Regulation by Carbon Dioxide of Wound-induced Ethylene Biosynthesis in the Peel of Citrus Fruit

2007 ◽  
Vol 13 (6) ◽  
pp. 497-504
Author(s):  
L. Zacarías ◽  
F. Alférez

The effect of carbon dioxide (CO2) on wound-induced ethylene biosynthesis in flavedo discs of mature orange fruits (Citrus sinensis L. Osbeck) is investigated. Wounding induced a marked and rapid increase on the rate of ethylene production, the content of 1-aminocyclopropane-1-carboxylic acid (ACC) and on the in vivo ACC oxidase (ACO) activity. Incubation of flavedo discs in a 15% CO2 atmosphere suppressed activation of these processes. Wound-induced ethylene production was inhibited by CO2 in a concentration-dependent manner but ACO activity was enhanced at concentrations between 1% and 5%. Kinetic analysis of the interaction between CO2 and ACO activity indicated that high CO2 acted as a noncompetitive inhibitor. Removal of CO2 after 24 h incubation did not restore normal rates of ethylene production. CO2 partially counteracted the increase in ethylene production and ACO activity induced by a pretreatment with an ethylene action inhibitor (STS, silver thiosulfate). This suggested that part of CO2 action on ethylene biosynthesis might be due to interfering ethylene action. Collectively, the results indicated that ACS activity appeared to be the major regulatory step by which CO2 suppresses wound-induced ethylene production. ACO was differentially modulated by CO2, which is being stimulated at low concentrations and inhibited at high concentrations.

Development ◽  
2002 ◽  
Vol 129 (8) ◽  
pp. 1925-1933 ◽  
Author(s):  
Baohe Shen ◽  
James L. Manley

The Drosophila Pelle kinase plays a key role in the evolutionarily conserved Toll signaling pathway, but the mechanism responsible for its activation has been unknown. We present in vivo and in vitro evidence establishing an important role for concentration-dependent autophosphorylation in the signaling process. We first show that Pelle phosphorylation can be detected transiently in early embryos, concomitant with activation of signaling. Importantly, Pelle phosphorylation is enhanced in a gain-of-function Toll mutant (Toll10b), but decreased by loss-of-function Toll alleles. Next we found that Pelle is phosphorylated in transfected Schneider L2 cells in a concentration-dependent manner such that significant modification is observed only at high Pelle concentrations, which coincide with levels required for phosphorylation and activation of the downstream target, Dorsal. Pelle phosphorylation is also enhanced in L2 cells co-expressing Toll10b, and is dependent on Pelle kinase activity. In vitro kinase assays revealed that recombinant, autophosphorylated Pelle is far more active than unphosphorylated Pelle. Importantly, unphosphorylated Pelle becomes autophosphorylated, and activated, by incubation at high concentrations. We discuss these results in the context of Toll-like receptor mediated signaling in both flies and mammals.


1992 ◽  
Vol 68 (05) ◽  
pp. 570-576 ◽  
Author(s):  
Mary A Selak

SummaryWe have previously demonstrated that human neutrophil cathepsin G is a strong platelet agonist that binds to a specific receptor. This work describes the effect of neutrophil elastase on cathepsin G-induced platelet responses. While platelets were not activated by high concentrations of neutrophil elastase by itself, elastase enhanced aggregation, secretion and calcium mobilization induced by low concentrations of cathepsin G. Platelet aggregation and secretion were potentiated in a concentration-dependent manner by neutrophil elastase with maximal responses observable at 200 nM. Enhancement was observed when elastase was preincubated with platelets for time intervals of 10–60 s prior to addition of a low concentration of cathepsin G and required catalytically-active elastase since phenylmethanesulphonyl fluoride-inhibited enzyme failed to potentiate cell activation. Neutrophil elastase potentiation of platelet responses induced by low concentrations of cathepsin G was markedly inhibited by creatine phosphate/creatine phosphokinase and/or indomethacin, indicating that the synergism between elastase and cathepsin G required the participation of ADP and thromboxane A2. On the other hand, platelet responses were not attenuated by the PAF antagonist BN 52021, signifying that PAF-acether did not play a role in elastase potentiation. At higher concentrations porcine pancreatic elastase exhibits similar effects to neutrophil elastase, demonstrating that the effect of elastase was not unique to the neutrophil protease. While neutrophil elastase failed to alter the ability of cathepsin G to hydrolyze a synthetic chromogenic substrate, preincubation of platelets with elastase increased the apparent affinity of cathepsin G binding to platelets. In contrast to their effect on cathepsin G-induced platelet responses, neither neutrophil nor pancreatic elasatse potentiated aggregation or dense granule release initiated by ADP, PAF-acether, arachidonic acid or U46619, a thromboxane A2 mimetic. Moreover, unlike its effect on cathepsin G, neutrophil elastase inhibited thrombin-induced responses. The current observations demonstrate that elastase can potentiate platelet responses mediated by low concentrations of cathepsin G, suggesting that both enzymes may function synergistically to activate platelets under conditions where neutrophil degranulation occurs.


2019 ◽  
Vol 26 (7) ◽  
pp. 494-501 ◽  
Author(s):  
Sameer Suresh Bhagyawant ◽  
Dakshita Tanaji Narvekar ◽  
Neha Gupta ◽  
Amita Bhadkaria ◽  
Ajay Kumar Gautam ◽  
...  

Background: Diabetes and hypertension are the major health concern and alleged to be of epidemic proportions. This has made it a numero uno subject at various levels of investigation. Glucosidase inhibitor provides the reasonable option in treatment of Diabetes Mellitus (DM) as it specifically targets post prandial hyperglycemia. The Angiotensin Converting Enzyme (ACE) plays an important role in hypertension. Therefore, inhibition of ACE in treatment of elevated blood pressure attracts special interest of the scientific community. Chickpea is a food legume and seeds contain carbohydrate binding protein- a lectin. Some of the biological properties of this lectin hitherto been elucidated. Methods: Purified by ion exchange chromatography, chickpea lectin was tested for its in vitro antioxidant, ACE-I inhibitory and anti-diabetic characteristic. Results: Lectin shows a characteristic improvement over the synthetic drugs like acarbose (oral anti-diabetic drug) and captopril (standard antihypertensive drug) when, their IC50 values are compared. Lectin significantly inhibited α-glucosidase and α-amylase in a concentration dependent manner with IC50 values of 85.41 ± 1.21 ҝg/ml and 65.05 ± 1.2 µg/ml compared to acarbose having IC50 70.20 ± 0.47 value of µg/ml and 50.52 ± 1.01 µg/ml respectively. β-Carotene bleaching assay showed antioxidant activity of lectin (72.3%) to be as active as Butylated Hydroxylanisole (BHA). In addition, lectin demonstrated inhibition against ACE-I with IC50 value of 57.43 ± 1.20 µg/ml compared to captopril. Conclusion: Lectin demonstrated its antioxidant character, ACE-I inhibition and significantly inhibitory for α-glucosidase and α-amylase seems to qualify as an anti-hyperglycemic therapeutic molecule. The biological effects of chickpea lectin display potential for reducing the parameters of medically debilitating conditions. These characteristics however needs to be established under in vivo systems too viz. animals through to humans.


Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 995
Author(s):  
Mohammad Darvish ◽  
Habib Shirzad ◽  
Mohammadreza Asghari ◽  
Parviz Noruzi ◽  
Abolfazl Alirezalu ◽  
...  

Ethylene is the most important factor playing roles in senescence and deterioration of harvested crops including cut flowers. Brassinosteroids (BRs), as natural phytohormones, have been reported to differently modulate ethylene production and related senescence processes in different crops. This study was carried out to determine the effects of different levels of 24-epibrassinolide (EBL) on ACC oxidase enzyme activity, the final enzyme in ethylene biosynthesis pathway, vase life, and senescence rate in lisianthus cut flowers. Harvested flowers were treated with EBL (at 0, 3, 6, and 9 µmol/L) and kept at 25 °C for 15 days. The ACC oxidase activity, water absorption, malondialdehyde (MDA) production and vase solution absorption rates, chlorophyll and anthocyanin contents, and the vase life of the flowers were evaluated during and at the end of storage. EBL at 3 µmol/L significantly (p ≤ 0.01) enhanced the flower vase life by decreasing the ACC oxidase activity, MDA production and senescence rates, and enhancing chlorophyll and anthocyanin biosynthesis and accumulation, relative water content, and vase solution absorption rates. By increasing the concentration, EBL negatively affected the flower vase life and postharvest quality probably via enhancing the ACC oxidase enzyme activity and subsequent ethylene production. EBL at 6 and 9 µmol/L and in a concentration dependent manner, enhanced the ACC oxidase activity and MDA production rate and decreased chlorophyll and anthocyanin accumulation and water absorption rate. The results indicate that the effects of brassinosteroids on ethylene production and physiology of lisianthus cut flowers is highly dose dependent.


Nutrients ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 123
Author(s):  
Natalia K. Kordulewska ◽  
Justyna Topa ◽  
Małgorzata Tańska ◽  
Anna Cieślińska ◽  
Ewa Fiedorowicz ◽  
...  

Lipopolysaccharydes (LPS) are responsible for the intestinal inflammatory reaction, as they may disrupt tight junctions and induce cytokines (CKs) secretion. Osthole has a wide spectrum of pharmacological effects, thus its anti-inflammatory potential in the LPS-treated Caco-2 cell line as well as in Caco-2/THP-1 and Caco-2/macrophages co-cultures was investigated. In brief, Caco-2 cells and co-cultures were incubated with LPS to induce an inflammatory reaction, after which osthole (150–450 ng/mL) was applied to reduce this effect. After 24 h, the level of secreted CKs and changes in gene expression were examined. LPS significantly increased the levels of IL-1β, -6, -8, and TNF-α, while osthole reduced this effect in a concentration-dependent manner, with the most significant decrease when a 450 ng/mL dose was applied (p < 0.0001). A similar trend was observed in changes in gene expression, with the significant osthole efficiency at a concentration of 450 ng/μL for IL1R1 and COX-2 (p < 0.01) and 300 ng/μL for NF-κB (p < 0.001). Osthole increased Caco-2 monolayer permeability, thus if it would ever be considered as a potential drug for minimizing intestinal inflammatory symptoms, its safety should be confirmed in extended in vitro and in vivo studies.


Blood ◽  
2001 ◽  
Vol 97 (9) ◽  
pp. 2648-2656 ◽  
Author(s):  
Juan A. Rosado ◽  
Else M. Y. Meijer ◽  
Karly Hamulyak ◽  
Irena Novakova ◽  
Johan W. M. Heemskerk ◽  
...  

Abstract Effects of the occupation of integrin αIIbβ3 by fibrinogen on Ca++signaling in fura-2–loaded human platelets were investigated. Adding fibrinogen to washed platelet suspensions inhibited increases in cytosolic [Ca++] concentrations ([Ca++]i) evoked by adenosine diphosphate (ADP) and thrombin in a concentration-dependent manner in the presence of external Ca++ but not in the absence of external Ca++ or in the presence of the nonselective cation channel blocker SKF96365, indicating selective inhibition of Ca++entry. Fibrinogen also inhibited store-mediated Ca++ entry (SMCE) activated after Ca++ store depletion using thapsigargin. The inhibitory effect of fibrinogen was reversed if fibrinogen binding to αIIbβ3 was blocked using RDGS or abciximab and was absent in platelets from patients homozygous for Glanzmann thrombasthenia. Fibrinogen was without effect on SMCE once activated. Activation of SMCE in platelets occurs through conformational coupling between the intracellular stores and the plasma membrane and requires remodeling of the actin cytoskeleton. Fibrinogen inhibited actin polymerization evoked by ADP or thapsigargin in control cells and in cells loaded with the Ca++ chelator dimethyl BAPTA. It also inhibited the translocation of the tyrosine kinase p60src to the cytoskeleton. These results indicate that the binding of fibrinogen to integrin αIIbβ3 inhibits the activation of SMCE in platelets by a mechanism that may involve modulation of the reorganization of the actin cytoskeleton and the cytoskeletal association of p60src. This action may be important in intrinsic negative feedback to prevent the further activation of platelets subjected to low-level stimuli in vivo.


Author(s):  
Kartika Arum Wardani ◽  
Kholida Nur Aini ◽  
Heny Arwati ◽  
Willy Sandhika

Abstract Sequestration of Plasmodium berghei ANKA-infected erythrocytes occurs in BALB/c mice as characteristic of  Plasmodium falciparum infection in humans. Animals’ bile has been widely used for centuries in Traditional Chinese Medicine. Goat bile has been used in healing infectious and non-infectious diseases; however, no report on the use of goat bile against malaria infection and sequestration. The purpose of this study was to analyze the correlation between parasitemia and sequestration in the liver of P.berghei ANKA-infected BALB/c mice treated with goat bile. This research was an in vivo experimental study using the post-test control group design. The male BALB/c mice aged ± 6 weeks, body weight 20-25 g were used. The mice were divided into five groups where Group 1-3 were mice treated with goat bile 25%, 50%, and 100%, respectively. Group 4-5 were negative (sterile water) and positive controls (DHP). Parasitemia was observed daily from each mouse and the number of sequestered infected erythrocytes on the endothelium of sinusoids. The data were analyzed using t independent test. Antimalarial activity of goat bile was shown by the lower parasitemia in goat bile-treated mice compared with the negative control. The average number of sequestration was goat bile concentration-dependent manner. The higher the concentration, the lower the number of sequestration. Sequestration was correlated with parasitemia (p=0,0001). Sequestration of P.berghei ANKA-infected erythrocytes correlated with parasitemia, and was goat bile concentration-dependent manner. Keywords: Malaria, parasitemia, sequestration, goat bileCorrespondence: [email protected]


Author(s):  
Edrees Khan Rahmatzada ◽  
Prof. Paras Nath Yadav ◽  
Dr. Yuba Raj Pokharel

Thiosemicarbazone have the antiviral, antibacterial, antifungal, and anticancer effects. 3-OH-Me-TSC inhibited the cell viability of HepG-2 cells by CV assay in a concentration dependent manner (control, 1μM, 3μM, 10μM, 30μM, and 100μM) with IC50 value of 9.587622μM. Further colony formation assay demonstrated that 3-OH-Me-TSC inhibits colony number and size of HepG-2. Wound healing assay exhibited that 3-OH-Me-TSC inhibit the migration of HepG-2 cells. DAPI staining showed that 3-OH-Me-TSC inhibited proliferation of HepG-2 cells in 30μM and 100μM concentrations respectively. 3-OH-Me-TSC inhibited VEGF, p38 alpha, C-JUN, BECN-1, ERK, NF-KB, in HepG-2 cells. We found that 3-OH-Me-TSC inhibit proliferation of HepG-2 cells by inhibiting MAPK signaling pathway, 3-OH-Me-TSC can be developed as future chemotherapeutic agent for treatment of hepatocellular carcinoma after the evaluation of this compounds in more cancer cells an in vivo model.


2008 ◽  
Vol 133 (6) ◽  
pp. 727-734 ◽  
Author(s):  
Hong Zhu ◽  
Eric P. Beers ◽  
Rongcai Yuan

Effects of naphthaleneacetic acid (NAA) and aminoethoxyvinylglycine (AVG) on young fruit abscission, leaf and fruit ethylene production, and expression of genes related to ethylene biosynthesis and cell wall degradation were examined in ‘Delicious’ apples (Malus ×domestica Borkh.). NAA at 15 mg·L−1 increased fruit abscission and ethylene production of leaves and fruit when applied at the 11-mm stage of fruit development, whereas AVG, an inhibitor of ethylene biosynthesis, at 250 mg·L−1 reduced NAA-induced fruit abscission and ethylene production of leaves and fruit. NAA also increased expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase genes (MdACS5A and MdACS5B), ACC oxidase gene (MdACO1), and ethylene receptor genes (MdETR1a, MdETR1b, MdETR2, MdERS1, and MdERS2) in fruit cortex and fruit abscission zones. However, AVG reduced NAA-induced expression of these genes except for MdERS2 in fruit abscission zones. NAA increased expression of the polygalacturonase gene MdPG2 in fruit abscission zones but not in fruit cortex, whereas AVG reduced NAA-enhanced expression of MdPG2 in fruit abscission zones. The expression of β-1,4-glucanase gene MdCel1 in fruit abscission zones was decreased by NAA but was unaffected by AVG. Our results suggest that ethylene biosynthesis, ethylene perception, and the MdPG2 gene are involved in young fruit abscission caused by NAA.


2002 ◽  
Vol 127 (6) ◽  
pp. 998-1005 ◽  
Author(s):  
Sastry Jayanty ◽  
Jun Song ◽  
Nicole M. Rubinstein ◽  
Andrés Chong ◽  
Randolph M. Beaudry

The temporal relationship between changes in ethylene production, respiration, skin color, chlorophyll fluorescence, volatile ester biosynthesis, and expression of ACC oxidase (ACO) and alcohol acyl-CoA transferase (AAT) in ripening banana (Musa L. spp., AAA group, Cavendish subgroup. `Valery') fruit was investigated at 22 °C. Ethylene production rose to a peak a few hours after the onset of its logarithmic phase; the peak in production coincided with maximal ACO expression. The respiratory rise began as ethylene production increased, reaching its maximum ≈30 to 40 hours after ethylene production had peaked. Green skin coloration and photochemical efficiency, as measured by chlorophyll fluorescence, declined simultaneously after the peak in ethylene biosynthesis. Natural ester biosynthesis began 40 to 50 hours after the peak in ethylene biosynthesis, reaching maximal levels 3 to 4 days later. While AAT expression was detected throughout, the maximum level of expression was detected at the onset of natural ester biosynthesis. The synthesis of unsaturated esters began 100 hours after the peak in ethylene and increased with time, suggesting the lipoxygenase pathway be a source of ester substrates late in ripening. Incorporation of exogenously supplied ester precursors (1-butanol, butyric acid, and 3-methyl-1-butanol) in the vapor phase into esters was maturity-dependent. The pattern of induced esters and expression data for AAT suggested that banana fruit have the capacity to synthesize esters over 100 hours before the onset of natural ester biosynthesis. We hypothesize the primary limiting factor in ester biosynthesis before natural production is precursor availability, but, as ester biosynthesis is engaged, the activity of alcohol acyl-CoA transferase the enzyme responsible for ester biosynthesis, exerts a major influence.


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