No abstract

Hydrogen peroxide production in reconstituted skim milk (10%) and low-fat milk by four strains lf Lactobacillus acidophilus was studied at 37 and 4 C. Strains A and B produced little, but strains C and D produced larger amounts, especially if agitated continuously during growth at 37 C or storage at 4 C. Continuous shaking was required at 4 C for C or D (4.0 X 10(8)/ml) to produce sufficient hydrogen peroxide to retard growth of Pseudomonas fragi. Flavin adenine dinucleotide stimulated the oxidation of reduced nicotinamide adenine dinucleotide by dialyzed cell-free extracts of C and D, which indicated that the reduced nicotinamide adenine dinucleotide oxidases of these strains produce hydrogen peroxide as an end product.

Pyruvate was shown to be the precursor of diacetyl and acetoin in Streptococcus diacetilactis, but dialyzed cell-free extracts of S. diacetilactis and Leuconostoc citrovorum that had been treated with anion-exchange resin to remove coenzyme A (CoA) formed only acetoin from pyruvate in the presence of thiamine pyrophosphate (TPP) and Mg++ or Mn++ ions. The ability to produce diacetyl was restored by the addition of acetyl-CoA. Acetyl-phosphate did not replace the acetyl-CoA. Neither diacetyl nor acetoin was formed when the otherwise complete reaction system was modified by using boiled extract or by omitting the extract, pyruvate, TPP, or the metal ions. Free acetaldehyde was not involved in the biosynthesis of diacetyl or acetoin from pyruvate, dialyzed cell-free extracts of the bacteria produced only acetoin (besides C02) from a-acetolactate, and acetoin was not involved in the biosynthesis of diacetyl. Only one of the optical isomers present in racemic a-acetolactate was attacked by the extracts, and there was no appreciable spontaneous decarboxylation of the a-acetolactate at the pH (4.5) used in experiments. 174 Vol. 95, No. I

Medical devices, such as contact lenses, bring bacteria in direct contact with human cells. Consequences of these host-pathogen interactions include the alteration of mammalian cell surface architecture and induction of cellular death that renders tissues more susceptible to infection. Gram-negative bacteria known to induce cellular blebbing by mammalian cells, Pseudomonas and Vibrio species, do so through a type III secretion system-dependent mechanism. This study demonstrates that a subset of bacteria from the Enterobacteriaceae bacterial family induce cellular death and membrane blebs in a variety of cell types via a type V secretion-system dependent mechanism. Here, we report that ShlA-family cytolysins from Proteus mirabilis and Serratia marcescens were required to induce membrane blebbling and cell death. Blebbing and cellular death were blocked by an antioxidant and RIP-1 and MLKL inhibitors, implicating necroptosis in the observed phenotypes. Additional genetic studies determined that an IgaA family stress-response protein, GumB, was necessary to induce blebs. Data supported a model where GumB and shlBA are in a regulatory circuit through the Rcs stress response phosphorelay system required for bleb formation and pathogenesis in an invertebrate model of infection and proliferation in a phagocytic cell line. This study introduces GumB as a regulator of S . marcescens host-pathogen interactions and demonstrates a common type V secretion system-dependent mechanism by which bacteria elicit surface morphological changes on mammalian cells. This type V secretion-system mechanism likely contributes bacterial damage to the corneal epithelial layer, and enables access to deeper parts of the tissue that are more susceptible to infection.

tococcus diacetilactis was unable to use citrate as a source of energy for growth, but the addition of citrate to a lactose-containing medium increased the specific growth rate 35%. Besides serving as the precursor of acetoin, some of the pyruvate formed from citrate was incorporated into cell material, primarily into lipids. A constant fraction of the weight of new cells was synthesized from the pyruvate formed from citrate. The rate of entry of citrate into cells was independent of the growth rate, and the usual result was that more pyruvate was formed from citrate than was required for cell synthesis. All excess pyruvate was converted to acetoin. Thus, acetoin formation acts as a detoxification mechanism, a means of removing intracellular pyruvate not required for synthesis of cell material.