Article

The antimycobacterial activity of a peptide preparation derived from calf thymus

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Abstract

A stable, water-soluble substance which possesses potent antimycobacterial activity under certain conditions in vitro has been prepared from calf thymus. This substance has been tentatively named thymus peptide. In final concentrations of 1 to 10 µg. per ml. of an albumin medium it inhibits the growth of various strains of mammalian mycobacteria, but manifests only little or no inhibitory activity against a variety of other microbial species. The ability of thymus peptide to inhibit the multiplication of tubercle bacilli diminishes when the inoculum is large, or when the medium is acidic. It is also markedly antagonized by addition of enzymatic hydrolysate of casein or beef heart infusion broth to the culture medium. Thymus peptide does not exert a rapid bactericidal action on tubercle bacilli, but organisms exposed to this compound for longer than 2 weeks could not be made to multiply in ordinary culture media. Substances similar or identical to the thymus peptide preparation could be extracted from calf spleen, sheep thymus, beef lymph nodes, and calf pancreas, but not from calf lung or calf liver.

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... 20 of tubercle bacilli by disrupted mononuclear cells obtained from normal and BCG-immunized guinea pigs, and the factor responsible for this inhibition was found to be a heat-labile nuclear fraction. This information reminds us of the former observation of Dubos and Hirsch (1954) that a peptide preparation derived from calf thymus had an :antimycobacterial activity. ...
... To answer this question, the research group of David demonstrated that some of these mediators can be distinguished from one another by their Ramseier and Suter (1964 a, b) observed in vitro inhibition of tubercle bacilli by disrupted mononuclear cells obtained from normal and BCG-immunized guinea pigs ; the factor responsible for the inhibition was found to be a heat-labile nuclear fraction. This information reminds us of the still earlier observation of Dubos and Hirsch (1954) that a peptide fraction from calf thymus had an antimycobacterial activity. Therefore, we think that basic proteins or peptides in general may have some antibacterial effect (Oshima, Myrvik and Leake, 1961;Hibbit, Cole and Reiter, 1969 ;Svihla, Dainko and Schlenk, 1969). ...
... Serpent is involved in cecropin gene expression by binding to the GATA site on cecropin gene promoter (Kadalayil et la., 1997). (Matanic, and Castilla, 2004), influenzavirus (Chernysh et la., 2002), cytomegalovirus (Andersen et la., 2001), TB (Mycobacterium tuberculosis) (Dubos and Hirsch, 1954;Miyakawa et la., 1996) (Mor et la., 1994;Lorin et la., 2005). Lactoferricin induces apoptosis in several human leukemia and carcinoma cell lines (Mader et la., 2005). ...
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A substance possessing antimycobacterial activity under certain conditions in vitro has been prepared from aqueous extracts of calf thymus. Chemical studies have demonstrated that the activity of this substance is due to a basic peptide or a mixture of basic peptides. Although this thymus fraction has been shown to be essentially free of compounds other than peptides, it has not been obtained in a homogeneous state. The thymus peptide preparation is soluble in water and in the lower alcohols. Its solubility is minimal between pH 10 and 11, suggesting that its isoelectric point may be in this vicinity. The microbiological activity of thymus peptide is destroyed by acid or alkaline hydrolysis and also by trypsin digestion, but is unaffected by pepsin digestion. Cellulose membranes are permeable to thymus peptide. The most noteworthy finding concerning the amino acid composition of thymus peptide is the preponderance of the basic amino acids lysine and arginine, which together account for about 40 per cent of the weight of this substance. No cystine, and only trace amounts of other amino acids containing sulfur, are present in the thymus peptide preparation.
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Chapter
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Sheep serum and bovine serum contain an enzyme which brings about a rapid oxidative deamination of certain biological amines. This enzyme differs from previously described amine oxidases in several regards and especially in its substrate specificity. Studies thus far indicate that only spermine and the closely related compound spermidine serve as substrates for the enzyme in sheep serum. For this reason, the enzyme has been named spermine oxidase. Spermine oxidase is active in a variety of fluids of various ionic strength and buffer composition. The reaction takes place between pH 6.0 and pH 8.0 with an optimal rate in the vicinity of neutrality. Under certain conditions, the rate of oxygen consumption during the initial phase of the reaction is independent of the concentration of substrate. The diminution in rate observed during the latter phase of the enzymatic attack appears to be due to an alteration in the kinetics at low concentrations of substrate, or to competitive inhibition by a product of the reaction. Carbonyl reagents almost completely block the action of spermine oxidase, while certain amines and the cyanide ion bring about partial inhibition. Thiol reagents and sequestering compounds do not alter the course of the oxidative process. In the presence of low concentrations of mercuric chloride, the sheep serum-spermine system consumes approximately twice as much oxygen as controls containing no mercuric ion. The mechanism by which the mercuric ion stimulates additional oxygen uptake is obscure.
Article
A substance possessing antimycobacterial activity under certain conditions in vitro has been prepared from aqueous extracts of calf thymus. Chemical studies have demonstrated that the activity of this substance is due to a basic peptide or a mixture of basic peptides. Although this thymus fraction has been shown to be essentially free of compounds other than peptides, it has not been obtained in a homogeneous state. The thymus peptide preparation is soluble in water and in the lower alcohols. Its solubility is minimal between pH 10 and 11, suggesting that its isoelectric point may be in this vicinity. The microbiological activity of thymus peptide is destroyed by acid or alkaline hydrolysis and also by trypsin digestion, but is unaffected by pepsin digestion. Cellulose membranes are permeable to thymus peptide. The most noteworthy finding concerning the amino acid composition of thymus peptide is the preponderance of the basic amino acids lysine and arginine, which together account for about 40 per cent of the weight of this substance. No cystine, and only trace amounts of other amino acids containing sulfur, are present in the thymus peptide preparation.
Article
The growth of tubercle bacilli in vitro was inhibited by the addition of the sodium salts of very low concentrations of certain organic acids to a variety of liquid and agar culture media containing whole serum or serum albumin. Capric acid was the most active of the compounds tested, but inhibition of growth occurred also with the shorter aliphatic acids. Lactic acid was also growth-inhibitory, whereas the keto and dicarboxylic acids tested were inactive in this respect. The inhibitory activity of the aliphatic acids and of lactic acid increased as the pH of the medium was lowered by addition of HCl. It was greater in media enriched with serum or with oleic acid-albumin complex, but was otherwise fairly independent of the composition of the medium. The inhibitory effect appears to be bacteriostatic rather than bactericidal and to depend upon a disturbance of the normal metabolic processes of the bacilli. Some of the long chain fatty acids caused a marked enhancement of growth when used in low concentrations and in admixture with enough serum albumin to overcome their toxicity. The significance of these findings is discussed with reference to the survival and multiplication of tubercle bacilli in vivo within inflammatory and caseous areas, which are known to be often acidic and to contain high concentrations of organic acids.
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