Presence of the Novel Pituitary Protein „7B2” in Bovine Chromaffin Granules: Possible Co-Release of 7B2 and Catecholamine as Induced by Nicotine

We observed the presence of the novel pituitary protein "7B2" and its release in the bovine adrenal medulla. The 7B2 concentration (mean +/- SEM) in extracts of the bovine adrenal medulla was 952 +/- 155 pg/mg tissue (n = 6). 7B2 was distributed in the chromaffin granule fraction prepared from the bovine adrenal medulla and was released by high K+ and/or nicotine from cultured cells of the bovine adrenal medulla. Co-release of 7B2 with catecholamine induced by nicotine from the cultured bovine chromaffin cells was also observed. In an analysis of the bovine adrenal medulla chromaffin granule fraction on gel permeation chromatography, there was a major peak with an apparent molecular weight of 45,000, whereas a major peak with an apparent molecular weight of 20,000 was found in that on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. On reverse-phase HPLC, a major peak with a retention time of 35 min was observed in the bovine chromaffin granule fraction and in the bovine anterior pituitary extract. These findings indicate that 7B2 is a secretory protein in the bovine adrenal medulla. The possibility that 7B2 might be released with catecholamine, possibly in response to stress, warrants investigation.     

Interspecific relationships ofCallithrix based on the dental characters

The dental structure of all species ofCallithrix, C. argentata, C. humeralifer, C. aurita, C. flaviceps, C. geoffroyi, C. penicillata, andC. jacchus is examined.Callithrix are divided intoC. jacchus group andC. argentata group, based on the analysis of dental characters.C. jacchus group consists ofC. jacchus, C. penicillata, C. geoffroyi, C. flaviceps, andC. aurita, whileC. argentata andC. humeralifer are assigned toC. argentata group. InC. jacchus group,C. aurita andC. flaviceps were differentiated from the original stock of their common ancestor, followed byC. geoffroyi, last byC. penicillata, and finallyC. jacchus. Based on the relationships among species ofCallithrix, it is possible to infer the relative age of the formation of the refuges which caused their speciation. First, the forests in southeastern Brazilian coast were split from Amazonia. In southeastern Brazilian coast, the Paulista center separated, followed by the Rio Doce center, the Bahia center, finally by the Pernambuco center.     

Fine structure of the dental enamel in the Family Callitrichidae (Ceboidea,primates)

The fine structure of the dental enamel was intensively examined in the Family Callitrichidae.Leontopithecus rosalia has the nonserial pattern, butCallimico goeldii andSaguinus midas appear more or less modified from the typical nonserial pattern asSaimiri sciureus has (see Figs. 1 & 5). On the other hand,Callithrix jacchus andCebuella pygmaea attain to the primitive stage of the multiserial pattern (see Figs. 4 & 6). So far as the structural patterns of the dental enamel are concerned, a serial trend is confirmed in the Callitrichidae; extending from the genusLeontopithecus through the generaCallimico andSaguinus to the generaCallithrix andCebuella. The genusSaguinus shows extremely wide interspecific variation in the fine structure of the dental enamel, ranging fromS. leucopus with the most primitive features toS. bicolor with the most advanced (see Figs. 2 & 3). The rows of enamel prisms are slightly twisted mesiodistally in the former species, but strongly folded in the latter. This reflects transitional stages from the nonserial pattern to the multiserial. As a whole, however, the genusSaguinus is still regarded as nonserial. Thus, there is a slight, but important gap between the genusSaguinus andCallithrix, as shown by the decussation of clusters (see Fig. 5), as well as by other characters.     

Effects of chaetoglobosin J on the G-F transformation of actin

It was shown that substoichiometric concentrations of chaetoglobosin J, one of the fungal metabolites belonging to cytochalasins, inhibited the elongation at the barbed end of an actin filament. Stoichiometric concentrations of chaetoglobosin J decreased both the rate and the extent of actin polymerization in the presence of 75 mM KCl, 0.2 mM ATP and 10 mM Tris-HCl buffer at pH 8.0 and 25 degrees C. In contrast, stoichiometric concentrations of cytochalasin D accelerated actin polymerization. Chaetoglobosin J slowly depolymerized F-actin to G-actin until an equilibrium was reached. Analyses by a number of different methods showed the increase of monomer concentration at equilibrium to depend on chaetoglobosin J concentrations. F-actin under the influence of stoichiometric concentrations of chaetoglobosin J only slightly activated the Mg2+-enhanced ATPase activity of myosin at low ionic strength. It is suggested that when the structure of the chaetoglobosin-affected actin filaments is modified, the equilibrium is shifted to the monomer side, and the interaction with myosin is weakened.     

Proposal of standardized methods and reference for assaying recombinant human tumor necrosis factor

Two assay methods for recombinant human tumor necrosis factor (rH-TNF) were developed, one a biological L-cell assay and the other an enzyme-linked immunosorbent assay. The accuracy and reproducibility of each and the correlation between the two were studied. As a result of this investigation, the two assay methods were found appropriate for standardization of rH-TNF. A freeze-dried reference was prepared, and examination of its potency and stability showed it to be suitable for use as a reference standard for rH-TNF assays.     

Purification of lectin induced in the hemolymph of Sarcophaga peregrina larvae on injury

A lectin was purified from the hemolymph of Sarcophaga peregrina larvae, obtained after injury of their body wall. This lectin agglutinated sheep red blood cells markedly and the hemagglutinating activity was inhibited by galactose and lactose. The active lectin was found to have a molecular weight of 190,000 and to consist of four alpha subunits and two beta subunits, with molecular weights of 32,000 and 30,000, respectively. During the early pupal stage, similar hemagglutinating activity in the hemolymph increased to several times than in larval hemolymph. This activity was completely inhibited by the antibody prepared against the lectin purified from the hemolymph of injured larvae. Thus, the same protein having lectin activity is apparently induced under two different physiological conditions: injury of the body wall of larvae and during pupation. The biological significance of this lectin is discussed.     

Molecular cloning of chick liver HMG 2a cDNA and developmental expression of HMG 2a mRNA.

The cDNA clone encoding HMG 2a of chick liver was isolated from a lambda gt11 expression library using polyclonal antibodies. DNA sequence analysis revealed an open reading frame of 201 amino acids. Comparison of the nucleotide sequences of cDNA coding for chick liver HMG 2a with pig thymus HMG 2 and human monocytic leukemia cell HMG 2 showed 70% homology. 2.0 kb and 1.2 kb mRNAs were found in newly hatched chick liver and decreased during postnatal development of chicks.     

Purification of a 29-kDa hemocyte proteinase of Sarcophaga peregrina.

Previously, we suggested the participation of a hemocyte proteinase in the dissociation of fat body of Sarcophaga peregrina (flesh fly) at metamorphosis. We have now purified this proteinase to near homogeneity from pupal hemocytes. It is a cysteine proteinase with a molecular mass of 29 kDa and has a unique substrate specificity hydrolyzing both Suc-Leu-Leu-Val-Tyr-MCA and Z-Phe-Arg-MCA (Suc, succinyl; MCA, methylcoumaryl-7-amide; Z, carbobenzoxy), which are substrates for chymotrypsin and cathepsin B, respectively. Partial similarity was found between the amino-terminal sequence of this proteinase and that of cathepsin B, including Pro, Glu and Arg residues conserved in the papain superfamily of enzymes.