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Biomedical subjects

F Vaughan

Publications and source records attributed to F Vaughan.

6 recordsLinked to original sources
PubMed1991

Production and isolation of guinea pig IgE antibody.

Immunoglobulins of the IgE and IgG classes have been causally associated with hypersensitivity reactions in man and in numerous animal species including mice, rats and guinea pigs. The use of the guinea pig as an animal model for both pulmonary and dermal hypersensitivity reactions, and the recent recognition of the importance of IgE antibodies in both early- and late-onset hypersensitivity responses, has heightened interest in production, separation, and isolation of this immunoglobulin class from the guinea pig. IgE antibodies were produced by treatment of strain 13 guinea pigs with cyclophosphamide followed by injection with S. aureus enterotoxin. Serum was obtained and the globulin fraction isolated by addition of caprylic acid then ammonium sulfate. Immunoglobulins were separated into classes using fast protein liquid chromatography (FPLC) employing a Mono Q column and a linear gradient of 0.01-0.3 M Na,K phosphate buffer, pH 7.5 (buffer B). IgG eluted in two major peaks. IgG2 was not retained on the column and emerged with the starting buffer; IgG1 was eluted with 15-20% buffer B. IgE, detected as heat labile homocytotropic antibody, was found in the fraction eluting with 30-35% buffer B. The elution profile of the guinea pig immunoglobulins was predicted from the pattern obtained with immunoglobulin classes from other species. This chromatographic procedure enabled rapid isolation of immunoglobulin classes from guinea pig sera and effectively separated IgG1 from IgE, the two classes associated with hypersensitivity reactions.

Animals↗
PubMed1982

Lactate acid inhibition of Salmonella typhimurium in yogurt.

We determined how lactic acid inhibits growth of Salmonella typhimurium in yogurt. This inhibition was demonstrated by microscopic examination not to be due to bacteriolysis. Neither growth nor metabolic activity could be initiated after cells were washed in phosphate buffer and exposed to 1.5% lactic acid for 1 h at 37 degrees C, indicating that lactic acid inhibition is irreversible. The growth rate of S. typhimurium at 37 degrees C, was computed at various combinations of pH and lactic acid concentrations, and the intracellular conditions (pH and lactic acid concentration) at bacteriostasis subsequently were extrapolated. Cellular death resulted when these intracellular bacteriostatic conditions were surpassed. Thus, growing cells could be used indirectly to determine intracellular conditions at the time of death. Intracellular pH (pHi) and inhibition of the growth rate were unrelated. Also, bacteriostasis was observed when hydrochloric acid was used to lower the pHi of Salmonella to 5.5 whereas a bactericidal effect was observed when the pHi was lowered to 5.5 with lactic acid. The lactate anion, rather than the hydrogen ion, exerted the inhibitory effect against S. typhimurium. When the pHi became less than 5.3, inhibition was from the hydrogen ion concentration. Thus, lactic acid inhibition was a complex and variable mechanism in relationship with pHi Lactic acid entered the cell in the undissociated state. Once inside the cell, it dissociated because the pHi was higher than the external pH. The dissociated moiety accumulated because it could not leave the cell in this form consequently lowering the pHi. Thus, inhibition of S. typhimurium in yogurt is from the intracellular dissociated moiety of lactic acid.

Culture Media↗
PubMed1979

Elucidation of the inhibitory factors of yogurt against Salmonella typhimurium.

The inhibitory nature of yogurt against contaminating microorganisms has been studied extensively. Nevertheless, the factors responsible for the death of Salmonella typhimurium in yogurt have not been elucidated. An understanding of these factors is important for the determination of yogurt's safety to consumer health. Yogurt fermented for 18 h at 42 C had a stable environment with the following conditions: pH 3.85, oxidation-reduction potential -80 mV, lactic acid concentration 158 mM, and acetic acid concentration 3.7 mM. Under these conditions, lactic acid was responsible for virtually all of yogurt's bactericidal activity against S. typhimurium at 37 C. Die-off rates were observed when these conditions were reproduced artificially in milk (artificial milk yogurt) and when lactic acid was added back to 18-h yogurt from which acids were removed by passage of the whey through a Dowex 1 (Cl-) anion exchange column (cationic yogurt). Factors that augmented lactic acid inhibition of S. typhimurium were low pH and low oxidation-reduction potential. The die-off rate of S. typhimurium was more rapid in yogurt whey (yogurt minus the casein fraction) than in whole yogurt, indicating that the casein fraction was partially able to protect Salmonella.

Acetates↗