‘Glucose 6-Phosphate-Dehydrogenase’ Activity and Thiol Content of Thymus Nuclei From Control and X-Irradiated Rats

Deakin, Helen; Ord, Margery G.; Stocken, L. A.

doi:10.1042/bj0890296

Cited by 90 publications

(32 citation statements)

References 21 publications

(17 reference statements)

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“…Rat CGA 7-57 peptide was synthesized on an ABI 431 A (Applied Biosystems) peptide synthesizer using the standard procedures of Fmoc (9-fluorenylmethoxycarbonyl) chemistry (29). Formation of the disulfide bridge between Cys 17 and Cys 38 was obtained in the presence of N-ethyldiisopropylamine at pH 8.5, leaving the mixture to stand under open atmosphere for 24 h until complete reaction, according to Ellman test (30). After reduction of the disulfide bridge in the presence of 8 M guanidine HCl in 1 M Tris-HCl, pH 8.5, containing 4 mM EDTA, S-pyridylation of cysteine residues was obtained by treatment with 4-vinylpyridine (31).…”

Section: Methodsmentioning

confidence: 99%

Antibacterial and Antifungal Activities of Vasostatin-1, the N-terminal Fragment of Chromogranin A

Lugardon¹,

Raffner²,

Goumon³

et al. 2000

Journal of Biological Chemistry

152

140

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Vasostatin-1, the natural N-terminal 1-76 chromogranin A (CGA)-derived fragment in bovine sequence, has been purified from chromaffin secretory granules and identified by sequencing and matrix-assisted laser desorption time-of-flight mass spectrometry. This peptide, which displays antibacterial activity against Gram-positive bacteria at micromolar concentrations, is also able to kill a large variety of filamentous fungi and yeast cells in the 1-10 M range. We have found that the C-terminal moiety of vasostatin-1 is essential for the antifungal activity, and shorter active peptides have been synthesized. In addition, from the comparison with the activity displayed by related peptides (human recombinant and rat synthetic fragments), we could determine that antibacterial and antifungal activities have different structural requirements. To assess for such activities in vivo, CGA and CGA-derived fragments were identified in secretory material released from human polymorphonuclear neutrophils upon stimulation. Vasostatin-1, which is stored in a large variety of cells (endocrine, neuroendocrine, and neurons) and which is liberated from stimulated chromaffin and immune cells upon stress, may represent a new component active in innate immunity.

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Section: Methodsmentioning

confidence: 99%

Antibacterial and Antifungal Activities of Vasostatin-1, the N-terminal Fragment of Chromogranin A

Lugardon¹,

Raffner²,

Goumon³

et al. 2000

Journal of Biological Chemistry

152

140

View full text Add to dashboard Cite

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“…The modified PPD eluting in the excluded volume was transferred to a stoppered vessel and to it was added dropwise, with stirring, 10 mg of LHRH-Gly-Cys-OH dissolved in 1 ml of buffer A previously degassed and purged with nitrogen. The mixture was stirred at 23°C under nitrogen for 2 h and the amount of LHRH which bound to the PPD determined by estimating the free thiol content of the mixture at regular intervals using 5,5'-dithiobis(2nitrobenzoic acid) (Sigma) according to Deakin et al [12]. A coupling efficiency of 65% was noted.…”

Section: Methodsmentioning

confidence: 99%

Computer‐aided design and physiological testing of a luteinising hormone‐releasing hormone analogue for ‘adjuvant‐free’ immunocastration

Morrison¹,

Fishleigh²,

Ward³

et al. 1987

FEBS Letters

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An analogue of LHRH containing an extension of Gly‐Cys at the carboxyl‐terminus has been designed to permit reproducible coupling to a suitably modified carrier via a thioether bond. Potential energy calculations indicated that this analogue adopted a conformation in solution virtually identical to the type II′ turn around Gly‐6—Leu‐7 predicted for native LHRH. Intradermal administration of a conjugate of this analogue with purified protein derivative of tuberculin to male rats previously primed with BCG vaccine rapidly led to complete testicular regression. This adjuvant‐free immunisation protocol may represent an alternative to castration for the veterinary control of reproductive function.

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“…12 mM 5,59-dithiobis-(2-nitrobenzoic acid) (DTNB; Sigma) in 0.1 mM potassium phosphate buffer (pH 7.0; Merck, Haarlem, the Netherlands) was added to the rest of the lysed cells to a final concentration of 6 mM DTNB. After 10 min the samples were measured at 405 nm [22]. The amount of free thiols was calibrated against a standard curve of L-cysteine (Sigma).…”

Section: Flow Cytometrymentioning

confidence: 99%

Resistance of quiescent and proliferating airway epithelial cells to H2O2 challenge

Vries

Toorn²,

Bischoff

et al. 2007

European Respiratory Journal

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Alveolar epithelial cell injury and recovery are important in the pathogenesis of oxidant-induced lung damage. The alveolar cell line A549 was used to study responses of proliferating and quiescent cells in culture to time-and dose-dependent hydrogen peroxide (H 2 O 2 ) challenges.Recovery was monitored after 24 h of incubation in fresh medium with 10% serum. The adherent cells were counted and the resistance and recovery of the attached cells was assessed by appearance, by measuring the number of viable, apoptotic and necrotic cells using fluorescentactivated cell sorting, and by determining the intracellular free thiol content.A549 cells recovered from a 1-h challenge with up to 1 mM H 2 O 2 but could not sustain a more prolonged challenge (6 or 24 h) with 0.5 mM or 1.0 mM H 2 O 2 . These more severe conditions resulted in: loss of cells by detachment from the plate surface; reduced numbers of viable cells primarily due to necrosis; and a strong reduction of the intracellular free thiol content.Quiescent cells proved to be more sensitive to oxidative stress than proliferating cells. Intracellular free thiol levels apparently play a decisive role in cell survival, preferentially protecting proliferating cells.

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‘Glucose 6-Phosphate-Dehydrogenase’ Activity and Thiol Content of Thymus Nuclei From Control and X-Irradiated Rats

Cited by 90 publications

References 21 publications

Antibacterial and Antifungal Activities of Vasostatin-1, the N-terminal Fragment of Chromogranin A

Antibacterial and Antifungal Activities of Vasostatin-1, the N-terminal Fragment of Chromogranin A

Computer‐aided design and physiological testing of a luteinising hormone‐releasing hormone analogue for ‘adjuvant‐free’ immunocastration

Resistance of quiescent and proliferating airway epithelial cells to H2O2 challenge

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