Effects of temperature and infection with Eimeria ochrogasteri on digestive organs of the prairie vole, Microtus ochrogaster

Prairie voles, Microtus ochrogaster, were infected with Eimeria ochrogasteri and exposed to 2 environmental temperatures, 5 and 22 C. Dry weights of the small and large intestines increased by 33% and 19%, respectively, in infected animals. Infected animals also exhibited a 14% decrease in cecal length compared to uninfected animals. The interaction between temperature and infection affected the length of the small intestine. Infected animals maintained at 5 C had longer small intestines than both infected animals housed at 22 C, and uninfected animals at 22 or 5 C. Furthermore, the dry weight of the small intestine was affected by a 3-way interaction (infection, temperature, and sex). Temperature affected stomach and liver dry weights, as well as lengths of the small intestine and cecum. Stomach and liver dry weights, as well as small intestine lengths, were greater in those animals held at 5 C, whereas cecum lengths decreased. Prepatency, patency, and total oocyst production were not affected by temperature; however, infected animals held at 5 C exhibited diarrhea during the patent period.     

Induction of transforming growth factor-alpha in activated human alveolar macrophages

The early monocyte infiltration observed in normal wound repair and in a number of pathologic processes precedes the epithelial and connective tissue proliferative responses, suggesting that the monocyte/macrophage may be an important source of growth factors for these tissues. In culture, activated macrophages secrete growth factors active on fibroblasts, smooth muscle, endothelium, and epithelium. This report demonstrates that activated human alveolar macrophages express the gene for transforming growth factor-alpha (TGF-alpha) in an inducible manner and secrete a factor into the culture medium that is functionally and immunologically identical to TGF-alpha. Two different molecular species of TGF-alpha activity (approximately 8,500-12,000 and 28,500 daltons) are identified in macrophage-conditioned medium. These observations establish the macrophage as a diploid human cell capable of synthesizing and secreting TGF-alpha. The activated macrophage therefore represents a cellular source of a mitogenic factor that is potentially important in epithelial proliferation and repair.     

Analysis of the mitogenic effect of fetuin preparations on arterial smooth muscle cells: the role of contaminant platelet-derived growth factor

Fetuin, a major protein of fetal calf serum, partially purified by the method of Pedersen, stimulated growth of aortic smooth muscle cells. More highly purified fetuin preparations stimulated growth less than Pedersen fetuin, as previously described for other cell types, suggesting that this activity is due to a contaminant. Recently bovine alpha 2-macroglobulin or "Embryonin" has been proposed as the mitogenic component of crude fetuin preparations. We found that active fetuin preparations did contain alpha 2-macroglobulin that stimulated smooth muscle cell growth. However, alpha 2-macroglobulin purified directly from platelet-poor bovine plasma or fetuin purified from Pedersen fetuin by gel filtration lacked appreciable mitogenic effect on smooth muscle cells. Since alpha 2-macroglobulin can bind platelet-derived growth factor (PDGF), and since highly acidic fetuin might bind the very basic PDGF molecule non-specifically, we measured the PDGF content of various fetuin preparations and found a good correlation between the PDGF content and mitogenic activity. Gel filtration experiments demonstrated that in Pedersen fetuin PDGF occurred both free, and in association with alpha 2-macroglobulin. We conclude that the principal mitogenic component for smooth muscle cells in crude fetuin preparations is PDGF, since purified bovine alpha 2-macroglobulin or fetuin do not appreciably affect growth of these cells. These results help to resolve a long-standing controversy regarding the nutrition of cultured cells. In addition, we suggest that before alpha 2-macroglobulin or "Embryonin" is accepted as a bona fide growth factor for a given cell type, the role of contamination with PDGF should be assessed.     

The B chain of PDGF alone is sufficient for mitogenesis.

The platelet-derived growth factor (PDGF) is a mitogen derived from human platelets consisting of two related polypeptides termed A and B chains. The entire B chain of PDGF is highly (96%) homologous to a portion of p28sis, the transforming protein of simian sarcoma virus. It has been suggested that p28sis exerts its transforming potential by mimicking the growth promoting activity of PDGF and stimulating the cell in an autocrine manner. We have directly examined the mitogenic potential of p28sis and the B chain homologous region by expressing these heterologous sequences in the yeast Saccharomyces cerevisiae. In our constructions, these proteins are encoded by portions of the v-sis gene. Expression and secretion from the yeast cell is achieved by using a yeast promoter and the alpha-factor pheromone secretory leader. The sis proteins thus expressed and secreted are immunoreactive with anti-PDGF antisera and are mitogenic for cultured fibroblasts. Furthermore, they mediate this mitogenic activity by specific binding to the PDGF cell surface receptor. Gel electrophoresis and cell binding analysis indicates that the mitogenic species is primarily a disulphide-bonded dimer. We are able to conclude that p28sis is a mitogen and that a polypeptide corresponding to the B chain alone is sufficient to account for the mitogenic activity attributed to PDGF.     

Cultural characteristics and fatty acid composition of propionibacteria

The cultural characteristics and cellular fatty acid composition of 40 strains representing 7 species of Propionibacterium and of 9 cultures of anaerobic corynebacteria were studied. The cultures were characterized by means of 23 separate cultural and biochemical tests. Cultures of the two genera differed consistently in only two reactions; the propionibacteria did not produce indole or liquefy gelatin, whereas the anaerobic corynebacteria were consistently positive with these tests. The fatty acids were extracted from whole cells and examined as methyl esters by gas-liquid chromatography. The most abundant acid in the seven Propionibacterium species was a C(15)-saturated branched-chain acid which was present in both the iso-and anteiso-form. Based on a comparison of the relative abundance of these isomers (i-C(15) and a-C(15)), the species were separated into two groups. P. freudenreichii and P. shermanii (group one) were similar and contained the a-C(15) isomer as the predominant acid. The i-C(15) isomer was the most abundant acid in the second group (P. arabinosum, P. jensenii, P. pentosaceum, P. thoenii, and P. zeae). The fatty acid profiles of the anaerobic corynebacteria were somewhat similar to those of the second group of propionibacteria, but were distinct from the profiles of P. freudenreichii and P. shermanii. The addition of branched-chain amino acids (l-leucine and l-isoleucine) to the growth medium increased the synthesis of the specific fatty acid(s) structurally related to the added amino acid.     

Wheat phosphoglycerate kinase: evidence for recombination between the genes for the chloroplastic and cytosolic enzymes.

We have isolated and sequenced cDNA clones containing the entire coding region of both the chloroplast and cytosolic versions of phosphoglycerate kinase from wheat. Comparison of these sequences reveals a higher than expected level of similarity between the nucleic acids and encoded proteins. Analysis of this data in relation to that for phosphoglycerate kinase sequences of mammals, prokaryotes and yeasts suggests that the wheat genes have recombined. This has resulted in the chloroplast and cytosolic kinases being more similar to each other than would be expected if the chloroplast enzyme had evolved directly from that of a prokaryotic progenitor.