An ultra rapid and sensitive high-performance liquid chromatographic method for determination of tissue and plasma free amino acids

Graser, Theodor; Godel, H.; Albers, Sabine; Földi, Péter; Fürst, Peter

doi:10.1016/0003-2697(85)90064-8

Cited by 149 publications

(43 citation statements)

References 17 publications

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“…Methods were derived from Graser et al (1985). Samples (20 l) were reacted with 1.0 l of o-phthaldialdehyde/3-mercaptoproprionic acid reagent (Sigma-Aldrich) for derivitization.…”

Section: Methodsmentioning

confidence: 99%

Protein Kinase C ϵ and γ: Involvement in Formalin-Induced Nociception in Neonatal Rats

Sweitzer

Wong²,

Peters³

et al. 2004

J Pharmacol Exp Ther

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The central nervous system undergoes dynamic changes as it matures. However, until recently, very little was known about the impact of these changes on pain and analgesia. This study tested the hypothesis that the ⑀ and ␥ isozymes of protein kinase C (PKC) contribute to formalin-induced nociception in an age-dependent manner. Expression of ⑀ and ␥ PKC and the contributions of these isozymes in formalin-induced nociception was examined in postnatal day 7, 15, and 21 rats. ⑀PKC expression in dorsal root ganglion neurons and ␥PKC expression in lamina II of the spinal cord increased from the first to the third postnatal week. Coupling immunohistochemical and Western analysis, translocation of ⑀PKC followed intraplantar formalin in all ages. In contrast, formalin-induced ␥PKC translocation was observed only in postnatal day 21 rats. Behaviorally, intrathecal administration of the ⑀PKC-specific inhibitor (⑀V1-2) attenuated phase 1 and phase 2 formalin behaviors at all ages. In contrast, intrathecal administration of the ␥PKC-specific inhibitor (␥V5-3) attenuated only phase 2 responses in postnatal day 15 and 21 rats. Functionally, inhibition of ⑀PKC decreased capsaicin-stimulated release of glutamate and calcitonin gene-related peptide in spinal cords isolated from postnatal day 7 rats. These results suggest that ⑀PKC age independently mediates inflammatory pain produced by intraplantar formalin. In contrast, ␥PKC contributes to formalin-induced nociception in an age-dependent manner. Identifying the molecular mechanisms responsible for age-specific patterns of nociception is necessary for the rational development of novel therapeutic strategies for treating pediatric pain.

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“…Methods were derived from Graser et al (1985). Samples (20 l) were reacted with 1.0 l of o-phthaldialdehyde/3-mercaptoproprionic acid reagent (Sigma-Aldrich) for derivitization.…”

Section: Methodsmentioning

confidence: 99%

Protein Kinase C ϵ and γ: Involvement in Formalin-Induced Nociception in Neonatal Rats

Sweitzer

Wong²,

Peters³

et al. 2004

J Pharmacol Exp Ther

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“…Plasma was deproteinized with 5% (wt/vol) sulfosalicylic acid containing 200 nmol/ml thienylalanine as internal standard and amino acid concentrations were measured in an automated amino acid analyzer (Beckman Amino Acid Analyzer; Beckman Instruments, Inc., Fullerton, CA). To measure tissue amino acids, the muscle biopsy specimens were homogenized in three volumes of 5% sulfosalicylic acid and after centrifugation at 5,000 g for 30 min, amino acids in the supernatant were measured by high performance liquid chromatography using a modification of the method of Graser et al (17) as described recently in a report from our laboratory (18).…”

Section: Methodsmentioning

confidence: 99%

Sepsis is associated with increased mRNAs of the ubiquitin-proteasome proteolytic pathway in human skeletal muscle.

Tiao¹,

Hobler²,

Wang³

et al. 1997

J. Clin. Invest.

236

104

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Previous studies provided evidence that sepsis-induced muscle proteolysis in experimental animals is caused by increased ubiquitin-proteasome-dependent protein breakdown. It is not known if a similar mechanism accounts for muscle proteolysis in patients with sepsis. We determined mRNA levels for ubiquitin and the 20 S proteasome subunit HC3 by Northern blot analysis in muscle tissue from septic ( n ϭ 7) and non-septic ( n ϭ 11) patients. Plasma and muscle amino acid concentrations and concentrations in urine of 3-methylhistidine (3-MH), creatinine, and cortisol were measured at the time of surgery to assess the catabolic state of the patients. A three-to fourfold increase in mRNA levels for ubiquitin and HC3 was noted in muscle tissue from the septic patients concomitant with increased muscle levels of phenylalanine and 3-MH and reduced levels of glutamine. Total plasma amino acids were decreased by ‫ف‬ 30% in the septic patients. The 3-MH/creatinine ratio in urine was almost doubled in septic patients. The cortisol levels in urine were higher in septic than in control patients but this difference did not reach statistical significance. The results suggest that sepsis is associated with increased mRNAs of the ubiquitin-proteasome pathway in human skeletal muscle. ( J. Clin. Invest. 1997. 99:163-168.)

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“…Samples of medium (180 l) were deproteinized with 20 l of 30% (wt/vol) sulphosalicylic acid (SSA). Amino acids in muscle were analyzed using the HPLC method of Graser et al (38) with modifications as previously described (39). All supplies for amino acid analysis using the Beckman Model 6300 amino acid analyzer (amino acid calibration standards, ion-exchange column, buffers, and ninhydrin) were purchased from Beckman Instruments Inc. Sulfosalicylic acid (SSA) was from Fluka Chemical Corp. (Ronkonkoma, NY).…”

Section: Methodsmentioning

confidence: 99%

Linkage of aerobic glycolysis to sodium-potassium transport in rat skeletal muscle. Implications for increased muscle lactate production in sepsis.

James¹,

Fang²,

Schrantz³

et al. 1996

J. Clin. Invest.

149

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Although a linkage between aerobic glycolysis and sodiumpotassium transport has been demonstrated in diaphragm, vascular smooth muscle, and other cells, it is not known whether this linkage occurs in skeletal muscle generally. Metabolism of intact hind-leg muscles from young rats was studied in vitro under aerobic incubation conditions. When sodium influx into rat extensor digitorum longus (EDL) and soleus muscles was facilitated by the sodium ionophore monensin, muscle weight gain and production of lactate and alanine were markedly stimulated in a dose-dependent manner. Although lactate production rose in both muscles, it was more pronounced in EDL than in soleus. Monensininduced lactate production was inhibited by ouabain or by incubation in sodium-free medium. Preincubation in potassium-free medium followed by potassium re-addition also stimulated ouabain-inhibitable lactate release. Replacement of glucose in the incubation medium with pyruvate abolished monensin-induced lactate production but exacerbated monensin-induced weight gain. Muscles from septic or endotoxin-treated rats exhibited an increased rate of lactate production in vitro that was partially inhibited by ouabain. Increases muscle lactate production in sepsis may reflect linked increases in activity of the Na ϩ , K ϩ -ATPase, consumption of ATP and stimulation of aerobic glycolysis. ( J. Clin. Invest. 1996. 98:2388-2397.)

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An ultra rapid and sensitive high-performance liquid chromatographic method for determination of tissue and plasma free amino acids

Cited by 149 publications

References 17 publications

Protein Kinase C ϵ and γ: Involvement in Formalin-Induced Nociception in Neonatal Rats

Protein Kinase C ϵ and γ: Involvement in Formalin-Induced Nociception in Neonatal Rats

Sepsis is associated with increased mRNAs of the ubiquitin-proteasome proteolytic pathway in human skeletal muscle.

Linkage of aerobic glycolysis to sodium-potassium transport in rat skeletal muscle. Implications for increased muscle lactate production in sepsis.

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