Leukocytic response in monkeys challenged with staphylococcal enterotoxin

Sugiyama, H. (University of Chicago, Chicago, Ill.), and E. M. McKissic, Jr. Leukocytic response in monkeys challenged with staphylococcal enterotoxin. J. Bacteriol. 92:349-352. 1966-The feeding of staphylococcal enterotoxin to monkeys elicited a leukocytosis which was evident within 0.5 hr of challenge. The peak neutrophilic leukocytosis was reached in 3 hr, and then subsided so that leukocyte counts were normal within 28 hr. Each of the three serological types of enterotoxin tested induced the same effects. Intravenous injection of enterotoxin slightly above the emetic ed(50) level produced an initial leukopenia followed by a neutrophilic leukocytosis which was maximal 9 or more hr postinjection. With smaller intravenous challenges, some animals responded with a leukopenia followed by a leukocytosis, some with only a leukocytosis, and others with no significant change in total leukocyte counts. The reversal of normal lymphocyte-to-neutrophile ratio toward a neutrophile-predominant white blood cell population occurred in all animals.     

In situ DNA-RNA hybridization using in vivo bromodeoxyuridine-labeled DNA probe

Summary An in vivo 5-bromodeoxyuridine (BrdUrd) labeled DNA probe was used for in situ DNA-RNA hybridization. BrdUrd was incorporated into plasmid DNA by inoculating E. coli with Luria-Bertani (LB) culture medium containing 500 mg/L of BrdUrd. After purification of the plasmid DNA, specific probes of the defined DNA fragments, which contained the cloned insert and short stretches of the vector DNA, were generated by restriction endonuclease. The enzymatic digestion pattern of the BrdUrd-labeled plasmid DNA was the same as that of the non-labeled one. BrdUrd was incorporated in 15%–20% of the total DNA, that is, about 80% of the thymidine was replaced by BrdUrd. Picogram amounts of the BrdUrd-labeled DNA probe itself and the target DNA were detectable on nitrocellulose filters in dot-blot spot and hybridization experiments using a peroxidase/diaminobenzidine combination. The BrdUrd-labeled DNA probe was efficiently hybridized with both single stranded DNA on nitrocellulose filters and cellular mRNA in in situ hybridization experiments. Through the reaction with BrdUrd in single stranded tails, hybridized probes were clearly detectable with fluorescent microscopy using a FITC-conjugated monoclonal anti-BrdUrd antibody. The in vivo labeling method did not require nick translation steps or in vitro DNA polymerase reactions. Sensitive, stable and efficient DNA probes were easily obtainable with this method.     

N-Acylation of stimulatory amino acids changes chiral recognition of the fleshfly labellar sugar receptor

Summary N-Acylation changed nonstimulatory Dvaline into a clear stimulant of the sugar receptor of the fleshfly,Boettcherisca peregrina. Of theN-acyl-D-valines, the most stimulatory wasN-acetyl-D-valine. Similar changes into stimulants were also observed in other aliphatic amino acids such as leucine and methionine. Dose-response curves ofN-acetyl-D-valine suggested an increase of binding affinity, compared with that ofN-acetyl-L-valine. By treatment experiment with pronase 10 mg/ml, stimulatoryN-acetyl-D-amino acids were suggested to react with the specific alkyl site (R site), which was presumed to discriminate between L- and D-forms of the amino acids through steric hindrance between its own spatial barrier and D-amino acids (Shimada and Isono 1978; Shimada and Tanimura 1981).This change of chiral recognition cannot be explained by simple steric hindrance at the R site. It means, instead, that a hydrophobic subsite rather than a spatial barrier must be postulated.