bluuebunny/crossref_metadata_2025 huggingface dataset:
Phylogenetic relationships among megabats, microbats, and primates.
Authors: Mindell D P , Dick C W , Baker R J
Published: May 2006

We present 744 nucleotide base positions from the mitochondrial 12S rRNA gene and 236 base positions from the mitochondrial cytochrome oxidase subunit I gene for a microbat, Brachyphylla cavernarum, and a megabat, Pteropus capestratus, in phylogenetic analyses with homologous DNA sequences from Homo sapiens, Mus musculus (house mouse), and Gallus gallus (chicken). We use information on evolutionary rate differences for different types of sequence change to establish phylogenetic character weights, and we consider alternative rRNA alignment strategies in finding that this mtDNA data set clearly supports bat monophyly. This result is found despite variations in outgroup used, gap coding scheme, and order of input for DNA sequences in multiple alignment bouts. These findings are congruent with morphological characters including details of wing structure as well as cladistic analyses of amino acid sequences for three globin genes and indicate that neurological similarities between megabats and primates are due to either retention of primitive characters or to convergent evolution rather than to inheritance from a common ancestor. This finding also indicates a single origin for flight among mammals.
Dominant male sterility in mice caused by insertion of a transgene.
Authors: Magram J , Bishop J M
Published: May 2006

While examining a series of transgenic mouse lines carrying the HCK protooncogene, we encountered one line in which males hemizygous for the transgene were sterile. The sterile males mated normally but failed to impregnate females. Light and electron microscopy revealed that spermatogenesis proceeds normally until nuclear condensation, which occurs but gives rise to a variety of abnormally shaped nuclei. Expression of the transgene was not detectable. Thus, the insertion itself probably caused the abnormal phenotype by disrupting a gene (or genes) important in spermatogenesis. The mutation is genetically dominant, causing an abnormal phenotype even though the sterile mice carry an ostensibly normal counterpart of the disrupted locus. The mutant phenotype is completely penetrant only in some genetic backgrounds, suggesting a modifying influence from a second locus. Junctions between the inserted transgene and adjoining cellular DNA were cloned, allowing us to confirm the heterozygous nature of the genetic disruption and to detect and associated deletion. We have designated the mutation Lvs (lacking vigorous sperm) and presume that it may define a previously undescribed locus important in spermatogenesis.
Construction of intertypic chimeric dengue viruses by substitution of structural protein genes.
Authors: Bray M , Lai C J
Published: May 2006

Dengue virus contains an 11-kilobase positive-strand RNA genome that codes for, in one open reading frame, three structural proteins (capsid, premembrane, and envelope), followed by seven nonstructural proteins. The structural protein genes of a full-length cDNA clone of type 4 dengue virus were replaced with the corresponding genes of dengue 1 or dengue 2 to create intertypic chimeric cDNA. The RNA transcripts made from these templates were infectious when transfected into permissive cells in culture. Progeny of chimeric cDNA produced apparently authentic dengue 1 or dengue 2 structural proteins, together with dengue 4 nonstructural proteins, and as a consequence exhibited type 1 or type 2 serological specificity. Both of the chimeras ultimately grew to the same titer as their type 1 or type 2 parent, but the type 2/type 4 chimera grew very slowly. This chimera also produced small plaques; in contrast, the type 1/type 4 chimera produced normal size plaques. The type 2/type 4 chimera retained the mouse neurovirulence of the dengue 2 virus, which was the source of its structural protein genes. Each of the mice inoculated intracerebrally with the chimera died, but survival time was prolonged. The retardation of replication of the type 2/type 4 chimeric virus suggests that this virus and possibly other intertypic dengue virus chimeras with similar properties should be examined for attenuation in primates and possible usefulness in a live dengue virus vaccine for humans.
Expanded distribution of mRNA for nerve growth factor, brain-derived neurotrophic factor, and neurotrophin 3 in the rat brain after colchicine treatment.
Authors: Ceccatelli S , Ernfors P , Villar M J , Persson H , Hökfelt T
Published: May 2006

The effect of intracerebroventricular injection of the mitosis inhibitor colchicine on expression of mRNA for nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin 3 was studied in the rat brain with in situ hybridization. Colchicine up-regulates mRNA for NGF and BDNF in many of the neuronal systems normally expressing these factors. In addition, after colchicine treatment NGF and BDNF mRNAs were localized in several brain areas where they normally cannot be detected. Thus, NGF mRNA was present, for example, in many motor nuclei and in the basal forebrain, and BDNF mRNA was seen in many nuclei in the brain stem and in catecholamine neurons, including dopamine neurons in the substantia nigra. The latter neurons have recently been shown to be sensitive to BDNF, and the present results show that these neurons can produce this factor themselves. A decrease in mRNA for BDNF and neurotrophin 3 was seen only in the granular-cell layer of the hippocampal formation. A strong hybridization signal for BDNF and neurotrophin 3 mRNA was also observed over several myelinated tracts in treated rats, supporting the hypothesis that glial cells as well as neurons can produce these trophic factors.
Geometric interpretation of vertex operator algebras.
Authors: Huang Y Z
Published: May 2006

In this paper, Vafa's approach to the formulation of conformal field theories is combined with the formal calculus developed in Frenkel, Lepowsky, and Meurman's work on the vertex operator construction of the Monster to give a geometric definition of vertex operator algebras. The main result announced is the equivalence between this definition and the algebraic one in the sense that the categories determined by these definitions are isomorphic.
Expression and assembly of spectrally active recombinant holophytochrome.
Authors: Wahleithner J A , Li L M , Lagarias J C
Published: May 2006

To develop an in vitro phytochrome assembly system, we have expressed an oat phytochrome cDNA in both the yeast Saccharomyces cerevisiae and the bacterium Escherichia coli. Analysis of soluble protein extracts showed that the recombinant apophytochromes were full-length and capable of covalently attaching the phytochrome chromophore analogue phycocyanobilin. Difference spectra indicated that in vitro-assembled holophytochrome species were photoreversible; however, maxima and minima difference absorption values were blue-shifted relative to those of the native photoreceptor. Extracts containing the recombinant apophytochromes were also incubated with phytochromobilin, the natural chromophore synthesized from biliverdin by cucumber etioplast preparations. In these experiments, the difference spectrum obtained was identical to that of native oat holophytochrome. These results suggest that the recombinant apophytochromes adopt a structure similar to that of the apoprotein biosynthesized in vivo. ELISAs were used to quantitate phytochrome expression levels in both yeast and E. coli extracts. These measurements show that 62-75% of the phytochrome apoprotein in the soluble protein extract was competent to assemble with bilins to form spectrally active holophytochrome.
The pharmacokinetics of, and humoral responses to, antigen delivered by microencapsulated liposomes.
Authors: Cohen S , Bernstein H , Hewes C , Chow M , Langer R
Published: May 2006

The feasibility of creating a s.c. depot for sustained protein delivery with the goal of enhancing antigen immunogenicity was investigated. The depot was designed as antigen-laden liposomes of hydrogenated egg phosphatidylcholine and cholesterol (1:1 molar ratio) encapsulated in alginate-poly(L-lysine) microcapsules and evaluated using iodinated bovine serum albumin (BSA) as a model antigen. The in vivo release behavior of the liposomes and microencapsulated liposomes (MELs) was evaluated from the BSA serum concentration profiles after s.c. injection into rats and the pharmacokinetic parameters of 125I-labeled BSA appearance after s.c. or i.v. injections of BSA in saline. Maximal BSA concentrations were detected 11 h after s.c. injection in all rats. The BSA serum concentrations decreased rapidly in rats injected with BSA in saline or Freund's adjuvant and less rapidly in rats injected with BSA in liposomes or MELs. Four to 5 weeks after injection, BSA-associated radioactivity was detected only in sera of rats injected with BSA in liposomes or MELs. Fifty days after injection, 50% of the originally injected BSA was recovered form the s.c. sites of rats injected with BSA in MELs; no radioactivity was recovered from the other three groups of rats. The antigen-reactive antibody levels induced in rats immunized with BSA in MELs were 2- to 3-fold higher than those obtained in rats immunized with BSA in liposomes, saline, or Freund's adjuvant. More significantly, high antibody levels were maintained for more than 150 days after a single injection of BSA in MELs, suggesting that MELs can serve as a long-term single-dose immunization vehicle.
Transcription factor requirement for multiple rounds of initiation by human RNA polymerase II.
Authors: Szentirmay M N , Sawadogo M
Published: May 2006

We have investigated conditions that allow multiple rounds of transcription initiation from the adenovirus major late promoter in an in vitro system derived from HeLa cell nuclear extracts. Templates containing guanine-free cassettes provided a direct assay for discriminating between reinitiated transcripts and transcripts generated by a first-round of transcription initiations. When reactions were reconstituted with the previously characterized class II transcription factors (TFIIA, TFIIB, TFIID, TFIIE/F), transcription by human RNA polymerase II from the adenovirus major late promoter was essentially restricted to a single round of initiations. Reinitiations at previously transcribed major late templates required an additional activity, designated reinitiation transcription factor (RTF). The RTF activity could be separated from the required transcription initiation factors. Semipurified human RTF also promoted transcription reinitiations at minimal promoters derived from the human c-myc, histone H4, and heat shock 70-kDa protein genes, indicating that the same reinitiation factor may be utilized by many, if not all, genes. The possible role of RTF in regulating the transcription rate of various class II genes is discussed.
A plausible model for reversal of neoplastic transformations in plants based on multiple steady states.
Authors: Hervagault J F , Ortoleva P J , Ross J
Published: May 2006

We offer a plausible interpretation of some experiments on the reversal of neoplastic transformations in plants. We suggest that normal cells and tumorous cells represent multiple stable-steady states corresponding to a reaction feedback mechanism. The (autocatalytic) feedback loop is constructed from observations on the role played by myo-inositol: it increases the permeability of ions through the membrane and the biosynthetic pathway to myo-inositol is activated by ions. Provided that the permeabilities of nutrients (sugars and salts) are a product-enhanced function of myo-inositol, then we have a (oversimplified) model that can exhibit multiple stationary stable states, one or two depending on the exogenous nutrients and myo-inositol concentrations, and reversible and irreversible transitions from one of these states to the other are possible. From this model, straightforward simple experiments are suggested. We also propose that recent models dealing with the intracellular calcium regulation by hormones, where one key step requires the hydrolysis of inositol phospholipids, take into account free myo-inositol and endogenous hormone concentrations (e.g., auxins).
DNA looping between the origin of replication of Epstein-Barr virus and its enhancer site: stabilization of an origin complex with Epstein-Barr nuclear antigen 1.
Authors: Su W , Middleton T , Sugden B , Echols H
Published: May 2006

Epstein-Barr nuclear antigen 1 (EBNA-1) is the only viral protein required to support replication of Epstein-Barr virus during the latent phase of its life cycle. The DNA segment required for latent replication, oriP, contains two essential binding regions for EBNA-1, termed FR and DS, that are separated by 1 kilobase pair. The FR site appears to function as a replicational enhancer providing for the start of replication at the DS site. We have used electron microscopy to visualize the interaction of EBNA-1 with its binding sites and to study the mechanism for communication between the FR and DS sites. We have found that DNA-bound EBNA-1 forms a DNA loop between the FR and DS sites. From these results, we suggest that EBNA-1 bound to the replicational enhancer acts by a DNA-looping mechanism to facilitate the initiation of DNA replication. Occupancy of the DS site alone is highly sensitive to competition with nonspecific DNA. In contrast, occupancy of the DS site by looping from FR is largely resistant to the competitor DNA. These experiments support the concept that enhancers act in cis from nearby sites to provide a high local concentration of regulatory proteins at their target sites and to stabilize regulatory interactions.
Magnetic field tomography of coherent thalamocortical 40-Hz oscillations in humans.
Authors: Ribary U , Ioannides A A , Singh K D , Hasson R , Bolton J P , Lado F , Mogilner A , Llinás R
Published: May 2006

This paper introduces the use of magnetic field tomography (MFT), a noninvasive technique based on distributed source analysis of magnetoencephalography data, which makes possible the three-dimensional reconstruction of dynamic brain activity in humans. MFT has a temporal resolution better than 1 msec and a spatial accuracy of 2-5 mm at the cortical level, which deteriorates to 1-3 cm at depths of 6 cm or more. MFT is used here to visualize the origin of a spatiotemporally organized pattern of coherent 40-Hz electrical activity. This coherence, initially observed during auditory input, was proposed to be generated by recurrent corticothalamic oscillation. In support of this hypothesis, we illustrate well-defined 40-Hz coherence between cortical-subcortical sites with a time shift that is consistent with thalamocortical conduction times. Studies on Alzheimer patients indicate that, while a similar activity pattern is present, the cortical component is reduced in these subjects.
Bag model for DNA migration during pulsed-field electrophoresis.
Authors: Chu G
Published: May 2006

A model for pulsed-field electrophoresis was developed by picturing large DNA as a deformable "bag" that (i) moves with limiting mobility in a continuous electric field, (ii) adopts an orientation aligned with the field direction, and (iii) reorients after a change in field direction in a size-dependent manner. The model correctly predicted the resolution of large DNA in a pulsed field including the surprising phenomena of mobility inversion, lateral band spreading, and improved resolution for obtuse angles. A simple parametrization agreed with observations of two completely different aspects of DNA behavior: bulk mobility as measured during gel electrophoresis and molecular reorientation as measured by linear dichroism. The model also provides quantitative guidelines for setting experimental parameters in pulsed-field electrophoresis experiments.
Cloning of the cocaine-sensitive bovine dopamine transporter.
Authors: Usdin T B , Mezey E , Chen C , Brownstein M J , Hoffman B J
Published: May 2006

A cDNA encoding the dopamine transporter from bovine brain substantia nigra was identified on the basis of its structural homology to other, recently cloned, neurotransmitter transporters. The sequence of the 693-amino acid protein is quite similar to those of the rat gamma-aminobutyric acid, human norepinephrine, and rat serotonin transporters. Dopamine transporter mRNA was detected by in situ hybridization in the substantia nigra but not in the locus coeruleus, raphe, caudate, or other brain areas. [3H]Dopamine accumulation in tissue culture cells transfected with the cDNA was inhibited by amphetamine, cocaine, and specific inhibitors of dopamine transport, including GBR12909.
RAM2, an essential gene of yeast, and RAM1 encode the two polypeptide components of the farnesyltransferase that prenylates a-factor and Ras proteins.
Authors: He B , Chen P , Chen S Y , Vancura K L , Michaelis S , Powers S
Published: May 2006

In the yeast Saccharomyces cerevisiae, mutations in either of two unlinked genes, RAM1 or RAM2, abolish the farnesyltransferase activity responsible for prenylation of Ras proteins and the a-factor mating pheromone. Here we report that the function of RAM1 and RAM2 genes is required for the membrane localization of Ras proteins and a-factor. The RAM2 gene was sequenced and can encode a 38-kDa protein. We examined the functional interaction of RAM2 and RAM1 by expressing the genes in Escherichia coli. Extracts derived from an E. coli strain that coexpressed RAM1 and RAM2 efficiently farnesylated a-factor peptide and Ras protein substrates. In contrast, extracts derived from E. coli strains that expressed either RAM gene alone were devoid of activity; however, when the latter extracts were mixed, protein farnesyltransferase activity was reconstituted. These results indicate that the yeast farnesyl-protein transferase is comprised of Ram1 and Ram2 polypeptides. Although Ram1 is a component of the enzyme, disruption of the RAM1 gene in yeast was not lethal, indicating that the Ram1-Ram2 farnesyltransferase is not essential for viability. In contrast, disruption of RAM2 was lethal, suggesting that Ram2 has an essential function in addition to its role with Ram1 in protein farnesylation.
Uterine expression of leukemia inhibitory factor coincides with the onset of blastocyst implantation.
Authors: Bhatt H , Brunet L J , Stewart C L
Published: May 2006

We have analyzed the expression of the cytokine leukemia inhibitory factor (LIF) during embryogenesis and in tissues of neonatal and adult mice. The site of the most abundant LIF expression is the uterine endometrial glands, specifically on day 4 of pregnancy. Analysis of LIF expression in pseudopregnant mice and in females undergoing delayed implantation showed that it is under maternal control and that its expression coincides with blastocyst formation and always precedes implantation. These results suggest that a principal function of LIF in vivo may be to regulate the growth and to initiate implantation of blastocysts.
Characterization of a protein that regulates the DNA-binding activity of NF-AT, the nuclear factor of activated T cells.
Authors: Granelli-Piperno A , McHugh P
Published: May 2006

NF-AT is a T-cell-restricted activity that regulates interleukin 2 expression by binding to positions -285 to -254 of the interleukin 2 gene. Prior studies have shown that NF-AT is found only in T cells that are appropriately stimulated with "two signals" and is sensitive to the immunosuppressive drugs cyclosporin A and FK506. However, NF-AT has yet to be characterized biochemically. Here we show that the activity of NF-AT in electrophoretic gel mobility-shift assays is greatly enhanced by a distinct protein with an apparent molecular mass of 28 kDa. This modulator has been enriched from the flow-through of NF-AT affinity columns and enhances by severalfold the activity of the factor that elutes from these columns. The modulator has a predominantly nuclear location, but unlike NF-AT, the modulator is found in nuclear extracts of many cell types, even unstimulated T cells and T cells immunosuppressed with cyclosporin A or FK506. This modulator protein may regulate NF-AT binding activity posttranscriptionally, and it should prove useful in the isolation and biochemical characterization of NF-AT.
Regulation of ferritin and heme oxygenase synthesis in rat fibroblasts by different forms of iron.
Authors: Eisenstein R S , Garcia-Mayol D , Pettingell W , Munro H N
Published: May 2006

Synthesis of the iron-storage protein ferritin is thought to be regulated at the translational level by the cytosolic content of chelatable iron. This response to iron is regulated by the iron-modulated binding to ferritin mRNAs of a repressor protein, the iron regulatory element-binding protein. From measurements made in a cell-free system, regulation of the iron regulatory element-binding protein has been recently suggested to involve direct interaction with hemin. The following observations on the synthesis of ferritin and of heme oxygenase (HO), the heme-degrading enzyme, in rat fibroblasts or hepatoma cells lead us to conclude that chelatable iron is a direct physiological regulator of ferritin synthesis in intact cells: (i) the inhibitor of heme degradation, tin mesoporphyrin IX, reduces the ability of exogenous hemin to induce ferritin synthesis but enhances HO synthesis; (ii) the iron chelator desferal suppresses the ability of hemin to induce synthesis of ferritin but not of HO; (iii) the heme synthesis inhibitor succinylacetone does not block iron induction of ferritin synthesis; (iv) there is no apparent relationship between the ability of various metalloporphyrins to inactivate the iron regulatory element-binding protein in cell-free extracts and their capacity to induce ferritin synthesis in intact cells; (v) administered inorganic iron significantly induces the synthesis of ferritin but not of HO; (vi) addition of delta-aminolevulinic acid to stimulate heme synthesis represses the ability of inorganic iron to induce ferritin synthesis while activating HO synthesis. Taken together, our results demonstrate that (i) release of iron by HO plays an essential role in the induction of ferritin synthesis by heme and (ii) chelatable iron can regulate ferritin synthesis independently of heme formation.
Drift, admixture, and selection in human evolution: a study with DNA polymorphisms.
Authors: Bowcock A M , Kidd J R , Mountain J L , Hebert J M , Carotenuto L , Kidd K K , Cavalli-Sforza L L
Published: May 2006

Accuracy of evolutionary analysis of populations within a species requires the testing of a large number of genetic polymorphisms belonging to many loci. We report here a reconstruction of human differentiation based on 100 DNA polymorphisms tested in five populations from four continents. The results agree with earlier conclusions based on other classes of genetic markers but reveal that Europeans do not fit a simple model of independently evolving populations with equal evolutionary rates. Evolutionary models involving early admixture are compatible with the data. Taking one such model into account, we examined through simulation whether random genetic drift alone might explain the variation among gene frequencies across populations and genes. A measure of variation among populations was calculated for each polymorphism, and its distribution for the 100 polymorphisms was compared with that expected for a drift-only hypothesis. At least two-thirds of the polymorphisms appear to be selectively neutral, but there are significant deviations at the two ends of the observed distribution of the measure of variation: a slight excess of polymorphisms with low variation and a greater excess with high variation. This indicates that a few DNA polymorphisms are affected by natural selection, rarely heterotic, and more often disruptive, while most are selectively neutral.
Heme-protein fission under nondenaturing conditions.
Authors: Smith M L , Paul J , Ohlsson P I , Hjortsberg K , Paul K G
Published: May 2006

Slow heme transfer from horseradish peroxidases C2 and A2, cytochrome c peroxidase, chloroperoxidase, and leghemoglobins to a heme acceptor protein, apomyoglobin, has been studied under mild conditions. The reaction is best described as heme release into water followed by quick engulfment by apomyoglobin. The energetics of the activated process are large and interpreted as connected to both polypeptide motions during release and the ordering of water around the heme during solvation. The free energy required to break the iron(III)-ligand 5 (L5) bond is a minor but crucial portion of the activation free energy. Donor-acceptor protein interactions are not involved in the transfer. Fast heme release from inactive protein has also been observed. Apoprotein recombination with porphyrins and hemes suggest that this lack of activity is a result of Fe-L5 bond breaking.
Seismic communication in a blind subterranean mammal: a major somatosensory mechanism in adaptive evolution underground.
Authors: Nevo E , Heth G , Pratt H
Published: May 2006

Seismic communication, through low-frequency and patterned substrate-borne vibrations that are generated by head thumping, and which travel long distances underground, is important in the nonvisual communication of subterranean mole rats of the Spalax ehrenbergi superspecies (2n = 52, 54, 58, and 60) in Israel. This importance pertains both intraspecifically in adaptation and interspecifically in speciation. Neurophysiologic, behavioral, and anatomic findings in this study suggest that the mechanism of long-distance seismic communication is basically somatosensory and is independent of the auditory mechanism. Seismic communication thus appears to be a channel of communication important in the evolution of subterranean mammals that display major adaptation to life underground.
Resolution of the African hominoid trichotomy by use of a mitochondrial gene sequence.
Authors: Ruvolo M , Disotell T R , Allard M W , Brown W M , Honeycutt R L
Published: May 2006

Mitochondrial DNA sequences encoding the cytochrome oxidase subunit II gene have been determined for five primate species, siamang (Hylobates syndactylus), lowland gorilla (Gorilla gorilla), pygmy chimpanzee (Pan paniscus), crab-eating macaque (Macaca fascicularis), and green monkey (Cercopithecus aethiops), and compared with published sequences of other primate and nonprimate species. Comparisons of cytochrome oxidase subunit II gene sequences provide clear-cut evidence from the mitochondrial genome for the separation of the African ape trichotomy into two evolutionary lineages, one leading to gorillas and the other to humans and chimpanzees. Several different tree-building methods support this same phylogenetic tree topology. The comparisons also yield trees in which a substantial length separates the divergence point of gorillas from that of humans and chimpanzees, suggesting that the lineage most immediately ancestral to humans and chimpanzees may have been in existence for a relatively long time.
Transformation of a partial nopaline synthase gene into tobacco suppresses the expression of a resident wild-type gene.
Authors: Goring D R , Thomson L , Rothstein S J
Published: May 2006

A portion of the nopaline synthase gene under the control of the cauliflower mosaic virus 35S promoter was used to transform a tobacco plant that had previously been transformed with a wild-type nopaline synthase (nos) gene. Unexpectedly, in all nine primary transformants tested the wild-type nos expression was virtually completely suppressed. In contrast, plants transformed with the control vector DNA, which differed only in the absence of the partial nos gene, did not show any inhibition of nos expression. Progeny plants were analyzed for the stability of the gene-silencing phenotype. All of the progeny that carried both the wild-type and partial nos genes had no detectable nopaline synthase activity. In addition, wild-type nos mRNA could not be detected in these plants. In most plants in which the wild-type gene was segregated away from the partial nos gene, wild-type levels of activity were detected. Although DNA methylation has been shown to be correlated with a decrease in promoter activity in plants, none of the progeny appeared to carry a methylated nos promoter. The underlying mechanism causing this gene suppression phenomenon is unclear at this time.
Unexpected, spontaneous conversion of a family of rats, from low-leukemic to high-leukemic inbred line.
Authors: Dreyfuss Y , Gross L
Published: May 2006

From a nucleus of Sprague-Dawley rats received in 1960 from the National Institutes of Health, we have raised, by brother-to-sister mating, a colony of these animals. The incidence of leukemia in 313 females and 316 males was 1.6% and 1.2%, respectively. About 3 years ago, we observed a relatively high incidence of leukemia in offspring of a healthy female, no. 1. Among the offspring of this female, observed through 12 successive generations, there were 17 leukemias among 44 females (38.6%) and 21 leukemias among 40 males (52.5%), developing at ages varying from 6 to 11.6 months. The most frequent form of leukemia observed was acute myeloid, with a high count of myeloblasts, promyelocytes, and myelocytes; lymphatic form was relatively rare but was observed occasionally; pronounced anemia was common. In most instances, on autopsy, the pathological picture was that of an enlarged spleen and liver, with the exception of those few animals that developed lymphatic leukemia, with thymic and mesenteric lymphoid tumors. We have no satisfactory explanation for this sudden, unexpected conversion of a part of our Sprague-Dawley rat colony from low-leukemic to high-leukemic inbred line. As a working hypothesis, the possibility of a spontaneous activation of a hypothetical oncogenic virus should be considered. The high leukemic C58 inbred line of mice originated in a similar, unexplained manner [MacDowell, E.C. & Richter, M.N. (1935) Arch. Pathol. 20, 709-724]. However, leukemia developing in mice was subsequently found to be caused by a transmissible virus, whereas, thus far at least, no evidence of a transmissible virus has been found in leukemia developing spontaneously in rats.
Laventiana annectens, new genus and species: fossil evidence for the origins of callitrichine New World monkeys.
Authors: Rosenberger A L , Setoguchi T , Hartwig W C
Published: May 2006

A previously unidentified middle Miocene primate from the La Venta deposits of Colombia is intermediate between squirrel monkeys (Saimiri) and callitrichines (marmosets and tamarins) in the morphology of the lower molars, mandible, and talus. Laventiana annectens is closely related to Saimiri and to Cebus (capuchin monkeys) yet resembles the probable callitrichine morphotype, demonstrating that archaic relatives of a Saimiri-like stock were suitable structural ancestors for the enigmatic callitrichines. Laventiana is also more primitive than Saimiri (= Neosaimiri) fieldsi from the same fauna, further increasing the likelihood that the latter is a lineal ancestor of modern squirrel monkeys.
Einstein's equations with an embedding-dependent energy-momentum tensor for the compactified Minkowski time space and their relationship to the conformal action of SO(4,4) on S3 x S3.
Authors: Biedrzycki W R
Published: May 2006

For a pseudoriemannian 4-manifold isometrically embedded in a pseudoriemannian 6-manifold an embedding-dependent energy-momentum tensor is proposed, as well as an embedding-dependent cosmological constant. Their behavior is examined under the conformal transformations. It is specialized to the case of S1 x S3 embedded in S3 x S3, where the latter has the metric tensor in the conformal class of the one inherited from sum from 4i=1 (dxi)2 - sum from 8i=5(dxi)2 on R8. Einstein's equations for the restricted metric are shown to be satisfied if and only if the scalar curvature of S3 x S3 vanishes on S1 x S3.
Escherichia coli DnaJ and GrpE heat shock proteins jointly stimulate ATPase activity of DnaK.
Authors: Liberek K , Marszalek J , Ang D , Georgopoulos C , Zylicz M
Published: May 2006

The products of the Escherichia coli dnaK, dnaJ, and grpE heat shock genes have been previously shown to be essential for bacteriophage lambda DNA replication at all temperatures and for bacterial survival under certain conditions. DnaK, the bacterial heat shock protein hsp70 analogue and putative chaperonin, possesses a weak ATPase activity. Previous work has shown that ATP hydrolysis allows the release of various polypeptides complexed with DnaK. Here we demonstrate that the ATPase activity of DnaK can be greatly stimulated, up to 50-fold, in the simultaneous presence of the DnaJ and GrpE heat shock proteins. The presence of either DnaJ or GrpE alone results in a slight stimulation of the ATPase activity of DnaK. The action of the DnaJ and GrpE proteins may be sequential, since the presence of DnaJ alone leads to an acceleration in the rate of hydrolysis of the DnaK-bound ATP. The presence of GrpE alone increases the rate of release of bound ATP or ADP without affecting the rate of hydrolysis. The stimulation of the ATPase activity of DnaK may contribute to its more efficient recycling, and it helps explain why mutations in dnaK, dnaJ, or grpE genes often exhibit similar pleiotropic phenotypes.
Early ingrowth of thalamocortical afferents to the neocortex of the prenatal rat.
Authors: Catalano S M , Robertson R T , Killackey H P
Published: May 2006

The initial ingrowth of thalamocortical afferents into the presumptive somatosensory cortex was examined in the fetal rat. Thalamic fibers were labeled in fixed brains with the carbocyanine dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI). On embryonic day 16, thalamocortical afferents arrive in the neocortex and course tangentially within the intermediate zone immediately underneath the cortical plate. By embryonic day 17, thalamocortical fibers have begun their radial growth into cortex and their arbors span the cell-sparse zone between layer VIb and the bottom of the cortical plate. By the day of birth (embryonic day 21), thalamocortical fibers from a dense plexus within layers VI and V below the dense cortical plate. Our observations indicate that in the rat thalamic afferents arrive in the cortex at a very early age and arborize within the forming cortical layers without an apparent "waiting" period.
Class II major histocompatibility complex molecules of murine dendritic cells: synthesis, sialylation of invariant chain, and antigen processing capacity are down-regulated upon culture.
Authors: Kämpgen E , Koch N , Koch F , Stöger P , Heufler C , Schuler G , Romani N
Published: May 2006

Dendritic cells (DCs), such as Langerhans cells (LCs) of the epidermis and the DCs of lymphoid organs such as spleen, are potent antigen presenting cells. DCs express high levels of major histocompatibility complex (MHC) class II molecules, but, partly because of the low numbers of primary DCs in any tissue, there has been no detailed study of the biochemistry of their class II molecules. This information may be needed to help explain recent findings that DCs process native protein antigens when freshly isolated from epidermis and spleen. Processing ceases during culture, yet a strong accessory function for activating resting T cells develops. We studied immunoprecipitates of DC class II and invariant chain (Ii) molecules by two-dimensional gel electrophoresis. We found that (i) freshly isolated LCs synthesize large amounts of class II and Ii polypeptides; (ii) Ii molecules that are known to be involved in antigen processing display an unusually large number of sialic acids in fresh LCs; (iii) with culture, class II and Ii synthesis decreases dramatically and has virtually ceased at 3 days; and (iv) the turnover of class II in pulse/chase experiments is slow, being undetectable over a 12- to 32-hr culture period, whereas the turnover of Ii is rapid. We conclude that MHC class II molecules of DCs do not seem to be qualitatively unique. However, the regulation of class II and Ii expression is distinctive in that biosynthesis proceeds vigorously for a short period of time and the newly synthesized class II remains stably on the cell surface, whereas Ii turns over rapidly. This may enable DCs to process and retain antigens in the peripheral tissues such as skin and migrate to the lymphoid organs to activate T cells there.
Replication deficiencies in priA mutants of Escherichia coli lacking the primosomal replication n' protein.
Authors: Lee E H , Kornberg A
Published: May 2006

The priA gene of Escherichia coli encodes the protein that initiates assembly of the promosome, the entity essential for the replication of phage phi X174 and ColE1-like plasmids in vitro. We have prepared a null priA mutant to assess its role in vivo in replication of phages, plasmids, and the host chromosome. Extracts of this mutant are inert in the initial conversion of the phi X174 viral strand to the duplex form, confirming the absence of the PriA activity. In vivo, the priA mutant fails to produce phi X174 phage and, remarkably, is unable to maintain plasmids that depend on the E. coli chromosome origin as well as those of ColE1. Deficiencies in cell growth and cell division are also manifest.
The role of the retinylidene Schiff base counterion in rhodopsin in determining wavelength absorbance and Schiff base pKa.
Authors: Sakmar T P , Franke R R , Khorana H G
Published: May 2006

Glu-113 serves as the retinylidene Schiff base counterion in bovine rhodopsin. Purified mutant rhodopsin pigments were prepared in which Glu-113 was replaced individually by Gln (E113Q), Asp (E113D), Asn (E113N), or Ala (E113A). E113Q, E113N, and E113A existed as pH-dependent equilibrium mixtures of unprotonated and protonated Schiff base (PSB) forms. The Schiff base pKa values determined by spectrophotometric titration were 6.00 (E113Q), 6.71 (E113N), and 5.70 (E113A). Thus, mutation of Glu-113 markedly reduced the Schiff base pKa. The addition of NaCl promoted the formation of a PSB in E113Q and E113A. An exogenously supplied solute anion replaced Glu-113 to compensate for the positive charge of the PSB in these mutants. The lambda max values of the PSB forms of the mutants in NaCl were 496 nm (E113Q), 506 nm (E113A), 510 nm (E113D), and 520 nm (E113N). To evaluate the effect of different types of solute anions on lambda max values, mutants were prepared in sodium salts of halides, perchlorate, and a series of carboxylic acids of various sizes and acidity. The lambda max values of E113Q and E113A depended on the solute anion present and ranged from 488 nm to 522 nm for E113Q and from 486 nm to 528 nm for E113A. The solute anion affected the lambda max values of E113N and E113D to lesser degrees. The reactivities of the mutants to hydroxylamine were also studied. Whereas rhodopsin was stable to hydroxylamine in the dark, E113N reacted slowly and E113Q reacted rapidly under these conditions, indicating structural differences in the Schiff base environments. The lambda max values and solute anion dependencies of the Glu-113 mutants indicate that interactions between Schiff base and its counterion play a significant role in determining the lambda max of rhodopsin.
Creation of a large genomic deletion at the T-cell antigen receptor beta-subunit locus in mouse embryonic stem cells by gene targeting.
Authors: Mombaerts P , Clarke A R , Hooper M L , Tonegawa S
Published: May 2006

Recently it has become possible to introduce predesigned mutations into a given gene in the mouse germ line by homologous recombination in embryonic stem cells. The mutations are usually introduced by inserting the neomycin phosphotransferase gene into an exon of a particular gene. Here we describe an extension of this method that can result in at least a 15-kilobase-long deletion. The deletion created in the present work encompasses one of the two diversity gene segments of the mouse T-cell receptor beta-subunit locus, 10 out of the 12 joining gene segments, and both constant gene segments. This strategy is a valuable alternative to sequential targeting of multiple genes forming a gene cluster, could simplify the construction of plasmids to be used for targeting, and could be the solution for inactivating small genes that have eluded conventional targeting approaches.
Antiestrogen can establish nonproductive receptor complexes and alter chromatin structure at target enhancers.
Authors: Pham T A , Elliston J F , Nawaz Z , McDonnell D P , Tsai M J , O'Malley B W
Published: May 2006

We describe in this report experiments in vivo that demonstrate that antiestrogens promote DNA binding of the estrogen receptor without efficiently inducing transcription. When the receptor is modified to carry a foreign unregulated transactivation domain, transcription can be induced efficiently by both estrogen and antiestrogens. Under apparent saturation conditions, antihormone-receptor complexes binding to responsive enhancer elements elicit only a low level of transcription. In addition, we show that both estrogen and an antiestrogen, nafoxidine, effect very similar alterations in chromatin structure at a responsive promoter. These results indicate that in vivo steroid receptor action can be regulated subsequent to the DNA binding step, by regulating interactions with the target transcriptional machinery. In this regard, antihormones can function by establishing receptor-DNA complexes that are transcriptionally nonproductive.
Feedforward lateral inhibition in retinal bipolar cells: input-output relation of the horizontal cell-depolarizing bipolar cell synapse.
Authors: Yang X L , Wu S M
Published: May 2006

Lateral inhibition is the ubiquitous strategy used by visual neurons for spatial resolution throughout the animal kingdom. It has been a puzzle whether lateral inputs in retinal bipolar cells are mediated by the horizontal cell (HC)-cone feedback synapse, by the HC-bipolar cell feedforward synapse, or by both. By blocking the central inputs of the depolarizing bipolar cells (DBCs) with L-2-amino-4-phosphonobutyrate, we were able to eliminate the contribution of the feedback synapse and to demonstrate the postsynaptic light response in DBCs mediated by the HC-DBC feedforward synapse. The HC-DBC feedforward synapse contributes roughly one-third of the surround response whereas the HC-cone-DBC feedback synapse probably contributes the rest.
Isolation and characterization of two peptides with prolactin release-inhibiting activity from porcine hypothalami.
Authors: Schally A V , Guoth J G , Redding T W , Groot K , Rodriguez H , Szonyi E , Stults J , Nikolics K
Published: May 2006

Two peptides with in vitro prolactin release-inhibiting activity were purified from stalk median eminence (SME) fragments of 20,000 pig hypothalami. Monolayer cultures of rat anterior pituitary cells were incubated with aliquots of chromatographic fractions and the inhibition of release of prolactin in vitro was measured by RIA in order to monitor the purification. The hypothalamic tissue extract was separated into 11 fractions by high-performance aqueous size-exclusion chromatography with one fraction showing a 4-fold increase in prolactin release-inhibiting factor (PIF) activity. This material was further purified by semipreparative reversed-phase (RP) HPLC. This process resulted in the separation of two distinct fractions that showed high PIF activity. These were further purified by semipreparative and analytical RP-HPLC to apparent homogeneity as judged by the UV absorbance profiles. Neither of the two peptides showed cross-reactivity with gonadotropin releasing hormone-associated peptide or with somatostatin-14 antibodies. Protein sequence analysis revealed that one of the PIF peptides was Trp-Cys-Leu-Glu-Ser-Ser-Gln-Cys-Gln-Asp-Leu-Ser-Thr-Glu-Ser-Asn-Leu-Leu- Ala-Cys - Ile-Arg-Ala-Cys-Lys-Pro, identical to residues 27-52 of the N-terminal region of the proopiomelanocortin (POMC) precursor (corresponding to amino acids 1-26 of the 16-kDa fragment). The sequence of the other PIF was Ala-Ser-Asp-Arg-Ser-Asn-Ala-Thr-Leu-Leu-Asp-Gly-Pro-Ser-Gly-Ala-Leu-Leu- Leu-Arg - Leu-Val-Gln-Leu-Ala-Gly-Ala-Pro-Glu-Pro-Ala-Glu-Pro-Ala-Gln-Pro-Gly-Val- Tyr, representing residues 109-147 of the vasopressin-neurophysin precursor. Synthetic peptides corresponding to the N-terminal region of POMC had significant PIF activity in vitro.
PC1 and PC2 are proprotein convertases capable of cleaving proopiomelanocortin at distinct pairs of basic residues.
Authors: Benjannet S , Rondeau N , Day R , Chrétien M , Seidah N G
Published: May 2006

A recombinant vaccinia virus vector was used to coexpress the two candidate mouse prohormone convertases, PC1 and PC2, together with mouse proopiomelanocortin (POMC) in the constitutively secreting cell line BSC-40 and in the endocrine tissue-derived cell lines PC12 and AtT-20, which exhibit regulated secretion. Monitoring of POMC processing demonstrated the distinct cleavage specificities of PC1 and PC2, since in the cell lines analyzed (i) PC1 cleaves POMC into corticotropin and beta-lipotropin, (ii) PC2 cleaves POMC into beta-endorphin, an N-terminally extended corticotropin containing the joining peptide, and either alpha MSH or desacetyl-alpha MSH, and (iii) PC2 cleaves POMC at the five pairs of basic residues analyzed, whereas PC1 cleaves two of them preferentially, suggesting that PC2 has a broader spectrum of activity than PC1. These data are consistent with our hypothesis on the physiological role of PC1 and PC2 as distinct proprotein convertases acting alone or together to produce a set of tissue-specific maturation products in the brain and in peripheral tissues.
Protein kinase C mediates transient spinule-type neurite outgrowth in the retina during light adaptation.
Authors: Weiler R , Kohler K , Janssen U
Published: May 2006

Light and dark adaptation of the teleost retina is accompanied by a remarkable morphological rearrangement of the synaptic connections between photoreceptors and second-order neurons: during light adaptation, numerous new neurites, the so-called spinules, arise from the terminal dendrites of horizontal cells invaginating the cone pedicle, and during dark adaptation, these spinules are retracted. The formation of these spinules is paralleled by the appearance of color opponency in horizontal and ganglion cells, which led to the suggestion that these spinules are the site of the inhibitory synapses in the negative feedback loop between cones and horizontal cells. The formation of the spinules in the light and their disappearance in darkness have a time course of minutes and are modulated by the neurotransmitters dopamine and glutamate, respectively. Neurotransmitters can modulate neuronal processing through a variety of second messengers that activate protein kinases, resulting most commonly in protein phosphorylation. Herein we report that activation of protein kinase C by phorbol esters promotes the formation of new horizontal-cell spinules in animals kept in the dark. Partial inhibition of protein kinase C activation with sphingosines prevents the formation of new spinules during light adaptation but does not affect established spinules. The spinule-forming effect of phorbol esters is not mediated by dopaminergic neurons, since the effect is also seen in retinas depleted of dopaminergic neurons. Phorbol esters also initiate the formation of spinules in synaptically isolated horizontal cells, demonstrating that they have a direct action on these cells. In addition, isolated horizontal cells have substrate proteins that are phosphorylated in a protein kinase C-dependent manner.