Comparative proteomic analysis of latex from Euphorbia kansui laticifers at different development stages with and without UV-B treatment via iTRAQ-coupled two-dimensional liquid chromatography–MS/MS

2020 ◽  
Vol 47 (1) ◽  
pp. 67 ◽  
Author(s):  
Xueyan Zhao ◽  
Yue Zhang ◽  
Meng Wang ◽  
Xiaoai Fang ◽  
Xia Cai
Keyword(s):  
Lysosomal Enzymes ◽  
Development Stage ◽  
Pcr Analysis ◽  
Absolute Quantitation ◽  
Protein Database ◽  
Development Stages ◽  
Refseq Protein ◽  
Euphorbia Kansui ◽  
Isobaric Tagging ◽  
Metabolite Synthesis

Euphorbia kansui Liou, an endemic species in China, is well-known in traditional Chinese medicine. All parts of E. kansui contain white latex, which is the protoplasm constituent of specialised cells known as laticifers. The latex contains many proteins with various biological functions. In the present study, isobaric tagging for relative and absolute quantitation (iTRAQ) and MS technology combined with western blot and quantitative real-time PCR analysis were used to identify latex proteins and analyse differentially accumulated proteins in laticifers at different development stages, with and without UV-B treatment according to the E. kansui transcriptome database and the NCBI Euphorbiaceae RefSeq protein database. A total of 322 latex proteins were successfully identified. Proteasome subunits, ubiquitinated proteins, vacuolar ATP synthase (V-ATPase) and lysosomal enzymes decreased, keeping the content at a higher level in laticifers in the early development stage. These results suggest that the ubiquitin–proteasome pathway and the lysosome autophagy pathway were involved in the partial degradation of laticifer cytoplasm. In addition, terpenoid biosynthesis-related proteins, 14–3–3 protein, V-ATPase and lysosomal enzymes increased under UV-B treatment, which showed that partial cytoplasmic degradation is positively correlated with secondary metabolite synthesis in the development of E. kansui laticifers. Besides, UV-B radiation can increase plant resistance by promoting laticifer development in E. kansui. This information provides a basis for further exploration of E. kansui laticifer development, and terpenoid synthesis and regulation.

scholarly journals Ethylene and 1-methylcyclopropene action over senescence of nasturtium flowers

2015 ◽  
Vol 33 (4) ◽  
pp. 453-458 ◽  
Author(s):  
Tania P Silva ◽  
Fernando L Finger
Keyword(s):  
Floral Development ◽  
Development Stage ◽  
Flower Senescence ◽  
Premature Senescence ◽  
Flower Opening ◽  
Development Stages ◽  
Post Harvest ◽  
The Third ◽  
Exogenous Ethylene ◽  
Floral Bud

ABSTRACT: This work describes ethylene and 1-methylcyclopropene (1-MCP) action on post-harvest shelf life of four development stages of nasturtium flowers. To reach this goal, we carried out three experiments. In the first and second experiments, we studied five ethylene (0; 0.1; 1; 10; 100 and 1000 μL/L) and three 1-MCP concentrations (0.25; 0.5 and 0.75 μL/L), respectively. In the third experiment, 1-MCP was followed by combined with ethylene (only 1-MCP; only ethylene; and 24 hours of exposure to 0.75 μL/L 1-MCP followed by 24 hours of exposure to 100 μL/L ethylene). All experiments had two control treatments, one keeping non-exposed flowers inside and another outside exposure chambers. Experiments were set in factorial design, in complete blocks at random, with four 10-flower replications each. Flower senescence was determined by a pre-established visual scale and by observing floral bud development. Ethylene dose above 10 μL/L induced flower wilting and premature senescence from the second floral development stage. Furthermore, higher concentrations of exogenous ethylene promoted irregular flower opening and/or morphological abnormalities in opened flowers. 1-MCP effectively extended post-harvest longevity of nasturtium flowers, independent of the concentration and even in the presence of exogenous ethylene.

scholarly journals Gene Microarray Integrated with High-Throughput Proteomics for the Discovery of Transthyretin in Rhabdomyolysis-Induced Acute Kidney Injury

2017 ◽  
Vol 43 (4) ◽  
pp. 1673-1688 ◽  
Author(s):  
Ou Li ◽  
Xiaodong Geng ◽  
Qian Ma ◽  
Weiwei Wang ◽  
Ran Liu ◽  
...  
Keyword(s):  
Acute Kidney Injury ◽  
Western Blotting ◽  
Kidney Injury ◽  
Physical Exertion ◽  
Renal Tissue ◽  
Gene Microarray ◽  
Absolute Quantitation ◽  
Gene And Protein Expression ◽  
Isobaric Tagging

Background/Aims: Rhabdomyolysis, one of the leading causes of acute kidney injury (AKI), develops after trauma, drug toxicity, infections, burns, and physical exertion. The aim of this study was to investigate differences in gene and protein expression to elucidate the pathogenesis of rhabdomyolysis (RM)-induced AKI. Methods: In this study, we used glycerol induced renal injury as a model of RM-induced AKI. Affymetrix U133 plus 2.0 microarrays were used to perform gene microarray analysis. Isobaric tagging with related and absolute quantitation (iTRAQ) labeling mass spectrometry (MS) was applied to screen and identify differentially expressed proteins between RM-induced AKI and normal murine renal tissue. Verification experiments included immunohistochemistry (IHC), real-time PCR, Western blotting, and the measurement of ATP and ROS production. HK-2 cells were incubated in vitro with ferrous myoglobin and pcDNA-TTR, followed by assays to detect cell proliferation, ROS and apoptosis. Results: According to gene microarray and iTRAQ-MS analysis, we screened 17 common elements. After multiple analyses, we selected transthyretin (TTR) as our focus and investigated TTR in the kidney. Verification experiments with IHC confirmed differential expression levels of TTR proteins. Furthermore, Western blotting showed a stepwise decrease in TTR in AKI renal tissues. Cell-based experiments showed that overexpression of TTR could improve HK-2 cell viability and inhibit apoptosis. TTR reduced apoptosis by decreasing the accumulation of reactive oxygen species (ROS). Conclusion: This study reports a possible mechanism for RM-induced AKI and suggests that reductions in TTR could increase the generation of ROS and induce apoptosis. TTR may be a potentially valuable target for RM-induced AKI.

scholarly journals Efecto de medicamentos homeopáticos en plantas de tomate (Solanum lycopersicum L.)

2020 ◽  
Vol 38 (1) ◽  
pp. 103
Author(s):  
Fernando Abasolo Pacheco ◽  
Boris Bonilla Montalván ◽  
Cesar Bermeo Toledo ◽  
Yarelys Ferrer Sánchez ◽  
Andy Jafet Ramirez Castillo ◽  
...  
Keyword(s):  
Root Biomass ◽  
Block Design ◽  
Development Stage ◽  
Stem Length ◽  
Control Treatment ◽  
Leaf Biomass ◽  
Control Group ◽  
Vegetative Development ◽  
Initial Development ◽  
Development Stages

Agrochemical use in horticultural cultivations generates negative effects, thus the need for searching to decrease or eliminate its use by means of other less toxic techniques. Agricultural homeopathy represents an alternative for ecological agriculture, impacting positively in cultivation development. The effect of four homeopathic medicines for human use were assessed in two centesimal dynamizations (7CH and 13CH) [(Silicea Terra (SiT), Natrum muriaticum (NaM), Zincum phosphoricum (ZiP) and Phosphoricum acidum (PhA)], and a control treatment (distilled water) on tomato plant germination, emergence, and initial development. The treatments were established under a randomized complete block design with three repiclates. Germination and emergence rate and percentage and morphometric variables (plant height, radicle length, dry and wet weight) were assessed, including the variables in stem diameter and wet and dry leaf weight, number of branches, leaves, and buds in the vegetative development stage. Signif icant differences were observed in all the morphometric variables assessed in function of the different development stages, homeopathic medicines, and their different dynamizations. During germination, greater growth in stem length was observed with ZiP-7CH (5.5 ± 0.98 cm) compared to the control group (4.3 ± 1.10 cm). During the emergence stage, the treatments SiT-7CH (6.6 ± 1.11 cm) and ZiP-7CH (5.9 ± 1.41 cm) increased stem length signif icantly whereas with PhA-7CH, the best effects were obtained in the variables assessed during the vegetative development stage, LT (94 ± 8.31 cm), leaf number (No hojas) (131 ± 27.71), fresh stem biomass (BFT) (17.20 ± 2.45 g), wet leaf biomass (BFH) (30 ± 7.72 g), dry leaf biomass (BSH) (2 ± 0.61 g), fresh root biomass (BFR) (10 ± 6.26 g), dry root biomass (BSR) (1 ± 0.43 g), and number of flower buds (No H) (6 ± 7.10). The homeopathic medicines applied impacted positively during the initial and vegetative development stages of tomato under controlled conditions. This research study represents and advance in the sustainable management of tomato cultivation.

Visceral larva migrans detection using PCR–RFLP in BALB/c mice infected with Toxocara canis

2019 ◽  
Vol 94 ◽  
Author(s):  
G. Özbakış ◽  
A. Doğanay
Keyword(s):  
Toxocara Canis ◽  
Development Stage ◽  
Clinical Syndrome ◽  
Larva Migrans ◽  
Pcr Analysis ◽  
Visceral Larva Migrans ◽  
Molecular Method ◽  
Affected Tissue ◽  
Test Specificity ◽  
Paratenic Hosts

Abstract Toxocara canis is an important zoonotic roundworm distributed worldwide. The infective larvae of T. canis are one of the causes of visceral larva migrans (VLM), a clinical syndrome in humans. Diagnosing VLM is difficult, and the differential diagnosis of the larval development stage is limited. Therefore, this experimental research aimed to diagnose T. canis larvae using a molecular method, not only in liver tissue, which is the most commonly affected tissue, but also in the limb muscles, lungs and brain tissues. For this purpose, 24 BALB/c mice were infected with 1000 embryonated T. canis eggs. Necropsies were performed on the second, fourth, seventh and 14th days post-infection. While a part of the samples were digested with pepsin-HCl, the molecular method was used for the remainder of the samples to replicate the mitochondrial DNA adenosine triphosphate (ATP) synthase subunit-6 gene region of T. canis. BbsI, a restriction endonuclease, was used to determine the specificity of the amplicons obtained from Polymerase chain reaction (PCR). The detection limit for embryonated eggs was recorded. The PCR results showed that the sensitivity of the PCR analysis was 83.3% in the liver (with 88.8% accuracy), 87.5% in the lungs (with 91.6% accuracy) and 75.0% in the brain, forelimb and hindlimb muscles (with 83.3% accuracy). In all tissues, the test specificity was determined to be 100%. In this study, the molecular method was applied to only experimentally infected BALB/c mice tissues; thus, it is suggested that it can be also employed in different paratenic hosts and materials possibly infected with T. canis.

scholarly journals iTRAQ-Based Proteomics Analysis of Autophagy-Mediated Responses against MeJA in Laticifers of Euphorbia kansui L.

2019 ◽  
Vol 20 (15) ◽  
pp. 3770
Author(s):  
Fang ◽  
Yao ◽  
Zhang ◽  
Tian ◽  
Wang ◽  
...  
Keyword(s):  
Developmental Process ◽  
Cysteine Proteinase ◽  
Defense Responses ◽  
Absolute Quantification ◽  
Pcr Analysis ◽  
Control Group ◽  
Proteomics Analysis ◽  
Meja Treatment ◽  
Euphorbia Kansui ◽  
Isobaric Tags

Autophagy is a well-defined catabolic mechanism whereby cytoplasmic materials are engulfed into a structure termed the autophagosome. Methyl jasmonate (MeJA), a plant hormone, mediates diverse developmental process and defense responses which induce a variety of metabolites. In plants, little is known about autophagy-mediated responses against MeJA. In this study, we used high-throughput comparative proteomics to identify proteins of latex in the laticifers. The isobaric tags for relative and absolute quantification (iTRAQ) MS/MS proteomics were performed, and 298 proteins among MeJA treated groups and the control group of Euphorbia kansui were identified. It is interesting to note that 29 significant differentially expressed proteins were identified and their associations with autophagy and ROS pathway were verified for several selected proteins as follows: α-L-fucosidase, β-galactosidase, cysteine proteinase, and Cu/Zn superoxide dismutase. Quantitative real-time PCR analysis of the selected genes confirmed the fact that MeJA might enhance the expression of some genes related to autophagy. The western blotting and immunofluorescence results of ATG8 and ATG18a which are two important proteins for the formation of autophagosomes also demonstrated that MeJA could promote autophagy at the protein level. Using the electron microscope, we observed an increase in autophagosomes after MeJA treatment. These results indicated that MeJA might promote autophagy in E. kansui laticifers; and it was speculated that MeJA mediated autophagy through two possible ways: the increase of ROS induces ATG8 accumulation and then aotophagosome formation, and MeJA promotes ATG18 accumulation and then autophagosome formation. Taken together, our results provide several novel insights for understanding the mechanism between autophagy and MeJA treatment. However, the specific mechanism remains to be further studied in the future.

scholarly journals A cellular and proteomic approach to assess proteins extracted from cryopreserved human amnion in the cultivation of corneal stromal keratocytes for stromal cell therapy

2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Beau J. Fenner ◽  
Nur Zahirah B. M. Yusoff ◽  
Matthias Fuest ◽  
Lei Zhou ◽  
Francisco Bandeira ◽  
...  
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Protein Extraction ◽  
Culture Media ◽  
Cell Metabolism ◽  
Absolute Quantitation ◽  
Human Amnion ◽  
Mesenchymal Transition ◽  
Proteomic Approach ◽  
Isobaric Tagging ◽  
Human Keratocytes

Abstract Background Human corneal stromal keratocytes propagated in culture media supplemented with human amnion extract (AME) can correct early corneal haze in an animal model. Clinical application of cultivated keratocytes is limited by infectious disease screening before amnion products can be used in humans. It remains unclear if AME from cryopreserved versus fresh human amnion can support human keratocyte propagation, and which components of the extract promote keratocyte growth. Methods Three placentas were collected for the preparation of fresh and cryopreserved amnion tissues followed by homogenization and protein extraction. AME protein profiles were studied using isobaric tagging for relative and absolute quantitation (iTRAQ) proteomics. Enriched gene ontology (GO) terms and functional classes were identified. Primary human keratocytes from 4 donor corneas were cultured in media supplemented with fresh AME (F-AME) or cryopreserved AME (C-AME). Cell viability, proliferation and keratocyte marker expression were examined by confocal immunofluorescence and flow cytometry. Results AME proteomics revealed 1385 proteins with similar expression levels (between 0.5- and 2-fold) between F- and C-AME, while 286 proteins were reduced (less than 0.5-fold) in C-AME. Enriched GO term and biological pathway analysis showed that those proteins with comparable expression between F-AME and C-AME were involved in cell metabolism, epithelial-mesenchymal transition, focal adhesion, cell-extracellular matrix interaction, cell stress regulation and complement cascades. Human corneal stromal keratocytes cultured with F-AME or C-AME showed similar morphology and viability, while cell proliferation was mildly suppressed with C-AME (P > 0.05). Expression of aldehyde dehydrogenase 3A1 (ALDH3A1) and CD34 was similar in both cultures. Conclusion AME from cryopreserved amnion had limited influence on keratocyte culture. It is feasible to use protein extract from cryopreserved amnion to propagate human keratocytes for potential translational applications.

scholarly journals Serum Proteome Alterations in Patients with Cognitive Impairment after Traumatic Brain Injury Revealed by iTRAQ-Based Quantitative Proteomics

2017 ◽  
Vol 2017 ◽  
pp. 1-13 ◽  
Author(s):  
Xin-gui Xiong ◽  
Qinghua Liang ◽  
Chunhu Zhang ◽  
Yang Wang ◽  
Wei Huang ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Cognitive Impairment ◽  
Brain Injury ◽  
Bioinformatic Analysis ◽  
Serum Samples ◽  
Absolute Quantitation ◽  
Proteomic Approach ◽  
Healthy Control ◽  
Isobaric Tagging ◽  
Experimental Findings

Background. Cognitive impairment is the leading cause of traumatic brain injury- (TBI-) related disability; however, the underlying pathogenesis of this dysfunction is not completely understood. Methods. Using an isobaric tagging for relative and absolute quantitation- (iTRAQ-) based quantitative proteomic approach, serum samples from healthy control subjects, TBI patients with cognitive impairment, and TBI patients without cognitive impairment were analysed to identify differentially expressed proteins (DEPs) related to post-TBI cognitive impairment. In addition, DEPs were further analysed using bioinformatic platforms and validated using enzyme-linked immunosorbent assays (ELISA). Results. A total of 56 DEPs were identified that were specifically related to TBI-induced cognitive impairment. Bioinformatic analysis revealed that a wide variety of cellular and metabolic processes and some signaling pathways were involved in the pathophysiology of cognitive deficits following TBI. Five randomly selected DEPs were validated using ELISA in an additional 105 cases, and the results also supported the experimental findings. Conclusions. Despite limitations, our findings will facilitate further studies of the pathological mechanisms underlying TBI-induced cognitive impairment and provide new methods for the research and development of neuroprotective agents. However, further investigation on a large cohort is warranted.

scholarly journals Characterization of Endogenous Gibberellins and Molecular Cloning of a Putative Gibberellin 3-Oxidase Gene in Bunching Onion

2011 ◽  
Vol 136 (6) ◽  
pp. 382-388 ◽  
Author(s):  
Nobutaka Shiraiwa ◽  
Kaori Kikuchi ◽  
Ichiro Honda ◽  
Masayoshi Shigyo ◽  
Hiroko Yamazaki ◽  
...  
Keyword(s):  
Reproductive Organs ◽  
Development Stage ◽  
Allium Fistulosum ◽  
Late Development ◽  
Oxidase Gene ◽  
Development Stages ◽  
Ga Biosynthesis ◽  
Bunching Onion

To clarify the role of gibberellin (GA) in the growth of bunching onion (Allium fistulosum), identification of endogenous GAs and expression analysis of a putative gibberellin 3-oxidase (AfGA3ox1) were conducted. GA1, GA3, GA4, GA9, GA20, and GA34 were identified with levels of GA4 and GA9 being higher than those of GA1, GA3, and GA20. The young seedlings were clearly elongated by exogenous GA4 treatment but not by GA3. These results indicate that the 13-non-hydroxylation pathway of GA biosynthesis may be predominant in shoots with GA4 playing an important role in the growth of bunching onion. Expression of AfGA3ox1 was higher in leaf sheaths than leaf blades during vegetative growth. In reproductive organs, expression of AfGA3ox1 was higher at early and middle development stages in the stalks but was detected at a late development stage in the umbels. AfGA3ox1 was mapped on chromosome 7A from shallot, a bunching onion-related species.

scholarly journals Identification and characteristics of postnatal development-related long non-coding RNAs under microbiota dependent condition

2020 ◽  
Author(s):  
Miaomiao Zheng ◽  
Binbin Zhang ◽  
Yidan Zhang ◽  
Tingting Sun ◽  
Baozhong Hu
Keyword(s):  
Epithelial Cells ◽  
Gut Microbiota ◽  
Postnatal Development ◽  
Intestinal Epithelial Cells ◽  
Development Stage ◽  
Intestinal Epithelial ◽  
Early Postnatal Development ◽  
Development Stages ◽  
Large Numbers ◽  
Non Coding Rnas

Abstract BackgroundThe interplay of long-non coding RNAs (lncRNAs) and the intestinal microbiota may serve as an essential role in intestinal development and homeostasis. Microbiota could regulate a large numbers of lncRNAs expression in intestinal epithelial cells. However, the associations between lncRNAs and microbiota during early postnatal development stages are still need to understand. MethodsIn present study, the microbial effects on lncRNA of intestinal epithelial cells (IECs) during postnatal development stage were investigated. ResultsWe identified gut microbiota-specific lncRNAs in diverse postnatal development stages including week 1, week 4 and week 12/16 of mice. A large proportion of gut microbiota-specific lncRNAs only were differential expressed in a single postnatal development stage. Up- and down-regulated gut microbiota-specific lncRNAs both showed consistent expression pattern. We also constructed gut microbiota-specific lncRNAs and coding genes interacted co-expressed networks. Functional analysis indicated that gut microbiota-specific lncRNAs were associated with ABC transporters. ConclusionsIn summary, the present study characterizes the landscape of lncRNAs associated with gut microbiota in different postnatal development stages. It provide assistance for exploring the relationships among lncRNAs, gut microbiota and postnatal development stages.

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