New embedding medium for sectioning undecalcified bone

Methylmethacrylate (MMA) is the most commonly used embedding medium for sectioning undecalcified bone; however, a number of problems exist with its use in a research laboratory. MMA requires a long infiltration time and temperature control, and it reacts with many polymers. We used Kleer Set resin™ as an alternative embedding medium for sectioning undecalcified bone specimens. Fluorochrome labeled bone specimens were sectioned transversely using a ground section technique and longitudinally on a sledge macrotome. The slides were viewed using both transmitted light and epifluorescence microscopy. High quality sections were obtained using Kleer Set resin™ for both sectioning techniques. We have shown that this new embedding medium is simpler, safer, quicker to use and does not interfere with visualization of fluorochromes.     

Noncytotoxic activation of neutrophils by eosinophil granule major basic protein. Effect on superoxide anion generation and lysosomal enzyme release

Eosinophil granule major basic protein (MBP) and neutrophils have each been implicated in the inflammatory late phase events of allergic disease. Based on this association and flow cytometric evidence presented in this report for MBP binding to neutrophils, we examined the ability of MBP to activate human neutrophils. Incubation of neutrophils with 0.5 to 3.0 microM MBP at room temperature produced a concentration-dependent chemiluminescence (CL) response that peaked after 50 to 70 min. Reduced-and-alkylated MBP, eosinophil cationic protein, and eosinophil-derived neurotoxin did not induce CL. MBP-induced CL was abrogated in the absence of Ca2+ and was absent in neutrophils isolated from two individuals with chronic granulomatous disease. MBP also stimulated release of superoxide anion (O2-) and lysozyme but not beta-glucuronidase or lactate dehydrogenase. Additionally, 1.5 microM MBP in combination with FMLP or platelet-activating factor stimulated a synergistic increase in O2- release from cytochalasin B-treated neutrophils. The degree of synergism with FMLP or platelet-activating factor was inversely related (p less than 0.005) to the level of MBP-induced O2- release. These results indicate that MBP activates neutrophils in a noncytolytic fashion and provide evidence that eosinophil-neutrophil collaboration may contribute to the pathogenesis observed in allergic late phase reactions.     

The amino terminal sequence of sea urchin sperm histone H1 and its phosphorylation by egg cytosol

1. The amino acid sequence of the first 34 residues of sperm histone H1 (SpH1) from Strongylocentrotus purpuratus shows striking similarity with sequences from three South African species. 2. Five contiguous repeats of the tetrapeptide SPBB (where B is R or K) occur between positions 10 and 29. 3. SpH1 was phosphorylated in vitro using egg cytosol as the source of protein kinase and approximately 4.2 mol phosphate were incorporated per mol H1. 4. Sequences of five phosphopeptides of SpH1 show the egg possesses protein kinase activity capable of phosphorylating multiple seryl residues including SPBB in the NH2-, and BBSP in the COOH-end of the protein.     

A-B-O blood groups in Kenya olive baboons (Papio anubis)

A feral population of 183 Kenyan olive baboons representing 5 troops was surveyed for salivary ABO-like antigens. Unlike previously reported populations, a high frequency of the O allele and low frequency of the B allele were detected. This observation may be the result of founder effect and/or genetic drift.