Activation of old cuticle chitin as a substrate for chitinase in the molt of Manduca

During the molt, chitin in the old cuticle of $\text{Manduca}$ is digested by chitinase taken up from molting fluid, but the chitin in intact (= premolt) cuticle is not accessible to chitinase. As a prerequisite of digestion, old cuticle chitin is rendered competent to serve as chitinase substrate in a reaction attributable to trypsin-like proteolytic activity of molting fluid.     

Solubilization of microsomal phosphoethanolaminetransferase by octyl glucoside

Phosphoethanolaminetransferase of high specific activity was solubilized from rat liver microsomes with the non-ionic detergent octyl glucoside. The solubilization method is fast and simple, allowing for processing of large amounts of material. The solubilized enzyme is stable. It contains virtually no phosphocholinetransferase activity. A preliminary characterization of the enzyme, with both diacyl- and alkylacyl-glycerol as substrate, is given. For the reaction, the lipid substrates were incorporated into artificial phospholipid bilayers (liposomes).     

New procedure for concentration and analytical isoelectric focusing of proteins.

Proteins in aqueous salt solutions (up to 4 mil) were adsorbed by hydrophobic interaction on phenyl-Sepharose gel (0.1 ml) in small columns. After washing out excess salt, gels were applied on the surface of flat bed polyacrylamide gels for isoelectric focusing, which resulted in efficient desorption and transport of protein out of the phenyl-Sepharose gel. There was no difficulty in obtaining a fifty-fold concentration. The following parameters at adsorption of protein were studied: (i) salt concentration in the protein solution; (ii) phenyl-Sepharose gel adsorptive capacity for protein; (iii) suitable volume of washing solution for the phenyl-Sepharose gel. Theoretical aspects on factors promoting adsorption and desorption of proteins on phenyl-Sepharose gel are discussed. Also discussed are earlier used procedures for concentration and/or dialysis. When dilute protein solutions are to be examined for analytical purposes, the proposed procedures seems to be a valuable aid, which does not require expensive equipment, and is quick and simple to perform.     

A new method for determining sulfoxides in peptide molecules using x-ray photoelectron spectroscopy

The problem of the quantitative determination of sulfoxide groups in peptide molecules has been re-examined. The approaches currently available for the estimation of δ-sulfoxide amino acids are limited in number and characterized by serious shortcomings; in addition, the choice of methods for the estimation of γ-sulfoxide amino acids is even more restricted. A new, rapid, and nondestructive direct method for determining quantitatively all types of sulfoxides in peptide molecules by using x-ray photoelectron spectroscopy is described.     

A novel type of endotoxin structure present in Bordetella pertussis. Isolation of two different polysaccharides bound to lipid A.

The endotoxin of Bordetella pertussis was cleaved by mild acidic hydrolysis to yield a polysaccharide (polysaccharide I, 15%), a glycolipid (63%) and lipid X (2%). Further treatment of the glycolipid with stronger acid released a second polysaccharide (polysaccharide II, 9%) and material similar to lipid A present in enterobacterial endotoxins. Both polysaccharides possess a single molecule of 3-deoxy-2-octulosonic acid as the reducing, terminal sugar. In polysaccharide II the octulosonic acid is phosphorylated in position 5 and presumably substituted in position 4; in polysaccharide I the octulosonic acid is not phosphorylated, but is substituted in position 5. Following treatment of the endotoxin with strong base, a fragment was isolated that contained bound, non-phosphorylated 3-deoxy-2-octulosonic acid, glucosamine phosphate and fatty acids. This indicated that polysaccharide I, like polysaccharide II, was bound to the lipid region of the endotoxin. The endotoxin structure thus defined is different from that proposed for the lipopolysaccharides of enterobacteria.     

Boophilus microplus: characterization of larval proteases.

Cercariae of Schistosoma mansoni were treated with undecenyl-pseudothiourea. After centrifugation, they agglutinated into a mass. Resuspended in water, they remained immobilized. When injected sub-cutaneously into mice, they produced bisexual infections. The immobilizing drug effect, together with a reduced worm recovery rate, are time and concentration dependent. The cercariae become avirulent (99.8%) only when the flame cell is affected. Immobilizing and “cercaricidal” effects are not necessarily related properties; the latter can be determined only by in vivo tests of infectivity. No protection against reinfection was noticed in mice injected with immobilized cercariae of reduced virulence. The immobilized cercariae produced infections with a 0.7% worm recovery rate by percutaneous exposure, compared to 2.2% by subcutaneous injection. Normal cercariae produced infections with average recovery rates of 11.1% subcutaneously and 45% percutaneously.     

Sesquiterpenes from Lactarius blennius

Six sesquiterpene lactones were isolated from Lactarius blennius. The structures of two new sesquiterpenes, blennin A and blennin B were determinated by spectroscopic methods and the structure of the seco-compound, blennin C, is revised. The two known furan sesquiterpenes and lactarorufin A were also identified.