Microdetermination of (−)carnitine and carnitine acetyltransferase activity

Parvin, Rokshana; Pande, Shri V.

doi:10.1016/0003-2697(77)90393-1

Cited by 190 publications

(71 citation statements)

References 23 publications

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“…Protein was determined by the method of Peterson [2] and phosphorus of ashed phospholipid according to Lanzetta et al [3]. The volume of the intraliposomal space was measured by determination of trapped carnitine assayed by the radioenzymatic method [4].…”

Section: Methodsmentioning

confidence: 99%

An essential requirement of cardiolipin for mitochondrial carnitine acylcarnitine translocase activity

Noël

Pande

1986

European Journal of Biochemistry

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The phospholipid requirement for the optimal solubilization of carnitine acylcarnitine translocase from the inner membrane vesicles of rat liver mitochondria and for its reconstitution in liposomes was investigated. At the octylglucoside-solubilization step, the presence of cardiolipin proved superior to the other lipids tested. For reconstitution, a mixture having phosphatidylcholine, phosphatidylethanolamine and cardiolipin was found to be particularly effective. The requirement of cardiolipin at this step was met less effectively by other anionic phospholipids. Moreover, in intact mitochondria of rat liver and heart, the translocase activity was markedly inhibited by micromolar concentrations of doxorubicin, a specific cardiolipin-binding agent.Carnitine acylcarnitine translocase, involved in the transmitochondrial transport of carnitine and its esters, has been solubilized from inner membrane vesicles of rat liver mitochondria with octyl glucoside and reconstituted into asolectin liposomes using the octyl glucoside dilution method [l]. Continuing studies on the optimization of these procedures have revealed an esstential requirement of cardiolipin for the translocase activity. These are being described herein.

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Section: Methodsmentioning

confidence: 99%

An essential requirement of cardiolipin for mitochondrial carnitine acylcarnitine translocase activity

Noël

Pande

1986

European Journal of Biochemistry

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“…The cell lysates were stored at −80°C until analysis. Carnitine was measured in triplicate by quantitative radioenzymatic assay slightly modified from that reported by Parvin and Pande [24]. Free carnitine was measured in untreated samples whereas total carnitine was determined after alkaline hydrolysis.…”

Section: Carnitine Assaymentioning

confidence: 99%

Carnitine and Dehydroepiandrosterone Sulfate Induce Protein Synthesis in Porcine Primary Osteoblast-Like Cells

et al. 1999

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Abstract. Age-related bone loss eventually leads to osteopenia in men and women. The etiology of age-related bone loss is currently unknown; however, decreased osteoblast activity contributes to this phenomenon. In turn, osteoblast proliferation and function is dependent on energy production, thus the loss of energy production that occurs with age may account for the deficient osteoblast activity. Carnitine and dehydroepiandrosterone-sulfate (DHEAS), both of which decline with age, promote energy production through fatty acid metabolism. Thus, we hypothesized that carnitine and DHEAS would increase osteoblast activity in vitro. Accordingly, we measured the effect of carnitine and DHEAS on palmitic acid oxidation as a measure of energy production, and alkaline phosphatase (ALP) activity and collagen type I (COL) as indices of osteoblast function in primary porcine osteoblast-like cell cultures. Carnitine (10 −3 and 10 −1 M) but not DHEAS (10 −9 , 10 −8 , and 10 −7 M) increased carnitine levels within the cells. Carnitine alone and in combination with DHEAS increased palmitic acid oxidation. Both carnitine and DHEAS alone and in an additive fashion increased ALP activity and COL levels. These results demonstrate that in osteoblast-like cells in vitro, energy production can be increased by carnitine and osteoblast protein production can be increased by both carnitine and DHEAS. These data suggest that carnitine and DHEAS supplementation in the elderly may stimulate osteoblast activity and decrease age-related bone loss. Key words:Osteoblast -Carnitine -Dehydroepiandrosterone sulfate -Alkaline phosphatase -Collagen Type I.Bone loss is often divided into two broad mechanisms: (1) postmenopausal bone loss, characterized by increased osteoclast activity, and (2) age-related bone loss, characterized by decreased osteoblast activity. Osteoblast activity is dependent on energy production. Thus, it is plausible that the loss of energy production, which accompanies age, may account for decreased osteoblast activity. Most cells use both glucose and fatty acid oxidation pathways for ATP synthesis, although some cell types show a substrate preference. For instance, some studies have suggested that glucose is the major fuel substrate for bone [1,2]. However, more recent work indicates that cell populations enriched with osteoblast-like cells generate 40-80% of the energy demands through fatty acid oxidation [3]. Osteoblasts are responsible for bone formation and it follows that they maintain a high rate of ATP utilization to support the requisite protein synthesis. Cells with a high rate of protein synthesis generate ATP from fatty acid oxidation, the most efficient metabolic process for energy production [4][5][6]. Thus, modulation of fatty acid oxidation may regulate the amount of energy available for protein synthesis in osteoblasts.A number of factors are required for fatty acid oxidation. For example, carnitine is the requisite carrier for the transport of long-chain fatty acids across the inner mitochondrial membrane into...

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“…10-12, 15-17, 21), renal failure (n 5 7, Nos. 13,14,18,20,22,23,25), cancer (n 5 1, No. 26), or thoracic bacterial infection (n 5 2, Nos.…”

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confidence: 99%

Characterization of a Degenerative Cardiomyopathy Associated with Domoic Acid Toxicity in California Sea Lions (Zalophus californianus)

et al. 2009

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Abstract. Domoic acid, produced by marine algae, can cause acute and chronic neurologic sequela in California sea lions (Zalophus californianus) from acute toxicity or sublethal exposure. Eight sea lions, representing acute and chronic cases, both sexes, and all age classes, were selected to demonstrate a concurrent degenerative cardiomyopathy. Critical aspects of characterizing the cardiomyopathy by lesion distribution and morphology were the development of a heart dissection and tissue-trimming protocol and the delineation of the cardiac conducting system by histomorphology and immunohistochemistry for neuron-specific protein gene product 9.5. Histopathologic features and progression of the cardiomyopathy are described, varying from acute to chronic active and mild to severe. The cardiomyopathy is distinguished from other heart lesions in pinnipeds. Based on histopathologic features, immunopositive staining for cleaved caspase-3, and comparison with known, similar-appearing cardiomyopathies, the proposed pathogenesis for the degenerative cardiomyopathy is the primary or at least initial direct interaction of domoic acid with receptors that are suspected to exist in the heart. LCarnitine, measured in the heart and skeletal muscle, and troponin-I, measured in serum collected at the time of death from additional animals (n 5 58), were not predictive of the domoic acid-associated cardiomyopathy. This degenerative cardiomyopathy in California sea lions represents another syndrome beyond central neurologic disease associated with exposure to domoic acid and may contribute to morbidity and mortality.

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Microdetermination of (−)carnitine and carnitine acetyltransferase activity

Cited by 190 publications

References 23 publications

An essential requirement of cardiolipin for mitochondrial carnitine acylcarnitine translocase activity

An essential requirement of cardiolipin for mitochondrial carnitine acylcarnitine translocase activity

Carnitine and Dehydroepiandrosterone Sulfate Induce Protein Synthesis in Porcine Primary Osteoblast-Like Cells

Characterization of a Degenerative Cardiomyopathy Associated with Domoic Acid Toxicity in California Sea Lions (Zalophus californianus)

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