Explant organ culture of hamster alimentary tract epithelium

Adult Syrian Golden hamster alimentary tract maintained as explants in organ culture was studied using the model system for hamster pancreas described by Resau et al. (1983a). Explants of esophagus, stomach, duodenum and colon were maintained in organ culture on Gelfoam® sponge rafts in a high-oxygen atmosphere with serum-supplemented CMRL-1066 medium. All of the tissues were observed to show evidence of sublethal acute cell injury during the first several days of culture. Subsequently, the epithelial tissues recovered from this injury, repopulated the denuded areas of the explants and replicated within the sponge matrix. Explants were maintained in a differentiated state for 30+ days and sampled for morphology to examine the process of cell injury, repair, differentiation and replication which occurs in mucosal epithelia. The percentage of basement membrane covered by epithelia in the explants from various tissues was compared to the level of LDH in the media to reveal the relationship between viability determined by biochemical and by morphological methods. Restitution of the mucosal surface occurred in all of the explants. We conclude that adequate populations of replicating cells are maintained within the epithelium of the hamster alimentary tract tissues in vitro so that restitution can occur through migration and subsequent differentiation of the epithelial cells within the mucosa of the explants.Abbreviations 4F-1G 4% formaldehyde 1% glutaraldehyde fixative - LDH lactate dehydrogenase - OsO₄ osmium tetroxide - PAS/PLH periodic acid, periodic acid Shiff lead hematoxylin stain     

Lamprey fibrinopeptide B is A glycopeptide

One of the peptides released from lamprey fibrinogen during its transformation into fibrin has been found to contain covalently bound carbohydrate. The peptide, which also contains tyrosine O-sulfate, corresponds to the mammalian fibrinopeptide B and is the amino-terminal segment of the lamprey fibrinogen β-chain. As noted previously, this peptide is the only one released when lamprey fibrinogen is coltted by mammalian thrombin. Of the more than fifty sets of fibrinopeptides characterized from various species, this is the first one found to contain carbohydrate.     

The Clara cell and pulmonary surfactant: A study using selective chemical ablation

Administration of 3-hydroxymethylfuran-N-ethylcarbamate (HFC) to female hamsters via the jugular vein under pentobarbitone anaesthetic at 20 mg per kg body weight produced pronounced necrosis of the Clara cells without apparent morphological effect on other cell types as judged by transmission electron microscope examination. The surfactant material recoverable by minimal lavage followed by purification by sucrose gradient ultracentrifugation increased, reaching a maximum around 48 h after treatment. At this time static pressure/volume measurements on isolated lungs indicated an increase in airway surface compliance. Lavageable surfactant phospholipid composition was examined by ³¹P nuclear magnetic resonance (n.m.r.). The distribution of phospholipids between the various classes was unchanged by HFC treatment. No change in the total lung surfactant pool size was seen. These results are discussed in relation to the possible roles of the Clara cell in influencing airway surfactant levels.     

Variations in mitochondrial DNA concentration as a function of different growth conditions in yeast

The relative concentration of mitochondrial DNA in the yeast, $\text{Saccharomyces}\text{cerevisiae}$ has been examined under a variety of different growth conditions by means of an isotope dilution procedure which is shown to yield accurate estimates of mitochondrial DNA content in small samples of this yeast. Under a derepression scheme in which only limited cell proliferation occurs, mitochondrial DNA exhibited nearly a doubling in relative amount. The concentration of mitochondrial DNA was also observed to fluctuate depending upon the strain, growth phase and carbon source included in the growth media. Our results indicate that the relative proportion of mitochondrial DNA does indeed vary according to a variety of different conditions that the cells are subjected to.     

Amino acid sequence studies on the alpha chain of human fibrinogen. Exact location of cross-linking acceptor sites

Human fibrinogen was clotted under conditions that promote latent factor XIII activity and in the presence of a radioactive substitute cross-linking donor ([14C]glycine ethyl ester). The labeled fibrin was reduced and alkylated in the presence of 6 M guanidinium chloride. After dialysis and freeze-drying, the preparation was separated into its constituent polypeptide subunits by chromatography on (carboxymethyl)cellulose in the presence of 8 M urea. Under the incorporation conditions used, the radioactivity was limited to gamma chains (one donor molecule/chain) and alpha chains (two donor molecules/chain). The labeled alpha chains were digested with cyanogen bromide and fractionated on Sephadex G-50. All the radioactivity was found in a fragment previously designated H alpha CNI, the largest of the cyanogen bromide fragments in the alpha chain. The fragment was further fragmented by digestion with plasmin, trypsin, chymotrypsin, and/or staphylococcal protease. The incorporated radioactivity was found to reside in equal amounts at two different sites located 38 residues apart. These were determined to be positions 88 and 126 in H alpha CNI, which correspond to glutamine-328 and glutamine-366 in the alpha chain.     

Hypoxia increases erythropoiesis and decreases thrombocytopoiesis in mice: a comparison of two mouse strains

Several previous studies have shown that hypoxia increases erythropoiesis and decreases thrombocytopoiesis in mice. It has been postulated that the thrombocytopenia is caused by stem cell competition between the erythrocytic and megakaryocytic cell lines. In the present work, we compared the effects of severe hypoxia (5.5-6.0% O2) in both male and female C3H and BALB/c mice by measuring their abilities to produce red blood cells and platelets. All mice had significant increases in packed cell volumes and marked decreases in platelet production after hypoxia; however, there were significant differences in the degree of stimulation in the two mouse strains. After 14 days of hypoxia, the percentage of 35S incorporation into platelets, total circulating platelet counts and total circulating platelet masses were lower in C3H mice than in BALB/c mice, but platelet sizes were larger. Also, hypoxia caused greater changes in male mice than in female mice, with male C3H mice showing the greatest increase in packed cell volumes and the lowest platelet counts of all mice tested. The least responses were observed in female BALB/c mice. BALB/c mice had higher P50 (right-shifted O2 dissociation curves) and lower erythrocyte 2,3-diphosphoglycerate values than C3H mice, indicating a lower hemoglobin O2 affinity for BALB/c mice. The results indicate that the effects of hypoxia are not direct upon platelet production, but that the thrombocytopenia is a result of stimulation of erythropoiesis. These data support the stem cell competition hypothesis and illustrate that the degree of the inverse relationship between red blood cells and platelet production of hypoxic mice is dependent, to a large degree, upon the sex and strain of mice that are used.