bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification and localization of amino acid substitutions between two phenobarbital-inducible rat hepatic microsomal cytochromes P-450 by micro sequence analyses.
Authors: Yuan P M , Ryan D E , Levin W , Shively J E
Published: May 2006

Two isozymes of rat liver microsomal cytochrome P-450--P-450b and P-450e--were compared by micro sequence analyses of their NH2 termini and tryptic fragments. These two phenobarbital-inducible hemoproteins, which are immunochemically indistinguishable with antibody against cytochrome P-450b, have extensive sequence homology. Automated Edman degradation of the native proteins revealed identical amino acids for the first 35 residues. Sequence determinations of the tryptic peptides, which constitute approximately 75% of each protein molecule, have thus far shown 10 amino acid differences between the two isozymes. Results of our amino acid sequence analyses established that two of the cDNAs, pcP-450pb1 and pcP-450pb4, reported by Fujii-Kuriyama et al. [Fujii-Kuriyama, Y., Mizukami, Y., Kamajiri, K., Sogawa, K. & Muramatsu, M. (1982) Proc. Natl. Acad. Sci. USA 79, 2793-2797] encode cytochrome P-450b whereas pcP-450pb2, a third cDNA whose nucleotide sequence differed slightly from that of the other two (six amino acid substitutions), encodes cytochrome P-450e. In addition to establishing the identity of these cloned cDNAs we provide direct evidence for seven additional amino acid differences between cytochromes P-450b and P-450e that occur beyond the region (Arg358) encoded by the cloned cDNA for cytochrome P-450e. Together, the amino acid sequences determined by micro sequence analysis and recombinant DNA techniques reveal 13 amino acid differences between these two isozymes. This report highlights the complementary nature of two different molecular approaches to elucidation of the amino acid sequences of isozymes with extensive structural homology.
Isolation of pancreatic trypsin inhibitor from bovine pituitary glands.
Authors: Li C H , Chung D
Published: May 2006

A trypsin inhibitor has been isolated from bovine pituitary extracts. From its amino acid composition, NH2-terminal residue, mobility in paper electrophoresis, and behavior in high-performance liquid chromatography and from the tryptic pattern of the performic acid-oxidized material, it appears that the inhibitor is identical to the Kunitz and Northrop pancreatic trypsin inhibitor.
Submitochondrial localization, cell-free synthesis, and mitochondrial import of 2-isopropylmalate synthase of yeast.
Authors: Hampsey D M , Lewin A S , Kohlhaw G B
Published: May 2006

2-Isopropylmalate synthase (EC 4.1.3.12) of yeast is a mitochondrial enzyme. We now provide evidence showing that a large part of the 2-isopropylmalate synthase activity that is associated with the mitochondria is located in the mitochondrial matrix. In vitro translation of total yeast RNA followed by immunoprecipitation with anti-2-isopropylmalate synthase antibody yields two polypeptides. The larger of these has an apparent molecular weight identical to that of purified 2-isopropylmalate synthase subunit (ca. 65,000). It is incorporated into isolated yeast mitochondria with no detectable change in molecular weight. The import requires energy. The smaller polypeptide migrates to a position corresponding to a molecular weight of 63,000-64,000. It is not taken up by mitochondria. Both polypeptides, which also can be obtained by immunoprecipitation of crude extracts, become labeled when in vitro translation is performed in the presence of N-formyl[35S]methionyl-tRNAf. Mutants with no detectable 2-isopropylmalate synthase activity are deficient in either one or both synthase-related polypeptides. These results are discussed in the light of recent evidence for two 2-isopropylmalate synthase-encoding genes in yeast.
Antibody-targeted liposomes: increase in specific toxicity of methotrexate-gamma-aspartate.
Authors: Heath T D , Montgomery J A , Piper J R , Papahadjopoulos D
Published: May 2006

Liposomes conjugated with anti-H2Kk antibody associate with L929 murine fibroblasts in 6- to 20-fold greater amount than do nonspecific liposomes. The ability of methotrexate-gamma-aspartate to inhibit L929 growth is increased 10-fold when encapsulated in targeted liposomes but is decreased to 50% when encapsulated in liposomes with no specificity for the target cells. Ammonium chloride inhibits the effects of the encapsulated but not the free drug. Soluble antibody does not inhibit the efficacy of targeted vesicles, but empty targeted vesicles do inhibit the efficacy. The compound in both targeted and nontargeted vesicles has a minimal effect on BALB/c3T6 fibroblasts. These results demonstrate the potential of antibody-targeted liposomes and the importance of selecting liposome-dependent cytotoxic agents.
Unusual structure of the chicken embryonic alpha-globin gene, pi'.
Authors: Engel J D , Rusling D J , McCune K C , Dodgson J B
Published: May 2006

We report the DNA sequence of the globin locus encoding the chicken embryonic alpha-globin, pi'. The structure differs significantly from that of the two chicken adult alpha-globin genes, alpha A and alpha D, as well as from that of previously studied adult alpha-globin genes in that the introns of the pi' gene are substantially larger than those in adult alpha-globin loci. In contrast, the pi' introns are structurally similar to the only other expressed embryonic alpha-globin gene reported to date, the human zeta gene. While completing the sequence of the pi' gene, we determined that only one chromosomal locus within the chicken genome hybridizes to a pi' central exon probe. These data lead to the conclusion that if the equimolar chicken embryonic alpha-globin polypeptides, called pi and pi', are indeed independently transcribed, then that transcription occurs from alleles of the same gene; however, we favor the possibility that the pi gene does not actually exist. This conclusion is drawn from the observation that the two chromosomal alleles of embryonic alpha-globins (represented by recombinant bacteriophage lambda CaG5 and lambda CaG7) both encode pi'.
Identification of a second class I antigen controlled by the K end of the H-2 complex and its selective cellular expression.
Authors: Tryphonas M , King D P , Jones P P
Published: May 2006

Immunoprecipitates obtained from [35S]methionine-labeled spleen cells by using monoclonal antibodies specific for H-2Kd and H-2Dd have been separated by two-dimensional polyacrylamide gel electrophoresis. Analysis of these gel patterns revealed the presence of an additional product of the K end of the H-2d complex, designated here as H-2K'. To determine whether H-2K' is a unique protein or a differentially glycosylated form of the previously characterized H-2Kd histocompatibility antigen, nonglycosylated molecules labeled in the presence of tunicamycin were examined. The results showed that both H-2K and H-2K' have distinct nonglycosylated polypeptide precursor forms. The approximate molecular weight differences between the fully glycosylated and nonglycosylated molecules also indicated the presence of three oligosaccharide side chains on H-2K', as is the case with H-2Kd, whereas H-2Dd has only two oligosaccharide units. The cellular expression of H-2K' was also investigated. Comparison of H-2 antigens immunoprecipitated from normal spleen cells and from thioglycollate-induced adherent peritoneal exudate cells cultured in the presence or absence of supernatant fluids from concanavalin A-stimulated spleen cells revealed that H-2K' was not expressed on the adherent peritoneal cells. This indicates that H-2K' is expressed in a tissue-specific manner, unlike the classical histocompatibility antigens H-2K and H-2D.
Effects of tetracaine on charge movements and calcium signals in frog skeletal muscle fibers
Authors: Vergara Julio , Caputo Carlo
Published: May 2006

Intramembrane charge movements in skeletal muscle fibers contain a tetracaine-sensitive component that can be isolated by the use of this drug. The time course and voltage dependence of this component, studied in relation to antipyrylazo III absorbance signals, suggest its direct involvement in the calcium release process in muscle.
In vivo carbon-13 nuclear magnetic resonance studies of heart metabolism.
Authors: Neurohr K J , Barrett E J , Shulman R G
Published: May 2006

Guinea pig heart metabolism was studied in vivo by 13C NMR at 20.18 MHz. High-quality proton-decoupled 13C NMR spectra with excellent signal-to-noise ratios and resolution could be obtained in 6 min. Natural-abundance spectra showed resonances that could be assigned to fatty acids, but glycogen was not seen. During intravenous infusion of D-[1-13C]glucose and insulin, the time course of myocardial glycogen synthesis was followed serially for up to 4 hr. Anoxia resulted in degradation of the labeled glycogen within 6 min and appearance of 13C label in lactic acid. Infusion of sodium [2-13C]acetate resulted in incorporation of label into the C-4, C-2, and C-3 positions of glutamate and glutamine, reflecting "scrambling" of the label expected from tricarboxylic acid cycle activity. Examination of the 31P NMR spectrum of the guinea pig heart in vivo demonstrated no change in the high-energy phosphates during the time periods of the 13C NMR experiments. Our studies indicate that 13C NMR is a unique non-destructive tool for the study of heart metabolism in vivo.
Dialyzable factor in human serum of platelet origin stimulates endothelial cell replication and growth
Authors: Clemmons David R. , Isley William L. , Brown M. Todd
Published: May 2006

Porcine aortic endothelial cells were isolated and maintained in Dulbecco's modified Eagle's medium (DME medium)/10% citrate-treated human plasma. They were stimulated by DME medium/10% human serum to grow from a density of 10,100 ± 500 per well to a final density of 83,000 ± 1,800 per well over a 9-day period. On the other hand these cells grew poorly (11% increase) in DME medium/10% human platelet-poor plasma prepared without chelating agents and containing platelet factor 4 at 18 ng/ml by radioimmunoassay. Dialysis of the human serum (
M
r
cutoff, 3,500) eliminated all the stimulatory activity. The activity recovered from the dialysate stimulated growth when added to endothelial cultures in conjunction with either dialyzed serum or platelet-poor plasma alone. The dialyzable factor could be obtained directly from platelets; both acetic acid extracts and boiled NaCl extracts stimulated porcine aortic endothelial cell replication. Gel filtration chromatography on Sephadex G-15 showed that the endothelial growth factor had a molecular weight of 700. Partially purified material induced a concentration-dependent stimulation of porcine aortic endothelial cell replication in the presence of DME medium alone; however, simultaneous incubation with platelet-poor plasma resulted in a much greater response. Fibroblast growth factor isolated from bovine brain was found to be mitogenic only in the presence of nondialyzed serum or of the dialyzable factor together with plasma. In the absence of this serum factor, fibroblast growth factor had no effect. We conclude that human serum contains a potent endothelial cell mitogen of platelet origin. Human plasma that is devoid of platelet content does not stimulate endothelial cell growth. This growth factor may be an important stimulant of the endothelial cell response to vascular wall injury.
Polymorphism of human erythrocyte C3b/C4b receptor.
Authors: Dykman T R , Cole J L , Iida K , Atkinson J P
Published: May 2006

The human erythrocyte receptor for the major activation fragments of the third and fourth components of complement (HuE-C3bR) was isolated from individual donors. Erythrocytes were surface labeled with 125I and solubilized in Nonidet P-40.HuE-C3bR was purified by using C3-Sepharose affinity chromatography and analyzed by autoradiography of NaDodSO4/polyacrylamide gels. Three distinct receptor patterns were demonstrated. Type a had a single major band with Mr of 190,000, type b had a single major band with Mr of 220,000, and type c had two major bands of Mr 190,000 and 220,000. In all three types, a minor band accounting for less than 25% of the total radioactivity was usually observed at a Mr 15,000 greater than that of each major band. Identical autoradiographic patterns were obtained by affinity chromatography using methylamine-inactivated C4-Sepharose or by immunoprecipitation of solubilized membranes with a monoclonal antibody against HuE-C3bR. All three types were distinct after reduction and alkylation, although the apparent Mr uniformly increased by approximately equal to 30,000. Characterization of HuE-C3bR types in 33 unrelated individuals demonstrated that 23 had type a, 1 had type b, and 9 had type c. Family studies provide evidence for transmission by two codominant alleles. Thus, in the normal population two alleles appear to control expression of HuE-C3bR phenotypes and account for the polymorphism of this integral membrane glycoprotein.
Homogeneously staining chromosomal regions contain amplified copies of an abundantly expressed cellular oncogene (c-myc) in malignant neuroendocrine cells from a human colon carcinoma.
Authors: Alitalo K , Schwab M , Lin C C , Varmus H E , Bishop J M
Published: May 2006

Two human neuroendocrine tumor cell lines derived from a colon carcinoma contain either numerous double minute chromosomes (COLO 320 DM) or a homogeneously staining marker chromosome (COLO 320 HSR). We found amplification and enhanced expression of the cellular oncogene c-myc in both COLO 320 DM and HSR cells, and we were able to show that the homogeneously staining regions of the COLO 320 HSR marker chromosome contain amplified c-myc. From previous and present karyotypes, it appears that the homogeneously staining regions reside on a distorted X chromosome. Therefore, amplification of c-myc has been accompanied by translocation of the gene from its normal position on chromosome 8 (8q24). Because double minute chromosomes were features of primary cultures from the original tumor, it seems reasonable to suspect that amplification of c-myc may have contributed to tumorigenesis.
Prostaglandin D2, a cerebral sleep-inducing substance in rats.
Authors: Ueno R , Honda K , Inoué S , Hayaishi O
Published: May 2006

We continuously monitored the circadian sleep patterns of unrestrained rats for more than 96 hr and infused various prostaglandins into their third ventricles for 10 hr to study the effects on inducing sleep. Prostaglandin D2 at 6 fmol/min had no effect on either slow wave sleep or paradoxical sleep. However, prostaglandin D2 at as little as 60 fmol/min caused a significant amount of excess slow wave sleep as compared with the control level during saline infusion. Paradoxical sleep was induced by prostaglandin D2 at doses greater than 600 fmol/min. Prostaglandin D2 (600 fmol/min) increased slow wave sleep by 33% and paradoxical sleep by 56%. Although prostaglandin F2 alpha (600 fmol/min) increased the amount of slow wave sleep, its activity was less than that of the same amount of prostaglandin D2. Prostaglandin E2 (600 fmol/min) had no effect on increasing the amounts of both slow wave sleep and paradoxical sleep. During the infusion of prostaglandin D2, rats were easily aroused by clap sound stimulation and their sleeping and waking postures remained normal. Further, their sleep was episodic, as observed in the physiological sleep of rats.
Degree of mapping for nonlinear mappings of monotone type
Authors: Browder Felix E.
Published: May 2006

A classical degree function is constructed for pseudomonotone mappings from a reflexive Banach space to its dual, using Galerkin approximations. This generalizes the Leray-Schauder degree when the Banach space is a Hilbert space and yields a flexible analytical tool for the study of nonlinear elliptic problems of higher order in divergence form.
Equivalence of partition properties and determinacy
Authors: Kechris Alexander S. , Woodin W. Hugh
Published: May 2006

It is shown that, within
L
(ℝ), the smallest inner model of set theory containing the reals, the axiom of determinacy is equivalent to the existence of arbitrarily large cardinals below Θ with the strong partition property κ → (κ)
κ
.
Modulation of the kinetics of cholesterol side-chain cleavage by an activator and by an inhibitor isolated from the cytosol of the cortex of bovine adrenals.
Authors: Warne P A , Greenfield N J , Lieberman S
Published: May 2006

Two modulators of sterol side-chain cleavage activity have been detected in the cytosol from the cortex of bovine adrenals. One is an inhibitor of side-chain cleavage which increases the Km of a purified and reconstituted mitochondrial side-chain cleavage system for both cholesterol and cholesterol sulfate. It also lowers the Vmax of cleavage when cholesterol sulfate is the substrate. The other modulator is a low molecular weight protein which in the reconstituted system increases the Vmax of cleavage for both substrates but does not affect the Km of either. Side-chain cleavage activity was also found in bovine adrenocortical cytosol and this appears to differ from that present in mitochondria.
Inhibitors of endocytosis perturb phospholipid metabolism in rabbit neutrophils and other cells.
Authors: Mato J M , Pencev D , Vasanthakumar G , Schiffmann E , Pastan I
Published: May 2006

Dansylcadaverine, amantadine, and rimantadine, which have been shown to inhibit the endocytosis of alpha 2-macroglobulin, epidermal growth factor, and vesicular stomatitis virus [Schlegel, R., Dickson, R. B., Willingham, M. C. & Pastan, I. (1982) Proc. Natl. Acad. Sci. USA 79, 2291-2295], were found to decrease phosphatidylcholine synthesis, chemotaxis, and internalization of a formylated peptide but to stimulate the incorporation of inositol into phosphatidylinositol in rabbit neutrophils. Dansylcadaverine decreased phosphatidylcholine synthesis by both the CDP-choline and transmethylation pathways and also inhibited the synthesis of phosphatidylethanolamine by the CDP-ethanolamine pathway. Dansylcadaverine had no effect on the phosphocholine, CDP-choline, or S-adenosyl-L-homocysteine pools but increased 2-fold the S-adenosyl-L-methionine pool. These results suggest that dansylcadaverine in some manner inhibited the condensation of CDP-choline with diacylglycerol to form phosphatidylcholine. Dansylcadaverine also inhibited phosphatidylcholine synthesis in human neutrophils, human fibroblasts, chicken embryo fibroblasts, rat hepatocytes, osteosarcoma cells, and neuroblastoma cells. It did not stimulate phosphatidylinositol synthesis in chicken embryo fibroblasts.
Stabilization of erythrocyte membranes by polyamines.
Authors: Ballas S K , Mohandas N , Marton L J , Shohet S B
Published: May 2006

Using a laser diffraction technique, we have studied the effects of putrescine, spermidine, and spermine, the three physiologic polyamines, on the deformability and mechanical stability of human erythrocyte membranes. Ghosts resealed with polyamines were subjected to high fluid shear stress in an ektacytometer. All polyamines increased the membrane shear modulus (decreased deformability) in a concentration- and time-dependent manner. The order of effectiveness was spermine greater than spermidine greater than putrescine. At 10 microM, spermine appreciably decreased membrane deformability. For the measurement of membrane mechanical stability, resealed ghosts were subjected to constant high shear stress in the ektacytometer and deformability was continuously recorded as the deformable ghosts fragmented into rigid spherical vesicles. Polyamines, especially spermine, caused a noticeable increase in the t1/2 for fragmentation. These effects could not be ascribed to proteolysis or Ca2+-induced transglutamination. That the effects of polyamines were specific and not simply due to their positive charge was demonstrated by the finding that Ca2+ and Mg2+ destabilized the erythrocyte membrane as evidenced by decreasing the t1/2 for fragmentation. Extracellular polyamines were not effective except under conditions that caused significant accumulation inside the cell. The data indicate that intracellular physiologic polyamines, especially spermine, decrease erythrocyte membrane deformability and stabilize the membrane skeleton, making it more resistant to fragmentation.
Intracisternal A-particle genes as movable elements in the mouse genome.
Authors: Kuff E L , Feenstra A , Lueders K , Smith L , Hawley R , Hozumi N , Shulman M
Published: May 2006

We analyzed two functionally defective mouse kappa light chain gene variants previously shown to contain novel insertions of repetitive DNA in their intervening sequences [Hawley, R. G., Shulman, M. J., Murialdo, H., Gibson, D. M. & Hozumi, N. (1982) Proc. Natl. Acad. Sci. USA 79, 7425-7429]. Heteroduplex analysis of the cloned genes shows that the insertions consist of intracisternal A-particle (IAP) genetic elements. Each insertion includes an IAP 5' long terminal repeat (LTR) sequence and extends to a characteristic IAP internal BamHI site where the IAP sequence is interrupted because the mutant genes were cloned from complete BamHI digests of the cellular DNAs. Restriction enzyme mapping indicates that the 5' LTR boundaries of the inserted IAP elements correspond closely to the previously determined rearrangement sites in the mutant genes. The IAP insertions in the two mutants can be distinguished by restriction-site differences and by the fact that one of them contains a deletion that is absent in the other. Nucleotide sequence data are presented for the LTRs of one full-length IAP gene copy randomly selected from a mouse genomic DNA library. These LTRs show many features typical of known integrated retroviral terminal repeat units, and the entire gene is bracketed by short direct repeats within the adjacent cellular DNA. Thus, the findings show that IAP genetic elements can appear in new locations in mouse cellular DNA and suggest that this may occur through a process of proviral insertion.
Paradox of the evolution of communication and of social interactivity.
Authors: Cavalli-Sforza L L , Feldman M W
Published: May 2006

Communication between individuals of a species is likely to increase the capacity to acquire skills useful for survival and propagation and thus may confer important selective advantages. Since interaction occurs between two or more individuals, the selective process is frequency dependent, and the analysis shows that communication cannot initially increase at a reasonable rate when it is limited to random unrelated individuals, so that it is likely to abort for stochastic reasons. However, this bottleneck is removed if the communication process takes place in the nuclear family or among close relatives or if aggregation of communicators occurs because of assortative mating or meeting. Use of the individual conditional fitnesses we have introduced earlier permits an exact analysis. We show that, in general, the initial rate of increase can be geometric if and only if, in the class of selective models considered, the conditional probability of a communicator interacting with another contains a positive constant term. In our discussion of communication, cost factors for the act of communication have been omitted. However, the model has been generalized to include cooperativeness, and also altruism, or competition, by introducing costs. There is a close relationship among these situations, and the same considerations about the initial bottleneck and its resolution also extend to them. The models given here are for haploids but they extend to diploids and the conclusions are similar.
Delayed hypersensitivity induced by anti-T-cell-line antisera is enhanced by pertussigen and not restricted by histocompatibility genes.
Authors: Gamble J R , Vadas M A , Munoz J J , Thomas W R , Miller J F
Published: May 2006

Anti-T-cell-line antisera were raised by repeatedly injecting mice with syngeneic, antigen-specific, delayed hypersensitivity (DH) inducing cells grown as continuous T-cell lines in vitro. Many of these antisera could induce antigen-specific DH responses which, in some cases, were rather slight. For example, the DH reaction to azobenzenearsonate induced in A/J mice by an A/J antiserum produced against the syngeneic azobenzenearsonate-specific cell line AA3 was only a weak response. Administration of pertussigen, a protein purified from Bordetella pertussis, markedly enhanced this response and prolonged its duration. Histologically the reaction showed the classical mononuclear infiltration of DH. It could be transferred to naive mice by lymph node cells that were anti-Thy-1.2 sensitive, and the transfer was restricted by genes of the major histocompatibility complex (MHC). The induction of DH by antiserum, however, was not influenced by MHC or non-MHC gene products.
An effect of nerve growth factor on parasympathetic neurite outgrowth.
Authors: Collins F , Dawson A
Published: May 2006

Addition of nerve growth factor (NGF) to dissociated parasympathetic ciliary ganglion neurons resulted, within 60 min of its addition, in a 2-fold increase in average neurite length and an accompanying enlargement and spreading of neuronal growth cones. These effects occurred over a concentration range of NGF of 0.1-10 ng/ml and were blocked by affinity-purified antibody to NGF. Epidermal growth factor, fibroblast growth factor, and angiotensin did not have these effects, although insulin at high concentrations was able to induce a response similar to that of NGF. Dissociated sympathetic chain neurons also responded to NGF with increased neurite lengths, and, in addition, NGF considerably extended the survival time of these neurons in culture. However, the effect of NGF on ciliary ganglion neurons was limited to neurite outgrowth, and NGF did not promote the survival of these parasympathetic neurons.
Cloning of cDNA encoding the pre-beta subunit of mouse thyrotropin.
Authors: Gurr J A , Catterall J F , Kourides I A
Published: May 2006

Double-stranded cDNA was synthesized from sucrose gradient-purified poly(A)+ mRNA from a mouse thyrotropic tumor, inserted into the Pst I site of plasmid pBR322 by using poly(dC) . poly(dG) homopolymeric extensions, and cloned in Escherichia coli RRI. Plasmids containing cDNA sequences coding for the beta subunit of thyrotropin (TSH) were identified by cell-free translation of hybrid-selected mRNA and immunoprecipitation with specific antibody to TSH beta subunit. Determination of the nucleotide sequence of one cDNA, 595 base pairs in length, allowed us to deduce the entire amino acid sequence of the mouse TSH beta subunit. The pre-beta subunit contains a 20-amino acid amino-terminal signal sequence followed by a 118-amino acid mature TSH beta subunit. There is 85-90% homology in amino acid sequence between mouse TSH beta subunit and subunits from man, pig, and cow; however, the mouse subunit contains an additional 5 or 6 amino acids at its carboxyl terminus compared to the bovine or human and pig subunits, respectively. TSH beta-subunit mRNA from mouse thyrotropic tumor was estimated to be 750 nucleotides in length by hybridization with labeled TSH beta-subunit cDNA.
Degree of mapping for nonlinear mappings of monotone type: Densely defined mapping
Authors: Browder Felix E.
Published: May 2006

The classical degree function constructed earlier for pseudomonotone mappings has been used to develop a broader degree theory of classical type for the sum of a maximal monotone map from a reflexive Banach space to its dual together with a bounded pseudomonotone map. The proof uses the generalized Yosida approximation of the maximal monotone mapping.
Characterization of integrated hepatitis B viral DNA cloned from a human hepatoma and the hepatoma-derived cell line PLC/PRF/5.
Authors: Dejean A , Brechot C , Tiollais P , Wain-Hobson S
Published: May 2006

Recombinant phage clones carrying integrated hepatitis B virus (HBV) DNA sequences have been isolated from two phage libraries made from human DNA of a hepatoma and a hepatoma-derived cell line. One clone from each library has been characterized both by restriction mapping and by electron microscopy. In one clone there is at least one complete and uninterrupted HBV genome, and in the other the HBV sequences are composed of two major subgenomic fragments inverted with respect to each other. The host-virus junctions are localized within the positions 1,700-2,600 base pairs on the physical map of the free viral genome. The pre-S/S (surface antigen gene) region is conserved between the two clones. The two clones do not have common cellular sequences nor do they contain cellular homologues to six retroviral oncogenes. For one clone, the hepatitis B surface antigen gene was found to be functional when introduced into mouse thymidine kinase-negative cells by transfection.
Bradykinin stimulates phospholipid methylation, calcium influx, prostaglandin formation, and cAMP accumulation in human fibroblasts.
Authors: Bareis D L , Manganiello V C , Hirata F , Vaughan M , Axelrod J
Published: May 2006

The biochemical events that lead to bradykinin stimulation of cAMP accumulation in human fibroblasts were examined. Treatment of human fibroblasts with bradykinin increases phospholipid methylation, Ca2+ influx, arachidonic acid release, prostaglandin formation, and cAMP content. The dose-response curves of bradykinin for the increase in the above changes were similar. In human fibroblasts, exogenous arachidonic acid was mainly incorporated into phosphatidylcholine, followed by phosphatidylserine, phosphatidylethanolamine, and phosphatidylinositol. Bradykinin caused a release of arachidonic acid from methylated phospholipids (phosphatidylcholine) and phosphatidylinositol. 3-Deazaadenosine, a methyltransferase inhibitor, almost completely inhibited bradykinin-stimulated phospholipid methylation and Ca2+ influx and partially reduced arachidonic acid release and prostaglandin formation but had no effect on cAMP formation. Mepacrine, a phospholipase inhibitor, blocked bradykinin-induced arachidonic acid release, prostaglandin release, and cAMP accumulation. Indomethacin, a cyclooxygenase inhibitor, blocked the effect of bradykinin on cAMP accumulation. Prostaglandins E1 and E2, but not F2 alpha, increased accumulation of cAMP. These observations indicate that bradykinin generates cAMP via arachidonic acid release and subsequent formation of prostaglandins. Our findings suggest that arachidonic acid can arise from either phosphatidylcholine synthesized by the methylation pathway or phosphatidylinositol.
Carbonic anhydrase II deficiency identified as the primary defect in the autosomal recessive syndrome of osteopetrosis with renal tubular acidosis and cerebral calcification.
Authors: Sly W S , Hewett-Emmett D , Whyte M P , Yu Y S , Tashian R E
Published: May 2006

The clinical, radiological, and pathological findings in three siblings affected with the autosomal recessive syndrome of osteopetrosis with renal tubular acidosis and cerebral calcification have been reported. In an effort to explain the pleiotropic effects of the mutation producing this disorder, we postulated a defect in carbonic anhydrase II (CA II), the only one of the three soluble isozymes of carbonic anhydrase that is known to be synthesized in kidney and brain. We report here biochemical and immunological evidence for the virtual absence of CA II in erythrocytes of patients affected with this condition, whereas CA I level is not reduced. Levels of CA II in erythrocyte hemolysates from asymptomatic obligate heterozygotes are about half of normal. These findings: (i) elucidate the basic defect in one form of inherited osteopetrosis; (ii) provide genetic evidence implicating CA II in osteoclast function and bone resorption; (iii) explain previous observations that carbonic anhydrase inhibitors block the normal parathyroid hormone-induced release of calcium from bone; (iv) clarify the role of renal CA II in urinary acidification and bicarbonate reabsorption; and (v) suggest a method to identify heterozygous carriers for the gene for this recessively inherited syndrome.
Nonlinear interactions in a dendritic tree: localization, timing, and role in information processing.
Authors: Koch C , Poggio T , Torre V
Published: May 2006

In a passive dendritic tree, inhibitory synaptic inputs activating ionic conductances with an equilibrium potential near the resting potential can effectively veto excitatory inputs. Analog interactions of this type can be very powerful if the inputs are appropriately timed and occur at certain locations. We examine with computer simulations the precise conditions required for strong and specific interactions in the case of a delta-like ganglion cell of the cat retina. We find some critical conditions to be that (i) the peak inhibitory conductance changes must be sufficiently large (i.e., approximately equal to 50 nS or more), (ii) inhibition must be on the direct path from the location of excitation to the soma, and (iii) the time course of excitation and inhibition must substantially overlap. Analog AND-NOT operations realized by satisfying these conditions may underlie direction selectivity in ganglion cells.
Location of amino acid alterations in mutants of aspartate transcarbamoylase: Structural aspects of interallelic complementation.
Authors: Schachman H K , Pauza C D , Navre M , Karels M J , Wu L , Yang Y R
Published: May 2006

Recent genetic studies of the pyrB locus of Escherichia coli resulted in the characterization of 29 mutant strains harboring defects in the structural gene that encodes the catalytic chains of aspartate transcarbamoylase (carbamoylphosphate: L-aspartate carbamoyltransferase, EC 2.1.3.2). Three alleles, pyrB554, pyrB730, and pyrB748, have been cloned, and their nucleotide sequences have been determined along with that of the wild-type pyrBI operon in order to locate the sites of the alterations in the catalytic chains. Missense mutation pyrB554 leads to replacement of serine-52 by phenylalanine, and the inactive mutant enzyme has properties similar to those of wild-type aspartate transcarbamoylase. The amber mutation pyrB730 results in unstable truncated polypeptide chains 27 amino acids shorter than wild-type chains. Deletion mutation pyrB748 causes the removal of 181 amino acids. Combining these results with knowledge of the crystallographic structure of the wild-type enzyme provides a basis for tentative structural mechanisms for the observed complementation behavior of the mutant proteins.
Reaction center and light-harvesting I genes from
Rhodopseudomonas capsulata
Authors: Youvan Douglas C. , Alberti Marie , Begusch Helmut , Bylina Edward J. , Hearst John E.
Published: May 2006

Five structural genes coding for the reaction center (RC) L, M, and H subunits and the two light-harvesting (LH) I polypeptides, B870α and B870β, have been mapped on two restriction fragments from the R-prime plasmid pRPS404. It has been recently shown that enhanced near-infrared fluorescence mutants of
Rhodopseudomonas capsulata
typically lack RC or LH I polypeptides and that these lesions are marker-rescued by two restriction fragments from the R-prime plasmid: the 7.5-kilobase-pair
Eco
RI F fragment and the 4.75-kilobase-pair
Bam
HI C-
Eco
RI fragment. We have now determined the nucleotide sequence of two restriction fragments and have found that the
Bam
HI C-
Eco
RI B fragment carries the structural genes for the RC L and M subunits and both LH I polypeptides. Forty kilobase pairs away from this locus, the
Bam
HI F fragment (within the
Eco
RI F fragment) carries the RC H subunit. The structural genes on the
Bam
HI C-
Eco
RI B fragment are probably transcribed as part of a polycistronic mRNA. All of the structural genes begin with a consensus Shine-Dalgarno sequence and separate AUG start codons, indicating that the structural polypeptides are not cleaved from larger precursor polypeptides.
tau, a repeated DNA sequence in yeast.
Authors: Chisholm G E , Genbauffe F S , Cooper T G
Published: May 2006

We have found a 371-base-pair (bp) repeated DNA element, tau, in Saccharomyces cerevisiae. The ends of tau are composed of a 5-bp inverted repeat, similar in sequence to those reported for the Ty, sigma, copia, and spleen necrosis virus elements. These inverted repeats are flanked by 5-bp direct repeats of a target sequence that occurs only once in an allele that lacks the tau element. This overall structure is characteristic of transposable elements. Like sigma, tau elements have been found (in both orientations) closely associated with tRNA genes (409 and 198 bp from the 5' end, respectively). It is noteworthy that one representative of tau was isolated in a concentric insertion of tau, delta, and sigma.
Uteroferrin has N-asparagine-linked high-mannose-type oligosaccharides that contain mannose 6-phosphate.
Authors: Baumbach G A , Saunders P T , Bazer F W , Roberts R M
Published: May 2006

Uteroferrin is an iron-containing, progesterone-induced, acid phosphatase that is secreted in large amounts by the uterine endometrium of pigs. During pregnancy, it transports iron across the chorioallantois (placenta) for use in fetal hematopoiesis. In this paper, it is reported that uteroferrin synthesized by cultured endometrial explants possesses N-linked, high-mannose, oligosaccharide chains that contain 6- phosphomannose units. The latter is regarded as a possible recognition marker whereby acid hydrolases are targeted to the lysosome. On uteroferrin, however, the majority of the phosphate is in single diester linkages between the mannose and a covering N-acetylglucosamine. It is suggested that uteroferrin is a lysosomal enzyme that has assumed a role in iron transport and metabolism and is secreted because the covering N-acetylglucosamine is not removed.
Occurrence of selenium-containing tRNAs in mouse leukemia cells.
Authors: Ching W M
Published: May 2006

Selenium incorporation into the polynucleotide structures of tRNAs has been documented in several microorganisms. In the present study, selenium-containing species were isolated from bulk tRNA preparations from 75Se-labeled mouse leukemia cells. The major 75Se-labeled species was similar in size and exhibited the same sensitivity to ribonuclease as did Escherichia coli tRNAs. The chromatographic properties of the intact major selenium-containing tRNA species indicated it to be very hydrophobic in character. The selenium component that is unstable at neutral-to-alkaline pH but is relatively stable at acid pH is not an esterified selenoamino acid. HPLC analysis of enzymic digests of the major selenium-containing species detected selenium-containing hydrophobic products (probably selenonucleosides ). These properties strongly suggest that the selenium in the mouse leukemia-cell tRNAs is present in the form of a selenium-modified nucleoside.
Tumor promoters block tyrosine-specific phosphorylation of the epidermal growth factor receptor.
Authors: Friedman B , Frackelton A R , Ross A H , Connors J M , Fujiki H , Sugimura T , Rosner M R
Published: May 2006

Tyrosine-specific phosphorylation of the epidermal growth factor (EGF) receptor in hormonally stimulated A431 cells is blocked by three chemically distinct classes of tumor promoters. Tumor-promoting esters of the diterpene phorbol (phorbol 12-myristate 13-acetate, beta-phorbol 12,13-dibutyrate, and beta-phorbol 12,13-didecanoate), indole alkaloids (teleocidin and lyngbyatoxin A), and polyacetates ( aplysiatoxin and debromoaplysiatoxin ) all inhibited EGF-stimulated phosphorylation of the receptor. Non-tumor-promoting analogs (beta-phorbol, alpha-phorbol 12,13-didecanoate, and hydrolyzed teleocidin) had no effect on the levels of receptor phosphorylation. The ED50 values of the inhibitory effect (0.1-3 ng/ml) reflected the relative tumor-promoting abilities of these compounds in vivo. None of the tumor promoters tested significantly decreased the overall specific binding of 125I-labeled EGF to A431 cells. Scatchard analysis, however, revealed two apparent EGF receptors in this cell type. The dose-responses for tumor-promoter inhibition of EGF receptor tyrosine phosphorylation and high-affinity EGF binding were similar, suggesting that the same initial event is responsible for both effects. This demonstrates a correlation between modulation of EGF receptor binding and phosphorylation of tyrosine by tumor promoters. The data suggest a possible role for protein kinase C, the putative cellular receptor for these tumor promoters, in the mechanism of action.
Potential use of biaromatic L-phenylalanyl derivatives as therapeutic agents in the treatment of sickle cell disease.
Authors: Votano J R , Altman J , Wilchek M , Gorecki M , Rich A
Published: May 2006

N-Phenylacetyl-L-phenylalanine (PAP) and L-phenylalanyl-3-aminopyridine ( PAPA ) are biaromatic agents with properties that make them suitable candidates for the development of a useful therapeutic agent for the treatment of sickle cell disease. PAP and PAPA are taken up by the erythrocyte to give intra-/extracellular concentration ratios of 2.2 and 1.5, respectively, after a 2-hr exposure period. The intracellular buildup of PAP and PAPA produces moderate decreases in the mean corpuscular hemoglobin concentration (MCHC) of 6 and 10%, respectively, at 3 mM and a further decline in MCHC with increased concentration. Both PAP and PAPA increase the deoxy-Hb S solubility, CS. If the solubility in the absence of the agent is COS, PAP and PAPA have CS/COS values of 1.21 and 1.14 at 20 mM, respectively, compared with a value of 1.06 for L-phenylalanine itself. Filterability assays of partially dexygenated homozygous sickle cells shows an increase in cell flexibility of 7 to 16 times more than that of untreated cells when these agents are present at 3-6 mM. These results are largely due to the reduction in the Hb S polymer content of the treated cells. At 3 mM or less, both PAP and PAPA delay the onset of gelation in reversible sickle cells for time periods that are likely to be therapeutically useful.
A natural representation of the Fischer-Griess Monster with the modular function
J
as character
Authors: Frenkel I. B. , Lepowsky J. , Meurman A.
Published: May 2006

We announce the construction of an irreducible graded module
V
for an “affine” commutative nonassociative algebra [unk]. This algebra is an “affinization” of a slight variant [unk] of the commutative nonassociative algebra
B
defined by Griess in his construction of the Monster sporadic group
F
1
. The character of
V
is given by the modular function
J
(
q
) =
q
-1
+ 0 + 196884
q
+.... We obtain a natural action of the Monster on
V
compatible with the action of [unk], thus conceptually explaining a major part of the numerical observations known as Monstrous Moonshine. Our construction starts from ideas in the theory of the basic representations of affine Lie algebras and develops further the calculus of vertex operators. In particular, the homogeneous and principal representations of the simplest affine Lie algebra
A
1
(l)
and the relation between them play an important role in our construction. As a corollary we deduce Griess's results, obtained previously by direct calculation, about the algebra structure of
B
and the action of
F
1
on it. In this work, the Monster, a finite group, is defined and studied by means of a canonical infinite-dimensional representation.
DNA and RNA virus species are inhibited by xanthates, a class of antiviral compounds with unique properties.
Authors: Sauer G , Amtmann E , Melber K , Knapp A , Müller K , Hummel K , Scherm A
Published: May 2006

Various DNA and RNA virus species are inhibited by xanthate compounds at concentrations that leave the mitotic activity of uninfected cells unimpaired. The concentration of tricyclodecan -9-yl- xanthogenate that reduces the yield of herpes simplex virus types 1 and 2 by 50% is between 4.5 and 33 microM. The replication of DNA viruses such as simian virus 40 can be blocked at the DNA and RNA level both early and late after infection. The xanthates are not incorporated into nucleic acids. Episomal bovine papilloma virus DNA replication and transcription are also inhibited in transformed cells. The treated cells revert to the normal phenotype by acquisition of contact inhibition and a flat morphology.
Generation of antibody activity from immunoglobulin polypeptide chains produced in Escherichia coli.
Authors: Cabilly S , Riggs A D , Pande H , Shively J E , Holmes W E , Rey M , Perry L J , Wetzel R , Heyneker H L
Published: May 2006

Plasmids have been constructed that direct the synthesis in Escherichia coli of heavy chains and/or light chains of an anti-carcinoembryonic antigen (CEA) antibody. Another plasmid was constructed for expression of a truncated form of heavy chain (Fd' fragment) in E. coli. Functional CEA-binding activity was obtained by in vitro reconstitution in E. coli extracts of heavy chain or Fd' fragment mixed with extracts containing light chain.
Mechanism of insertion of diphtheria toxin: peptide entry and pore size determinations.
Authors: Zalman L S , Wisnieski B J
Published: May 2006

Diphtheria toxin ( DTx ) is an extremely potent inhibitor of protein synthesis. It is secreted as a linear polypeptide, which is cleaved to produce disulfide-linked A and B fragments. Fragment A, the inhibitor of protein synthesis, requires fragment B, the recognition subunit, for entry into intact cells. Fragment B has been proposed to form a transmembrane channel through which A gains access to the cytosol. If it were demonstrated that the B subunit had an exclusive association with membrane lipid acyl chains, this might indicate that A is secluded in a proteinaceous B channel. However, our results from intramembranous photolabeling studies show that both subunits of DTx enter the hydrocarbon domain of the bilayer. Toxin cleavage is not required for penetration. Decreasing pH leads to increased binding and hence indirectly to increased penetration. Parallel permeability studies indicate that cleaved DTx does indeed form pores (24 A in diameter) and they are larger than those previously reported (5 A) with native toxin. The data suggest that these are dimeric structures. Cleaved DTx is much more effective than intact DTx at pore formation. Thus, we conclude that, while pore formation is a feature of toxin-membrane interaction, the pore structure does not protect A from contact with lipid side chains and may in fact consist of both the A and B domains in a dimeric configuration, (AB)2.
Extensive homology exists between Marek disease herpesvirus and its vaccine virus, herpesvirus of turkeys.
Authors: Gibbs C P , Nazerian K , Velicer L F , Kung H J
Published: May 2006

Marek disease is a lymphomatous disease of chickens caused by infection of a herpesvirus, Marek disease virus (MDV). Marek disease is the only neoplastic disease for which a successful vaccine has been developed. The vaccine virus, herpesvirus of turkeys (HVT), is non-oncogenic in chickens. Despite the strong antigenic relationship between these viruses, previous studies showed that the two viral DNAs share little or no homology. Using less stringent hybridization conditions and methods that greatly improve the reassociation kinetics, we have reexamined the sequence homology between MDV and HVT DNA. We report here that HVT and MDV are far more closely related than previously reported. The homology between these two viral DNAs ranges between 70% and 80% at the nucleotide level and appears to extend over 90-95% of the respective genomes. Under the low stringency conditions used, MDV DNA fails to cross-hybridize with DNA from feline rhinotracheitis virus, an antigenically unrelated herpesvirus with a G-C content identical to that of MDV and HVT.
Conformational analysis of 6-cis- and 6-trans-leukotriene B4-calcium complexes.
Authors: Brasseur R , Deleers M
Published: May 2006

We present a computational description of the conformation of a pair of two isomeric molecules (6-cis-and 6-trans-leukotriene B4) forming a complex with one calcium ion. Our theoretical prediction of the membrane-water interface conformation and of the bulk lipid phase conformation of the two different isomeric complexes are in excellent agreement with experimental data on the leukotriene-mediated calcium ionophoresis in liposomes. The two isomers lead to vastly different conformations in the presence of Ca2+, and the most probable conformation of the permeant species shows a globular conformation able to cross a lipid membrane.