 s intervals ; these values were later averaged over the 3 biological replicates to increase accuracy. To minimize the effects of diurnal changes in temperature and light intensity, measurements of sg and E were performed alternatively between the HPS and LED compartment .All physiological measurements took place in weeks 3–4 after the start of treatments. The outermost three leaflets of the most fully expanded leaf were used  per plant and three plants per genotype were measured. In all experiments, −2s−1) to leaf internal CO2 partial pressure  was determined during experiment 1, 3 and 4. The LI-6400, equipped with the 6400-40 fluorescence cuvette , was used. The measurements were done on one leaf per plant for three plants per genotype for each bench. Leaves were enclosed in the cuvette at 1500 µmol m−2s−1 PPFD, 101.5 ± 0.05 kPa atmospheric pressure, 23 ± 0.2 °C cuvette temperature, 70 ± 5% RH, a flow rate of 400 µmol air s−1 and 400 ± 2 µbar CO2 partial pressure. After waiting for A to stabilize (∼15 min), CO2 partial pressure was changed in steps to 300, 100, 50, 400, 400, 600, 850, 1100 and 1400 µbar, while all other environmental variables were kept constant. At each CO2 step (2–3 min duration), after A was stabilized, values of CO2 and H2O measured by the infrared gas sensor of the sample cell were calibrated against those of the reference cell (“matching”), and A was logged. A and Ci values were corrected for leaks of CO2 into or out of the cuvette, according to Long and Bernacchi [2 response curves, Vc,max (maximum carboxylation rate), J1500 (electron transport rate at 1500 μmol m−2 s−1) and TPU (maximum triose phosphate utilization rate) were determined according to Sharkey [2 response curves, mesophyll conductance was assumed to be 0.189 mol m−2s−1, as determined for young tomato leaves [−2 s−1 PPFD (determined during experiment 1) were used to calculate day respiration (Rd). A negative-exponential function was fitted to A/PPFD, and the resulting intercept value was used as Rd. Since Rd was not significantly different between treatments (p = 0.14) or genotypes (p = 0.23), a global value of Rd = 2.1 ± 0.1 µmol m-2s−1 was used for fitting CO2 response curves.The response of net photosynthesis rate .In experiments 3 and 4, on 20 DAT, one stomatal imprint per biological replicate  was taken on the leaf abaxial side using the silicon rubber impression technique . Stomata−2, Astrobrights by Wausau papers, Mosinee, WI, USA) was also measured. Leaf temperature was measured with (i) an infrared non-contact thermometer  in experiment 3, and (ii) using a thermal camera  in all four experiments. Measurements were conducted in weeks 3–4 after transplanting. For (i) and (ii), the temperature of the outermost three leaflets of the most fully expanded leaf, which was approximately perpendicular to the lamps, was measured. Measurements with the infrared thermometer took place one hour before sunrise, while lamps were switched on, to avoid the influence of solar radiation on leaf temperature. To correct for the different temperatures at plant level in the HPS and LED compartments, the temperature of a reference leaf  and an average temperature was calculated. Leaf heat emissivity was assumed to be 0.95 [Using the thermal camera, leaf temperature readings from thermal images were taken over a circle with an area of 6 cm be 0.95 . To miniG) [dry as dry reference, Tleaf as leaf and Twet as wet reference temperature, respectively. The value of IG can vary from 0 (no transpiration) to ∞ ; and the crop water stress index (CWSI) [From the thermal images, the thermal index (IG)  was calcx (CWSI)  was calcCWSI can vary from 0 (no transpiration) to 1 .Each experiment was considered as an independent activity. Three replicate plants per genotype were evaluated per experiment. Data were first tested for homogeneity (Levene’s test) and for normal distribution of variances (Shapiro–Wilk test). The experimental factors were fixed in a two-way analysis of variance (ANOVA). To determine differences between means, a Tukey test was used. Correlations were tested using Pearson’s correlation coefficient. Analyses were performed using R . Packages used for the analysis were: agricolae , multcomE, sg, stomatal number and photosynthetic capacity. However, no genotypic differences were identified for leaf temperature. At higher ratios between natural light and SL (experiment 4), differences between HPS and LED were smaller. Finally, our results demonstrate that thermography is a potentially rapid and noninvasive technique to study plant water status and predict sg. However, further studies on, e.g., validation under a wide range of different conditions will be necessary to improve the accuracy of this prediction.Leaves of young tomato plants under LED showed higher gas exchange and photosynthetic capacity compared to HPS; this may be related to the increased leaf temperature under HPS, causing subtle but detectable differences in environmental acclimation. The fact that leaves were warmer under HPS lamps could be a result of the higher heat radiation (NIR light) relative to LED lamps. Moreover, the presence of green light in HPS lamps may have led to stomatal closure and/or reduced stomatal density, in turn reducing transpiration and increasing leaf temperature. Genotypic differences were observed for


Although combination therapy is the gold standard for patients with pulmonary arterial hypertension (PAH), some of these patients are still being treated with monotherapy.We conducted a retrospective analysis at four German PH centres to describe the prevalence and characteristics of patients receiving monotherapy.We identified 131 incident PAH patients, with a mean age of 64 ± 13.8 years and a varying prevalence of comorbidities, cardiovascular risk factors and targeted therapy. As in other studies, the extent of prescribed PAH therapy varied with age and coexisting diseases, and younger, so-called “typical” PAH patients were more commonly treated early with combination therapy (48% at 4–8 months). In contrast, patients with multiple comorbidities or cardiovascular risk factors were more often treated with monotherapy (69% at 4–8 months). Survival at 12 months was not significantly associated with the number of PAH drugs used  and was not different between “atypical” and “typical” PAH patients (89% vs. 85%).Although “atypical” PAH patients with comorbidities or a more advanced age are less aggressively treated with respect to combination therapy, the outcome of monotherapy in these patients appears to be comparable to that of dual or triple therapy in “typical” PAH patients. These iThe presented analysis aimed to answer the following questions: (1) what is the proportion of PAH patients treated with monotherapy in daily routine at German PH centres; (2) do PAH patients with comorbidities receive monotherapy more frequently; and (3) do PAH patients receiving monotherapy have poorer outcomes?Out of 782 PH patients treated at four German PH centres between 2016 and 2018, 158 were classified as having PAH. In this group, complete data, including data on comorbidities, cardiovascular risk factors and PAH medications, were available for 131 incident PAH patients, representing the group analysed.2), diastolic dysfunction (by echocardiography) or atrial fibrillation , interstitial lung disease (ILD), cancer and obstructive sleep apnoea.The four contributing university PH centres are considered representative of German PH centres, as they are well-established institutions with documented expertise in diagnosing and treating PH patients. Furthermore, they regularly participate in clinical studies and maintain good collaboration. The number of PH patients treated with monotherapy at these four centres is comparable to data on the German registry .The following data were collected retrospectively from medical records: age, sex, weight, height, secondary diagnoses, selected echocardiographic parameters, spiroergometric parameters, 6-min walking distance and haemodynamic parameters. The prescribed PAH drugs were documented for the entire observation period, and the vital status  was recorded at the end of follow-up on September 30, 2019.Follow-up data were collected at 0—3 months (baseline), 4—8 months (1st follow-up) and 9—15 months (2nd follow-up).Continuous data are presented as the mean (± standard deviation), and categorical variables are presented as absolute frequencies and percentages. The t-test was used to compare selected parameters between patients with “typical” or “atypical” PAH. Categorical variables were compared by the chi-square test, Fisher’s exact test, or the McNemar test. Survival was evaluated using Kaplan–Meier analysis, and differences between groups were assessed by the log-rank test. Analyses were performed with SAS 9.4 .The study was approved by the ethics committee of the University of Greifswald .The study included 131 patients , of whom 48 (36.6%) were classified as having idiopathic pulmonary arterial hypertension (IPAH) and 83 (63.4%) were classified as having pulmonary arterial hypertension (PAH). At baseline, the mean age was 64 ± 13.8 years, and the functional class (FC) was III in 90 (68.7%) and IV in 23 patients (17.6%)  patients were classified as having “typical” PAH and 45 (34.4%) were classified as having “atypical” PAH. Comparing “typical” and “atypical” PAH patients, significant differences between the two groups were found in terms of age, sex, diastolic dysfunction, arterial hypertension, coronary heart disease, atrial fibrillation, diabetes mellitus, chronic kidney disease and peak oxygen uptake Table .At baseline, 117/131 (89.3%) patients were treated with targeted PAH therapy, with 83 (70.9%) receiving monotherapy, 27 (23.1%) receiving dual therapy and 7 (6.0%) receiving triple therapy. At the first follow-up, 125/131 patients (95%) were treated with PAH therapy, of whom 72 (57.6%) continued to receive monotherapy, while 53 (42.4%) were on dual or triple therapy. At the second follow-up, 111/131 patients (85%) were available, of whom 50 (45.0%) continued to receive monotherapy, while 61 (55.0%) were on dual or triple therapy , a pattern that persisted, although no longer significant, at the first and second follow-ups .Regarding the “atypical” or “typical” phenotype, we found significant differences in the distribution of monotherapy vs. dual/triple therapy at baseline (ps Table . In “atyp = 0.016). This difference was no longer significant at the first or second follow-up .With increasing age, the proportion of patients treated with combination therapy decreased. At baseline, older patients received more monotherapy (up Table . Howeverp = 0.411) in survival with respect to the number of PAH drugs prescribed . This subgroup of patients was described as having “atypical” PAH to distinguish them from “classical” IPAH patients with few comorbidities [Only 36.6% of our patients were classified as having IPAH; however, this number reached up to 75% in clinical trials  and rangbidities . This tebidities  and in tbidities . The incbidities . Our finbidities . Among tbidities , 35 and bidities  are progbidities , 38. Morbidities . It is nbidities .Table 4CSimilar to other chronic diseases, such as chronic heart failure  or COPD It remains an important goal to treat PAH patients according to the current guidelines and reduce the gap between patients who do and do not receive aggressive combination therapy, when appropriate , 49. AccDespite these differences, the outcome of patients remaining on monotherapy during the entire observation period was not different from that of patients receiving dual or triple therapy. This was true for “atypical” as well as for “typical” PAH patients, although the number of patients was too small for a reliable survival analysis within each of these groups.Considering these results, upfront combination therapy for “atypical” PAH patients may not be needed when PAH is complicated by advanced age and multiple comorbidities, since the outcome of monotherapy in these patients appears to be comparable to that of dual or triple therapy in “typical” PAH patients.


Network analysis is an effective approach for examining complex relationships between psychiatric symptoms. This study was designed to examine item-level relationships between depressive and anxiety symptoms using network analysis in an adolescent sample and identified the most central symptoms within the depressive-anxiety symptoms network model. Depressive and anxiety symptoms were assessed using the Patient Health Questionire-9 (PHQ-9) and Generalized Anxiety Disorder Screener (GAD-7), respectively. The structure of depressive and anxiety symptoms was characterized using “Strength” and “Bridge Strength” as centrality indices in the symptom network. Network stability was tested using a case-dropping bootstrap procedure. Finally, a Network Comparison Test (NCT) was conducted to examine whether network characteristics differed on the basis of gender, school grade and residence. Network analysis revealed that nodes PHQ2 (“Sad mood”), GAD6 (“Irritability”), GAD3 (“Worry too much”), and PHQ6 (“Guilty”) were central symptoms in the network model of adolescents. Additionally, bridge symptoms linking anxiety and depressive symptoms in this sample were nodes PHQ6 (“Guilty”), PHQ2 (“Sad mood”), and PHQ9 (“Suicide ideation”). Gender, school grade and residence did not significantly affect the network structure. Central symptoms  and key bridge symptoms  in the depressive and anxiety symptoms network may be useful as potential targets for intervention among adolescents who are at risk for or suffer from depressive and anxiety symptoms. Adolescence is a crucial period in life, characterized by many unique changes and challenges. One in six people fall within this age group i.e., 10–19 years) and half of all psychiatric conditions start before 14 years of age , of whic–19 yearsComorbid depression and anxiety have been widely examined with traditional conceptualizations of psychopathology that rely on total scale scores to describe symptom severity. Unfortunately, such approaches may obscure meaningful associations between individual symptoms . NetworkN = 1409) revealed that “Self-hatred”, “Loneliness”, “Sadness”, and “Pessimism” were the most central symptoms [Network analysis has been used to understand symptom-symptom relationships in psychiatric comorbidities. For example, a network analysis of the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) study revealed the importance of “Sad mood” and “Anhedonia” in non-psychotic depressive disorder . In anotsymptoms . Anothersymptoms .Clinical features of depression and anxiety are closely associated with sociocultural and economic factors . TherefoThis cross-sectional study was conducted between 8 August 2020 and 12 March 2021, using snowball sampling method through the collaborative research network of the National Clinical Research Center for Mental Disorders, China. Eligible participants were (1) secondary school students residing in China during the COVID-19 pandemic, who were (2) able to understand the purpose and contents of the assessment. To avoid contagion during the COVID-19 pandemic, following previous studies , 32 dataSeverity of the depressive symptoms was assessed using the Chinese version of the 9-item Patient Health Questionnaire (PHQ-9) , 34 eachAll analyses were conducted using R (Version 4.0.3) . Means, To estimate and visualize the network, R-package “qgraph” (Version 1.6.5)  and “booz scores”).Following previous studies , 47, theAccording to recommendations of Epskamp et al. , the robCorrelation stability coefficients (CS-C) assessed the stability of centrality indexes  using subset bootstraps . If the Considering the moderating effects of gender, school grade, and residence on anxiety and depressive symptoms among adolescents , depressAltogether, 1183 adolescents were invited to participate in this study; of these 1057 met study inclusion criteria and completed the assessment. A majority was female (60.3%) and their mean age was 16.30 years (SD = 3.61 years). The sample mean PHQ-9 and GAD-7 total scores were 3.98 (SD = 5.44) and 2.67 (SD = 4.40), respectively (Table MSD = 0.44 ± 0.04); as such, none of the items were poor vis a vis informativeness. Moreover, no items were redundant with any other items . Therefore, all PHQ-9 and GAD-7 items were retained in the analyses.Tests of item informativeness and redundancy indicated no item ratings were <2.5 SD from the mean level for informativeness  had the most direct connection with node GAD2 (“Uncontrollable Worrying”), followed by the connection between nodes GAD4 (“Trouble relaxing”) and GAD5 (“Restlessness”), the connection between nodes GAD3 (“Worry too much”) and GAD7 (“Feeling afraid”) and the connection between nodes GAD4 (“Trouble relaxing”) and GAD6 (“Irritability”). In the depressive and anxiety symptoms network model of Chinese adolescents, node GAD5 (“Restlessness”) was most strongly associated with node PHQ8 (“Motor”) (average edge Weight=1.09), followed by connections between nodes GAD1 (“Nervousness”) and PHQ6 (“Guilty”) (average edge weight = 0.84), and nodes PHQ1 (“Anhedonia”) and GAD6 (“Irritability”) (average edge Weight=0.79) Fig. .In terms of strength, node PHQ2 (“Sad mood”) had the highest strength. Nodes GAD6 (“Irritability”), GAD3 (“Worry too much”), and PHQ6 (“Guilty”) were also statistically stronger than most other symptoms in the network Fig. . TherefoFollowing previous studies , 54, briFor stability of the network analysis, strength had an excellent level of stability , indicating that 51% of the sample could be dropped without significant changes in the network structure Fig. . BootstrS = 0.08, p = 0.484), but there was a significant difference in edge weights  or the distribution of edge weights  or the distribution of edge weights , GAD6 (“Irritability”), GAD3 (“Worry too much”) and PHQ6 (“Guilty”) were central symptoms in the network model. Additionally, bridge symptoms linking anxiety and depressive symptoms in this sample were nodes PHQ6 (“Guilty”), PHQ2 (“Sad mood”) and PHQ9 (“Suicide ideation”). Gender, school grade and residence did not significantly affect the overall network structure.“Sad mood” (PHQ2) was one of the most central symptoms to emerge within the depression-anxiety network of Chinese adolescents. Similar findings have been reported in previous studies on depressive and anxiety symptom networks of adults with psychiatric disorders , 55, aduThe symptom, “worry too much” (GAD3), was another prominent central symptom in the depressive-anxiety symptom network of Chinese adolescents as indicated by its strength. This finding is also consistent with previous findings of depressive and anxiety symptoms network in an adult psychiatric sample , 55 and “Irritability” (GAD3) was another prominent central symptom in this network analysis, and is also a key criterion, both for depression in children and adolescents and anxiety disorders in the DSM-5 . In addiGuilt or negative self-referential thinking has also been implicated in the onset and/or maintenance of depression among adolescents , 75. TheComorbid depressive and anxiety symptoms are often associated with poor treatment efficacy, and increased rates of hospitalization and disability . We founPrevious meta-analysis on depressive and anxiety symptoms in children and adolescents found that female sex was associated with increased risk of depressive and anxiety symptoms during COVID-19 pandemic . HoweverIdentifying central symptoms and bridge symptoms within the depressive-anxiety symptom network model has possible clinical significance; targeting these symptoms may contribute to prevention among at-risk adolescents and improve the effectiveness of treatments targeting co-occurring depressive and anxiety symptom . For exaPossible implications aside, the main limitations of this study should be noted. First, findings may not be generalized to adolescents in clinical settings, as previous studies have suggested differences in network connectivity may exist in different populations or samples . FurtherIn conclusion, this study is the first to identify central symptoms  and key bridge symptoms  within the depression-anxiety network of Chinese adolescents. Findings may provide an impetus for future studies examining symptom networks in other groups of adolescents as a means of clarifying key symptoms that extend across adolescents from different cultures and those that are unique to particular groups. In addition, central symptoms and bridge symptoms identified in this study may be useful targets for interventions designed to prevent these disturbances among at-risk Chinese adolescents and treat those who are currently suffering from co-occurring depressive and anxiety symptoms.supplementary material


Burn injuries from exploding electronic cigarettes have been well documented in the medical literature and lay press, as have injuries from smoking conventional cigarettes while on supplemental oxygen. However, there is a paucity of literature regarding injuries from smoking electronic cigarettes while on supplemental oxygen. Empirically, it might seem safe to use e-cigarettes while on oxygen, but it can result in explosion. The electronic cigarette coil can heat up to 350°C and serve as a source of ignition. The purpose of this study is to describe and characterize this relatively novel and uncommon mechanism of injury.This study was a descriptive review of 2013-2016 National Burn Repository (NBR) Data. Injury description fields were queried for “oxygen”, “O2”, “electronic cigarettes”, and various abbreviations and misspellings. Demographics, burn size, length of stay, expected length of stay using 1.1 days per %TBSA, and LOS index were reported. In addition, a Google search for lay press articles using the terms “smoking,” “’oxygen,” and “electronic (or e) cigarettes was performed, and these cases were tabulated.Of approximately 60,000 NBR entries, only 8 records of injury while smoking e-cigs on oxygen were found. Patients were predominantly male (63%) with a mean age of 63±9 years old, burn size of 3.4%±4 TBSA and LOS of 5.8±7 days. Inpatient stay averaged 1.65 days per %TBSA, which was 150% of expected LOS for the cohort. Two patients sustained third degree burns of 0.5% and 11%TBSA. Three patients were intubated, and had a mean of 3.33 ventilator days. Most injuries occurred at home (88%), while one occurred in a hospital. All 8 patients had Medicare and none of them suffered mortality. A google search revealed three distinct lay press articles regarding this topic.Although an uncommon mechanism of injury, smoking electronic cigarettes on supplemental oxygen can result in injury and a significant hospital stay. Limitations in abstraction from the database and/or reporting might have resulted in an under capture of the true number of injuries. Circumstances resulting in this phenomenon warrant further investigation, as it is unclear if it occurs from malfunction or a hot coil alone. Patients who are prescribed home O2 should be warned about this potential danger.


Escherichia coli bioparticles, and (c) mitogen-induced leukocyte proliferation and cytokine release. In men, lower childhood socioeconomic status predicted decrements in immunological performance across functional assays, along with greater spontaneous cytokine release from PBMCs. These changes co-occurred with elevations in plasma testosterone levels. Similar effects were not observed for other sources of stress, nor were they found in women (with the exception of spontaneous cytokine release). These findings provide evidence that low childhood socioeconomic status has a lasting negative impact on multiple aspects of immune function, particularly in men.Early life stress increases one’s risk for health problems later in life, and many studies find that these effects are sex-differentiated. Here, we examined relationships between multiple sources of early life stress and adult immune function in humans across several functional assays. Adult participants provided retrospective information about their childhood (a) socioeconomic status, (b) household unpredictability, and (c) exposure to adverse experiences. Participants’ peripheral blood mononuclear cells (PBMCs) were then isolated for use in functional assays of immune performance: (a) tumor cell lysis by natural killer cells, (b) phagocytosis of  This propensity is found to operate independently of an individual’s adult economic circumstances7, suggesting that the impact of early life environments on health is long-lasting and not easily reversed. For example, one study found that exposure to childhood socioeconomic disadvantage predicted an increased risk of cardiovascular disease in a cohort of male physicians, despite their achieved adult socioeconomic status far exceeding that of the average American6.Over the last three decades, a considerable body of research has demonstrated that individuals exposed to stressors during childhood exhibit an increased risk for a number of chronic health problems later in life, including metabolic disorders7, the myriad environmental and biological pathways through which one’s early life experiences impact adult health outcomes are only beginning to be understood. Initial clues about contributors to this relationship have surfaced in research demonstrating a link between early life adversity and the tendency to exhibit an exaggerated inflammatory response to immunological stimulation8. Individuals exposed to abuse and parental absence during childhood also tend to have higher peripheral levels of inflammatory biomarkers in adulthood, such as C-reactive protein, interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α)9. Moreover, many have proposed that chronic inflammation plays a mediating role in the relationship between early life trauma and adult disease states . Additional clues are found in research demonstrating that adults who report lower childhood socioeconomic status (SES) exhibit increased susceptibility to an experimentally-induced viral infection relative to those reporting higher childhood SES4. However, little is known about the broader immunological context in which such tendencies emerge  or the extent to which individual-level factors buffer or exacerbate the impact of adverse childhood environments.Despite the well-established link between childhood conditions and adult health7, many studies find these effects are sex-differentiated16. For example, early life exposure to infection and nutritional stress are often found to have an asymmetrically negative impact on men’s physical development and health compared to women’s18. Additionally, intervention programs aimed at improving health in the context of poverty are often disproportionately beneficial for boys compared to girls21. On the other hand, research also finds that certain early life stressors, such as maltreatment during childhood, more negatively impact the physical and mental health of women than men (e.g.16). Together, such results suggest that sex differences exist in health-related developmental plasticity, for better and for worse.One individual-level factor that is likely to moderate the relationship between early life environments and immunological function is biological sex. Indeed, although much research finds that developmental experiences can have a lasting impact on both men’s and women’s health12) and stronger canalization of bodily growth in women than men14. Others have proposed that sex differences in behavior play a role in this context, such as the greater tendency for men to engage in health-harming behaviors in stressful environments compared to women22. Further, research in the evolutionary sciences suggests that such differences reflect sex-differentiated energetic investments in mating effort vs. somatic development and repair34.Existing explanations for sex differences in susceptibility to the health consequences of early life stress include factors such as sex differences in hormone levels  What are the critical features of stressful early life environments that drive the association between childhood stress and adult immune function?, (b) How do the effects of these environmental stressors manifest themselves across qualitatively distinct aspects of immunological function?, and (c) Are these patterns sex-differentiated?a, the relative impact of three related, but distinct, sources of childhood stress on immune function were examined, including: (1) resource availability , (2) unpredictability36, and (3) adverse childhood experiences (ACEs37). Isolating the unique impact that each of these environmental correlates of stress has on adult immune function will yield needed insights into the types of environmental interventions that are likely to have a lasting positive impact on adult health outcomes.To address question b, much of the previous work linking childhood environments to adult immune function has focused primarily on inflammatory processes10. In the current research, data were collected on a variety of immunological endpoints, many of which were measured at multiple time-points or at multiple effector-to-target (E:T) cell ratios. These measures included: (1) proliferation of peripheral blood mononuclear cells (PBMCs) in response to, and in the absence of, stimulation by lipopolysaccharide (LPS), phytohemagglutinin (PHA), and polyinosinic:polycytidylic acid (poly (I:C)) measured at 24, 48, and 72 h post-plating; (2) natural killer (NK) cell lysis of target tumor cells at 100:1, 50:1, 25:1, and 12.5:1 E:T ratios; (3) phagocytosis of opsonized Escherichia coli (E. coli) bioparticles; and (4) PBMC release of the pro-inflammatory cytokines interleukin-1β (IL-1β), IL-6, and TNF-α in response to LPS stimulation, as well as in the absence of stimulation , at 2, 24, 48, and 72 h post-plating. Additionally, plasma levels of total testosterone were also assayed to test whether diminished immune function co-occurs with increased investment in mating effort.Regarding question c, in line with the National Institute of Health’s recent policy on accounting for sex as a biological variable38, a number of steps were taken during study design, data collection, and data analysis to allow for targeted tests of sex differences in the impact of early life stress on immune function. These included recruiting equal numbers of men and women , testing interactions between each predictor and sex in all analyses , and clearly presenting sex-differentiated results , sex, and each dependent measure were tested in separate models for each environmental predictor. The results of these models are reported below as the primary analyses  were regressed on the childhood environmental factor, sex, and the interaction between sex and the childhood environment factor. Per convention, if no significant interaction was found, main effects were reported without inclusion of the interaction term. Where significant interactions were found, direct effects of the childhood factor(s) on the dependent immunological measures within each sex  were examined. Sex differences at high and low levels  of childhood factors involved in a significant interaction with sex in the initial models  were also probed. Given that the primary objective of the current study was to test whether the impact of childhood environmental factors on immune function differed by sex, only the simple slopes within each sex are reported in the main text. Regions of significance analysis can be found in the Supplementary Information and Figs. Given that the proliferation (time collected), NK cell cytotoxicity (E:T ratios), and cytokine release (time collected) data all contained nested structures, complex multilevel modeling (cytokine release data) and latent curve models  were used to analyze these data. First, unconditional models for each of the dependent measures  were tested to assess model fit to linear change across repeated measures . Note that while neither of these two childhood variables significantly predicted any immunological outcome in the primary models or models controlling for covariates, results of the partial effects model revealed significant interactions between each of these variables and sex in predicting PHA-stimulated proliferation. Further, there was a main effect of childhood unpredictability on spontaneous cytokine release by PBMCs. These results were not included in the main text as they only emerged in the partial effects model; full results can be found in the Supplementary Information. Additionally, no factors or interactions emerged as significant predictors of unstimulated PBMC proliferation or stimulated cytokine release in vitro.Results of all models revealed good model fit , phagocytosis of E. coli bioparticles, β =  − 0.64, SE = 0.27, t =  − 2.35, p = 0.02, and testosterone levels, β = 0.38, SE = 0.14, t = 2.73, p = 0.006.As predicted based on research indicating greater male susceptibility to adverse health effects of early life stress, results revealed significant interactions between childhood SES and sex in predicting NK cell cytotoxicity , and lower levels of testosterone, β =  − 0.24, SE = 0.06, t =  − 3.79, p < 0.001. For women, childhood SES did not significantly predict phagocytosis, β =  − 0.09, SE = 0.12, t =  − 0.22, p = 0.82, NK cell cytotoxicity , or testosterone levels, β =  − 0.11, SE = 0.13, t =  − 0.87, p = 0.38. Notably, although only reaching statistical significance at the highest E:T ratio , β = 0.21, SE = 0.09, t = 2.44, p = 0.02, but not the slopes of proliferation over time in these conditions (ps > 0.27). These effects were no longer significant after controlling for covariates  and should thus be interpreted with caution. Lastly, there was a main effect of childhood SES on the latent factor of spontaneous in vitro proinflammatory cytokine release by PBMCs , β =  − 0.22, SE = 0.07, t =  − 3.12, p = 0.002, but not stimulated cytokine release, β = 0.04, SE = 0.05, t = 0.86, p = 0.39. Specifically, higher childhood SES predicted a diminished tendency to release proinflammatory cytokines in the absence of overt immunological stimulation. These results remained significant when controlling for both covariates and the effects of the other childhood environmental factors.In addition to these sex-differentiated results, significant main effects of childhood SES  were found predicting the intercept of PBMC proliferation  in response to stimulation by LPS, p-values in ascending order and comparing each p-value to the product of the alpha value  and the quotient of the rank divided by the total number of tests yielding a significant result . If the ranked p-value is less than the above product, the null hypothesis is rejected. This analysis revealed that all significant p-values passed this test  procedure. This method involves rank ordering the significant 10, particularly for men21. The current research sought to extend this work by addressing three key questions that have been heretofore unexplored in the context of a single investigation: (a) What are the critical features of early life stress that drive the association between childhood stress and adult health outcomes?, (b) How do these factors manifest themselves across qualitatively distinct aspects of immunological function?, and (c) Are these patterns sex-differentiated?See Supplementary Information for additional discussion. Considerable research indicates that early life experiences have a lasting impact on adult healthResults revealed that men exposed to lower childhood SES exhibited a diminished immune response across each measure of immunological function, as well as increased spontaneous pro-inflammatory cytokine release by PBMCs, compared to men with a higher childhood SES. Conversely, with the exception of the effects of childhood SES on stimulated PBMC proliferation and spontaneous cytokine release (both effects not sex-differentiated), the measured childhood environmental factors did not significantly predict immunological function in women. Further, with the exception of the relationship between childhood SES and stimulated PBMC proliferation, significant effects were not attenuated when controlling for adult SES , suggesting that these differences are primarily the result of developmental processes rather than current resource availability.10, as well as research demonstrating sex differences in key immunological outcomes28. Further, consistent with research and theory suggesting that androgens play an important in mediating sex-differentiated patterns of immunological investment44, results revealed that, in men, reduced immune performance co-occurs with higher testosterone levels. The results of the current research suggest that men who grow up in conditions of low SES may be more vulnerable to poor health outcomes later in life due to diminished investment in immune function. Moreover, the current results suggest that these differences might emerge in response to differential resource access, per se, as there was little evidence of a negative impact of childhood unpredictability or other adverse experiences  on immune function in either sex. It is important to note that others have found that the relationship between early life disadvantage and markers of poorer health  are driven by alternative features of such environments, such as abuse and parental neglect11, and that these factors may even more negatively impact the physical and mental health of girls relative to boys in certain contexts (see e.g.16). Accordingly, additional research is needed to understand the conditions in which each type of developmental stress is likely to negatively impact immunological development in each sex. Future studies may find, for example, that the timing of stress exposure influences both the magnitude of decrements in immune function, as well as differences in the extent to which men and women are affected. These questions could not be answered in the current study given its cross-sectional design and reliance on retrospective accounts of stress exposure across a wide period of early life.The present results are consistent with a growing body of research demonstrating a negative relationship between early life adversity on adult health45 and levels of these hormones are impacted by early life stress46.There are limitations of the current research that should be considered. First, although several components of immune function were measured in the current study, together they capture only a narrow range of the tremendous complexity that comprises immune function. Accordingly, future research might find that, in women, early life experiences play an important role in influencing other aspects of immune function, such as adaptive immunity. Further, the current research did not experimentally examine the proximate biological mechanisms driving the observed relationships between early life stress and adult immune function. One possibility is that glucocorticoids and other adrenal hormones ) play an important role in this context, as they have immunomodulatory propertiesIt is also important to consider that the majority of the participants in the current study were healthy college students. As a consequence, the low amount of variability in the adult SES variable may have limited our ability to detect effects of participants’ current socioeconomic circumstances on the key immunological outcomes. The present sample’s exposure to certain types of early life stress may also have been lower than that of the general population. Moreover, all of measures of early life stress used in this study were retrospective. Thus, a key direction for future research will be to examine, prospectively, the effects of childhood SES and multiple types of early life adversity on adult immune function in a more diverse sample using a longitudinal design. Such research would minimize the interpretability issues that arise from cross-sectional research using retrospective accounts of early life environments, which is a weakness of the current work. Future research may also benefit from utilizing longitudinal designs to track sex differences in the impact of early life conditions on immune function across multiple stages of development.The current research provides important evidence of sex differences in the impact of early life SES on multiple facets of immune function. Together, these findings suggest that, for men in particular, early life experiences become immunologically embedded in ways that may have a lasting impact on health. These results have important implications for interventions targeted at reducing the negative impact of early life disadvantage on health, and lay the groundwork for future research examining sex differences in relationships between childhood environments and immune function across the lifespan.Mage = 20.17 years, SD = 2.75). Restriction criteria were: (1) participants must have no history of chronic medical problems, including mental illness, (2) must be of healthy weight  < 30), (3) must be free from acute illness for at least two weeks prior to participation, (4) women must not be on hormonal contraceptives, (5) must be willing to abstain from steroidal and non-steroidal anti-inflammatory medications, exercise, and alcohol use for at least 2 days before participating, and (6) must fast the morning of the session. Additional characteristics of the sample are published elsewhere47. Women participated 4–7 days after the start of their most recent menstrual cycle . In exchange for participation, participants were given the choice of partial course credit or a $50 gift card.A total of 159 individuals from Texas Christian University or the surrounding community, participated in exchange for $25 or experimental research credit . The study was conducted in accordance with the protocol approved by the review board.On the day of their sessions, participants arrived at the laboratory at 7:30 AM, provided informed consent, and answered study compliance questions . Next, for the purpose of a larger study, participants completed a series of behavioral tasks and questionnaires. Finally, participants were led into an adjoining room, in which 85 mL of blood was collected by venipuncture into heparinized (or EDTA-containing) Vacutainer tubes .Escherichia coli (E. coli) bioparticles, (3) PBMC proliferation in response to, and in the absence of, mitogen/toll-like receptor stimulation, and (4) PBMC release of proinflammatory cytokines in response to, and in the absence of, mitogen stimulation.Peripheral blood mononuclear cells (PBMCs) were immediately isolated from whole blood using density gradient centrifugation in Ficoll Paque Plus ). PBMCs were used in four functional immunoassays: (1) natural killer (NK) cell lysis of target tumor cells, (2) PBMC phagocytosis of fluorescent dye-labelled l-glutamine, 1 mM sodium pyruvate, 100 U of penicillin/mL, 100 µg of streptomycin/mL, and 0.25 µg of amphotericin B/mL . In addition, plasma was collected at the time of blood processing and frozen at − 80 °C. Additional information about each assay follows, below.For each assay, PBMCs were brought to the plating density appropriate for the assay in RPMI-1640 supplemented with 10% heat-inactivated fetal bovine serum (FBS), 2 mM 48. This measure was chosen to capture the extent to which participants felt they did or did not have access to resources during their childhood. Participants indicated their childhood SES by responding to three statements about their life before age 12  using a 7-point Likert scale . The items together yielded acceptable reliability (α = 0.81) and were formed into a mean composite, with a higher score indicating greater resource availability/a higher SES during childhood .First, participants’ childhood socioeconomic status (SES) was assessed using a previously validated scale36, consisting of three statements about chaos and uncertainty within the home . Participants reported agreement with how well each statement described their lives before age 12 using a 7-point scale . Together, the items yielded acceptable reliability (α = 0.74) and were formed into a mean composite with a higher score indicating greater experience with unpredictability during childhood .Next, childhood unpredictability was measured with a scale used in previous research37. This questionnaire assesses experience with adversity during childhood across five domains: abuse, family dysfunction, and peer, community, and collective violence/war. Given the nature of the present sample, which was primarily composed of college students, a truncated version of the survey was administered, removing the collective violence, sexual abuse, and secondary household abuse questions, and leaving a total of 20 items. Continuous items were recoded as binary, per convention, and all items were then summed into a composite measure of cumulative exposure to adverse experiences during childhood .Finally, participants’ experience with other types of adversity during childhood were also measured using the well-validated Adverse Childhood Experience International Questionnaire (ACE-IQ), developed by research teams from the World Health Organization51Cr-release assay. K-562 target tumor cells  were grown inside T-25 flasks  in RPMI-1640 growth medium supplemented with 10% FBS, 2 mM l-glutamine, 1 mM sodium pyruvate, 100 U of penicillin/mL, 100 µg of streptomycin/mL, and 0.25 µg of amphotericin B/mL  and incubated at 37 °C, 5% CO2, and 100% humidity.NK cell cytotoxicity was measured using the classic 51Cr  for one hour before plating them at a density of 1 × 104 cells/well in a 200 µL final volume into Corning V-bottom plates . PBMCs were plated with target tumor cells, in triplicate, at the following effector:target (E:T) cell ratios: 100:1 (1 × 106 PBMCs/well), 50:1 (5 × 105 PBMCs/well), 25:1 (2.5 × 105 PBMCs/well), and 12.5:1 (1.25 × 105 PBMCs/well). Spontaneous/background lysis controls  and maximal lysis controls ) were plated in sextuplicate. Plates were incubated for four hours at 37 °C, 5% CO2, and 100% humidity.Target tumor cells were labeled with 1 µCi 51Cr release on a CAPRAC-t gamma counter . Percent maximal lysis of target tumor cells by participant NK cells was calculated at each E:T ratio by dividing 51Cr release by maximal release after subtracting spontaneous release from both values and multiplying by 100.After brief centrifugation of the V-bottom plate, supernatants were collected into glass scintillation vials and quantified E. coli BioParticles  that were opsonized using 1 mg/mL of manufacturer-provided opsonization buffer. PBMCs were plated in triplicate with E. coli bioparticles into BrandTech black, flat-bottom microplates  at a density of 5 × 105 cells/well in a 200 µL final volume. Negative controls (bioparticles plated with media only in 200 µL volume) and positive controls ) were plated in triplicate. Plates were incubated for two hours at 37 °C, 5% CO2, and 100% humidity before being read on a fluorescence plate reader  at FITC dye settings of 490 nm excitation/520 nm emission. Percent maximal fluorescence was computed by dividing experimental fluorescence by maximal fluorescence , after subtracting fluorescence in the negative controls from both, and multiplying by 100.The phagocytic capability of participants’ PBMCs was assessed using fluorescent pHrodo Green 5 cells/well in a 200 µL final volume for four conditions: (1) with media only , (2) with 1 µg/mL of lipopolysaccharide (LPS) obtained from E. coli , (3) with 5 µg/mL of phytohemagglutinin , and (4) with 50 µg/mL of polyinosinic:polycytidylic acid (poly (I:C); high molecular weight; InvivoGen, San Diego, CA). Plates were incubated at 37 °C, 5% CO2, and 100% humidity.PBMCs were isolated as described above, and plated into Falcon 96-well tissue culture plates . PBMCs were plated in triplicate at a final density of 2.5 × 10Approximate cell density was measured at three time-points: 24, 48, and 72 h post-plating using the CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay . Each plate was read on a plate reader  at 490 nm.5 cells/well, in a 200 µL final volume. PBMCs were plated both in media only (unstimulated condition), as well as with 1 µg/mL of LPS, obtained from E. coli , and were incubated for up to three days at 37 °C, 5% CO2, and 100% humidity. Cell culture supernatants were collected at 2, 24, 48, and 72 h post-plating, and then stored them at − 80 °C until assays were conducted.PBMC release of proinflammatory cytokines was measured in vitro both in response to LPS stimulation, as well as in the absence of stimulation. After isolation, PBMCs were plated in triplicate at a density of 2.5 × 10Cell culture supernatants were later thawed and assayed in duplicate for levels of a trio of proinflammatory cytokines: interleukin-1beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) using a MILLIPLEX MAP Human Cytokine Panel magnetic bead kit , and read using a Luminex MAGPIX fluorescent detection system  and xPONENT software . Intra-assay coefficients of variation (CVs) were 8.20% (IL-6), 6.97% (IL-1β), and 5.98% (TNF-α). Inter-assay coefficients of variation (CVs) were 17.27% (IL-6), 10.53% (IL-1β), and 11.62% (TNF-α).47). No compromised samples were assayed, and thus data from these samples were not included in any analysis. All other biological samples from these participants  were stored elsewhere and were thus unaffected by the freezer failure.Due to a freezer failure on November 5, 2016, cell culture supernatant samples were compromised for 32 participants . Plates were read on a plate reader  at a wavelength of 450 nm. The intra-assay CV was 2.48% and the inter-assay CV was 13.45%.Several variables that may influence relationships between early life conditions, sex, and immune function were measured. These included race, age, exercise, sleep, BMI, stress, loneliness, recent illness, season, and adult SES. See Supplementary Information for more details.Supplementary Information.


Foeniculum vulgare essential oil (EO) emitter, within an active package system, to delay quality decay, thus extending shelf life; (iii) to estimate the impact of EO emitter on the sensory profile of the RTC slices after cooking. Results revealed that both globe artichoke genotypes possess a good attitude to be processed as RTC product. “Opera F1” showed the best performances for color parameters, texture and chemical indexes, while “Spinoso sardo” showed lower mass loss (ML) over the storage time. The addition of EO emitter slowed down the consumption of O2, better preserved texture when compared to the control and more effectively control polyphenol oxidase (PPO) activity and antioxidants’ retention during the cold storage. Microbial counts in control globe artichoke RTC slices were significantly higher than those packed with EO emitter, confirming the inhibiting role played by EO of F. vulgare. In addition, the EO emitter did not influence negatively the sensory profile of RTC globe artichoke slices after microwave cooking.Two globe artichoke genotypes, “Spinoso sardo” and “Opera F1”, have been processed as ready-to-cook (RTC) slices and refrigerated at 4 °C for 12 days (i) to evaluate the suitability to be processed as RTC slices; (ii) to evaluate the effect of a  Cynara cardunculus L. var. scolymus (L.) Fiori = C. scolymus L.), whose production for a long time was limited to the native Mediterranean region, is becoming increasingly popular and desired by consumers all over the world for its taste and functional properties [Food market demand is increasingly focused on food products with a high level of healthy compounds. In this context, globe artichoke , which generates dark pigments called melanoidins, unacceptable in terms of safety, since they could support the microbial growth. Additionally, although moderate, stress conditions or short storage times could be due to the loss of sensory and nutritional quality of the processed globe artichokes ,10,11. Til, etc. . The posil, etc. .An important role in plant defense is played by EOs, secondary metabolites, some of which exert powerful antimicrobial activities ,17,18, aFoeniculum vulgare seeds, which is rich in trans-anethole, has been demonstrated to possess antioxidant activities [Shigella dysenteriae [Escherichia coli [F. vulgare EO in preserving quality of RTC globe artichoke slices of “Spinoso sardo” during refrigerated storage for 11 days. However, although EO-based packaging has the ability to increase food shelf life, the selection of EO-based materials should be done on the basis of the compatibility with foods in terms of flavor to avoid any negative impact of EOs on the sensory profile.EOs have been deeply studied also as active components in bio-based emulsified films and coatings. The EO from tivities , good anenteriae , Escherihia coli , and othhia coli ,23,24. Whia coli  demonstrF. vulgare EO emitter, within an active package system, as a strategy to slow down quality decay, and thus increasing shelf life; (iii) to estimate the impact of EO emitter within the package on the sensory profile of the RTC slices after microwave cooking.The aims of this study, performed on RTC globe artichoke slices stored at 4 °C for 12 days, were therefore: (i) To evaluate the suitability of two globe artichoke genotypes, “Spinoso sardo” and “Opera F1”, to be processed as RTC slices; (ii) to evaluate the effect of a −1 of available P2O5 and 280 mg kg−1 of exchangeable K2O. The climate of the area is semiarid-Mediterranean, with mean long-term monthly maximum and minimum temperatures from 14.8 (January) to 30.6 °C (July) and from 7.8 (January) to 22.3 °C (August), respectively [Experimental trial was performed during 2014–2015 on Siracusa plain ), one of the main Italian site for globe artichoke crop. The soil, classified as calcixerollic xerochrepts , had theectively .−2. Crop management was carried out in agreement with the usual commercial practice of the cultivation area.“Spinoso sardo”, an early reflowering multiclone globe artichoke genotype, with conical green heads characterized by purple shades and big yellow spines , and “Op3/m2/24 h at 23 °C and 0% relative humidity (RH). Totally 80 trays were filled with “Opera F1” and “Spinoso sardo” slices respectively. Inside a half of the totally package for each genotype (40) was placed a small square (1 × 1 cm) of sterilized TNT textile, soaked with 150 µL of F. vulgare EO (0.75% w/w) produced by Rao Erbe  immediately before the hermetical sealing of trays with SPP plastic bags (20 × 15 cm) by a sealing bar . Samples were kept at 4 ± 0.5 °C and 90–95% RH up to 12 days and analysis were done at the processing day (T0), and after 5, 8 and 12 days of refrigerated storage  and immediately weighed before further analysis. Mass loss (ML) was expressed as % of the initial sample weight at T0. The headspace gas composition, to quantify carbon dioxide and oxygen, was measured using a CheckPoint portable gas analyzer (MOCON Europe A/S (Dansensor), Ringsted, Denmark). Analysis were done on three replicates every sampling time.2 + Δa*2 + Δb*2)1/2 pointing at the total color difference, to better assess the overall color changes during storage as previously done by Licciardello et al. [Surface color of RTC globe artichoke slices was assessed according to CIE L*a*b* scale as stated by Rizzo et al. , using ao et al. .The texture was measured by applying a 500 N nominal force cell and a stainless-steel probe (length 5 mm) and measuring the maximum shear force, using instruments and data elaboration as reported by Rizzo et al.  Results −1 chloramphenicol , incubated at 25 °C for 48–72 h; Enterobacteria (TEB) on violet red bile glucose agar , incubated at 37 °C for 24 h; Escherichia coli on brilliance E. coli selective agar , incubated at 37 °C for 24 h; Pseudomonas spp. on Pseudomonas Agar Base , supplemented with Pseudomonas CFC selective agar supplement  and incubated at 25 °C for 48 h. All microbiological analyses were performed in triplicate and expressed as average log10 CFU g−1.Microbiological determinations were performed on 25 g of RTC artichokes, aseptically sampled from each package and homogenized in a Lab-Blender 400  for 3 min with 225 mL of Ringer solution . The determined microorganisms were: Total aerobic mesophilic bacteria (MB) and total aerobic psychrotrophic bacteria (PB) on plate count agar  with cycloheximide 0.1% solution , incubated at 30 °C for 48 h and at 7 °C for 5–10 days, respectively; at; yeasts and molds (YM) on Sabouraud Dextrose Agar  supplemented with 0.1 g LAn amount of RTC globe artichoke slices from each package was freeze dried  for performing the following chemical determinations.−1 of DM (dry matter).L-ascorbic acid content (AsAC) was evaluated following the HPLC method proposed by Lombardo et al.  and expr−1 of DM, using chlorogenic acid as a standard; in the same extracts used for TPC analysis, the antioxidant activity (AA), expressed in terms of DPPH  percentage of inhibition, was determined using the method reported by Brand-Williams et al. [A modified Folin-Ciocalteu method  was adops et al.  and calc−1.The determination of PPO activity was spectrophotometrically performed ,33, usinAll these chemical determinations were conducted by using bi-distilled water and analytical or HPLC grade reagents and solvents obtained from Sigma-Aldrich .The UNI EN ISO 13,299  method wAccording to Rizzo et al. ,17 the fThe panel evaluated the randomized samples adopting a scale from 1 (absence of sensation) to 9 (extremely intense) in the sensory laboratory  of the DF-test was significant (p ≤ 0.05). Limitedly to AsAC, TPC, and AA, the correlation analysis was carried out to estimate their relationship. The software package Statgraphics® CenturionXVI  was used to perform ANOVA.Bartlett’s test was adopted to verify the homoscedasticity, and then the data  were subjected to a three-way analysis of variance (ANOVA) as a factorial combination of “genotype (2) × postharvest treatment (2) × storage time (4)”. For each sensory attribute a two-way ANOVA as a factorial combination of “genotype (2) × postharvest treatment (2)” was performed. Means were separated by LSD test, when the It is assumed that fresh globe artichoke heads are cooked prior to consumption. The USDA National Nutrient Database for Standard Reference reports that about 40% of a fresh head is edible, while the USDA Food and Nutrient Database for Dietary Studies indicates that about 6% of this edible share is further lost through cooking. Therefore, while the consumers have to consider only the weight lost through cooking of RTC globe artichoke slices, producers have to consider also the whole mass loss to define their profit. As previously reported by Licciardello et al. , the mosML was slightly higher in “Opera F1” at the end of the experimental trial (12 day) respect to “Spinoso sardo”, while samples with EO”s pad began with higher values that were maintained until 8 days . It is e2 and CO2 were not influenced by G, as expected, thus confirming previous results [2 and CO2 for p ≤ 0.001 and p ≤ 0.01, respectively  and “G × T × St” (p ≤ 0.01), as reported in In RTC globe artichoke slices texture is doubly important because it has to satisfy the consumer at package’s opening, but also after cooking. Texture was significantly influenced by interactions “G × St”, “T × St”  and AsAC (r = 0.775 ***). As reported previously [F. vulgare.The AA (measured by DPPH assay) had a good correlation with both TPC  were all individually significant for the PPO parameter, but only “G × St” and “T × St” interactions were significant .−1 g−1) respect to “Spinoso sardo” (2.36 mmol min−1 g−1). In addition, “Spinoso sardo” shows a maximum of activity after the 5th day of storage, while in “Opera F1” it appeared delayed at 8 days, but lower than “Spinoso sardo” in the same sampling time. This tendency should be linked with the browning reactions activated by specific polyphenolic substrates. As previously observed, the extent of the peroxidase  isoenzymes in “Opera F1” is really poorer if compared to other globe artichoke genotypes or other crops, such as tomato, melon and strawberry [O- and 3,5-O-dicaffeoylquinic acids, which are the most representative phenolic compounds in this crop [Based on the significance of the “G × St” interaction, a similar trend was observed for the PPO in both the selected genotypes a, althourawberry . In addirawberry . The subrawberry , as wellhis crop ,48.Referring to the interaction “T × St” b, both tp ≤ 0.001). Regarding G, higher average microbial counts were recorded for “Spinoso sardo” than “Opera F1”  moderately exceeded the relative limit of 5 log CFU g−1 recommended by CNERNA-CNRS [E. coli plate count was 2.91 log CFU g−1 at the end of the considered period, in conformity with European Regulation EC 1441/2007 [−1 was observed for all microbial groups. As reported in −1, respectively) and for Pseudomonas spp. in “Opera F1” samples (0.76 log CFU g−1).pera F1” , probablpera F1” . In partRNA-CNRS , of 7.7 getables  and of 7ed meals . InsteadRNA-CNRS , but notRNA-CNRS , being i441/2007 . RegardiThe sensory attributes that significantly differentiated, at each sampling time, the RTC globe artichoke slices of the two studied genotypes were shown in At 0, 8, and 12 days, samples were perceived as similar by panelists, except for fennel odor and flavor, while after 5 days of refrigerated storage the samples were significantly different also for bitter attribute. The latter was more intense in the control C as especially observed for “Opera F1”. As expected, the samples packed with EO emitter showed the highest intensities of fennel odor and flavor, regardless of genotype .F. vulgare EO emitter into the packaging system did not influence ML, color appearance and texture, while its presence slowed down the consumption of O2, better preserved texture when compared to the control and more effectively controlled PPO activity and antioxidants (AsAC and TPC) retention during the cold storage.The present findings revealed that both globe artichoke genotypes possess a good attitude to be processed as RTC slices, although with a difference response to the applied technological conditions: “Opera F1” showed the best performances for color appearance, thanks to the lower isoenzymes activity, texture and chemical indexes , while “Spinoso sardo” showed lower ML over the storage time. The addition of F. vulgare. In addition, the EO emitter did not influence negatively the sensory profile of RTC globe artichoke slices after microwave cooking. Control and active-packaged samples were considered similar by panelists, with the exception of fennel odor and flavor that of course were revealed in samples packed with EO emitter.As regards the microbiological quality, all the considered microbial groups gradually increased until the end of the storage period, with a significant higher extent in control RTC globe artichoke slices than in those packed with EO emitter, confirming the inhibiting role played by EO of F. vulgare EO emitter, within an active package system, as a promising technological strategy for preserving quality attributes and reducing microbial spoilage of RTC globe artichoke slices.Our findings demonstrate the potential application of a


Avena sativa L.), ‘Triple Crown’ and ‘Qinghai 444’, with yellow and black bracts, respectively, were found to both have green bracts at the heading stage, but started to turn black at the flowering stage and become blackened at the milk stage for ‘Qinghai 444’. Their photosynthetic characteristics were analyzed and compared, and the key genes, proteins and regulatory pathways affecting photosynthetic physiology were determined in ‘Triple Crown’ and ‘Qinghai 444’ bracts. The results show that the actual PSII photochemical efficiency and PSII electron transfer rate of ‘Qinghai 444’ bracts had no significant changes at the heading and milk stages but decreased significantly (p < 0.05) at the flowering stage compared with ‘Triple Crown’. The chlorophyll content decreased, the LHCII involved in the assembly of supercomplexes in the thylakoid membrane was inhibited, and the expression of Lhcb1 and Lhcb5 was downregulated at the flowering stage. During this critical stage, the expression of Bh4 and C4H was upregulated, and the biosynthetic pathway of p-coumaric acid using tyrosine and phenylalanine as precursors was also enhanced. Moreover, the key upregulated genes  of anthocyanin biosynthesis might complement the impaired PSII activity until recovered at the milk stage. These findings provide a new insight into how photosynthesis alters during the process of oat bract color transition to black.The color of bracts generally turns yellow or black from green during cereal grain development. However, the impact of these phenotypic changes on photosynthetic physiology during black bract formation remains unclear. Two oat cultivars ( Bracts, the outer protective structure of flower and seed organs in Gramineae, consist of lemma and palea . Most brOryza sativa L.) and oat (Avena sativa L.). The formation of black bracts is related to the accumulation of melanin. Melanin is a strong antioxidant, which can endow seeds with more vitality [Fusarium spp. [p-coumaric acid, one of the key intermediates in the anthocyanin biosynthesis pathway with phenylalanine as a precursor, which was catalyzed by cinnamate 4-hydroxylase (C4H) [Bh4 (black hull 4) and Phr1 (phenol reaction 1)—encoding a tyrosine transporter and a polyphenol oxidase, respectively, which are involved in melanin biosynthesis [Hordeum vulgare L.) bracts occurred in chloroplasts [Bract color is among the important morphological characteristics in Gramineae plants ,9. Durinvitality . Melaninvitality . It can ium spp. ,13,14. Vium spp.  found thse (C4H) ,17. In wynthesis ,19,20,21roplasts . The dev6fb (Cyt 6fb) and ATP synthase (ATPase). PSII and PSI bind to different light harvesting pigment protein complexes  [β-carotene [+ to NADPH, together with ATP production [The thylakoid membrane complex system of photosynthetic organisms mainly includes photosystem II (PSII), photosystem I (PSI), cytochrome nd LHCI) ,24,25. Gcarotene ,27. LHCIcarotene . LHCII acarotene . At prescarotene . LHCs aroduction ,32,33. Aoduction . The devOat, an important annual cereal crop grown worldwide, can provide proteins, soluble fibers (β-glucans), unsaturated fatty acids, vitamins, minerals, and antioxidants ,36. It hTwo cultivars, ‘Triple Crown’ and ‘Qinghai 444’, show different colors in bracts from heading to the mature seed stage A. The brp < 0.05) at the flowering and milk stage, while carotenoids only decreased significantly (p < 0.05) at the flowering stage. These data indicated that the total chlorophyll and carotenoid contents in bracts of ‘Qinghai 444’ were lower than ‘Triple Crown’, with bracts color gradually turning dark at flowering stage.Furthermore, the photosynthetic pigment content of bracts at the heading, flowering and milk stages were measured for ‘Triple Crown’ and ‘Qinghai 444’ B. The renP), transpiration rate (E), stomatal conductance (sG) and substomatal CO2 concentrations (iC) were measured for the oat bracts during development, from the heading to milk stages, and significant differences were observed between ‘Triple Crown’ and ‘Qinghai 444’ (p > 0.05) for nP, E and sG in the bracts at the heading stage between ‘Triple Crown’ and ‘Qinghai 444’. nP, E and sG of ‘Qinghai 444’ bracts were significantly (p < 0.05) lower than those of ‘Triple Crown’ at the flowering and milk stages (iC in the bracts between ‘Triple Crown’ and ‘Qinghai 444’ during development from the heading to milk stages (Photosynthetic parameters including net photosynthetic rate (hai 444’ . Althougk stages A–C. Furtk stages D.F0), actual PSII photochemical efficiency (φPSII), PSII electron transfer rate (ETR), PSII potential photosynthetic activity (Fv/F0), and PSII maximum photochemical efficiency (Fv/Fm)  lower than that in ‘Triple Crown’  higher than those in ‘Triple Crown’, but Fv/F0, and Fv/Fm were significantly (p < 0.05) lower (F0 in ‘Qinghai 444’ maintained a significantly (p < 0.05) lower level, whereas Fv/F0, Fv/Fm and NPQ in ‘Qinghai 444’ were all significantly (p < 0.05) higher than those in ‘Triple Crown’ , minimum fluorescence ( (Fv/Fm) . The chaar trend A–E. At te Crown’ F. At the5) lower A–E. Up te Crown’ A–C,F.Analysis of the transcriptome sequence and data comparison between the cultivars of ‘Triple Crown’ and ‘Qinghai 444’ were carried out for the bracts sampled at the heading, flowering and milk stages. The results of principal component analysis showed that there were differences in gene expression of ‘Triple Crown’ vs. ‘Qinghai 444’ in the three stages, and a significant difference in gene expression occurred in the flowering stage, i.e., TCf (‘Triple Crown’ in the flowering stage) vs. QHf (’Qinghai 444’ in the flowering stage) . Data anKEGG enrichment analysis revealed a difference in the metabolism pathways in bracts in the different developing stages between ‘Qinghai 444’ and ‘Triple Crown’ B. RibosoO-glycosyl compounds and cell wall were downregulated; and all these processes in ‘Qinghai 444’ bracts were enriched in the heading stage. In particular, the upregulated and downregulated chloroplast thylakoid membrane were both enriched in the TCh vs. QHh. Ribosome and cytosolic large ribosomal subunit processes were upregulated; chloroplast thylakoid membrane, photosynthesis, photosystem I and II processes were downregulated; and all these processes in ‘Qinghai 444’ bracts were enriched in the flowering stage. Only the downregulated chloroplast thylakoid membranes were both enriched in the TCf vs. QHf. Nucleosome and protein heterodimerization activity processes were upregulated; oxylipin biosynthetic protein folding was downregulated; and all these processes in ‘Qinghai 444’ bracts were enriched in the milk stage. Similarly, the upregulated and downregulated chloroplast thylakoid membranes were both enriched in the TCm vs. QHm.GO enrichment analysis  revealedLhca4 (gene ID: avena_sativa_T6406 and avena_sativa_T33623) of LHCI and Lhcb1 of LHCII were significantly (p < 0.05) downregulated in all three stages of ‘Qinghai 444’. Lhca3 of LHCI was significantly (p < 0.05) downregulated in the flowering stage of ‘Qinghai 444’, whereas Lhcb5 of LHCII exhibited no significant differences in all three stages. Additionally, oxygen-evolving complex gene PsbO in PSII was significantly (p < 0.05) downregulated at the flowering and milk stages (Lhca4 (gene ID: avena_sativa_T6406) with the fold change ranked the second in the list of top 50 downregulated DEGs between ‘Qinghai 444′ and ‘Triple Crown’. Lhca4 (gene ID: avena_sativa_T33623) was ranked as fifteenth at the flowering stage and fourteenth at the milk stage, respectively. Lhcb1 was found at the flowering stage as the twentieth  and chlorophyllide reductase (POR) were significantly (p < 0.05) downregulated at the flowering stage of ‘Qinghai 444’ bracts compared with ‘Triple Crown’ , chalcone isomerase (CHI) and flavanone3-hydroxylase (F3H) were all significantly (p < 0.05) upregulated at the flowering stage of ‘Qinghai 444’ bracts compared with ‘Triple Crown’. However, the expression of CHS and F3H was significantly (p < 0.05) downregulated at the milk stage for ‘Qinghai 444’ bracts  upregulated at the flowering stage (Bh4 in ‘Qinghai 444’ was significantly (p < 0.05) upregulated at the flowering and milk stages (p > 0.05) difference in Phr1 expression was observed between ‘Triple Crown’ and ‘Qinghai 444’ , although iC increased with no significant difference, indicating that the decrease in nP might be caused by non-stomatal factors. Weng et al. [nP during development from the heading to the mature grain stage. Varone et al. [nP. The chlorophyll fluorescence parameters measurement in ‘Qinghai 444’ bracts showed that there was no significant difference for F0, Fv/F0, Fv/Fm, ETR, and φPSII, and only NPQ decreased significantly (p < 0.05) at the heading stage (p < 0.05), indicating a loss of PSII activity (D2 (Triticum aestivum L.), an increase in D2 transcription level could enhance PSII activity and promote photosynthesis. In addition, the downregulated expression of oxygen-evolving complex gene PsbO in ‘Qinghai 444’ corresponded to the decrease in nP  upregulated at the flowering and milk stages  increased at the flowering and milk stages in ‘Qinghai 444’  upregulated at the flowering stage in ‘Qinghai 444’ compared to ‘Triple Crown’  upregulated  downregulated at the milk stage, while ETR, φPSII went back to a normal level without significant differences  enriched at the milk stage  at the flowering and milk stages, indicating a decreasing pigment content in light harvesting antenna complexes. Meanwhile, the LHC genes, including Lhcb1, Lhcb5, Lhca3 and Lhca4, exhibited a significant decrease in transcription at the flowering stage  in September 2019. The oat seedling facility consisted of a substrate ratio of Pindstrup substrate and vermiculite equal to 2:1 in a plastic flowerpot of 23 cm height and 16cm width; four oat seeds were randomly planted in each pot. Bract color was observed at the heading stage , flowering stage , milk stage  and mature stage  in both ‘Triple Crown’ and ‘Qinghai 444’; 4 g oat bracts at the heading, flowering and milk stages were cut for subsequent experiments.The seeds of ‘Triple Crown’ (TC) and ‘Qinghai 444’ (QH) used in the experiments were from the Forage Seed Laboratory of China Agricultural University, and harvested (20 kg) at Yuershan farm in Hebei Province, China , in 2018. The bract color of ‘Triple Crown’ and ‘Qinghai 444’ was yellow and black, respectively, at the mature stage. The seeds were planted in an artificial climate chamber  in the dark until oat bracts turned completely white. The absorbance values at 663 nm, 645 nm and 470 nm were measured with 80% acetone as the blank sample. The concentrations of chlorophyll and carotenoid were calculated using the following formula :ChlorophnP), stomatal conductance (sG), transpiration rate (E) and intercellular CO2 concentrations (iC) of intact bracts were measured using a CIRAS-3 portable photosynthesis system  at the conditions of 25 °C, 1200 μmol m−2 s−1 PPFD, 50–60% relative humidity and 380 μmol mol−1 CO2. The well-developed bracts of three oat spikelets in a plastic pot were randomly selected and placed in the PLC-3 (7 × 25 mm) leaf chamber for determination, and then the data were recorded once. The whole measurement process was recorded three times, with a total of nine repeats.The gas exchange in oat bract at different developmental stages was measured according to Jin et al. . The netF0) was recorded by measuring light, and maximum fluorescence yield (Fm) was recorded with a saturating flash, while variable fluorescence yield (Fv) was calculated under dark adaptation. After 15 min of illumination, maximum fluorescence of light-adapted leaves (Fm’), steady-state fluorescence (Fs) and ground fluorescence (F0’) were recorded under light adaption. The following equations were used for calculating photosynthetic parameters.The method described by Wingler et al.  was adopFv/Fm = (Fm − F0)/Fm; PSⅡ potential photochemical activity Fv/F0; actual photochemical efficiency, φPSII = (Fm′ − Fs)/Fm′; non-photochemical quenching NPQ = (Fm − Fm′)/Fm′; photosystem II electron transfer rate, ETR = φPSII × 0.84 × 0.5.Maximum photochemical rate, 3), the sample was filtered into a three-layer Miracloth. The pellets were resuspended three times in 1 mL medium II  after centrifuging at 4 °C and 4200× g for 5min; in 1mL medium III  after centrifuging at 4 °C and 8000× g for 2 min; and in 500 μL medium III after centrifuging at 4 °C and 8000× g for 2 min. The total thylakoid membrane protein was quantified with a Pierce® BCA Protein Assay Kit .The method described by Li et al.  was usedg) were gently washed twice with buffer containing 25 mM Bis-Tris/HCl (pH 7.0) and 20% glycerol, and were solubilized in buffer containing 25 mM Bis-Tris/HCl (pH 7.0), 20% glycerol, and 1% DM, at a final protein concentration of 10 mg/mL. After incubation on ice for 30 min and centrifugation at 12,000× g for another 10 min, the supernatants were supplemented with 1/10 volume of BN sample buffer . Thylakoid protein complexes were separated by 5–12% gradient BN-PAGE in 0.75-mm thick gels. For two dimensional SDS-urea-PAGE/Western blotting analysis, excised BN-PAGE lanes were soaked in SDS sample buffer  for 30 min at room temperature and then layered onto 1-mm thick 15% gels.A slightly modified method by Peng et al.  was usedw/v) acrylamide gels containing 6M urea and transferred to nitrocellulose membranes. Proteins were bound with twelve specific antibodies belonging to each thylakoid membrane complex  in TTBS buffer  with 1% skimmed milk, and immunoblot signals were measured by the chemiluminescence method in an AlphaImager HP multifunctional imaging analysis system, FluorChemR .For normal immunoblotting, thylakoid membranes were resuspended in SDS sample buffer containing 1% bromophenol blue. Protein samples were separated by SDS-urea-PAGE using 15% . Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase. Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. The cDNAs were fragmented for a preferred size of 240 bp, and the library preparations were sequenced on an Illumina Hiseq 2000 platform with paired-end mode.Total RNA was extracted from fresh intact oat bracts at different developmental stages according to the manufacturer’s instructions for TRIzol reagent . RNA concentration was measured using a NanoDrop 2000 . RNA integrity was assessed using the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system . A total amount of 1μg RNA per sample with three replicates was used for library construction using the NEBNextp value ≤ 0.05. GO and KEGG enrichment analyses on the DEGs were performed in R package clusterProfiler [We created an oat transcriptome reference (unpublished) for RNA-seq analysis in this study. Full-length transcriptome sequencing was performed based on an RNA pool of embryos, endosperm, seedlings, leaves, stems, roots and florets. After de-redundancy, 67,184 high-quality transcript sequences were obtained. RNA-seq data were analyzed with trimming by Trimmomatic-0.39 , mappingProfiler . The RNATMRT reagent kit . cDNA was diluted 10-fold for qRT-PCR analysis. Each sample was amplified three times using SYBR Premix Ex Taq  on the Bio-Rad CFX96 real-time PCR detection system , with Actin-2 as the internal control (−ΔΔCT) of target gene expression was calculated using the comparative cycle threshold method [All gene-specific primers used in qRT-PCR experiments were designed by Primer Premier 5  Table S. The RNA control . The reld method .p < 0.05) at the flowering stage compared with ‘Triple Crown’ and the bract color began to change at this time. The chlorophyll content decreased, LHCII involved in the assembly of supercomplexes in the thylakoid membrane was inhibited, and the expression of Lhcb1 and Lhcb5 was downregulated at the flowering stage. During this critical stage, the expression of Bh4 and C4H was upregulated, and the biosynthetic pathway of p-coumaric acid using tyrosine and phenylalanine as precursors was enhanced. The key genes of anthocyanin biosynthesis  were also upregulated, indicating their role in complementation for PSII activity. Then, the activity of PSII could be recovered to the original level at the milk stage. However, the mechanism of gene regulating remains unclear for the transition from green to black in bract color change during oat development. The regulatory network among the key genes of melanin, anthocyanin and chlorophyll biosynthesis and the subunit genes of the light harvesting antenna complex needs to be studied further. These results provided a foundation for better understanding of the molecular mechanism of photosynthesis limitation at the flowering stage during the development of black bract, and also established a theoretical basis for using genetic modification to improve the photosynthetic potential of non-foliar tissues.Based on the above results, we mapped a molecular regulatory scheme of photosynthesis in response to color changes during oat bract development. . The φPS