Probing single molecule dynamics

The room temperature dynamics of single sulforhodamine 101 molecules dispersed on a glass surface are investigated on two different time scales with near-field optics. On the 10(-2)- to 10(2)-second time scale, intensity fluctuations in the emission from single molecules are examined with polarization measurements, providing insight into their spectroscopic properties. On the nanosecond time scale, the fluorescence lifetimes of single molecules are measured, and their excited-state energy transfer to the aluminum coating of the near-field probe is characterized. A movie of the time-resolved emission demonstrates the feasibility of fluorescence lifetime imaging with single molecule sensitivity, picosecond temporal resolution, and a spatial resolving power beyond the diffraction limit.     

Chitin in sea anemone shells

Chitin, which is widely distributed among life forms, is well documented in the coelenterate class Hydrozoa and is contained in one member of class Scyphozoa. In class Anthozoa, hard corals synthesize it but soft corals do not. Chitin was identified by infrared spectrophotometry in the trochoid shell of the actinian Stylobates. It constitutes 1.7 percent of the shell by weight, the rest probably being protein. The ability of sea anemones to synthesize chitin is there by confirmed.     

Identification of genes associated with prophage-like gene transfer agents in the pathogenic intestinal spirochaetes Brachyspira hyodysenteriae, Brachyspira pilosicoli and Brachyspira intermedia

VSH-1 is an unusual prophage-like gene transfer agent (GTA) that has been described in the intestinal spirochaete Brachyspira hyodysenteriae. The GTA does not self-propagate, but it assembles into a virus-like particle and transfers random 7.5kb fragments of host DNA to other B. hyodysenteriae cells. To date the GTA VSH-1 has only been analysed in B. hyodysenteriae strain B204, in which 11 late function genes encoding prophage capsid, tail and lysis elements have been described. The aim of the current study was to look for these 11 genes in the near-complete genomes of B. hyodysenteriae WA1, B. pilosicoli 95/1000 and B. intermedia HB60. All 11 genes were found in the three new strains. The GTA genes in WA1 and 95/1000 were contiguous, whilst some of those in HB60 were not-although in all three strains some gene rearrangements were present. A new predicted open reading frame with potential functional importance was found in a consistent position associated with all four GTAs, located between the genes for head protein Hvp24 and tail protein Hvp53, overlapping with the hvp24 sequence. Differences in the nucleotide and predicted amino acid sequences of the GTA genes in the spirochaete strains were consistent with the overall genetic distances between the strains. Hence the GTAs in the two B. hyodysenteriae strains were considered to be strain specific variants, and were designated GTA/Bh-B204 and GTA/Bh-WA1 respectively. The GTAs in the strains of B. intermedia and B. pilosicoli were designated GTA/Bint-HB60 and GTA/Bp-95/1000 respectively. Further work is required to determine the extent to which these GTAs can transfer host genes between different Brachyspira species and strains.     

Stability of Native Folate and Added Folic Acid in Micronutrient‐Fortified Corn Masa and Tortillas

Degradation of added folic acid and native folates in micronutrient‐fortified corn masa and tortillas was evaluated using masa prepared from either nixtamalized corn flour or fresh nixtamal. Variations in masa pH, masa holding time at an elevated temperature, and iron source failed to show significant differences in folate loss in corn flour masa prepared in the laboratory. Masa was subsequently prepared from fresh nixtamal in a commercial mill in Mexico, and fortified with one of two different micronutrient premixes containing iron, zinc, B‐vitamins, and either unencapsulated or lipid‐encapsulated folic acid. Folate loss in commercial masa increased significantly with prebake masa holding time for both premixes. Unencapsulated folic acid showed a 73% loss after 4 hr of holding, compared to 60% loss for encapsulated. The difference was statistically significant, indicating a protective effect from the lipid coating. No significant differences in folate levels were found between prebake masa and baked tortillas. Holding baked tortillas for up to 12 hr also had no effect on folate levels. Native folate showed no significant losses throughout the process. Results from the commercial tortilla mill indicate that most of the loss in added folic acid occurs during prebake holding of masa, possibly from microbial degradation.