scholarly journals Metabolism of inositol 1,4,5-trisphosphate and inositol 1,3,4-trisphosphate in rat parotid glands

1985 ◽  
Vol 229 (2) ◽  
pp. 505-511 ◽  
Author(s):  
R F Irvine ◽  
E E Anggård ◽  
A J Letcher ◽  
C P Downes
Keyword(s):  
High Performance ◽  
Inositol Trisphosphate ◽  
Significant Proportion ◽  
Complete Separation ◽  
Parotid Glands ◽  
Phosphatidylinositol Bisphosphate ◽  
Metabolic Characteristics ◽  
Inositol 1,4,5 Trisphosphate ◽  
Rat Parotid ◽  
And Control

A complete separation of myo-inositol 1,4,5-[4,5-(32)P]trisphosphate prepared from human erythrocytes, and myo-[2-3H]inositol 1,3,4-trisphosphate prepared from carbachol-stimulated rat parotid glands [Irvine, Letcher, Lander & Downes (1984) Biochem. J. 223, 237-243], was achieved by anion-exchange high-performance liquid chromatography. This separation technique was then used to study the metabolism of these two isomers of inositol trisphosphate in carbachol-stimulated rat parotid glands. Fragments of glands were pre-labelled with myo-[2-3H]inositol, washed, and then stimulated with carbachol. At 5s after stimulation a clear increase in inositol 1,4,5-trisphosphate was detected, with no significant increase in inositol 1,3,4-trisphosphate. After this initial lag however, inositol 1,3,4-phosphate rose rapidly; by 15s it predominated over inositol 1,4,5-trisphosphate, and continued to rise so that after 15 min it was at 10-20 times the radiolabelling level of the 1,4,5-isomer. In contrast, after the initial rapid rise (maximal within 15s), inositol 1,4,5-trisphosphate levels declined to near control levels after 1 min and then rose again very gradually over the next 15 min. When a muscarinic blocker (atropine) was added after 15 min of carbachol stimulation, inositol 1,4,5-trisphosphate levels dropped to control levels within 2-3 min, whereas inositol 1,3,4-trisphosphate levels took at least 15 min to fall, consistent with the kinetics observed earlier for total parotid inositol trisphosphates [Downes & Wusteman (1983) Biochem. J. 216, 633-640]. Phosphatidylinositol bisphosphate (PtdInsP2) from stimulated and control cells were degraded chemically to inositol trisphosphate to seek evidence for 3H-labelled PtdIns(3,4)P2. No evidence could be obtained that a significant proportion of PtdInsP2 was this isomer; in control tissues it must be less than 5% of the total PtdInsP2 radiolabelled by myo-[2-3H]inositol. These data indicate that, provided that inositol 1,4,5-trisphosphate is studied independently of inositol 1,3,4-trisphosphate, the former shows metabolic characteristics consistent with its proposed role as a second messenger for calcium mobilization. The metabolic profile of inositol 1,3,4-trisphosphate is entirely different, and its function and source remain unclear.

scholarly journals Inositol 1,3,4,5-tetrakisphosphate and not phosphatidylinositol 3,4-bisphosphate is the probable precursor of inositol 1,3,4-trisphosphate in agonist-stimulated parotid gland

1986 ◽  
Vol 238 (2) ◽  
pp. 501-506 ◽  
Author(s):  
C P Downes ◽  
P T Hawkins ◽  
R F Irvine
Keyword(s):  
Parotid Gland ◽  
Prolonged Exposure ◽  
Inositol Trisphosphate ◽  
Erythrocyte Membranes ◽  
Head Group ◽  
Inositol Polyphosphate ◽  
Parotid Glands ◽  
Phosphatidylinositol Bisphosphate ◽  
Inositol Tetrakisphosphate ◽  
Rat Parotid

When [3H]inositol-prelabelled rat parotid-gland slices were stimulated with carbachol, noradrenaline or Substance P, the major inositol trisphosphate produced with prolonged exposure to agonists was, in each case, inositol 1,3,4-trisphosphate. Much lower amounts of radioactivity were present in the inositol 1,4,5-trisphosphate fraction separated by anion-exchange h.p.l.c. Analysis of the inositol trisphosphate head group of phosphatidylinositol bisphosphate in [32P]Pi-labelled parotid glands showed the presence of phosphatidylinositol 4,5-bisphosphate, but no detectable phosphatidylinositol 3,4-bisphosphate. Carbachol-stimulated [3H]inositol-labelled parotid glands contained an inositol polyphosphate with the chromatographic properties and electrophoretic mobility of an inositol tetrakisphosphate, the probable structure of which was determined to be inositol 1,3,4,5-tetrakisphosphate. Since an enzyme in erythrocyte membranes is capable of degrading this tetrakisphosphate to inositol 1,3,4-trisphosphate, it is suggested to be the precursor of inositol 1,3,4-trisphosphate in parotid glands.

scholarly journals Inositol trisphosphates in carbachol-stimulated rat parotid glands

1984 ◽  
Vol 223 (1) ◽  
pp. 237-243 ◽  
Author(s):  
R F Irvine ◽  
A J Letcher ◽  
D J Lander ◽  
C P Downes
Keyword(s):  
Parotid Gland ◽  
Inositol Trisphosphate ◽  
Erythrocyte Membranes ◽  
Parotid Glands ◽  
Phosphatidylinositol Bisphosphate ◽  
Rat Parotid Gland ◽  
Periodate Treatment ◽  
Human Erythrocyte Membranes ◽  
Rat Parotid ◽  
Stimulation Of

Carbachol stimulation of rat parotid gland fragments prelabelled with myo-[3H]-inositol results in a large accumulation after 15 min of [3H]inositol trisphosphate. Only some of this is the D-1,4,5 isomer which would be expected to be derived from the known phosphatidylinositol bisphosphate. The predominant inositol trisphosphate is not susceptible to hydrolysis by human erythrocyte membranes. It yields altritol after periodate treatment followed by reduction and dephosphorylation, and, from partial dephosphorylation experiments, does not have a phosphate in the 2 position; the most likely structure of this inositol trisphosphate is therefore (D/L)-myo-inositol 1,3,4-trisphosphate. The possible origin and significance of this compound are discussed.

scholarly journals Rapid formation of inositol 1,3,4,5-tetrakisphosphate and inositol 1,3,4-trisphosphate in rat parotid glands may both result indirectly from receptor-stimulated release of inositol 1,4,5-trisphosphate from phosphatidylinositol 4,5-bisphosphate

1986 ◽  
Vol 238 (2) ◽  
pp. 507-516 ◽  
Author(s):  
P T Hawkins ◽  
L Stephens ◽  
C P Downes
Keyword(s):  
Initial Rate ◽  
Parotid Glands ◽  
Inositol 1,4,5 Trisphosphate ◽  
Rapid Formation ◽  
Inositol Tetrakisphosphate ◽  
Rat Parotid ◽  
Phosphatidylinositol 4 ◽  
Hydrolysis Of ◽  
Direct Hydrolysis ◽  
Stimulation Of

Addition of 1 mM-carbachol to [3H]inositol-labelled rat parotid slices stimulated rapid formation of [3H]inositol 1,3,4,5-tetrakisphosphate, the accumulation of which reached a peak 20 s after stimulation, and then declined rapidly towards a new steady state. The initial rate of formation of inositol 1,3,4,5-tetrakisphosphate was slower than that for inositol 1,4,5-trisphosphate. The radioactivity in [3H]inositol 1,3,4,5-tetrakisphosphate fell quickly in carbachol-stimulated and then atropine-blocked parotid slices, suggesting that it is rapidly metabolized during stimulation. Parotid homogenates rapidly dephosphorylated inositol 1,4,5-trisphosphate, inositol 1,3,4,5-tetrakisphosphate and, less rapidly, inositol 1,3,4-trisphosphate. Inositol 1,3,4,5-tetrakisphosphate was specifically hydrolysed to a compound with the chromatographic properties of inositol 1,3,4-trisphosphate. The only 3H-labelled phospholipids that we could detect in parotid slices labelled with [3H]inositol for 90 min were phosphatidylinositol, phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. Parotid homogenates synthesized inositol tetrakisphosphate from inositol 1,4,5-trisphosphate. This activity was dependent on the presence of ATP. We suggest that, during carbachol stimulation of parotid slices, the key event in inositol lipid metabolism is the activation of phosphatidylinositol 4,5-bisphosphate-specific phospholipase C. The inositol 1,4,5-trisphosphate thus liberated is metabolized in two distinct ways; by direct hydrolysis of the 5-phosphate to form inositol 1,4-bisphosphate and by phosphorylation to form inositol 1,3,4,5-tetrakisphosphate and hence, by hydrolysis of this tetrakisphosphate, to form inositol 1,3,4-trisphosphate.

scholarly journals Breakdown of polyphosphoinositides and not phosphatidylinositol accounts for muscarinic agonist-stimulated inositol phospholipid metabolism in rat parotid glands

1983 ◽  
Vol 216 (3) ◽  
pp. 633-640 ◽  
Author(s):  
C P Downes ◽  
M M Wusteman
Keyword(s):  
Molecular Mechanisms ◽  
Inositol Phosphates ◽  
Inositol Phosphate ◽  
Muscarinic Agonist ◽  
Parotid Glands ◽  
Muscarinic Agonists ◽  
Inositol 1 ◽  
Inositol 1,4,5 Trisphosphate ◽  
Rat Parotid ◽  
Phosphatidylinositol 4

The molecular mechanisms underlying the ability of muscarinic agonists to enhance the metabolism of inositol phospholipids were studied using rat parotid gland slices prelabelled with tracer quantities of [3H]inositol and then washed with 10 mM unlabelled inositol. Carbachol treatment caused rapid and marked increases in the levels of radioactive inositol 1-phosphate, inositol 1,4-bisphosphate, inositol 1,4,5-trisphosphate and an accumulation of label in the free inositol pool. There were much less marked changes in the levels of [3H]phosphatidylinositol, [3H]phosphatidylinositol 4-phosphate and [3H]phosphatidylinositol 4,5-bisphosphate. At 5 s after stimulation with carbachol there were large increases in [3H]inositol 1,4-bisphosphate and [3H]inositol 1,4,5-trisphosphate, but not in [3H]inositol 1-phosphate. After stimulation with carbachol for 10 min the levels of radioactive inositol 1,4-bisphosphate and inositol 1,4,5-trisphosphate greatly exceeded the starting level of radioactivity in phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate respectively. When carbachol treatment was followed by addition of sufficient atropine to block all the muscarinic receptors the radioactive inositol phosphates rapidly returned towards control levels. The carbachol-evoked changes in radioactive inositol phosphate and phospholipid levels were blocked in the presence of 2,4-dinitrophenol (an uncoupler of oxidative phosphorylation). The results suggest that muscarinic agonists stimulate a polyphosphoinositide-specific phospholipase C and that these lipids are continuously replenished from the labelled phosphatidylinositol pool. [3H]Inositol 1-phosphate in the stimulated glands probably arises via hydrolysis of inositol 1,4-bisphosphate and not directly from phosphatidylinositol.

scholarly journals Inositol 1,2-cyclic 4,5-trisphosphate is not a product of μscarinic receptor-stimulated phosphatidylinositol 4,5-bisphosphate hydrolysis in rat parotid glands

1987 ◽  
Vol 243 (1) ◽  
pp. 211-218 ◽  
Author(s):  
P T Hawkins ◽  
C P Berrie ◽  
A J Morris ◽  
C P Downes
Keyword(s):  
Parotid Gland ◽  
Inositol Phosphates ◽  
Inositol Phosphate ◽  
Water Soluble ◽  
Parotid Glands ◽  
Inositol 1 ◽  
Inositol 1,4,5 Trisphosphate ◽  
Cyclic Phosphate ◽  
Rat Parotid ◽  
Cyclic Compounds

We have employed a neutral-pH extraction technique to look for inositol 1,2-cyclic phosphate derivatives in [3H]inositol-labelled parotid gland slices stimulated with carbachol. The incubations were terminated by adding cold chloroform/methanol (1:2, v/v), the samples were dried under vacuum and inositol phosphates were extracted from the dried residues by phenol/chloroform/water partitioning. Water-soluble inositol metabolites were separated by h.p.l.c. at pH 3.7. 32P-labelled inositol phosphate standards (inositol 1-phosphate, inositol 1,2-cyclic phosphate, inositol 1,4,5-trisphosphate and inositol 1,2-cyclic 4,5-trisphosphate) were quantitively recovered through both extraction and chromatography steps. Treatment of inositol cyclic phosphate standards with 5% (w/v) HClO4 for 10 min prior to chromatography resulted in formation of the expected non-cyclic compounds. [3H]Inositol 1-phosphate and [3H]inositol 1,4,5-trisphosphate were both present in parotid gland slices and both increased during stimulation with 1 mM-carbachol. There was no evidence for significant quantities of [3H]inositol 1,2-cyclic phosphate or [3H]inositol 1,2-cyclic 4,5-trisphosphate in control or carbachol-stimulated glands. Parotid gland homogenates rapidly converted inositol 1,4,5-trisphosphate to inositol bisphosphate and inositol tetrakisphosphate, but metabolism of the inositol cyclic trisphosphate was much slower. The results suggest that inositol 1,4,5-trisphosphate, but not inositol 1,2-cyclic 4,5-trisphosphate, is the water-soluble product of muscarinic receptor-stimulated phospholipase C in rat parotid glands.

scholarly journals Mealtime Environment and Control of Food Intake in Healthy Children and in Children with Gastrointestinal Diseases

Children ◽  
2021 ◽  
Vol 8 (2) ◽  
pp. 77
Author(s):  
Katerina Sdravou ◽  
Elpida Emmanouilidou-Fotoulaki ◽  
Athanasia Printza ◽  
Elias Andreoulakis ◽  
Athanasios Evangeliou ◽  
...  
Keyword(s):  
Food Intake ◽  
Gastrointestinal Disease ◽  
Significant Proportion ◽  
Gastrointestinal Diseases ◽  
Healthy Children ◽  
Cross Sectional ◽  
Frequent Use ◽  
Child Autonomy ◽  
And Control ◽  
Mealtime Environment

Parental feeding practices and mealtime routine significantly influence a child’s eating behavior. The aim of this study was to investigate the mealtime environment in healthy children and children with gastrointestinal diseases. We conducted a cross-sectional case–control study among 787 healthy, typically developing children and 141 children with gastrointestinal diseases, aged two to seven years. Parents were asked to provide data on demographics and describe their mealtime environment by answering to 24 closed-ended questions. It was found that the majority of the children had the same number of meals every day and at the same hour. Parents of both groups exerted considerable control on the child’s food intake by deciding both when and what their child eats. Almost one third of the parents also decided how much their child eats. The two groups differed significantly in nine of the 24 questions. The study showed that both groups provided structured and consistent mealtime environments. However, a significant proportion of children did not control how much they eat which might impede their ability to self-regulate eating. The presence of a gastrointestinal disease was found to be associated with reduced child autonomy, hampered hunger cues and frequent use of distractions during meals.

scholarly journals Reduction of Polycyclic Aromatic Hydrocarbons (PAHs) in Sesame Oil Using Cellulosic Aerogel

Foods ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 644
Author(s):  
Do-Yeong Kim ◽  
Boram Kim ◽  
Han-Seung Shin
Keyword(s):  
Polycyclic Aromatic Hydrocarbons ◽  
Aromatic Hydrocarbons ◽  
High Performance ◽  
Oil Quality ◽  
Acid Value ◽  
Quality Parameters ◽  
Sesame Oil ◽  
Miscanthus Sinensis ◽  
Polycyclic Aromatic ◽  
And Control

The effect of cellulosic aerogel treatments used for adsorption of four polycyclic aromatic hydrocarbons (PAHs)—benzo[a]anthracene, chrysene, benzo[b]fluoranthene, and benzo[a]pyrene [BaP])—generated during the manufacture of sesame oil was evaluated. In this study, eulalia (Miscanthus sinensis var. purpurascens)-based cellulosic aerogel (adsorbent) was prepared and used high performance liquid chromatography with fluorescence detection for determination of PAHs in sesame oil. In addition, changes in the sesame oil quality parameters (acid value, peroxide value, color, and fatty acid composition) following cellulosic aerogel treatment were also evaluated. The four PAHs and their total levels decreased in sesame oil samples roasted under different conditions (p < 0.05) following treatment with cellulosic aerogel. In particular, highly carcinogenic BaP was not detected after treatment with cellulosic aerogel. Moreover, there were no noticeable quality changes in the quality parameters between treated and control samples. It was concluded that eulalia-based cellulosic aerogel proved suitable for the reduction of PAHs from sesame oil and can be used as an eco-friendly adsorbent.

Advanced high-performance processing tools for diagnostics and control in fusion devices

2021 ◽  
Vol 170 ◽  
pp. 112529
Author(s):  
N. Cruz ◽  
A.J.N. Batista ◽  
J.M. Cardoso ◽  
B.B. Carvalho ◽  
P.F. Carvalho ◽  
...  
Keyword(s):  
High Performance ◽  
Diagnostics And Control ◽  
And Control

scholarly journals Detection of Antimicrobial Residues in Poultry Litter: Monitoring a Risk through a Selective and Sensitive HPLC–MS/MS Method

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1399
Author(s):  
Karina Yévenes ◽  
Ekaterina Pokrant ◽  
Lina Trincado ◽  
Lisette Lapierre ◽  
Nicolás Galarce ◽  
...  
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
Poultry Litter ◽  
Chromatography Tandem Mass Spectrometry ◽  
Antimicrobial Residues ◽  
Field Samples ◽  
Liquid Chromatography Tandem Mass ◽  
Commercial Poultry ◽  
Agriculture Fertilizer ◽  
And Control

Tetracyclines, sulphonamides, and quinolones are families of antimicrobials (AMs) widely used in the poultry industry and can excrete up to 90% of AMs administrated, which accumulate in poultry litter. Worryingly, poultry litter is widely used as an agriculture fertilizer, contributing to the spread AMs residues in the environment. The aim of this research was to develop a method that could simultaneously identify and quantify three AMs families in poultry litter by high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS). Samples of AMs free poultry litter were used to validate the method according to 657/2002/EC and VICH GL49. Results indicate that limit of detection (LOD) ranged from 8.95 to 20.86 μg kg−1, while limits of quantitation (LOQ) values were between 26.85 and 62.58 µg kg−1 of tetracycline, 4-epi-tetracycline, oxytetracycline, 4-epi-oxytetracycline, enrofloxacin, ciprofloxacin, flumequine, sulfachloropyridazine, and sulfadiazine. Recoveries obtained ranged from 93 to 108%. The analysis of field samples obtained from seven commercial poultry flocks confirmed the adequacy of the method since it detected means concentrations ranging from 20 to 10,364 μg kg−1. This provides us an accurate and reliable tool to monitor AMs residues in poultry litter and control its use as agricultural fertilizer.

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