TIMAHAC: Streamlined Tandem IMAC-HILIC Workflow for Simultaneous and High-Throughput Plant Phosphoproteomics and N-glycoproteomics.

Protein post-translational modifications (PTMs) are crucial in plant cellular processes, particularly in protein folding and signal transduction. N-glycosylation and phosphorylation are notably significant PTMs, playing essential roles in regulating plant responses to environmental stimuli. However, current sequential enrichment methods for simultaneous analysis of phosphoproteome and N-glycoproteome are labor-intensive and time-consuming, limiting their throughput. Addressing this challenge, this study introduces a novel tandem S-Trap-IMAC-HILIC (S-Trap: suspension trapping; IMAC: immobilized metal ion affinity chromatography; HILIC: hydrophilic interaction chromatography) strategy, termed TIMAHAC, for simultaneous analysis of plant phosphoproteomics and N-glycoproteomics. This approach integrates IMAC and HILIC into a tandem tip format, streamlining the enrichment process of phosphopeptides and N-glycopeptides. The key innovation lies in the use of a unified buffer system and an optimized enrichment sequence to enhance efficiency and reproducibility. The applicability of TIMAHAC was demonstrated by analyzing the Arabidopsis phosphoproteome and N-glycoproteome in response to abscisic acid (ABA) treatment. Up to 1954 N-glycopeptides and 11,255 phosphopeptides were identified from Arabidopsis, indicating its scalability for plant tissues. Notably, distinct perturbation patterns were observed in the phosphoproteome and N-glycoproteome, suggesting their unique contributions to ABA response. Our results reveal that TIMAHAC offers a comprehensive approach to studying complex regulatory mechanisms and PTM interplay in plant biology, paving the way for in-depth investigations into plant signaling networks.

Luftsichel sign.

Here we present a case of a patient with breathlessness and cough admitted to the COVID ward. Chest radiography demonstrates findings consistent with lobar collapse, giving rise to the 'Luftsichel sign'. This sign has been described in the literature and highlights the importance of recognition and prompt further investigation.

Analysis of Pathogenesis and Treatment of Chronic Coronary Syndrome Based on Vessel-collateral Theory / 中国实验方剂学杂志

Based on the syndrome and treatment system of collateral disease， and inheriting the development of the bloodline theory of traditional Chinese medicine （TCM）， academician WU Yi-ling systematically constructed the vessel-collateral theory of TCM and proposed that its core theory was the theory of Yingwei， that is， "Ying in the vein， and Wei outside the vein" （Huangdi Neijing·Lingshu·Yingwei Shenghui）， "obstructing of Yingwei， congelation of blood" （Treatise on Febrile Diseases· Pulse Differentiation Method）， "pathogen transferring through blood vessels， obstructed by blocking" （Jinkui Yaolue·Zangfu Jingluo Xianhou Bingmaizheng First）， and "damage of the heart， adjust its Yingwei". Based on the consistency of vessel-collateral as the channel of blood circulation in TCM with the blood vessels in western medicine， and guided by the Yingwei theory of vessel-collateral theory， the diagnostic criteria of syndrome differentiation of disease of vessel-collateral and vascular system represented by coronary heart disease， arrhythmia， heart failure and others was established to guide the prevention and treatment of vascular diseases. Based on the above analysis， guided by the Yingwei theory of vessel-collateral theory， and on the basis of related researches of vessel-collateral and vascular system， this paper discussed the etiology and pathogenesis of chronic coronary syndrome （CCS） in TCM. Taking useful collateral with unblocking as the treatment principle， the representative Tongluo prescription （Tongxinluo） was constructed， the research progress of Tongxinluo from various aspects such as animal experiment research， pharmacological research and clinical evidence-based research was summarized， a comprehensive system from etiology， pathogenesis， syndrome differentiation to treatment was formed， in order to provide new ideas for the clinical treatment of CCS.

Effects of Outdoor Temperature on Blood Pressure in a Prospective Cohort of Northwest China / 生物医学与环境科学（英文）

Objective@#The relationship between outdoor temperature and blood pressure (BP) has been inconclusive. We analyzed data from a prospective cohort study in northwestern China to investigate the effect of outdoor temperature on BP and effect modification by season.@*Methods@#A total of 32,710 individuals who participated in both the baseline survey and the first follow-up in 2011-2015 were included in the study. A linear mixed-effect model and generalized additive mixed model (GAMM) were applied to estimate the association between outdoor temperature and BP after adjusting for confounding variables.@*Results@#The mean differences in systolic blood pressure (SBP) and diastolic blood pressure (DBP) between summer and winter were 3.5 mmHg and 2.75 mmHg, respectively. After adjusting for individual characteristics, meteorological factors and air pollutants, a significant increase in SBP and DBP was observed for lag 06 day and lag 04 day, a 0.28 mmHg (95% @*Conclusions@#This study demonstrated a significant negative association between outdoor temperature and BP in a high-altitude environment of northwest China. Moreover, BP showed a significant seasonal variation. The association between BP and temperature differed by season and individuals' demographic characteristics (age, gender, BMI), unhealthy behaviors (smoking and alcohol consumption), and chronic disease status (CVDs, hypertension, and diabetes).

Pathogenesis of Metabolic Syndrome from "Qi of five Internal Organs" / 中国实验方剂学杂志

Metabolic syndrome (MS) is a group of syndromes caused by the disorder of metabolism of various substances in the body. The main clinical manifestations are dyslipidemia, central obesity, hypertension, abnormal glucose tolerance and insulin resistance. With the changes of diet structure and habits, the incidence rate of MS is increasing, and the patients are younger. It is an important factor in many diseases, such as diabetes, atherosclerosis, coronary heart disease, hyperlipidemia, cirrhosis and some cancers. MS has seriously affected people's lives and health. Central obesity and insulin resistance are recognized as important pathogenic factors. At present, the pathogenesis of MS and its components has not been fully understood. The clinical manifestations of metabolic syndrome are complex and diverse. Traditional Chinese medicine (TCM) believes that the occurrence of metabolic syndrome is related to such factors as proper diet, emotional disorders, excessive escape and little movement, old age and physical deficiency. TCM scholars have studied the pathogenesis of MS in such pathological factors as phlegm and blood stasis, such visceral functions as liver, spleen and kidney, roles of Qi and blood, and emotional factors. As the basic substance of organism, Qi is closely related to the process of metabolism. The occurrence of MS is closely related to the rise and fall of Qi moving to and from the body as well as the abnormal gasification function of the transformation of Qi. Qi is derived from the five internal organs, which are respectively called Heart Qi, liver Qi, spleen Qi, lung Qi and kidney Qi. The "Qi of the five internal organs" is involved in the whole process of the generation, transportation and excretion of the essence of the body. Based on the "Qi of five internal organs", this paper discusses the pathogenesis of MS with phlegm, blood stasis and water drink as pathological factors.

Effect of nurse-led multiple disciplinary team-based intervention in the prevention of venous thromboembolism in paitents undergoing general surgery / 中国实用护理杂志

Objective:To investigate the effect of nurse-led multiple disciplinary team-based intervention in the prevention of venous thromboembolism in paitents undergoing general surgery.Methods:A total of 118 patients who underwent general surgery in the Tianjin First Central Hospital from May 2017 to October 2018 were divided into study group and control group by random digits table method, with 59 cases in each group. The control group received routine thrombosis prevention nursing, the study group carried out nurse-led multiple disciplinary team-based intervention. The condition of lower limbs deep venous hemodynamic was detected by color Doppler ultrasonography at 3 days after surgery, the levels of D-dimer, thrombelastograph coagulation analyzer (TEG) coagulation parameters were also measured at after 24 hours of admission and postoperative day 3, respectively.Results:The vein blood stasis rate was 94.9% (3/59) in the study group, 79.7% (12/59) in the control group, the venous blood flow of the lower 1imbs in the study group was better than that in the control group ( Z value was 2.477, P<0.05). At 3 days after surgery, the levels of D-dimer were (5.26±1.42) mg/L in the study group, (6.36±1.58) mg/L in the control group, D-dimer was decreased in study group compared to the control group, the difference was statistically significant ( t value was 3.991, P<0.05). Coagulation reaction time(R) value and solidification angle(Angel), maximum thrombus intensity(MA), composite coagulation index(CI) levels were (5.30±0.91) min, (69.64±21.93) deg, (65.40±13.76) mm and (1.23±0.20) in the study group, those index were (4.41±0.75) min, (76.64±16.02) deg, (70.98±13.39) mm, (2.09±0.36) in the control group, R value were increased and Angel, MA, CI levels were decreased in the study group compared to the control group ( t value was 2.001-15.997, P<0.05). Conclusions:Nurse-led multiple disciplinary team-based intervention improves the lower limbs deep venous hemodynamic and coagulation function, as well as reduce the incidence of venous thromboembolism.

Perioperative nursing experience of intravascular interventional treatment of patients with acute lower extremity deep venous thrombosis complicated with pulmonary embolism / 中国中西医结合急救杂志

Objective To observe the impact of enhancing perioperative care on the clinical efficacy of the patients with intravascular intervention for treatment of patients with acute lower extremity deep venous thrombosis (LEDVT) complicated with pulmonary embolism (PE). Methods Twenty-nine patients with LEDVT complicated with PE admitted to the Tianjin First Center Hospital from May 2015 to February 2019 were enrolled, the pulmonary angiography + pulmonary thrombolysis was applied for all the patients, and 25 cases used inferior vena cava filter implantation, 1 patient underwent catheter-directed thrombolysis (CDT), 4 patients underwent mechanical endovascular aspiration of thromboembolism (PTM), etc intravascular interventional treatment, the peri-operative care was strengthened, including closely observation on the patient's condition, correct use of anticoagulant and thrombolytic drugs, in the mean time, after operation, the nursing care and observation on thrombolysis catheters, involved limbs, complications should be well performed, and health education and discharge guidance ought to be carried out. Before the patient was discharged from the hospital, the improvement of clinical symptoms and the changes of coagulation indexes D-dimer, fibrinogen (Fib), prothrombin time (PT), and activated partial thromboplastin time (APTT) were observed; the Bartel index evaluation quantitative scale was used to assess the patient's activity of daily living (ADL) score; WeChat was used to investigate patient's satisfaction. Results In this group of 29 patients, postoperative chest tightness, shortness of breath symptoms were improved significantly, no fatal PE etc complications occurred. The pulmonary artery computed tomographic angiography (CTA) showed that the pulmonary artery and its branches PE basically disappeared or the measured range or area was obviously decreased. Re-examination of blood gas analysis showed that the arterial oxygen partial pressure (PaO2) was increased. Echocardiography revealed the improvement in pulmonary hypertension. 12 patients with pulmonary infection were improved after anti-inflammatory treatment, the swelling of the affected limb was significantly relieved, and the patients had no distending pain after activities on the ground. After treatment, the coagulation indexes D-dimer and fibrinogen (Fib) were significantly lower than those before treatment [D-dimer (μg/L):2 389.45±2 061.02 vs. 6 093.45±2 586.99, Fib (g/L): 3.18±0.91 vs. 4.07±1.49, both P < 0.01], there were no significant differences in PT and APTT before and after treatment [PT (s): 13.12±2.27 vs. 12.33±2.06, APTT (s):34.51±11.79 vs. 31.84±3.05, both P > 0.05]. After treatment, the ADL score of patients was obviously higher than that before treatment (scores: 79.66±6.26 vs. 17.24±8.30, P < 0.01). WeChat was used to investigate the patients' satisfaction, the nursing score was 95-100 points with an average of (97.38±1.37) points. Conclusion High quality nursing care can minimize the suffering to the largest extent in the patients with pulmonary thrombosis, elevate the quality of life, and reduce the disability and recurrence rates.

The Clinical Analysis of Necrotizing Enterocolitis / 昆明医科大学学报

Objective From May 2010 to May 2013 and from June 2013 to June 2016, we compared the clinical analysis of patients with necrotizing enterocolitis (NEC) in the Department of Pediatrics, studied incidence rate of NEC and the relationship between stages of NEC and prognosis, and compared the prognosis between the two three-year periods to provide the experience of diagnosis and treatment for clinical practice in the future. Methods The clinical data of patients diagnosed with NEC were retrospectively analyzed according to the diagnostic criteria of modified Bell staging classification using SPSS statistical software package. Early diagnosis rate and prognosis were compared between the two three-year periods. P＜0.05 showed that statistical significance was found. Results From May 2010 to May 2013, 40 cases of NEC were diagnosed in our hospital, among which 8 was in the first stage (20%),24 in the second stage (60%),and 8 in the third stage (20%) . The number of premature infants was 32, accounting for 80% , among which 25 cases were infants with very low birth weight accounting for 62.5% and 8 cases were full-term infants, accounting for 20% (P＜0.05) . In the study, 32 cases (80%) received traditional treatment, which was effective in stage Ⅰ. During the second stage, two abandoned the treatment was confirmed death in the follow-up, two underwent surgery successfully due to the progression of the disease and four (25%) died in the hospital. Statistic significance of treatment of internal medicine was found when comparing stage Ⅰ with stage Ⅱ (P＜0.05) . Ten children underwent surgery, among which one gave up treatment after the surgery because of financial problems and was found dead in the follow-up and four (60%) died in the hospital. The comparison of surgical treatment in patients in stage Ⅱ and Ⅲ showed no statistical significance (P＞0.05) . Fatality rates were statistically significant (P＜0.05) . From June 2013 to June 2016, 41 children were diagnosed with NEC in our hospital, including 24 cases in stage Ⅰ (59%), 14 in stage Ⅱ (34%), and 3 in stage Ⅲ (7%) . The number of premature infants was 33, accounting for 80%, among which 26 cases were infants with very low birth weight accounting for 62.5% and 8 cases were full-term infants, accounting for 20% (P＜0.05) . In the study, 35 cases (85.3%) received traditional treatment, which was effective in stage Ⅰ. During the second stage, one underwent surgery successfully due to the progression of the disease. Statistic significance of treatment of internal medicine was found when comparing stage Ⅰ with stage Ⅱ (P＜0.05) . Four children underwent surgery, among which one gave up treatment after the surgery because of financial problems and was found dead in the follow-up and the surgery was successful in other three. The comparison of surgical treatment in patients in stage Ⅱ and Ⅲ showed statistical significance (P＜0.05) . Fatality rates were statistically significant comparing the two three-year periods (P＜0.05) . The comparison of the rates of early diagnosis in the two three-year periods showed statistically significance (P＜0.001) . Conclusion Early diagnosis of NEC is the key to reduce mortality. Intestinal fatty acid binding protein (I-FABP) is a serological marker for early diagnosis of NEC. As an important factor, the reduction of the incidence of premature birth will result in the drop in the incidence of NEC.

NKX2.5 and TBX5 gene mutations in in vitro fertilization children with congenital heart disease / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the differences of NKX2.5 and TBX5 gene mutations between in vitro fertilization (IVF) children with congenital heart disease (CHD) and naturally conceived children with CHD.</p><p><b>METHODS</b>Blood samples from 68 IVF children with CHD and 98 naturally conceived children with CHD were collected. The mutations in coding regions 1 and 2 of the NKX2.5 gene, and coding regions 4, 5, and 8 of the TBX5 gene were examined by polymerase chain reaction (PCR) and DNA sequencing.</p><p><b>RESULTS</b>An A-to-G mutation at nucleotide 63 (c.63A>G) in coding region 1 of the NKX2.5 gene was found in both IVF and naturally conceived children with CHD. There were no significant differences in genotype and allele frequencies at c.63A>G locus of the NKX2.5 gene between the two groups. No mutations were detected in coding region 2 of the NKX2.5 gene and coding regions 4, 5 and 8 of the TBX5 gene.</p><p><b>CONCLUSIONS</b>There is no difference in NKX2.5 and TBX5 gene mutations between IVF and naturally conceived children with CHD. Therefore, it is presumed that assisted reproductive technology may not lead to mutations in the NKX2.5 and TBX5 genes.</p>

Involvement of type VI secretion system in secretion of iron chelator pyoverdine in Pseudomonas taiwanensis.

Rice bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most destructive rice diseases worldwide. Therefore, in addition to breeding disease-resistant rice cultivars, it is desirable to develop effective biocontrol agents against Xoo. Here, we report that a soil bacterium Pseudomonas taiwanensis displayed strong antagonistic activity against Xoo. Using matrix-assisted laser desorption/ionization imaging mass spectrometry, we identified an iron chelator, pyoverdine, secreted by P. taiwanensis that could inhibit the growth of Xoo. Through Tn5 mutagenesis of P. taiwanensis, we showed that mutations in genes that encode components of the type VI secretion system (T6SS) as well as biosynthesis and maturation of pyoverdine resulted in reduced toxicity against Xoo. Our results indicated that T6SS is involved in the secretion of endogenous pyoverdine. Mutations in T6SS component genes affected the secretion of mature pyoverdine from the periplasmic space into the extracellular medium after pyoverdine precursor is transferred to the periplasm by the inner membrane transporter PvdE. In addition, we also showed that other export systems, i.e., the PvdRT-OpmQ and MexAB-OprM efflux systems (for which there have been previous suggestions of involvement) and the type II secretion system (T2SS), are not involved in pyoverdine secretion.

Genome mining reveals the biosynthetic potential of the marine-derived strain Streptomyces marokkonensis M10.

Marine streptomycetes are rich sources of natural products with novel structures and interesting biological activities, and genome mining of marine streptomycetes facilitates rapid discovery of their useful products. In this study, a marine-derived Streptomyces sp. M10 was revealed to share a 99.02% 16S rDNA sequence identity with that of Streptomyces marokkonensis Ap1T, and was thus named S. marokkonensis M10. To further evaluate its biosynthetic potential, the 7,207,169 bps of S. marokkonensis M10 genome was sequenced. Genomic sequence analysis for potential secondary metabolite-associated gene clusters led to the identification of at least three polyketide synthases (PKSs), six non-ribosomal peptide synthases (NRPSs), one hybrid NRPS-PKS, two lantibiotic and five terpene biosynthetic gene clusters. One type I PKS gene cluster was revealed to share high nucleotide similarity with the candicidin/FR008 gene cluster, indicating the capacity of this microorganism to produce polyene macrolides. This assumption was further verified by isolation of two polyene family compounds PF1 and PF2, which have the characteristic UV adsorption at 269, 278, 290 nm (PF1) and 363, 386 and 408 nm (PF2), respectively. S. marokkonensis M10 is therefore a new source of polyene metabolites. Further studies on S. marokkonensis M10 will provide more insights into natural product biosynthesis potential of related streptomycetes. This is also the first report to describe the genome sequence of S. marokkonensis-related strain.

Value of combined measurement of intestinal fatty acid-binding protein and fecal calprotectin in diagnosis of necrotizing enterocolitis in full-term neonates / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the value of combined measurement of intestinal fatty acid-binding protein (I-FABP) and fecal calprotectin (FC) in the diagnosis of necrotizing enterocolitis (NEC) in full-term neonates.</p><p><b>METHODS</b>A total of 36 full-term neonates with NEC (case group) and 39 neonates without digestive system diseases (control group) were enrolled as study subjects. ELISA was used to measure the serum I-FABP level and fecal FC level, and the clinical value of I-FABP combined with FC in the diagnosis of NEC was evaluated.</p><p><b>RESULTS</b>The case group had significantly higher I-FABP and FC levels than the control group (P<0.05). In the case group, serum I-FABP level was positively correlated with fecal FC level (r=0.71, P<0.05). In the diagnosis of NEC, I-FABP alone, FC alone, and I-FABP/FC combination had sensitivities of 83.3%, 81.5%, and 79.5%, specificities of 72.5%, 75.8%, and 86.3%, and areas under the ROC curve (AUCs) of 0.82, 0.81, and 0.88. The combined measurement showed significantly higher specificity and AUC than single measurement (P<0.05).</p><p><b>CONCLUSIONS</b>Children with NEC have significant increases in I-FABP and FC levels, and there is a correlation between them. Combined measurement of I-FABP and FC can increase the specificity of the diagnosis of NEC.</p>

Effect of advanced maternal age on birth defects and postnatal complications of neonates / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of advanced maternal age on birth defects and postnatal complications of neonates.</p><p><b>METHODS</b>Among the 1 109 neonates who were born at The First People's Hospital of Yunnan Province between January 2014 and December 2015, 536 neonates whose mothers were aged ≥35 years were enrolled as advanced age group and 573 neonates whose mothers were aged <35 years were enrolled as appropriate-age group. The incidences of the comorbidities in pregnancy, fetal intrauterine distress, neonatal birth defects, and postnatal complications were compared between the two groups. A univariate logistic regression analysis was performed to analyze the effect of advanced maternal age on neonatal comorbidities during perinatal period.</p><p><b>RESULTS</b>Compared with the appropriate-age group, the advanced age group had significantly higher rate of caesarean section and incidence rates of multiple birth, gestational diabetes, pregnancy-induced hypertension, in vitro fertilization, and fetal intrauterine distress (P<0.01). The neonates in the advanced age group had a significantly higher incidence rate of cleft lip and palate and a significantly lower rate of skeletal dysplasia than in the appropriate-age group (P<0.05). Advanced maternal age was the risk factor for fetal intrauterine distress (OR=2.27, 95%CI: 1.33-3.88, P=0.003), neonatal resuscitation (OR=1.66, 95%CI: 1.19-2.31, P=0.003), and intracranial hemorrhage (OR=2.70, 95%CI: 1.21-6.04, P=0.02).</p><p><b>CONCLUSIONS</b>The women of maternal advanced age have higher incidence rates of pregnancy comorbidities than those of appropriate age, and the neonates born to the mothers of advanced maternal age have a higher incidence rate of cleft lip and palate. Advanced maternal age may increase the risks of fetal intrauterine distress, neonatal resuscitation, and intracranial hemorrhage.</p>

Vitroprocines, new antibiotics against Acinetobacter baumannii, discovered from marine Vibrio sp. QWI-06 using mass-spectrometry-based metabolomics approach.

A robust and convenient research strategy integrating state-of-the-art analytical techniques is needed to efficiently discover novel compounds from marine microbial resources. In this study, we identified a series of amino-polyketide derivatives, vitroprocines A-J, from the marine bacterium Vibrio sp. QWI-06 by an integrated approach using imaging mass spectroscopy and molecular networking, as well as conventional bioactivity-guided fractionation and isolation. The structure-activity relationship of vitroprocines against Acinetobacter baumannii is proposed. In addition, feeding experiments with (13)C-labeled precursors indicated that a pyridoxal 5'-phosphate-dependent mechanism is involved in the biosynthesis of vitroprocines. Elucidation of amino-polyketide derivatives from a species of marine bacteria for the first time demonstrates the potential of this integrated metabolomics approach to uncover marine bacterial biodiversity.

Neonatal complications and birth defects in infants conceived by in vitro fertilization / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the survival quality of infants conceived by in vitro fertilization (IVF) and to identify the factors that cause birth defects and neonatal complications in IVF infants.</p><p><b>METHODS</b>The study included 150 IVF infants (IVF group) and 200 naturally conceived infants (control group). Indicators such as birth situation, gestational disease, birth defects, and neonatal complications were compared between groups. The influencing factors for birth defects and neonatal complications were analyzed by non-conditional logistic regression analysis.</p><p><b>RESULTS</b>Compared with the control group, the IVF group had increased incidences of twin pregnancy and low birth weight (P<0.01) but decreased average birth weight (P<0.05). In the IVF group, the mother's age was elder, with higher incidence of cesarean section, premature rupture of membranes, and pregnancy complications, as compared with the control group (P<0.05). There was no significant difference in the incidence of birth defects between the two groups (P>0.05). The IVF group had higher incidence rates of low birth weight and neonatal scleroderma (P<0.05), with a longer hospital stay (P<0.01), as compared with the control group. The non-conditional logistic regression analysis indicated that IVF, prematurity, twin pregnancy, and pregnancy complications were risk factors for low birth weight (P<0.05).</p><p><b>CONCLUSIONS</b>There is no significant difference in the incidence of birth defects between IVF and naturally conceived infants. However, IVF infants have higher incidences of twin pregnancy and low birth weight, with a longer hospital stay, as compared with naturally conceived infants. Natural conceiving, avoiding prematurity, twin pregnancy, and pregnancy complications will reduce the incidence of low birth weight.</p>

Compositional and proteomic analyses of genetically modified broccoli (Brassica oleracea var. italica) harboring an agrobacterial gene.

Previously, we showed improved shelf life for agrobacterial isopentenyltransferase (ipt) transgenic broccoli (Brassica oleracea var. italica), with yield comparable to commercial varieties, because of the protection mechanism offered by molecular chaperones and stress-related proteins. Here, we used proximate analysis to examine macronutrients, chemical and mineral constituents as well as anti-nutrient and protein changes of ipt-transgenic broccoli and corresponding controls. We also preliminarily assessed safety in mice. Most aspects were comparable between ipt-transgenic broccoli and controls, except for a significant increase in carbohydrate level and a decrease in magnesium content in ipt-transgenic lines 101, 102 and 103, as compared with non-transgenic controls. In addition, the anti-nutrient glucosinolate content was increased and crude fat content decreased in inbred control 104 and transgenic lines as compared with the parental control, "Green King". Gel-based proteomics detected more than 50 protein spots specifically found in ipt-transgenic broccoli at harvest and after cooking; one-third of these proteins showed homology to potential allergens that also play an important role in plant defense against stresses and senescence. Mice fed levels of ipt-transgenic broccoli mimicking the 120 g/day of broccoli eaten by a 60-kg human adult showed normal growth and immune function. In conclusion, the compositional and proteomic changes attributed to the transgenic ipt gene did not affect the growth and immune response of mice under the feeding regimes examined.

Analysis of the biosynthesis of antibacterial cyclic dipeptides in Nocardiopsis alba.

Nocardiopsis alba is frequently isolated from environment and has recently been suggested as a casual symbiotic actinobacterium of diverse invertebrates. Using activity-guided fractionation, we purified two antibacterial cyclic dipeptides, cyclo(ΔPhe-ΔLeu) (albonoursin) and cyclo(ΔmTyr-ΔLeu), from a culture of Nocardiopsis alba ATCC BAA-2165. Analysis of N. alba genome revealed genetic information similar to albonoursin biosynthetic gene cluster, albABC. An albABC gene deletion mutant of N. alba was generated. Liquid chromatography-mass spectrometry analysis showed that the mutant could not produce the cyclic dipeptides. Cyclic dipeptide production in the mutant was restored by genetic complementation with the albABC cloned in a native plasmid of Nocardiopsis. ß-Glucuronidase reporter assays with a second mutant construct, in which albABC promoter is transcriptionally fused to the reporting gene gusA, indicated that albABC gene expression was subject to osmoregulation. The system presented will be used to study the metabolic and genetic control of cyclic dipeptide biosynthesis in Nocardiopsis.

Bringing microbial interactions to light using imaging mass spectrometry.

Covering: up to the end of September 2013 Microorganisms are a plentiful resource for natural products research. Traditionally, natural products discovery from microbial sources depends on the screening of target-mediated inhibition. The natural products identified through this strategy usually correlate to significant microbial phenotypes. However, the target-mediated transcriptions deduced from low concentrations of natural products sometimes do not generate an obvious phenotype. The better understanding of the true biological roles of those microbial natural products will permit the application of rational approaches to the more effective exploitation of their use. Imaging mass spectrometry (IMS) has been developed and applied in many fields for decades. However, the applications of IMS on microbial natural products research have just been recently reported. IMS is one of few tools capable of revealing both phenotype and relevant and irrelevant chemotypes of microorganisms. In this review, we summarize the latest applications of IMS technologies. The challenges and prospect of improvement and application of IMS to microbial natural products research are discussed as well.

Proteomics and transcriptomics of broccoli subjected to exogenously supplied and transgenic senescence-induced cytokinin for amelioration of postharvest yellowing.

Previously, we investigated transgenic broccoli harboring senescence-associated-gene (SAG) promoter-triggered isopentenyltransferase (ipt), which encodes the key enzyme for cytokinin (CK) synthesis and mimics the action of exogenous supplied CK in delaying postharvest senescence of broccoli. Here, we used proteomics and transcriptomics to compare the mechanisms of ipt-transgenic and N(6)-benzylaminopurine (BA) CK treatment of broccoli during postharvest storage. The 2 treatments conferred common and distinct mechanisms. BA treatment decreased the quantity of proteins involved in energy and carbohydrate metabolism and amino acid metabolism, and ipt-transgenic treatment increased that of stress-related proteins and molecular chaperones and slightly affected levels of carbohydrate metabolism proteins. Both treatments regulated genes involved in CK signaling, sugar transport, energy and carbohydrate metabolism, amino acid metabolism and lipid metabolism, although ipt-transgenic treatment to a lesser extent. BA treatment induced genes encoding molecular chaperones, whereas ipt-transgenic treatment induced stress-related genes for cellular protection during storage. Both BA and ipt-transgenic treatments acted antagonistically on ethylene functions. We propose a long-term acclimation of metabolism and protection systems with ipt-transgenic treatment of broccoli and short-term modulation of metabolism and establishment of a protection system with both BA and ipt-transgenic treatments in delaying senescence of broccoli florets. BIOLOGICAL SIGNIFICANCE: Transgenic broccoli harboring senescence-associated-gene (SAG) promoter-triggered isopentenyltransferase (ipt), which encodes the key enzyme for cytokinin (CK) synthesis and N(6)-benzylaminopurine (BA) CK treated broccoli both showed retardation of postharvest senescence during storage. The mechanisms underlying the two treatments were compared. The combination of proteomic and transcriptomic evidences revealed that the 2 treatments conferred common and distinct mechanisms in delaying senescence of broccoli florets. We propose a long-term acclimation of metabolism and protection systems with ipt-transgenic treatment of broccoli and short-term modulation of metabolism and establishment of a protection system with both BA and ipt-transgenic treatments in delaying senescence of broccoli florets. This article is part of a Special Issue entitled: Translational Plant Proteomics.

Molecular and functional characterization of broccoli EMBRYONIC FLOWER 2 genes.

Polycomb group (PcG) proteins regulate major developmental processes in Arabidopsis. EMBRYONIC FLOWER 2 (EMF2), the VEFS domain-containing PcG gene, regulates diverse genetic pathways and is required for vegetative development and plant survival. Despite widespread EMF2-like sequences in plants, little is known about their function other than in Arabidopsis and rice. To study the role of EMF2 in broccoli (Brassica oleracea var. italica cv. Elegance) development, we identified two broccoli EMF2 (BoEMF2) genes with sequence homology to and a similar gene expression pattern to that in Arabidopsis (AtEMF2). Reducing their expression in broccoli resulted in aberrant phenotypes and gene expression patterns. BoEMF2 regulates genes involved in diverse developmental and stress programs similar to AtEMF2 in Arabidopsis. However, BoEMF2 differs from AtEMF2 in the regulation of flower organ identity, cell proliferation and elongation, and death-related genes, which may explain the distinct phenotypes. The expression of BoEMF2.1 in the Arabidopsis emf2 mutant (Rescued emf2) partially rescued the mutant phenotype and restored the gene expression pattern to that of the wild type. Many EMF2-mediated molecular and developmental functions are conserved in broccoli and Arabidopsis. Furthermore, the restored gene expression pattern in Rescued emf2 provides insights into the molecular basis of PcG-mediated growth and development.