Lamellar bodies of cultured human fetal lung: content of surfactant protein A (SP-A), surface film formation and structural transformation in vitro

Lamellar bodies were isolated from dexamethasone and T3-treated explant cultures of human fetal lung, using sucrose density-gradient centrifugation. We examined their content of surfactant apoprotein A (SP-A), and their ability to form surface films and to undergo structural transformation in vitro. SP-A measured by ELISA composed less than 2% of total protein within lamellar bodies; this represented, as a minimum estimate, a 2-12-fold enrichment over homogenate. One- and two-dimensional gel electrophoresis also suggested that SP-A was a minor protein component of lamellar bodies. Adsorption of lamellar bodies to an air/water interface was moderately rapid, but accelerated dramatically upon addition of exogenous SP-A in ratios of 1:2-16 (SP-A:phospholipid, w/w). Similar adsorption patterns were seen for lamellar bodies from fresh adult rat and rabbit lung. Lamellar bodies incubated under conditions that promote formation of tubular myelin underwent structural rearrangement only in the presence of exogenous SP-A, with extensive formation of multilamellate whorls of lipid bilayers (but no classical tubular myelin lattices). We conclude that lamellar bodies are enriched in SP-A, but have insufficient content of SP-A for structural transformation to tubular myelin and rapid surface film formation in vitro.     

A rapid method for the analysis of N tau-methylhistidine in human urine

A novel, automated method is described for the determination of N^τ-methylhistidine in human urine. The method uses a modification (H. Nakamura and J. J. Pisano (1976) Arch. Biochem. Biophys.177, 334–335) of the reaction of fluorescamine with amines, which renders it specific for certain imidazoles. Interference due to histidine and histamine is selectively removed by prior reaction with aldehydes. The fluorescence yield for N^τ-methylhistamine is 280, 440, 50, and 1.7, respectively. The concentrations of N^τ-methylhistidine in human urine as determined by this technique correlate well (r = 0.99) with those determined by ion-exchange chromatography. Furthermore, the technique is rapid (6–7 samples/h net throughout), is reproducible (coefficient of variation 1.8%), requires no prior treatment of the sample, and is implemented with widely available equipment.     

Structure of tendon organs in fishes of the genus Polymixia

Summary Modified branchiostegal rays 1 through 3 support the proximal end of the paired hyoid barbels in the beardfish (Beryciformes: Polymixiidae). The polymixiid barbel is unusual in that it has an unique intrinsic muscular system. Using silver impregnation and electron microscopic techniques, unencapsulated, free nerve endings were located within the tendon of the third modified branchiostegal ray. Branchiostegal rays 1 and 2 do not have any free nerve endings associated with their tendons, however. It is suggested that the free nerve endings are proprioceptors acting as stretch-sensitive mechanoreceptors, and that branchiostegal ray 3 acts as part of a sensory apparatus for monitoring the positional state of the barbel. Branchiostegal rays 1 and 2 merely provide support for the barbel.Abbreviations used in Figures BA barbel - br r 3 branchiostegal ray 3 - IM intermandibularis - IOP interoperculum - LIM interoperculomandibular ligament - MD mandible - MX maxilla - OP operculum - PM premaxilla - POP preoperculum - SOP suboperculum     

Transposition of the kanamycin-resistance transposon Tn903

Summary The insertion of the kanamycin-resistance transposon, Tn903, into the Escherichia coli chromosome was studied. Tn903 is similar in structure to the well known transposons Tn5 and Tn10 in that it has a unique central sequence flanked by inverted repeat sequences extending more than a thousand base pairs. However, the central region of Tn903 has enough single-frame coding capacity only for the drug modifying enzyme, whereas Tn5 and Tn10 carry multigenic unique sequences. In this paper we demonstrate that two different classes of insertion event occur: (1) the first class is a complex event in which all or part of the genome of the bacteriophage lambda vector is co-inserted near the purE locus on the E. coli chromosome (11.7 min); (2) the second class appears to be a simple transposition event in which the transposon alone is inserted at relatively nonspecific sites in the chromosome, as has been described for Tn5 and Tn10. Furthermore both classes show dependency on homology-requiring recombination systems. We suggest that Tn903 transposes infrequently because it must utilize a recA-controlled host function, whereas Tn5 and Tn10 are recA-independent and encode similar but more active functions on the transposon DNA.     

Lectin induction of lymphokines in cultured murine leukocytes

Antigens induce sensitized lymphocytes to undergo mitosis and to secrete soluble products, termed lymphokines, which modulate the immune response. Plant lectins are known to act as polyclonal lymphocyte mitogens and, in some cases, stimulate lymphocytes to produce lymphokines. In an effort to explore the relationship of specific cell surface glycoconjugates to the induction of mitosis and the production of lymphokine activities we have examined the ability of the mitogenic lectins, concanavalin A and Wistaria floribunda mitogen, and the nonmitogenic hemagglutinin from Wistaria floribunda seeds to stimulate the production of macrophage migration inhibition factor (MIF), macrophage chemotactic factor (CF), and lymphotoxin (LT). Concanavalin A causes lymphocytes to produce MIF and LT but no detectable CF activities. W. floribunda mitogen induces lymphocytes to produce soluble substances which exhibit all three lymphokine activities. The nonmitogenic W. floribunda agglutinin causes lymphocytes to produce MIF and CF but no observable LT activity. Within the sensitivity of the assays employed, the results indicate that mitogenesis is not a corequisite of the expression of either macrophage migration inhibition factor or lymphocyte-derived chemotactic factor but it may be associated with the induction of lymphotoxin. It is also apparent that the expression of each lymphokine activity is independent of the expression of the other lymphokines studied.     

Systematic studies of Pinus balfouriana based on volatile terpenoids from wood and needles and on seed morphology

Essential oil from wood and from needles of Pinus balfouriana, growing in six geographically well-separated locations in California, was analysed by GLC. Several monoterpenoid components, in particular -pinene from needles, were found to be usable for distinguishing between trees from the northern and southern parts of the geographic range. Similarity coefficients were calculated and dendrograms constructed. These demonstrated the distinct separation of the northern from the southern populations, and thus substantiated the proposal by Mastrogiuseppe to regard the southern populations as a subspecies. While the northern populations exhibited a tendency to produce larger seeds with longer wings, the difference was of only moderate diagnostic value.     

Evaluation of three E-rosette assays in melanoma patients

Summary The percentage of E-rosette-forming cells (E-RFC) was determined repeatedly in the peripheral blood of 43 patients with malignant melanoma. Three methods of E-rosette assaying were used: total E-rosette test, active E-rosette test, and 29° C E-rosette assay.Depressed E-RFC values were found in fewer assays than normal values. The mean values of E-RFC and the proportion of depressed E-RFC in patients in stage I did not differ from the values in healthy control persons whatever method of assay was used.The frequency of depressed E-RFC values was higher in advanced stages. Significant differences were demonstrated between stage I and stages II and III combined in the values for total and for active E-rosettes (P<0.01).The active E-rosette test and the 29° C E-rosette assay gave no more positive results (i.e., depressed E-RFC values) than the total E-rosette test in melanoma patients.     

Adjacent repeating units of xenopus laevis 5S DNA can be heterogeneous in length

The distribution of length heterogeneity in adjacent repeating units of X. laevis 5S DNA has been examined by “cloning” 5S DNA in bacteria. Fragments of 5S DNA produced by partial digestion with Hind III and containing 1, 4, and 5 repeating units have been inserted at the single Hind III site of the tetracycline-resistance plasmid, pSC101, and the hybrid plasmids cloned in E. coli. Adjacent 5S DNA repeats in the cloned multi-repeat fragments can differ in length. This finding rules out some mechanisms which have been proposed to account for the parallel evolution of tandem repeated DNAs. The results are consistent with an unequal crossing-over mechanism and place some constraints on the molecular processes in this recombinatory event.     

Inactivation of phosphoenolypyruvate carboxykinase (GTP) by liver extracts.

1. The inactivation of phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) in liver extracts was catalysed by the microsomal fraction, and led to the enzyme becoming bound to the microsomal membranes. 2. Inactivation by microsomal fraction, typsin or heating at 48degreesC was accelerated by L-cystine, D-cystine and oxidized glutathione and decreased by dithiothreitol. 3. MnC1(2) and CoC1(2) protected the enzyme from inactivation by heat or microsomal fraction, but did not affect the inactivation caused by trypsin. 4. Several proteinase inhibitors had no effect on the microsomal inactivation reaction, suggesting that proteolysis was not involved. 5. It is argued that the initial step in the degradation of phosphoenolpyruvate carboxykinase (GTP) is an inactivation reaction, perhaps involving oxidized thiol compounds.     

Regulation of phosphoenolpyruvate carboxykinase (GTP) in adipose tissue in vivo by glucocorticoids and insulin.

1. The regulation of the synthesis of phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) in epididymal adipose tissue, liver and kidney in vivo was studied immunochemically. 2. Phosphoenolpyruvate carboxykinase (GTP) synthesis in adipose tissue is increased by starvation, diabetes and noradrenaline, and decreased by re-feeding and insulin. These changes were also seen in adrenalectomized rats and are qualitatively similar to those observed for the liver enzyme. This indicates the involvement of cyclic AMP as an inducer and insulin as a de-inducer in the regulation of phosphoenolpyruvate carboxykinase (GTP) in both tissues. (Induction and de-induction are defined as selective increase and decrease respectively in the rate of enzyme synthesis, regardless of the mechanism involved.)3. Adrenalectomy had little effect on phosphoenolpyruvate carboxykinase (GTP) synthesis in liver and kidney, but increased the synthesis rate of the adipose-tissue enzyme. Starvation and adrenalectomy had additive effects in increasing the synthesis rate of adipose-tissue phosphoenolpyruvate carboxykinase (GTP). In adrenalectomized diabetic rats glucocorticoids increased phosphoenolpyruvate carboxykinase (GTP) synthesis in liver and kidney while decreasing enzyme synthesis in adipose tissue. De-induction of adipose tissue phosphoenolpyruvate carboxykinase (GTP) is therefore regulated independently by glucocorticoids and insulin. 4. Although liver, kidney and adipose-tissue phosphoenolpyruvate carboxykinases (GTP) are seemingly identical, there is an apparent tissue-specific differentiation in regulatory systems for the enzyme.