Repair mechanisms involved in muscle regeneration following partial excision of the rat gastrocnemius muscle

The sequential cytological events of the regeneration process, after partial excision of the gastrocnemius muscle in the rat, were followed by light and electron microscopy. During the first 2 days after injury leukocytes and macrophages infiltrate into the traumatized area. Myogenic regeneration is then characterized by mainly two repair mechanisms. Mononucleated cells, that populate the excised area, most probably fuse together to give rise to newly formed multinucleated myotubes that further develop to striated myofibers. Another mechanism involves the repair of injured muscle fibers by the possible fusion of mononucleated cells with their necrotic cut ends. Consequently, by addition of nuclei and new muscular material, sarcoplasmic outgrowths from the injured fibers are formed. It is concluded that mainly two repair mechanisms are involved in the regeneration process following partial excision of a muscle: addition of new muscle fibers in a process similar to that of embryonic myogenesis and also meristic growth from the injured fibers.     

Expression patterns of the T antigen and the cryptic T antigen in rat fetuses: detection with the lectin amaranthin

The lectin amaranthin, purified from the seeds of Amaranthus caudatus, has been shown to react specifically with the Gal beta 1,3GalNAc-alpha and the NeuAc alpha 2,3Gal beta 1,3GalNAc-alpha sequence which represent the T antigen and the cryptic T antigen, respectively. We report here the development of labeling techniques that apply amaranthin to stain paraffin sections from rat fetuses. Amaranthin staining was inhibited by pre-incubation of lectin-gold complexes with 10 mM Gal beta 1,3GalNAc-alpha-O-benzyl (synthetic T antigen) or 10 mM Gal beta 1,3GalNAc-alpha-O-aminophenylethyl-human serum albumin (T antigen neoglycoprotein), asialoglycophorin, asialofetuin, and asialomucin. The beta-elimination reaction also abolished the lectin staining demonstrating specificity for O-glycosidically linked structures. A comparison with monoclonal anti-T antigen antibody immunostaining demonstrated that amaranthin detects the T antigen and its cryptic form in tissue sections. Application of the galactose oxidase-Schiff sequence abolished amaranthin (and anti-T antibody) binding to the T antigen but not to its cryptic form, and therefore permitted their differentiation in tissue sections. Histochemical evidence was obtained indicating that amaranthin is a more specific anti-T reagent than peanut lectin. Data are presented that show the differential expression of the T antigen and the cryptic T antigen in organs and cells of rat fetuses late in gestation. Therefore, amaranthin can be used for histochemical detection of the T antigen and the cryptic T antigen, and facilitates discrimination between them.     

New series of lipoxins isolated from human eosinophils

Granulocytes from human eosinophilic donors were incubated with arachidonic acid or 15-hydroxyeicosatetraenoic acid (15-HETE) and stimulated with the ionophore A23187. The eicosanoids were extracted with reversed-phase cartridges and subjected to RP-HPLC analysis. When extracts from eosinophil-enriched populations were analysed and compared with extracts from human neutrophils, three additional peaks were detected which coeluted with 15-hydroxy-delta 13-trans-15H derivatives of leukotriene C4, D4 and E4 in different HPLC systems. The recorded absorbance spectra of the eluted compounds and the standards were identical and showed a maximum at 307 nm which is characteristic for a conjugated tetraene system with a bathochromic shift by the sulfur moiety in alpha-position to the tetraene system. The compound which coeluted with the 15-hydroxy-LTC4 standard was treated with gamma-glutamyltransferase and converted to the corresponding leukotriene D4 derivative. The results indicate that interaction between the 5- and 15-lipoxygenase pathways leads to the formation of a new series of arachidonic acid metabolites in human eosinophils. Since the biosynthetic route is similar to that of lipoxin A4 and lipoxin B4, we suggest the trivial names lipoxin C4, D4 and E4.     

Gene assignment by means of somatic cell hybrids: a new method for the analysis of data

Summary A statistical approach to the interpretation of data from gene assignment with somatic cell hybrids is presented. The observed data are analysed under a variety of hypotheses. The fit to the hypotheses is compared by means of the likelihood obtained under a given hypothesis. Two of these hypotheses are related to fundamental questions: is a gene responsible for the enzyme observation and if so, is that gene located on a specific chromosome or could it change its position and be sometimes on chromosome j and, in another hybrid line, on chromosome k? The other hypotheses concern the assignment of the gene to just one of the chromosomes.To improve the traditional data analysis approach we considered additional information: the uncertainties and possible errors of laboratory methods in all our calculations and the length of the donor chromosomes in connection with one specific hypothesis.This method allows us to account for the reliability of the investigation methods and the nature of the hybrid lines involved. Data can be evaluated at different error probabilities within a realistic range in order to compare and discuss results.     

Characterization of an endogenous substrate related to insulin and insulin-like growth factor-I receptors in lizard brain

Lizard insulin receptors are evolutionarily highly conserved. Wheat germ agglutinin-purified brain membranes demonstrate the presence of an endogenous substrate (pp 105) for both the insulin and insulin-like growth factor-I receptors. Both insulin and I-insulin-like growth factor-I stimulate the phosphorylation of this endogenous substrate in a dose-dependent manner. Following insulin-stimulated autophosphorylation of the beta subunit, there is a lag period of about 5 min prior to observable phosphorylation of the endogenous substrate. Phosphoamino acid analysis of both the beta subunit as well as pp 105 reveal primarily phosphotyrosine in both the basal as well as the stimulated state.