COMPARISON OF DNA RENEWAL IN GERM-FREE AND CONVENTIONAL MICE USING [125I]IODODEOXYURIDINE AND [3H]THYMIDINE

Germ-free (GF) and conventional (CV) C3H mice received a single injection of 1 μCi [³H]thymidine and 3 μCi [¹²⁵I]iododeoxyuridine to provide simultaneous labeling of DNA with the two precursors. Thymus, spleen, mesenteric lymph nodes, bone marrow (femora), small intestine, colon and skin were examined for total organ activity and rate of DNA renewal 1–8 days after injection. Precursor incorporation, assayed on day 1, was lower in the thymus, mesenteric lymph nodes and femora (and, to a lesser extent, in the spleen and colon) of GF mice as compared to CV animals. The opposite was observed in the small intestine and skin, i.e. total organ activity was higher in GF animals. Differences in precursor incorporation were partly due to differences in organ weights between the two groups of mice. In comparison to CV animals, DNA renewal rates were diminished in the mesenteric lymph nodes, bone marrow, colon (following a 3-day plateau) and spleen of GF mice. Little, if any, difference was observed between the two groups with respect to the rate of DNA turnover in the thymus and skin. Radioactivity of the small intestine remained constant for 2 days. Thereafter intestinal activity in GF mice declined at an initial slow rate between days 2 and 5 followed by a rapid decrease between days 5 and 8. In CV mice the first phase of activity loss was short with the rapid decline in intestinal activity beginning on day 3. From the slopes of the regression lines, the percentage thymidine reutilization was estimated. Reutilization varied from 0 to 63% in the various organs examined, with the greatest difference between GF and CV mice occurring in the mesenteric lymph nodes.     

Post-translational heterogeneity of the human vitamin D-binding protein (group-specific component)

The vitamin D-binding protein in human serum (the group-specific component) is an alpha 2-globulin which is genetically polymorphic in all populations studied. Previous work (J. Svasti and B. H. Bowman (1978) J. Biol. Chem. 253, 5188-5194, and J. Svasti, A. Kurosky, A. Bennett, and B. H. Bowman (1979) Biochemistry 18, 1611-1617) has shown that the electrophoretic variations of the proteins controlled by two allelic genes, Gc1 and Gc2, are due to at least three amino acid substitutions between Gc1 and Gc2 (Svasti et al. (1979] and to heterogeneity in the Gc1 phenotype arising from carbohydrate dissimilarities. Gc1 migrates electrophoretically as two protein bands, while Gc2 migrates cathodally as a single band. This study demonstrates a post-translational glycosylation difference occurring in a single area of the Gc1 sequence which accounts for the heterogeneity observed previously. The glycosylation site, a threonine residue, appears to be in a sequence which differs between Gc1 and Gc2. The O-glycosidic bond, which is typical of mucins, is rare in plasma proteins. The cyanogen bromide fragment containing the galactosamine-containing carbohydrate in Gc1 was partially sequenced through 20 residues from the amino terminus. No detectable galactosamine could be found in the homologous cyanogen bromide fragment in Gc2. A new purification procedure for the vitamin D-binding protein in human plasma has been developed. Three chromatographic steps provide purified protein.     

Separation of peptides by reverse-phase high-performance liquid chromatography using propyl- and cyanopropylsilyl supports

The separation of peptides and proteins by reverse-phase high-performance liquid chromatography with cyanopropylsilyl and large-pore propylsilyl supports, together with aqueous trifluoroacetic acid/acetonitrile gradients, was studied. Operating parameters (trifluoroacetic acid concentration, flow rate, and gradient slope) were evaluated using different enzymatic digests of horse cytochrome c and bovine serum albumin. Peptides ranging in size from five amino acids to 68 kDa could be separated on the propylsilyl column in a single chromatographic run. The cyanopropylsilyl column is suitable as a supplement to the use of the large-pore column for medium size (5-20 amino acids) peptides. The chromatographic supports and conditions presented here offer a simple, sensitive, and rapid separation system for a wide size range of peptides and proteins. They extend the versatility of separation methodology for these molecules.     

Population Changes of Heterodera glycines and Soybean Yields Resulting from Soil Treatment with Alachlor,Fenamiphos, and Ethoprop

The population dynamics of Heterodera glycines as influenced by alachlor, fenamiphos, and ethoprop alone and in herbicide-nematicide combinations were studied in the field. Numbers of H. glycines juveniles and eggs were higher at midseason and harvest where nematicides were applied. Fenamiphos alone or in combination with alachlor provided better control of H. glycines and greater seed yields than treatments with ethoprop. Numbers of H. glycines eggs at harvest in 1980 were positively correlated with numbers of juveniles at planting in 1981 and negatively related to seed yield in 1981.     

Immunomorphological analysis of G2-chalone localization during the process of rat epidermis histogenesis

The appearance of G2-chalone in the cytoplasm of the intermediate cell layer and partly in the periderm of 17-day-old rat embryo epidermis has been demonstrated by the indirect method of Coons using a monospecific antiserum. G2-chalone was absent from the basal cell layer of 17--21-day-old embryos and of the newborn rats. It was found in all the epidermal layers in 2--5-day-old postnatal rats, while in 6--9-day-old animals it was primarily detected in the cytoplasm of spinous and basal cells. Thus the localization of epidermal G2-chalone typical for defined tissue becomes stabilized at the end of epidermis histogenesis.     

Factor 390 chromophores: phosphodiester between AMP or GMP and methanogen factor 420

Two chromophores with absorbance maxima at 390 nm (factors 390) have been isolated from oxidized cells of Methanobacterium thermoautotrophicum delta H. The isolation procedure included anion-exchange chromatography of the soluble cofactor pool followed by reverse-phase chromatography. The factor 390 species are novel derivatives of methanogen coenzyme factor 420 in which the 5-deazaflavin 8-hydroxy group is in a phosphodiester linkage to adenosine 5'-phosphate or guanosine 5'-phosphate. The structural assignments were based, in part, on the UV-visible and 1H NMR spectra. In addition, the results from amino acid analysis, phosphate determination, 31P NMR spectroscopy, and fast atom bombardment mass spectrometry were consistent with the proposed structures. Confirmation of the factor 390 structures was made following phosphodiesterase release of the nucleotide monophosphates from factor 420. The nucleotide monophosphates were identified as AMP and GMP by UV-visible spectra and based on elution position by using reverse-phase and anion-exchange high-performance liquid chromatography. The presence of AMP was further demonstrated by using adenylate-5'-phosphate kinase which induced a spectral shift during conversion of the sample to IMP. In addition, the presence of GMP was established by a specific enzymatic assay.     

The phospholipid-bound fatty acids of fowl and turkey spermatozoa

The composition of the phospholipid-bound fatty acids in the spermatozoa of the turkey, Meleagris gallopavo, and fowl, Gallus domesticus, was studied. The types of fatty acids were similar in the two birds. The ratio of polyunsaturated : saturated fatty acids was generally low but slightly higher in the turkey than in the fowl. The significance of the findings in relation to the origin of the semen collected in these gallinaceous birds and the greater difficulty of freezing turkey spermatozoa was discussed.     

The distribution of living benthic foraminifera on the continental slope and rise off southwest Norway

Surface sediment samples taken by ? corer from 45 stations on the Norwegian continental margin and in the Norway Basin have been investigated for their benthic foraminiferal content. Unlike previous studies, the living benthic foraminiferal fauna was differentiated from empty tests comprising the foraminiferal death assemblage. Factor analysis of both the living and dead faunal data reveals six living species assemblages and five corresponding dead assemblages. The additional living assemblage is characterized by the arenaceous speciesCribrostomoides subglobosum that dominates between 1400 and 2000 m water depth, but is rare in the dead faunal data.Trifarina angulosa and, to a lesser extent,Cibicides lobatulus characterize the shallowest foraminiferal assemblage from 200 to 600 m water depth, where it is associated with strong bottom currents and warm, saline Atlantic water of the North Atlantic Drift. On the slope between 600 and 1200 m water depth, theMelonis zaandami Species Assemblage dominates, particularly in areas characterized today by rapid sedimentation of terrigeneous material. Between 1000 and 1400 m depth, where the slope is covered by fine grained, organic-rich, terrigeneous mud, the living foraminiferal assemblage is characterized byCassidulina teretis andPullenia bulloides. Below 1400 m, three foraminiferal assemblages are found:C. subglobosum is found from 1400 to 2000 m,Cibicidoides wuellerstorfi andEpistominella exigua predominantly live from 2000 to 3000 m water depth, and below 3000 m,Oridorsalis umbonatus andTriloculina frigida dominate the fauna.All of theElphidium excavatum tests found in this study and theCassidulina reniforme tests found above 500 m water depth were found to be reworked.Analysis of the sediment grain-size distribution and the organic carbon content in surface samples from the deepest stations suggest that the abundance ofC. wuellerstorfi andE. exigua is positively correlated to relatively coarse (caused by planktic foraminifera) and organic-rich sediments, whereas high frequencies ofO. umbonatus andT. frigida coincide with low organic carbon content. We suggest thatC. wuellerstorfi is adapted to deep-sea environments with relatively high food supply, tolerating relatively low interstitial water oxygen content, whereasO. umbonatus may tolerate lower food supply prefering well-oxygenated interstitial waters.     

A subclass of proteins and sulfated macromolecules secreted by AtT-20 (mouse pituitary tumor) cells is sorted with adrenocorticotropin into dense secretory granules

The AtT-20 cell, a mouse pituitary tumor line that secretes adrenocorticotropin and beta-endorphin, sorts the proteins it externalizes into two exocytotic pathways. Cells that are labeled with [35S]methionine or [35S]sulfate can be shown to transport three acidic polypeptides (65,000, 60,000, and 37,000 mol wt) and at least two sulfated macromolecules into storage secretory granules. When the cells are stimulated by the secretagogue 8-bromo-cAMP, these polypeptides are coordinately secreted with mature adrenocorticotropin into the culture medium. In contrast, a completely different set of secreted polypeptides and sulfated macromolecules does not enter a storage form and is transported to the cell surface more rapidly. Their secretion from the cells is constitutive and does not require the presence of secretagogues. These molecules, like a viral membrane glycoprotein described previously (Gumbiner, B., and R. B. Kelly, 1982, Cell, 28:51-59) are not found in isolated secretory granules and therefore must reach the cell surface in a different exocytotic vesicle. The segregation of a subclass of secretory macromolecules into the secretory granules, despite the existence of another potential secretory pathway, suggests that these molecules have specific functions related to regulated hormone secretion or storage. Presumably all of the proteins secreted by the regulated secretory granule pathway share some common property that targets them to the secretory granule.