bluuebunny/crossref_metadata_2025 huggingface dataset:
Association of salmonella mutants with germfree rats: site specific model to detect carcinogens as mutagens.
Authors: Wheeler L A , Carter J H , Soderberg F B , Goldman P
Published: May 2006

An association of the histidine auxotroph of Salmonella typhimurium (strain TA1538) within the gastrointestinal tract of otherwise germfree Sprague-Dawley rats is maintained during periods of observation lasting as long as 7 months. The bacteria are found at levels exceeding 10(7) per g in the forestomach and at levels greater than 10(8) per g in the lower bowel and in the feces. Only approximately 10(4) bacteria per g are found in the posterior stomach and in the upper small intestine. The association of the salmonella mutants is maintained when the bacterial association is increased by the addition of other bacteria characteristic of the gastrointestinal flora. Carcinogenic amines, which cause strain TA1538 to revert to histidine independence in Ames' in vitro assays, increase the number of revertants in the feces when fed to the salmonella-associated rats. In contrast, the number of revertants in the feces does not increase when the rats are fed structurally related compounds which are not mutagenic to the bacteria in vitro and for which no evidence of carcinogenicity exists. Sacrifice of rats after feeding the carcinogen 2-nitrofluorene indicates that the number of revertants is increased in the cecum and colon as well as in the feces. The apparent proximity of the bacterial mutagenic response to the location of the tumor response in the colon suggests that the rat associated with the histidine auxotroph may provide a useful model for further investigation of the possible association between bacterial mutagenesis and carcinogenesis within the gastrointestinal tract. In addition, with this model it may be possible to evaluate selectively the effects of various constituents of the flora on the activation of compounds provoking the revertant response.
Isolation and characterization of two acidic proteins of 60s ribosomes from Artemia salina cysts.
Authors: Möller W , Slobin L I , Amons R , Richter D
Published: May 2006

60S ribosomes from encysted gastrulae of the brine shrimp Artemia salina contain two acidic proteins, which are homologous to the Escherichia coli proteins L7 and L12. The proteins were purified and characterized with respect to molecular weight, amino-acid composition, peptide maps, and their functional requirement in the elongation factor dependent binding of aminoacyl transfer RNA to the ribosome.
Architecture of the Escherichia coli ribosome as determined by immune electron microscopy.
Authors: Tischendorf G W , Zeichhardt H , Stöffler G
Published: May 2006

Binding sites for antibodies specific to nineteen of the twenty-one ribosomal proteins from the 30S subunit of E. coli ribosomes have been localized on the surface of the 30S ribosomal subunit by immune electron microscopy. The locations of 13 ribosomal proteins from the 50S subunit were similarily assessed. The arrangement of these proteins is illustrated in three-dimensional models of the 30S and 50S ribosomal subunits and of 70S ribosomes. With specific antibodies to six proteins of the 30S subunit we found only one attachment point for each protein. Antibodies against each of nine of the proteins attached at two separate sites that were separated by various distances. Four further proteins were exposed at three or four sites for antibody binding. Altogether eight to ten of the 19 proteins of the 30S subunit have shown antibody attachment sites at remote points on the surface of the ribosome, at distances which are incompatible with globular shapes; these proteins must therefore have elongated or fibrous structures within the ribosome. On the other hand, only two proteins of the 50S subunit, namely L11 and L18, have so far revealed two separated antibody binding sites; proteins L7/L12 occurred, however, at multiple sites.
Effect of rotation on the diffusion-controlled rate of ligand-protein association.
Authors: Hill T L
Published: May 2006

The rate of binding a fairly large ligand molecule to a protein is reduced below the usual diffusion-controlled rate by the requirement of a certain rotational orientation. A simple, approximate treatment of this effect is given for special cases of spherical and ellipsoidal ligands. As the center of an ellipsoidal ligand approaches a protein surface, there is an effective repulsive potential between ligand and surface owning to restricted rotation of the ligand. The frequency factor kT/h of the Eyring rate theory is replaced in these reactions involving diffusion in solution by D/Rlambda, where D = diffusion coefficient of ligand, lambda = thermal deBroglie wavelength of ligand, and R = "capture" distance around the binding site on the protein.
Functional interactions of viral and histocompatibility antigens at tumor cell surfaces.
Authors: Schrader J W , Cunningham B A , Edelman G M
Published: May 2006

Several lines of evidence are presented to suggest that histocompatibility antigens can be physically associated on the cell surface with viral antigens and possibly other foreign antigens. The lysis of the murine tumor cells EL4 and P388 by syngeneic cytotoxic lymphocytes was inhibited by antisera directed against the H-2 antigens on the tumor cells, consistent with the hypothesis that H-2 antigens are part of the target of the cytotoxic lymphocytes. Moreover, it was found that patching and capping of the H-2 antigens on EL4 cells resulted in the co-patching and co-capping of viral antigens as detected by antisera against Rauscher leukemia virus. Capping of H-2 antigens also resulted in co-capping of determinants detected by an antiserum to the viral protein gp69/71. On the basis of these and other observations, we propose the hypothesis that the H-2 molecules serve as adaptors that combine with viral antigens on the cell surface to form hybrid antigens containing elements of self (H-2) and non-self (virus). The adaptor-antigen complex may then be recognized by a subclass of thymus-derived (T) lymphocytes that possesses a repertoire of receptors directed against hybirds of foreign and H-2 antigens. This raises the possibility that other products of the major histocompatibility complex may have analogous functions.
Specific human B lymphocyte alloantigens linked to HL-A.
Authors: Mann D L , Abelson L , Henkart P , Harris S D , Amos D B
Published: May 2006

Sera, previously found to react specifically with B lymphoid cultured cells, were tested on isolated T and B peripheral blood lymphocytes in a microcytotoxicity assay. Studies were performed on lymphocytes obtained from several large Amish families. The sera used in these studies were cytotoxic to peripheral blood, B lymphocytes, but not cytotoxic to T lymphocytes. The antigens detected followed the inheritance pattern of HL-A haplotypes. The strong linkage disequilibrium with HL-A antigens suggests that genes controlling the expression of B lymphocyte antigens are linked to genes controlling HL-A alloantigens.
Alterations in envelope structure of heptose-deficient mutants of Escherichia coli as revealed by freeze-etching.
Authors: Bayer M E , Koplow J , Goldfine H
Published: May 2006

The surface of freeze-etched E. coli strain GR467, a heptose-deficient ("deep rough") mutant derived from CR34, was studied by electron microscopy. The outer membrane of GR467 has an increased ratio of phospholipid to protein, mainly due to a decreased protein content. Freeze-etched CR34 showed structural features indistinguishable for wild-type E. coli, i.e., the primary cleavage occurring in the inner membrane with only minor appearance of cleavage within the outer membrane. In contrast to this, in mutant GR467 most of the freeze-cleavages had taken place along a new plane, presumably in a hydrophobic region of the outer membrane. In this cleavage plane numerous particles were seen. Often the cleavage extended over the entire exposed cell surface; occasionally only a few large plateaus were visible, around which the next deeper cleavage plane, that of the protoplasmic or inner membrane, was discernible. Two spontaneous revertants (R11 and R16) with protein and lipid A levels similar to wild-type cells showed mostly freeze fractures with wild-type characteristics, and only a few cells had retained fracturing properties of GR467. A partial revertant revealed intermediate characteristics. Thus, there appears to be a morphological correlation with the chemical data relating the amount of outer membrane protein with the heptose content of the lipopolysaccharide.
A novel murine oncornavirus with dual eco- and xenotropic properties.
Authors: Fischinger P J , Nomura S , Bolognesi D P
Published: May 2006

Infection of Swiss mouse 3T3FL cells with a clonal isolate of Moloney leukemia virus (MLV-IC) resulted in virus progeny composed of at least three different murine helper oncornaviruses. Each entity was purified in appropriate cells by several sequential terminal dilution isolations and was grouwn to high titers. Besides ecotropic MLV-IC there was a pure xenotropic virus and a third novel virus with properties of both eco- and xenotropic viruses. The purified xenotropic virus had a wide host range, was restricted in mouse cells, and was inactivated by normal mouse sera like other xenotropic isolates. The purified virus with hybrid properties (HIX) could infect a wide range of mammalian cells, which included both N and B mouse cells. HIX gave single-hit titrations with equal titers on both mouse and cat indicator cells. Envelope properties of HIX were examined by virus preinfection interference, by interference involving viral glycoprotein, and by neutralization with specific antisera. Both xenotropic and MLV-IC type ecotropic determinants were found on the virus coat. The origins of HIX and the xenotropic virus were investigated in detail. The original MLV-IC stock had HIX type virus in low titer but no detectable pure xenotropic virus. Infection of mouse cells with a single infectious unit of the ecotropic virus from the MLV-IC virus stocks could at times give rise to HIX type virus. HIX type virus, passed once through heterologous rat cells, was subjected to long-term passage either in infected mouse or cat cells. After several months HIX type virus disappeared from some mouse and cat cell systems. The possible hybrid nature of HIX and the origins of newly appearing xenotropic viruses are discussed.
Biosynthesis of ribosylthymine in the transfer RNA of Streptococcus faecalis: a folate-dependent methylation not involving S-adenosylmethionine.
Authors: Delk A S , Rabinowitz J C
Published: May 2006

Ribosylthymine is not present in tRNA of Streptococcus faecalis grown in the absence of folic acid, although this methylated residue does occur in the tRNA of this organism when it is grown in the presence of folate. We have found that, unlike other methylated residues in RNA of S. faecalis and other organisms, the methyl moiety of ribosylthymine of the tRNA of folate-sufficient S. faecalis is not derived from S-adenosylmethionine. Preliminary evidence suggests that a folate derivative serves as the methyl donor in this methylation.
Regulation of hepatic regeneration in rats by synergistic action of insulin and glucagon.
Authors: Bucher M L , Swaffield M N
Published: May 2006

Rats were subjected to resection of the gastrointestinal tract, pancreas, and spleen, and maintained by c-ntinuous intravenous infusion. Such animals, receiving only electrolytes and glucose, and deprived of a portal blood supply, responded to 68% hepatectomy with a significant rise in hepatic DNA synthesis, which was, however, greatly delayed and diminished compared to normal controls. The activity was restored to normal by infusion of insulin and glucagon (supplemented with glucose and amino acids), but not by either hormone alone; it was not decreased by delaying the start of hormone treatment for 6-7 hr after partial hepatectomy. Infusion of insulin and glucagon into a small series of rats with intact livers did not appreciably elevate DNA synthesis. In normal rats partial hepatectomy is followed by an abrupt fall in portal vein insulin concentration. These findings are all consistent with the suggestion that agents other than insulin and glucagon may serve to initiate hepatic regeneration, but that these two hormones acting synergistically are major regulators of the rate and perhaps also the extent of the regenerative process.
RNA synthesis in isolated brian nuclei after administration of d-lysergic acid diethylamide (LSD) in vivo.
Authors: Brown I R
Published: May 2006

RNA synthesis in isolated brain nuclei was analyzed 2.5 hr after the intravenous administration of d-lysergic acid diethylamide (LSD) to young rabbits. The drug stimulated transcription by 54% in brain stem nuclei and by 13% in cerebral hemisphere nuclei expressed over saline controls. Both nucleoplasmic and nucleolar RNA synthesis were increased. The main activity in the isolated nuclei assay was due to nucleoplasmic RNA polymerase, since alpha-amanitin reduced synthesis by over 70% in either drug or control treatments.
Structure-function relationships and mode of replication of animal rhabdoviruses.
Authors: Sokol F , Koprowski H
Published: May 2006

Recently accumulated knowledge allows more precise comparison of the structural (and possibly evolutionary) relationships of several different animal rhabdoviruses: vesicular stomatitis virus, rabies virus, Kern Canyon virus, and spring viremia of carp virus. Each virus is composed primarily of a glycoprotein, an RNA-associated nucleoprotein, and one or two membrane proteins. Vesicular stomatitis virus group viruses contain lesser amounts of two additional distinct polypeptides, NS and L. The separate viruses undergo structural polypeptide phosphorylation in vivo according to characteristic patterns. In vesicular stomatitis virus the NS protein is selectively phosphorylated. In rabies group viruses and in spring viremia of carp virus, the nucleoprotein is the predominant phosphoprotein; in these viruses only the phosphorylated moiety is selectively cleaved off with trypsin. In Kern Canyon virus, only membrane protein and glycoprotein are weakly phosphorylated. Each virus possesses a virion-bound protein kinase. Vesicular stomatitis virus group viruses, Kern Canyon virus, and spring viremia of carp virus only contain virion-bound transcriptases of respectively decreasing levels of activity demonstrable in vitro. Vesicular stomatitis and Kern Canyon viruses replicate efficiently in enucleated cells; rabies virus does not. Based upon these observations, it is suggested that vesicular stomatitis virus may represent the most highly evolved of these rhabdoviruses, whereas spring viremia of carp and Kern Canyon viruses may represent "evolutionary links" between the vesicular stomatitis and rabies virus groups.
Patients with Ehlers-Danlos syndrome type IV lack type III collagen.
Authors: Pope F M , Martin G R , Lichtenstein J R , Penttinen R , Gerson B , Rowe D W , McKusick V A
Published: May 2006

One of the genetically distinct collagens (type III) normally found in skin, aorta, and intestine is missing from the tissues of patients with the Ehlers-Danlos syndrome type IV. While skin fibroblasts from other individuals synthesize both types I and III collagen. Ehlers-Danlos syndrome IV cells synthesize only type I. These results suggest that the fragile skin, blood vessels, and intestines of Ehlers-Danlos syndrome IV patients result from an absence of type III collagen.
Chemical nature of bioluminescence systems in coelenterates.
Authors: Shimomura O , Johnson F H
Published: May 2006

Analysis of substances involved in light-emitting reactions among bioluminescent coelenterates has revealed a pronounced uniformity in the structural features of initial reactants, i.e., "luciferins" and photo-protein chromophores, as well as the light-emitter product. This product is structurally identical among the different classes of coelenterates: Hydrozoa (the jellyfish, Aequorea), Anthozoa (the sea cactus, Cavernularia; sea pansy, Renilla; and sea pen, Leioptilus), and very likely also the Scyphozoa (the jellyfish, Pelagia). In each of these instances the reaction product, namely, 2-(p-hydroxy-pnenylacetyl)amino-3-benzyl-5-(p-hydroxyphenyl) pyrazine, is the actual light-emitter, whether it occurs in a Ca2+-triggered photoprotein type of luminescence, or in a "luciferin-luciferase" type. The evidence indicates that in certain coelenterates, e.g., Cavernularia, these two types are equally significant, whereas in others (Renilla and Leioptilus) the "luciferin-luciferase" type predominates over the Ca-triggerable photoprotein type, and finally that only the photoprotein type functions in the luciferaseless jellyfish, Aequorea. In all instances investigated, the structure of the light-emitter prior to the luminescence reaction appears to be essentially the same as that of the chromophore of unreacted aequorin. The product of the luminescence reaction is absent in extracts of non luminous species. However, a product very similar to that of luminescent coelenterates occurs also in representatives of other phyla, including the cephalopod molluscs, e.g., the "firefly squid" Watasenia and probably various ctenophores as well.
Conjugated polyene fatty acids as membrane probes: preliminary characterization.
Authors: Sklar L A , Hudson B S , Simoni R D
Published: May 2006

The use of fluorescent conjugated polyenoic fatty acids as probes of membrane structure is introduced. alpha- and beta-parinaric acid (cis, trans, trans, cis-, and all trans-9,11,13,15-octadecatetraenoic acid) and synthetic lecithins containing an alpha-parinaric acid chain in position 2 are prepared and their absorption and fluorescence properties are determined. Phase transitions are detected as fluorescence changes at characteristic temperatures when either the free fatty acid probes or the labeled phospholipid probe are included in sonicated aqueous dispersions of L-alpha-dimyristoyl lecithin, L-alpha dipalmitoyl lecithin, or L-alpha-distearoyl lecithin. The phase transitions are detected at about 23 degrees C (dimyristoyl), 44 degrees C (dipalmitoyl), and 53 degrees C (distearoyl lecithin). Binding of alpha-parinaric acid to bovine serum albumin is measured by shifts in the absorption spectrum and enhanced quantum yield of the fatty acid upon binding and by energy transfer between 2 tryptophyl residues in bovine serum albumin and alpha-parinaric acid. Approximately six binding sites are detected. Other applications of these probe molecules, including phase transitions of phospholipid/cholesterol dispersions, induced circular dichroism of parinaric acid bound to albumin, and biosynthetic incorporation of parinaric acid into biological membranes, are discussed.
Distribution of end-to-end distances of oligopeptides in solution as estimated by energy transfer.
Authors: Haas E , Wilchek M , Katchalski-Katzir E , Steinberg I Z
Published: May 2006

A homologous series of oligopeptides each containing at its ends a donor and an acceptor of electronic excitation energy was synthesized by the solid-phase method. N-5-(2-Hydroxyethyl)-L-glutamine was the repeating unit, and peptides containing 4, 5, 6, 7, 8, and 9 of these amino-acid residues were prepared. The chromophores naphthalene and dansyl, which were used as donor and acceptor, respectively, fulfil the conditions necessary for energy transfer according to the Förster mechanism. A distance corresponding to 50% efficiency of energy transfer, tro = 22 plus or minus 1 A, was calculated. The kinetics of fluorescence decay of an oligomer containing the naphthalene chromophore only could be described precisely by a monoexponential function. In contrast, the kinetics of the decay curves of the fluorescence of the donor of all of the oligomers containing both donor and acceptor, as measured in viscous solution, deviated markedly from monoexponential behavior. The deviation was interpreted in terms of the great number of different conformations that the various molecules of each of the oligomers attain in solution, leading to characteristic end-to-end distribution functions between the donor and acceptor. Numerical adjustment of the parameters of some of the previously proposed expressions to describe the end-to-end distribution enabled the reconstruction of the kinetics of the fluorescence decay of the donor with great precision. The end-to-end distribution functions for the various oligopeptides were thus evaluated.
Folding of the DNA double helix in chromatin-like structures from simian virus 40.
Authors: Germond J E , Hirt B , Oudet P , Gross-Bellark M , Chambon P
Published: May 2006

Relaxed circular, covalently closed simian virus 40 DNA molecules were associated with the four histones that are present in virions. In electron micrographs the resulting complexes appear twisted, with globular structures (nucleosomes) along the DNA. Incubation with an untwisting extract converts the twisted complexes to relaxed structures. Extraction of the DNA from the relaxed complexes yields supercoiled molecules. The number of superhelical turns in these molecules corresponds to the number of nucleosomes per DNA molecule in the complexes.
Interrelationship between poly (ADP-Rib) synthesis, intracellular NAD levels, and muscle or cartilage differentiation from mesodermal cells of embryonic chick limb.
Authors: Caplan A I , Rosenberg M J
Published: May 2006

Mesodermal cells of embryonic chick limbs have the capacity to differentiate into either muscle or cartilage. Previous reports from this laboratory show a correlation between pyridine nucleotide levels and this differentiation, and thus suggest that fluctuations in the cellular NAD levels play a role in the control of muscle versus cartilage development. Poly(ADP-Rib) is chromatin-associated and forms from the polymerization of NAD with the excision of nicotinamide. The studies reported here show that: (A) the rate of net synthesis of poly(ADP-Rib) is correlated with the differentiation of chondrogenic cells from stage 24 limb mesodermal cells grown in vitro; (B) inhibition of chondrogenic expression caused by exposure to nicotinamide or BrdUrd is correlated with maintenance of basal levels of poly(ADP-Rib) synthesis, and this inhibition is dependent on the concentration of nicotinamide or BrdUrd exogenously supplied; (C) potentiation of chondrogenic expression caused by exposure of limb mesodermal cells in vitro to 3-acetylpyridine is correlated with stimulation of the rate of poly(ADP-Rib) synthesis if corrected for the specific activity of the ATP pool or compared to untreated cultures on a per cell basis; (D) isolated chromatin from mesodermal cells has the enzymatic machinery for synthesizing poly(ADP-Rib); (E) this machinery is inhibited by nicotinamide, thymidine, and 3-acetylpyridine; and (F) newly synthesized poly(ADP-Rib) is either associated with a discrete fraction of chromatin or is completely extracted from chromatin by the high column salts, which result in an aggregation with eventual elution at the exclusion volume position of the agarose column. Taken together, these observations provide a possible explanation for how fluctuations in cellular NAD levels can communicate with or be "sensed" by genomic related machinery and eventually result in differtial phenotypic expression.
Isolation of a glycogen synthase I kinase that is independent of adenosine 3':5'-monophosphate.
Authors: Schlender K K , Reimann E M
Published: May 2006

Three protein kinases (ATP:protein phosphotransferase, EC 2.7.1.37) were detected when the soluble fraction of rabbit kidney medulla was chromatographed on DEAE-cellulose with a linear NaC1 gradient. The first two kinases eluted (Peak 1 and Peak II) were cyclic-AMP-dependent, wheras Peak III was cyclic-AMP-independent. A procedure was developed to separate the catalytic subunit of Peak II cyclic-AMP-dependent protein kinase (representing the bulk of the histone kinase activity) from Peak III protein kinase. In contrast to the catalytic subunit, Peak III protein kinase phosphorylated casein more rapidly than histone. Peak III was insensitive to the heat-stable protein inhibitor of cyclic-AMP-dependent protein kinases and appeared to have a higher requirement for ATP than did the catalytic subunit. Peak III catalyzed the conversion of glycogen synthase (UDPglucose:glycogen alpha-4-glucosyltransferase, EC 2.4.1.11) from the I (glucose-6-phosphate-independent) to the D (glucose-6-phosphate-dependent) form. This conversion was dependent on Mg-2+ and ATP and was unaffected by cyclic AMP, cyclic GMP, or the protein inhibitor. Glycogen synthase I in the soluble fraction of kidney medulla could be converted to the D form by endogenous glycogen synthase I kinase if Mg-2+ and ATP were added. Most of this glycogen synthase I kinase activity was unaffected by cyclic AMP or by the protein inhibitor, suggesting that Peak III may be of major importance in the regulation of glycogen synthase in vivo.
On the prokaryotic nature of red algal chloroplasts.
Authors: Bonen L , Doolittle W F
Published: May 2006

The sequences of oligonucleotides released by T1 ribonuclease digestion of 32-P-labeled 16S (chloroplast) and 18S (cytoplasmic) ribosomal RNAs from a marine species of Porphyridium (Rhodophyta) have been determined. The resultant catalogs have been compared to those obtained for three prokaryotes:Escherichia coli, Bacillus subtilis, and Anacystis nidulans (a blud-green alga). There is extensive sequence homology between the Porphyridium chloroplast 16S ribosomal RNA and each of the prokaryotic 16S ribosomal RNAs, but little homology between the Porphyridium cytoplasmic 18S ribosomal RNA and any of the 16S species. These data provide a measure of the evolutionary distance separating existing chloroplasts from contemporary bacteria and blue-green algae, and are discussed in terms of the hypothesis that these organelles evolved from endosymbiotic photosynthetic prokaryotes.
Isolation from the asian mouse Mus caroli of an endogenous type C virus related to infectious primate type C viruses.
Authors: Lieber M M , Sherr C J , Todaro G J , Benveniste R E , Callahan R , Coon H G
Published: May 2006

Treatment of a cell line derived from the Asian feral mouse Mus caroli with 5-bromodeoxyuridine induces an infectious, xentropic type C virus. This virus shares strongly cross-reactive reverse transcriptase (RNA-dependent DNA polymerase) and p30 antigens and crossinterferes with type C viruses isolated from a woolly monkey (SSAV) and gibbon apes (GALV). By similar criteria, the caroli virus is much less related to previously described type C viruses of the laboratory mouse, Mus musculus. Induction of virus from 10 of 13 single cell clones indicates that the virus is endogenous in Mus caroli cells. Thre results suggest that infectious primate type C viruses arose by trans-species infection(s) of certain primates with endogenous type C viruses from MUs caroli or a closely related Mus species.
X-ray absorption spectroscopy using synchrotron radiation for structural investigation of organometallic molecules of biological interest.
Authors: Kincaid B M , Eisenberger P , Hodgson K O , Doniach S
Published: May 2006

The technique of x-ray absorption spectroscopy using tuneable, very intense x-rays from a high energy electron storage ring has been applied to study of the estended x-ray absorption fine structure for Cu and Ni tetraphenylporphyrin and methemoglobin. Preliminary analysis shows that the spectra may be interpreted as a super-position of modulations arising from the nearest neighbor nitrogen and pyrrole alpha-carbon coordination sheels of the metal atoms. We estimate that with the observed magnitude of noise to modulation amplitude, relative shifts of 0,5% in the metal-nitrogen to metal-carbon bond distances in the prophyrins should be observable using extended x-ray absorption fine structure and that this technique may provide a method of observing these types of structural changes in solution.
An intercistronic region and ribosome-binding site in bacterial messenger RNA.
Authors: Platt T , Yanofsky C
Published: May 2006

A messenger RNA fragment about 220 nucleotides long has been isolated from 32-P-labeled tryptophan operon mRNA of Escherichia coli. When point mutations at the end of trpB and the beginning of trpA were introduced, the resulting nucleotide changes were found; hence the mRNA fragment must include the trpB-trpA intercistronic region. Most of the nucleotide sequences can be assigned to specific locations in the structural genes, based on the amino-acid sequences of the trpB and trpA proteins. In vitro, ribosomes bind to this piece of mRNA and protect from nuclease attack a region about 40 nucleotides long, containing a central AUG codon. The triplet codons to the 3' side of this AUG correspond to the first seven amino acids of the trpA protein; the codons to the 5' side correspond to the last six amino acids of the trpB protein. Translation of trpB is terminated by single UGA codon, which overlaps the trpA AUG initiation codon: UGAUG. Thus the untranslated "intercistronic" region consists of only two nucleotides. The RNA sequence spanning this region undoubtedly fulfills two functions, specifying ribosome recognition signals as well as encoding amino-acid sequences.
Cytoplasmic control of nuclear DNA synthesis during early development of Xenopus laevis: a cell-free assay.
Authors: Benbow R M , Ford C C
Published: May 2006

Nuclei isolated from nondividing cells were induced to synthesize DNA by incubation with cytoplasm from early embryos of Xenopus laevis. Numerous replication eyes were formed in the nuclear DNA molecules, and high levels of [3-H]dTTP were incorporated. With this assay a protein(s) which appears to initiate DNA synthesis was found at high levels in the cytoplasm of eggs, blastulae, or gastrulae, but only at low levels in the cytoplasm of oocytes, hatched embryos, or adult tissues.
Infinitesimal generators and quasi-units in potential theory
Authors: Arsove Maynard , Leutwiler Heinz
Published: May 2006

The underlying structure is taken as a strongly superharmonic cone [unk], defined as a partially ordered abelian semigroup with identity 0 which admits a multiplication by nonnegative scalars and satisfies two fundamental axioms of a potentialtheoretic character. In terms of a fixed nonzero element
e
there is introduced on [unk] a one-parameter family of nonlinear operators
S
λ
(λ ≥ 0) closely connected with the abstract theory of quasibounded and singular elements. The semigroup {
S
λ
} admits an infinitesimal generator
A
, and the elements invariant under
A
, called quasi-units, generalize the Yosida quasi-units in the theory of Riesz spaces. Quasi-units in [unk] are studied, both from a potentialtheoretic and a function-alanalytic viewpoint, culminating in a spectral representation theorem for quasi-bounded elements which extends the classical Freudenthal spectral theorem of Riesz space theory.
Interaction of cholera toxin and membrane GM1 ganglioside of small intestine.
Authors: Holmgren J , Lönnroth I , Månsson J , Svennerholm L
Published: May 2006

Ganglioside GM1 was isolated from the small intestinal mucosa of man, pig, and beef and amounted to 0.1, 2.0, and 43 nmol per g fresh weight, respectively. These differences in GM1 content were associated with a quantitatively differing ability of the mucosal cells to bind cholera toxin. Human cells bound about 15,000 toxin molecules when saturated with the toxin, porcine cells 120,000, and bovine cells 2,600,000 molecules. The association constant (KA) of the cholera toxin binding was, for cells of all three species, about 10(9) liters/mol. Exogenously added GM1 ganglioside was incorporated in intestinal mucosal cells as well as in intact rabbit small bowel. The increment in GM1 was associated with a correspondingly increased number of binding sites for cholera toxin, whereas KA was unchanged. GM1 incorporation increased the sensitivity of the rabbit small bowel to the diarrheogenic action of cholera toxin. Vibrio cholerae sialidase hydrolyzed isolated intestinal diand trisialogangliosides to GM1. However, the enzyme did not change the ganglioside pattern of intestinal mucosa, had very little influence on the number of toxin binding sites on intestinal cells, and did not alter the sensitivity of the small bowel to the diarrheogenic action of the toxin. These results demonstrate a relationship in the intestinal mucosa between the GM1 ganglioside concentration, the number of binding sites for cholera toxin, and the sensitivity to the biologic action of the toxin. Thus, the study strongly supports the concept that the GM1 ganglioside is the intestinal binding receptor for cholera toxin.
Release of (oligo) peptidyl-tRNA from ribosomes by erythromycin A.
Authors: Otaka T , Kaji A
Published: May 2006

Erythromycin A released peptidyl-tRNA in the in vitro polypeptide synthesis system with bacterial components programmed by synthetic polynucleotide. This is consistent with our hypothesis that erythromycin A inhibits translocation by preventing proper situation of oligopeptidyl-tRNA in the donor (D) site on ribosomes.
Follicle-stimulating hormone stimulates estradiol-17beta synthesis in cultured Sertoli cells.
Authors: Dorrington J H , Armstrong D T
Published: May 2006

Sertoli cells isolated from testes of 20-day-old rats and maintained in primary culture synthesized estradiol-17beta [1,3,5(10)-estratriene-3,17beta-diol] (measured by specific radioimmunoassay) when testosterone (17beta-hydroxy-4-androsten-3-one) 0.5 muM, was added to the culture medium. No detectable estradiol synthesis occurred when cells were incubated in medium containing pregnenolone (3beta-hydroxypregn-5-en-20-one), 0.5 muM, or containing no added steroid substrate. Follicle-stimulating hormone (FSH) (NIH-FSH-S10, 5 mug/ml) stimulated estradiol synthesis 12- to 80-fold when added to medium containing testosterone, but not when added to medium containing pregnenolone or no exogenous steroid substrate. A highly purified FSH preparation, with FSH potency 50 times that of the NIH-FSH, caused a similar stimulation at a concentration of 0.25 mug/ml of medium, whereas luteinizing hormone (NIH-LH-S18, 5 MUG/ML) Caused only marginal stimulation. Dibutyryl-adenosine 3':5' cyclic monophosphate, 0.1 mM, caused a 30-fold increase in estradiol synthesis by Sertoli cells cultured in medium containing testosterone. These studies provide direct demonstration of estradiol-17beta production by Seroli cells from normal animals, and offer evidence that the synthesis of this steroid is regulated at the level of the aromatizing enzyme system by FSH and adenosine 3':5' cyclic monophosphate.
Microfibril assembly by granules of chitin synthetase.
Authors: Ruiz-Herrera J , Sing V O , Van der Woude W J , Bartnicki-Garcia S
Published: May 2006

Purified preparations of chitin synthetase (EC 2.4.1.16; UDP-2-acetamido-2-deoxy-D-glucose:chitin 4-beta-acetamidodeoxyglucosyltransferase), capable of forming microfibrils in vitro, were isolated from yeast cells of Mucor rouxii. Chitin synthetase was obtained either by substrate-induced liberation of bound enzyme (54,000 x g pellet) or by isolation of unbound enzyme present in the 54,000 x g supernatant of a cell-free extract. Both preparations contained ellipsoidal granules from about 350 to 1000 A diameter. Many granules exhibited a marked depression. No typical unit membrane profiles appeared in thin sections of glutaraldehyde/OsO4-fixed samples. Upon incubation with substrate and activators, chitin microfibrils were produced. The microfibrils were often found intimately associated with granules. The most common configurations were: a microfibril with a granule at one end, or two microfibrils "arising" from the same granule. These findings lend support to the granule hypothesis for the elaboration of cell wall microfibrils by end-synthesis.
Bovine factor X1 (Stuart factor): amino-acid sequence of heavey chain.
Authors: Titani K , Fujikawa K , Enfield D L , Ericsson L H , Walsh K A , Neurath H
Published: May 2006

The amino-acid sequence of the heavy chain of bovine blood coagulation factor X1 (Stuart factor) isolated before and after activation has been determined. Sequence analysis was performed on fragments obtained by cleavage with cyanogen bromide and by tryptic digestion. Comparison of the complete sequence with those of other hepatic and pancreatic serine proteases demonstrates homology of the heavy chain of activated factor X1 (factor X1a) with the B chain of bovine thrombin as well as with bovine trypsin, chymotrypsins A and B, and porcine elastase. The activation peptide cleaved near the amino terminus by a protease from Russell's viper venom differs in both size and sequence from those of other serine proteases. With three exceptions, all of the residues which are important in the catalytic functions of trypsin and chymotrypsin occur in corresponding loci in the heavy chain of factor Xa. These finding suggest that the three-dimensional structure of the heavy chain is similar to that of the pancreatic serine proteases and that these enzymes have evolved from a common ancestral gene.
Brownian motion in biological membranes.
Authors: Saffman P G , Delbrück M
Published: May 2006

Brownian motion (diffusion) of particles in membranes occurs in a highly anisotropic environment. For such particles a translational mobility (independent of velocity) can be defined if the viscosity of the liquid embedding the membrane is taken into account. The results of a model calculation are presented. They suggest that for a realistic situation translational diffusion should be about four times faster in relation to rotational diffusion than in the isotropic case.
The ovary as a source of alpha-ecdysone in an adult mosquito.
Authors: Hagedorn H H , O'Connor J D , Fuchs M S , Sage B , Schlaeger D A , Bohm M K
Published: May 2006

The ovaries of the mosquito Aedes aegypti cultured in vitro secrete material that behaves like ecdysone in a radioimmunoassay. The material was identified as alpha-ecdysone by high-resolution liquid and gas-liquid chromatography. Secretion reached a maximum 16 hr after a blood meal as shown by bioassay and direct determination. Ovariectomy reduced the concentration of ecdysone in the adult after a blood meal. Qualitative analysis of whole-body extracts indicated beta-ecdysone to be the principal species present. Thus the ovaries appear to secrete a prohormone, alpha-ecdysone, which is converted to beta-ecdysone. Beta-ecdysone plays a significant role in stimulating egg development in the adult mosquito and may have reproductive roles in other insects.
Chinese hamster ovary cells selected for resistance to the cytotoxicity of phytohemagglutinin are deficient in a UDP-N-acetylglucosamine--glycoprotein N-acetylglucosaminyltransferase activity.
Authors: Stanley P , Narasimhan S , Siminovitch L , Schachter H
Published: May 2006

Several clones of Chinese hamster ovary cells resistant to the cytotoxicity of the phytohemagglutinin from Phaseolus vulgaris show decreased binding of 125I-labeled phytohemagglutinin and contain decreased levels of a UDP-N-acetylglucosamine--glycoprotein N-acetylglucosaminyltransferase (EC 2.4.1.51; UDP-2-acetamido-2-deoxy-D-glucose:glycoprotein 2-acetamido-2-deoxy-D-glucosyltransferase) activity when compared to wild-type cells. The decrease in transferase activity varies from 45% to 96%, depending on the exogenous acceptor used in the enzyme assay. No differences between lectin-resistant and wild-type cells were noted for several other glycosyltransferases. The absence of a particular N-acetylglucosaminyltransferase in the lectin-resistant cells apparently results in defective glycosylation of lectin-binding glycoproteins on the cell surface. A phytohemagglutinin-resistant clone which shows decreased binding of 125I-labeled phytohemagglutinin but does not exhibit the enzyme deficiency has also been isolated.
Regulation of binding properties of the nicotinic receptor protein by cholinergic ligands in membrane fragments from Torpedo marmorata.
Authors: Weber M , David-Pfeuty T , Changeux J P
Published: May 2006

Exposure of receptor-rich membrane fragments from Torpedo marmorata to carbamylcholine causes a slow (half-time of 5--10 min) and reversible change of properties of the cholinergic receptor protein manifested by a decrease of the initial rate of Naja nigricollis alpha-[3H]toxin binding in the presence of carbamylcholine. This change corresponds to a 5- to 20-fold increase of affinity for carbamylcholine. Other agonists, acetylcholine, phenyltrimethylammonium, show the same effect but not the antagonists d-tubocurarine and flaxedil. Decamethonium and hexamethonium show little, if any, agonistic effect in vitro on the same membrane fragments but cause the affinity change. This regulatory property can be lost after aging of the preparation of membrane fragments. Since the affinity increase progresses with a similar time course as the decrease of amplitude of the permeability response consecutive to agonist preincubation, it is proposed that, in the membrane at rest, the receptor protein is present under a state of low affinity for agonists and that the reversible stabilization by the agonists of a high affinity state corresponds to the "pharmacological desensitization" of the system as predicted by one of the models of Katz and Thesleff.
Nuclear coupling of 33S and the nature of free radicals in irradiated crystals of cysteine hydrochloride and N-acetyl methionine.
Authors: Hadley J H , Gordy W
Published: May 2006

The 33S hyperfine structure has been observed in the electron spin resonance of irradiated crystals of cysteine-HCl-H2O and of N-acetyl DL-methionine. An analysis of the results shows that in both substances the free radicals that are stable at room temperature are disulfide radicals, RCH2SS. A possible mechanism for formation of these radicals is proposed.
Quasi-elastic behavior of solutions of viral capsid and RNA at very low shearing stresses.
Authors: Hodgins M G , Hodgins O C , Kupke D W , Beams J W
Published: May 2006

By the application of shearing stresses on the order of 10(-3) dyne cm-2 (10(-2) muN cm-2), via the magnetic viscodensimeter, extremely high relative viscosities (greater than 500) were observed when turnip yellow mosaic virus was degraded in alkali into its capsid and RNA. The solutions, however, possessed a watery consistency at this stage and exhibited a quasi-elastic character by rotor-recoil experiments. The development of this curious behavior was concentration and temperature dependent; it was not seen less than 0.5% nor at 8 degrees, and appeared sooner at 30 degrees than at 20 degrees. The time of appearance was delayed as the pH was lowered; however, the effect was still observed when the pH was as low as 9. Whereas reversibility was demonstrated when the shearing stresses exceeded the elastic resistance [0.17 dyne cm-2 (1.7 muN CM-2)], thorough mixing usually resulted in a normal behavior of the solutions thereafter. Values for the modulus of rigidity at 20 degrees for about 1% virus concentration was less than 2 X 10(-2) dyne cm-2 rad-1 (0.2 muN cm-2 rad-1), which, while extremely small, was reproducible. A porous structure, possibly involving a capsid and RNA complex, is envisioned.
Surface marker characteristics and Epstein-Barr virus studies of two established North American Burkitt's lymphoma cell lines.
Authors: Epstein A L , Henle W , Henle G , Hewetson J F , Kaplan H S
Published: May 2006

Tumor cell lines have been established in continuous culture from two North American Burkitt's lymphomas. The SU-AmB-1 line, derived from a patient with low serum antibody titers to Epstein-Barr virus (EBV), was devoid of EBV genomes by the reaction for EBV-associated nuclear antigen (EBNA), could not be induced to express EBV antigens, and was highly refractory to EBV superinfection. Conversely, the SU-AmB-2 cell line, derived from a patient with "African type" serology, yielded a positive EBNA reaction and was readily inducible and superinfectable. Although both cell lines possessed B (bone-marrow-derived) cell characteristics, they had different surface marker patterns. It is postulated that two different classes of undifferentiated B cell lymphomas exist, one of which is positive for the presence of EBV genomes and occurs endemically in Africa and New Guinea and sporadically in other parts of the world, the other of which is EBV-negative and occurs sporadically throughout the world, including the endemic areas.
Coherent anti-Stokes Raman scattering (CARS) spectra, with resonance enhancement, of cytochrome c and vitamin B12 in dilute aqueous solution.
Authors: Nestor J , Spiro T G , Klauminzer G
Published: May 2006

Coherent anti-Stokes Raman scattering (CARS) spectra have been obtained for ferrocytochrome c and cyano cobalamin in aqueous solution at millimolar concentrations, using a pair of tunable dye lasers pumped by a pulsed nitrogen laser. Resonance enhancement was obtained by tuning the omega1 laser to the visible absorption bands of the samples. The spectral features correspond to those observed in the conventional resonance Raman spectra. It appears that CARS spectroscopy, with its advantageous fluorescence rejection, can be usefully applied to biological samples by exploiting resonance enhancement. While the background scattering from water is 10 times higher than that of benzene and other aromatic solvents, it is actually at the low end of the scale for most liquids. The anomalously low background of aromatic liquids is thought to result from competition by the unusually efficient stimulated Raman scattering which they display. Off-resonance spectra for both cobalamin and cytochrome c contain negative peaks, i.e., absorption bands in the background. These are interpreted as inverse Raman processes induced by the omega1 photons in the presence of the continuum provided by the background scattering. While both CARS and the inverse Raman effect are subject to resonance enhancement, the wavelength dependence of CARS is evidently steeper.
lac repressor: 3-fluorotyrosine substitution for nuclear magnetic resonance studies.
Authors: Lu P , Jarema M , Mosser K , Daniel W E
Published: May 2006

This paper describes the isolation of 3-fluorotyrosine-substituted lac repressor, and its 19F nuclear magnetic resonance spectrum. From the spectrum, one can conclude that for each of the four identical subunits of the repressor there are four or five surface tyrosines, two buried or internal tyrosines, and one tyrosine with an phenolic group ionized or involved in a hydrogen bond. Conditions are described that can be used for the 3-fluorotyrosine substitution of a variety of Escherichia coli proteins for 19F nuclear magnetic resonance studies.
Structural heterogeneity of mitochondrial DNA molecules within the genus Drosophila.
Authors: Fauron C M , Wolstenholme D R
Published: May 2006

We have determined by electron microscopy the molecular weight of circular mitochondrial DNA (mtDNA) molecules from 39 species representing 13 groups of five subgenera of the genus Drosophila. mtDNA molecules of all species examined, other than members of the melanogaster group, had, with one exception, molecular weights in the rather narrow range 9.90 X 10(6). The one exception was D. robusta, which had a molecular weight of 10.61 X 10(6). In contrast, mtDNA molecules from species within the melanogaster group had molecular weights covering the considerably greater range 9.92 X 10(6) to 12.35 X 10(6). Applying the electron microscope denaturation mapping technique of Inman to mtDNA molecules of eight species of the melanogaster group, we found each of them to contain a region [presumably rich in adenine and thymine (A+T)] which denatured at a specific temperature (40 degrees) at which most of the remainder of the molecule remained undenatured. The size of the A+T-rich region was constant for mtDNA molecules of a species, but varied from 0.62 X 10(6) to 3.41 X 10(6) for mtDNA molecules of different species. It was demonstrated that the differences in molecular weights of the A+T-rich regions can almost completely account for the differences in total molecular weights of the mtDNA molecules from species of the melanogaster group.