Macromolecular synthesis in organ cultures of neonatal rat lung

Organ cultures of newborn rat lungs synthesize and accumulate DNA, RNA, collagen and noncollagenous proteins almost at a linear rate for at least 5 days. During this period the synthesis of collagen consistently exceeds the synthesis of noncollagenous proteins in a pattern similar to neonatal lung growth in vivo. Although some morphological characteristics of lung architecture are distorted after culture, fundamental structural similarities to lungs growing in intact animals are retained. When these cultures are maintained in atmospheres rich in oxygen, increased collagen synthesis is observed, a response similar to that of lungs in intact animals exposed to high oxygen concentrations in vivo. Our studies suggest that lung organ cultures may be a suitable system for investigating the biochemical aspects of lung tissue-environmental interaction. These studies were supported in parts by NIH Grant HL-19668, a contract (68-03-2005) from the U.S. Environmental Protection Agency, and grants from the California Lung Association.     

Palaeocommunity Dynamics and Behavioral Analysis of Conichnus: Bhuj Formation (Lower Cretaceous),Kachchh-India

A palaeocommunity of large Conichnus conicus, a conical, cone-in-cone shaped burrow, created by sea anemones, occurs in medium-grained, crossbedded, well-sorted sandstone in the middle part of the Cretaceous Guneri Member of the Bhuj Formation in India. The trace fossil Conichnus is considered to be a common element of the Skolithos ichnofacies and is interpreted to reflect equilibrium movement in response to substrate aggradation. In the present study, three different varieties of Conichnus conicus are distinguished based on morphology and internal fabric. Community dynamics and burrowing behavior are revealed by inter-burrow relationships, burrow initiation levels and sedimentology. Three types of behavior are envisaged: retrusive equilibrium, protrusive equilibrium response, and escape behavior. Palaeocommunity dynamics show that the tracemakers consisted of only adult organisms that initiated burrows during neap tides and are adapted to feed effectively during weak flow conditions. The occurrence of Conichnus palaeocommunity in the Guneri Member indicates the tidal conditions in a fully marine setting. Results presented herein may aid in the understanding of palaeocommunity dynamics in other shallow marine sequences.     

Aldimine to ketoamine isomerization (Amadori rearrangement) potential at the individual nonenzymic glycation sites of hemoglobin a: Preferential inhibition of glycation by nucleophiles at sites of low isomerization potential

The relative roles of the two structural aspects of nonenzymic glycation sites of hemoglobin A, namely the ease with which the amino groups could form the aldimine adducts and the propensity of the microenvironments of the respective aldimines to facilitate the Amadori rearrangement, in dictating the site selectivity of nonenzymic glycation with aldotriose has been investigated. The chemical reactivity of the amino groups of hemoglobin A forin vitro reductive glycation with aldotriose is distinct from that in the nonreductive mode. The reactivity of amino groups of hemoglobin A toward reductive glycation (i.e., propensity for aldimine formation) decreases in the order Val-1(), Val-1(), Lys-66(), Lys-61(), and Lys-16(). The overall reactivity of hemoglobin A toward nonreductive glycation decreased in the order Lys-16(), Val-1(), Lys-66(), Lys-82(), Lys-61(), and Val-1(). Since the aldimine is the common intermediate for both the reductive and nonreductive modification, the differential selectivity of protein for the two modes of glycation is clearly a reflection of the propensity of the microenvironments of nonenzymic glycation sites to facilitate the isomerization reaction (i.e., Amadori rearrangement). A semiquantitative estimate of this propensity of the microenvironment of the nonenzymic glycation sites has been obtained by comparing the nonreductive (nonenzymic) and reductive modification at individual glycation sites. The microenvironment of Lys-16() is very efficient in facilitating the rearrangement and the relative efficiency decreases in the order Lys-16(), Lys-82(), Lys-66(), Lys-61(), Val-1(), and Val-1(). The propensity of the microenvironment of Lys-16() to facilitate the Amadori rearrangement of the aldimine is about three orders of magnitude higher than that of Val-1() and is about 50 times higher than that of Val-1(). The extent of nonenzymic glycation at the individual sites is modulated by various factors, such as thepH, concentration of aldotriose, and the concentration of the protein. The nucleophiles—such as tris, glycine ethyl ester, and amino guanidine—inhibit the glycation by trapping the aldotriose. The nonenzymic glycation inhibitory power of nucleophile is directly related to its propensity to form aldimine. Thus, the extent of inhibition of nonenzymic glycation at a given site by a nucleophile directly reflects the relative role ofpK _a of the site in dictating the glycation at that site. The nonenzymic glycation of an amino group of a protein is an additive/synergestic consequence of the propensity of the site to form aldimine adducts on one hand, and the propensity of its microenvironment to facilitate the isomerization of the aldimines to ketoamines on the other. The isomerization potential of microenvironment plays the dominant role in dictating the site specificity of the nonenzymic glycation of proteins.     

Cellular retinoic acid-binding protein and its relationship to the biological activity of four synthetic retinoids in hamster tracheal organ culture

Summary The mechanism of action of retinoid in reversing keratinization in hamster trachea is yet unknown. The purpose of this study was to determine if cellular retinoic acid binding protein (CRABP) is present in tracheal epithelium following incubation in serum-free, vitamin A-deficient culture medium for 10 days, and if the effectiveness of a retinoid in reversing keratinization in organ culture is correlated with its ability to compete for CRABP sites. The cytosol prepared from tracheal cultures contained CRABP at a concentration of 2.61 pmoles per mg protein. Of the four retinoids with carboxyl end group selected for the study, two of the biological active retinoids competed for the CRABP sites. However, no correlation was observed between the biological activity of the inactive retinoids and their ability to associate with the CRABP sites. These results indicate that even though the action of retinoid may be mediated by retinoid binding protein, it cannot be used as a sole predicator of retinoid response in hamster trachea. This investigation was supported by Contract N01-CP-31012 and U. S. P. H. Grants CA30512 and CA32428, which were awarded by the Division of Cancer Etiology, National Cancer Institute, DHHS. Editor's Statement Tracheal organ cultures provide a useful model for the study of epithelial differentiation and carcinogenesis. Much attention has been given to the action of retinoids in this process. Mehta et al. demonstrate a lack of correlation between biological activity and specific cytosolic binding of members of this class of compounds, pointing out the need for a more complete biochemical understanding of the mechanism of action and active forms of retinoids in this and other systems in vivo and in vitro. David W. Barnes     

On the carbohydrate—protein linkage group in vitreous humor hyaluronate

Fractional molar ratios of serine, threonine and aspartic acid to neutral sugars in the purified bovine vitreous humor hyaluronate, and a 4–5-fold increase in the percentage of these amino acids and the absence of sugar alditols in hyaluronate reduced with NaBH₄---PdCl₂ after alkali treatment indicated the absence of a carbohydrate—protein linkage. Gel filtration behavior, a decrease in intrinsic viscosity of reduced hyaluronate to about one-half and a significant decrease in its specific rotation suggested that the two antiparallel chains of the hyaluronate double helix may come apart upo reduction. The vitreous humor hyaluronate contained 109.2 ppm of “bound” silicon. It is suggested that the bound silicon may bridge the two antiparallel chains through the neutral sugars and/or through the hydroxyl group of the uronic acid moiety.     

Developmental changes in the secondary xylem ofAcer rubrum induced by various auxins and 2,3,5-tri-iodobenzoic acid

Summary TIBA has been applied laterally to actively growing stems of uprightAcer rubrum seedlings. The frequency of initiation of tracheary elements is reduced and a complete ring of tension wood is developed in the stem locally below the TIBA application site. Rings of tension wood were never formed above the TIBA treatment site. In regard to anatomy, lignin distribution and peroxidase activity, the tension wood fibers formed as a result of TIBA treatment are identical to those which can be induced by bending.In the region of the stem above the site of TIBA application there is a particularly strong alteration in the development of tracheary elements.Application of IAA, NAA, or 2,4-D to the TIBA treatment site suppresses the capacity of TIBA to induce the development of tension wood and at the same time generally increases the frequency of initiation of tracheary elements.The effect of auxin alone on theAcer rubrum system has been studied. The secondary xylem formed during treatment with auxins (especially 2,4-D and NAA) at the stated concentrations is generally characterized by large groups of tracheary elements with a conspicuous angular outline in transverse section.The evidence suggests that auxins are involved in the regulatory systems which bring about the orderly development of the secondary xylem in arborescent angiosperms.This material was included in a doctoral thesis submitted by P. R.Morey to the graduate school of Yale University, New Haven.     

Dry-matter production and recovery of fertilizer nitrogen by rice as affected by nitrification retarders ‘N-Serve’ and ‘AM’

Summary In a pot-culture experiment simulating semi low-land rice field conditions 5 to 11 per cent increase in dry matter yield and 27 to 43 per cent increase in recovery of applied N was obtained by the use of N-Serve and AM nitrification retarders.Although the term frequently used is 'nitrification inhibitors, the term nitrification retarders is proposed since under field conditions these chemicals only partially control the nitrification.Trade name of The Dow Chemical Company, Midland, Michigan, U.S.A. for 2-chloro-6-(trichloromethyl) pyridine.Trade name of Toyo Koatsu Industries, Inc., Tokyo, Japan for 2-amino-4chloro-6methyl pirimidine.     

Action of Endo-(1 → 6)-α-d-glucanases on the soluble dextrans produced by three extracellular α-d-glucosyltransferases of Streptococcus sobrinus

Four different α-d-glucosyltransferases (GTF) have been obtained from culture filtrates of Streptococcus sobrinus strains grown in the chemostat at pH 6·5 in complex medium supplemented with Tween 80. Three of the enzymes, GTF-S1, GTF-S3 and GTF-S4, converted sucrose into soluble glucans. Their limit of hydrolysis with endodextranase, the proportion of linear to branched oligosaccharides among the end products of enzymic degradation, and methylation analysis, all supported the view that the glucans were dextrans. The S1-dextrans were highly branched (32% of α-(1 → 3)-branch points), S3-dextrans were linear, and the branching of S4-dextrans was intermediate in value (9%). The enzymes that catalyze the synthesis of three such diverse dextrans were thus proved to be three different GTF, each with a characteristic specificity. Conditions of growth in the chemostat could be varied to provide maximum yields of either GTF-S1, -S3 or -S4.