The human fibroblast growth factor receptor genes: a common structural arrangement underlies the mechanisms for generating receptor forms that differ in their third immunoglobulin domain.

To determine the mechanisms by which multiple forms of fibroblast growth factor (FGF) receptors are generated, we have mapped the arrangement of exons and introns in the human FGF receptor 1 (FGFR 1) gene (flg). We found three alternative exons encoding a portion of the third immunoglobulin (Ig)-like domain of the receptor. One of these alternatives encodes a sequence that is part of a secreted form of FGFR 1. The other two encode sequences that are likely part of transmembrane forms of FGFR 1. One of these forms has not been previously reported in published cDNAs. Also, we have determined the structural organization of a portion of the human FGFR 2 gene (bek) and found a similar arrangement of alternative exons for the third Ig-like domain. The arrangement of these genes suggests that there are conserved mechanisms governing the expression of secreted FGF receptors as well as the expression of at least two distinct membrane-spanning forms of the FGF receptors. The diverse forms appear to be generated by alternative splicing of mRNA and selective use of polyadenylation signals.     

Morphometry of feline neutrophil granule genesis

During neutrophil granule genesis, the formation of primary granules is generally thought to be limited to the promyelocyte stage; whereas synthesis of secondary granules is thought to occur only at the myelocyte stage. This hypothesis was tested morphometrically in feline neutrophils that are known to contain both granule types. Marrow specimens obtained from six cats were stained with peroxidase for identification of neutrophil primary granules and counterstained with periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) for identification of secondary granules. By regression analysis using arithmetic models, numbers of cytoplasmic granules in 311 cells were correlated with the degree of nuclear chromatin condensation, which was shown to be an adequate parameter for cell maturation. Promyelocytes and myelocytes had similar mean numbers of peroxidase-positive granules per unit area. A significant increase (p less than or equal to 0.0001) in the numbers of peroxidase-positive granules was noted between the metamyelocyte and the mature neutrophil stage, despite the lack of peroxidase activity in endoplasmic reticulum and Golgi lamellae. By contrast, a significant increase of peroxidase-negative granules between the metamyelocyte and the mature neutrophil stage was not clearly established with these methods. The increase in peroxidase-positive granules may indicate continued production of peroxidase-containing granules and/or redistribution of peroxidase among lysosomal organelles in late feline neutrophils.     

Two-dimensional 1H-NMR studies of the solution structure of plasminogen kringle 4

Native kringle 4 from human plasminogen has been studied by two-dimensional 1H-NMR methods in order to obtain new structural information about the kringle fold. Two-dimensional scalar correlated spectroscopy (COSY), two-dimensional dipolar correlated spectroscopy (NOESY) and two-dimensional relayed coherance transfer spectroscopy (RCT) experiments were recorded, allowing most resonances arising from the aromatic and methyl-containing residues to be assigned in the spectrum. From an analysis of NOE data, a small segment of double-stranded beta-sheet has been identified near residues Phe63 and Thr64. Further analysis of the NOESY spectrum has allowed detailed study of the conformation of sidechains located in regions near Leu45 and Val69. A model has been constructed of the polypeptide segment comprising residues 40-49 which accounts for the observed NOE interactions.     

Identification of a cysteine residue as the binding site for the dipyrromethane cofactor at the active site of Escherichia coli porphobilinogen deaminase

The dipyrromethane cofactor of Escherichia coli porphobilinogen deaminase was specifically labelled with 13C by growth of the bacteria in the presence of 5-amino[5-13C]levulinic acid. Using 13C-NMR spectroscopy, the structure of the cofactor was confirmed as a dipyrromethane made up of two linked pyrrole rings each derived from porphobilinogen. The chemical shift data indicate that one of the pyrrole rings of the cofactor is covalently linked to the deaminase enzyme through a cysteine residue. Evidence from protein chemistry studies suggest that cysteine-242 is the covalent binding site for the cofactor.     

Effect of low-dose irradiation upon T cell subsets involved in the response of primed A/J mice to SaI cells

A/Jax (A/J) mice primed to Sarcoma I (SaI) exhibit an augmented response in association with low-dose (0.15 Gy) irradiation. This phenomenon is best demonstrated in tumour neutralization (Winn assay) or cell transfer experiments utilizing mice depleted of thymus-derived (T) cells. It is particularly dependent upon the duration of priming and the growth characteristics of the tumour in the primary host. The importance of these two variables appears to relate to their influence upon the cell types responsible for the host response, and includes both an effector and a suppressor component. Radiation-induced inhibition of the suppressor component appears responsible for low-dose augmentation and results in injury to a T cell of the Lyt-1-2+ phenotype. In Winn assays employing equal numbers of immune spleen cells and SaI cells, the smallest tumours are associated with Lyt-1-positive (Lyt-1+2- and Lyt-1+2+) cells and exposure to 0.15 Gy markedly inhibits their anti-SaI activity. Thus, even though the effect is in the opposite direction, both the effector and suppressor components of the anti-SaI response in A/J mice are exceedingly radiosensitive.     

Metabolism of sulfur-containing amino acids by pregnant merino ewes

The availability and utilization of cystine and methionine were measured in single-bearing Merino ewes on three occasions, approximately 90, 110 and 130 days after mating, and the effects on these traits of sulfur amino acids (SAA) infused into the abomasum were also measured. Two levels of SAA were infused containing 0.5 or 1.0 g day-1 organic sulfur with DL-methionine contributing two-thirds and L-cystine one-third of the supplementary sulfur. The quantity of the diet offered was increased at each occasion so as to maintain maternal liveweight. The rates of irreversible loss of both cystine and methionine from plasma increased as pregnancy advanced, but the ratios between the rates of irreversible loss and intake of digestible organic matter (DOMI) did not vary with stage of pregnancy. The average daily rates of irreversible loss of cystine and methionine by the ewes consuming the diet alone were 13.6 and 119 mmol kg-1 DOMI respectively. The average rates of irreversible loss of methionine (Im, mmol h-1) and of cystine (Ic, mmol h-1) were both linearly (P less than 0.05) related to the rate of infusion of organic sulfur into the abomasum (s, g day-1): Im = 2.44 (+/- 0.33) s + 1.28 (+/- 0.13); and Ic = 0.16 (+/- 0.02) s + 0.30 (+/- 0.01). Five per cent of the rate of irreversible loss of cystine arose from trans-sulfuration of methionine by ewes consuming the ration only, but greater percentages (14 and 22%) were observed when the ration was supplemented with SAA (P less than 0.05). These transfer quotients were not influenced by stage of pregnancy. The stage of pregnancy did not influence the concentration of cystine or methionine in the plasma, but the abomasal infusions of SAA significantly increased the concentration of both SAA. The ewes consuming the basal diet were in positive balance for both nitrogen and sulfur. The retention of nitrogen did not vary with stage of pregnancy (average (s.e.), 5.8 (0.9) g day-1), but that of sulfur increased from 0.6 to 1.0 and 1.3 g day-1 in periods 1, 2 and 3, respectively (P less than 0.05). The retentions of nitrogen (N, g day-1) and of sulfur (S, g day-1) were linearly and significantly related to the rate of infusion of organic sulfur into the abomasum (s, g day-1): N = 2.7 (+/- 0.7)s + 4.4 (+/- 0.3); and S = 0.49 (+/- 0.03)s + 0.72 (+/- 0.01).(ABSTRACT TRUNCATED AT 400 WORDS)