Evolutionary significance of the renal excretion of transferrin half-molecule fragments

It is generally thought that the duplicated structure of serum transferrin in vertebrates arose by gene duplication and fusion from a small ancestral protein. We have found that the isolated domains of transferrin are rapidly lost from the bloodstream via the kidneys. Therefore we suggest that the ancestral transferrin was not a serum protein or, alternatively, that it was not as small as the half-molecule.     

Thin-layer separation and quantification of bile acids

A method has been developed for quantification of total free and conjugated bile acids separated on silica gel HR coated thin-layer chromatography plates. Aliquots of bile acid solutions are applied to channeled plates which are developed with either ethyl acetate: isooctane: glacial acetic acid 10:10:2 v/v for free bile acid separation, or chloroform:methanol:glacial acetic acid:water 130:50:4:8 v/v for conjugated bile acid separation. Bile acids are determined directly in serial areas of silica gel by treating gel areas suspended in tris buffer with resazurin reagent. The method is quantitative and as little as 0.1 μg of bile acid is readily determined. Application of the method to determinations of bile acids in crude fecal extracts is described.     

Chromosomal and isozyme studies of Nicotiana tabacum — Glycine max hybrid cell lines

Summary The chromosomal stability of a number of somatic hybrids derived from soybean (Glycine max (L.) Merr.) and Nicotiana tabacum var. Xanthi were investigated. Several of the hybrid cell lines retained more than half the complement of N. tabacum chromosomes after 7 months of culturing. A number of chromosomal abnormalities were observed. The hybrids were positively identified by employing isozyme analysis of several dehydrogenases and aspartate aminotransferase.NRCC No. 20130     

Degradation of Arachidonoyl-Labeled Phosphatidylinositols by Brain Synaptosomes

Abstract: Incubation of synaptosomes together with 1-acyl-2-[¹⁴C]arachi-donoyl-sn-glycerophosphoinositols (GPI) and sodium deoxycholate yielded diacylglycerols and free arachidonic acid. Diacylglycerol formation is attributed to hydrolysis by the diacyl-GPI-specific phospholipase C (EC 3.1.4.10), and this reaction requires sodium deoxycholate for optimal activity. The free arachidonic acid formed is attributed to hydrolysis of diacyl-GPI by phospholipase A (EC 3.1.1.5). Free fatty acid release was observed during incubation, even in the absence of bile salts, but this process was preferentially stimulated by sodium taurocholate. The release of fatty acids was not specific for diacyl-GPI, as similar release was obtained during incubation with other phosphoglycerides. In the presence of deoxycholate (2 mg/ml), the release of diacylglycerols was maximal at a diacyl-GPI concentration around 1.0 mM. However, the free fatty acid release was linear with respect to the substrate at least up to 1.4 mM. The rate of diacylglycerol release from diacyl-GPI was more rapid in the initial 30 min, whereas the free fatty acid release was linear with time up to 2 h. Under this incubation condition, calcium was found to stimulate both types of hydrolytic action, although the concentration needed to achieve this stimulation was rather high. This type of labeled precursor is potentially useful for studies of the different modes of diacyl-GPI degradation by enzymes in brain subcellular membranes.     

Retardation of leaf senescence by inhibitors of RNA and protein synthesis

Effects of actinomycin-D (ACT), cycloheximide (CH), rifampicin (RIF) and chloramphenicol (CAP) on senescence of soybean leaf discs were investigated. All inhibitors tested are effective in retarding senescence of soybean leaf discs. However, CH is more effective than ACT, RIF and CAP, suggesting that activation of preexisting, latent metabolic systems present in the cytoplasm piays predominant role in the initiating of leaf senescence. However, the possibility that events taking place in the nucleus or chloroplast are essential for the initiation of leaf senescence cannot be excluded.     

Identification of the diol associated with variations in wax ultrastructure on Rhus cotinus leaves

The diol constituent of Rhus cotinus leaf epicuticular wax has been identified as nonacosane-5,10-diol from chemical investigations of the free compound, the TMSi ether and the nonacosane-5,10-dione prepared from the diol by oxidation. The form and distribution of the crystalline waxes changed as the leaves expanded, dense clusters of short tubes covering the thin ribbons formed during the initial stages of growth. The diol content of the wax decreased by more than 50% over the same period.     

Kinetics for the secretion of nonhelical procollagen by freshly isolated tendon cells

Fibroblasts isolated by enzymic digestion of chick embryo tendons have previously been used to examine the kinetics for the secretion of procollagen (Kao, W. W.-Y., Berg, R. A., and Prockop, D. J. (1977) J. Biol. Chem. 252, 8391-8397). The results indicated that the kinetics approximated the sum of two first order processes with half-times of 14 and 115 min. Here, the same fibroblasts were incubated in the presence of 1.53 mM cis-4-hydroxyproline, an analogue of proline, or in the presence of 0.3 mM alpha,alpha'-dipyridyl, an inhibitor of prolyl hydroxylase, so that the cells synthesized procollagen which could not assume a triple helical conformation characteristic of procollagen. Measurements of the secretion of nonhelical procollagen indicated that the kinetics for secretion differed from the kinetics for the secretion of procollagen and approximated a single first order process with a half-time of approximately 130 min. The nonhelical procollagen synthesized and secreted in the presence of either cis-4-hydroxyproline or alpha,alpha'-dipyridyl consisted of disulfide-bonded pro gamma chains of type I procollagen. The results suggested that the intracellular nonhelical procollagen was present in a single metabolic pool and secretion from this pool occurred with a different rate-limiting step than for helical procollagen. Further results indicated that nonhelical procollagen had a high affinity for prolyl hydroxylase and the affinity for the enzyme was greatly reduced if the procollagen was allowed to assume the triple helical conformation characteristic of normal procollagen. The results are consistent with the hypothesis that the secretion of procollagen is influenced by its conformation-dependent interaction with prolyl hydroxylase or other post-translational enzymes.