bluuebunny/crossref_metadata_2025 huggingface dataset:
The differential effects of ionizing radiation on the circadian oscillator and other functions in the eye of Aplysia.
Authors: Woolum J C , Strumwasser F
Published: May 2006

Ionizing radiation has been used to selectively separate the circadian oscillator function of the eye of Aplysia from some of its other functions--synchronous compound action potential (CAP) generation, the light response, synaptic transmission between photoreceptors and output neurons, and the bursting pacemaker mechanism. Doses of 4-krad (50 kV peak) x-rays have a minimal effect on the circadian rhythm of CAP frequency, measured from the otpic nerve, whereas irradiation with a 40-krad dose abolishes the rhythm without affecting any of the four other functions of this eye (1 rad = 0.01 J/kg = 0.01/Gy). We estimate a 50% survival of the oscillator function at doses of about 6 krad. The oscillators of irradiated eyes are not merely desynchronized when the rhythm is abolished, because in vitro light-dark entrainment does not restore free-running rhythmicity. The results, including those from selective irradiation of the anterior or posterior poles of the eye, suggest that there are a number of circadian oscillators in the eye--most of them in the posterior portion near the optic nerve. An approximate target size has been obtained from target theory approximately equal to 10(8) A3, which is somewhat larger than the target size for viral infectivity function, as one example. There are reservations about estimating target size in a complex organ such as the eye. However, this approximate target size and the fact that recovery or repair can occur in vivo suggest that the oscillator may involve nucleic acid molecules.
Mitochondrial carbonic anhydrase.
Authors: Dodgson S J , Forster R E , Storey B T , Mela L
Published: May 2006

We have assayed carbonic anhydrase activity (carbonate dehydratase, carbonate hydro-lyase, EC 4.2.1.1) and bicarbonate permeability in suspensions of broken and intact guinea pig mitochondria by monitoring the disappearance of C16O18O. We found significant activity in preparations from liver and skeletal muscle, but not in preparations from heart muscle, brain, and kidney. Intact mitochondria containing carbonic anhydrase produce a two-phase acceleration of the disappearance of the labeled CO2, which indicates that the enzyme is located in a region more accessible to CO2 than to HCO3-. Acetazolamide inhibits the enzyme activity instantly in broken mitochondria but only after a delay in intact mitochondria, indicating that the enzyme is in a region not immediately accessible to the inhibitor. Sonication of mitochondria containing carbonic anhydrase activity releases the enzyme, which remains in the supernatant after sedimentation of the submitochondrial particles. This shows that mitochondrial carbonic anhydrase is in the matrix compartment and not in, or bound to, the inner membrane. The activity of the enzyme increases markedly with increasing pH. The enzyme activity of intact mitochondria is greater than that of the broken mitochondria at the same pH of the suspending fluid, corresponding to an intramitochondrial pH that is 0.2-0.5 unit more alkaline.
Stereoacuity of human infants.
Authors: Held R , Birch E , Gwiazda J
Published: May 2006

Stereograms were presented in a two-choice preference procedure. The mean age at which stereopsis was first demonstrable was 16 weeks. By a mean age of 21 weeks, infants had achieved stereoacuity of 1 minute of arc or better. In comparison with the relatively slow development of visual acuity, the time course for the development of stereoacuity is extremely rapid.
Measurement of mammalian 25-hydroxyvitamin D3 24R-and 1 alpha-hydroxylase.
Authors: Tanaka Y , DeLuca H F
Published: May 2006

An in vitro assay of mammalian 25-hydroxyvitamin D3 1 alpha- and 24R-hydroxylases in kidney has been developed. It had been suggested that 25-hydroxyvitamin D binding protein present in mammalian blood and tissues inhibits the enzyme activities in cell-free preparations by binding the substrate 25-hydroxyvitamin D3 more strongly than the hydroxylases bind it. This inhibitory effect is overcome by the addition of substantial amounts of unlabeled 25-hydroxyvitamin D3 to saturate the binding sites of this protein. The resulting metabolites produced in vitro by rat kidney homogenates were isolated and firmly identified by ultraviolet absorption spectrometry and mass spectrometry as 1,25-dihydroxyvitamin D3 and (24R)-24,25-dihydroxyvitamin D3. Maximal 1 alpha-hydroxylation of 25-hydroxyvitamin D3 could be demonstrated in kidney homogenates prepared from vitamin D-deficient rats. Thyroparathyroidectomy of these rats resulted in total suppression of the 1 alpha-hydroxylase. Homogenates of kidney from rats given vitamin D showed little or no 1 alpha-hydroxylase and substantial 24R-hydroxylase activity. Thyroparathyroidectomy of these rts markedly increased the 24R-hydroxylase activity.
Replacement of riboflavin by an analogue in the blue-light photoreceptor of
Phycomyces
Authors: Otto M. K. , Jayaram M. , Hamilton R. M. , Delbrück M.
Published: May 2006

Under suitable conditions, roseoflavin [7-methyl-8-dimethylamino-10-(1′-D-ribityl)isoalloxazine] replaces riboflavin to about 80% in the photoreceptor of
Phycomyces
. The substitute-bearing photoreceptor functions with an efficiency of about 0.1% of that of the normal receptor. The substitution is proven by (
i
) a decrease of the effective light flux by a factor of 4.7, expressed as a corresponding increase in threshold, and (
ii
) an increase of the effectiveness of 529-nm light relative to 380-nm light. It has also been shown that roseoflavin is taken up by the mycelium, translocated to the sporangiophore, and effectively phosphorylated by the riboflavin kinase of
Phycomyces
.
Demonstration, by somatic cell genetics, of coordinate regulation of genes for two enzymes of purine synthesis assigned to human chromosome 21.
Authors: Patterson D , Graw S , Jones C
Published: May 2006

A method for determining coordinate genetic regulation is proposed for mammalian cells. The method involves (i) isolation of a set of mutants defective in the relevant pathway; (ii) complementation analysis of these mutants to determine dominance and to categorize the mutants into various different complementation groups; (iii) determination of the biochemical blocks in the mutants; (iv) identification of individual mutants that fail to complement the members of at least two distinct complementation groups that complement each other, such mutants being said to show coordinate regulation of the affected functions; (v) biochemical and reversion analysis of the relevant cell types to confirm the basis for the observed coordinate regulation; (vi) assignment of the individual genes to particular human chromosomes; (vii) mapping of the genes to determine contiguity on the genome; and (viii) examination of the structure of the relevant gene products. This method has allowed the demonstration of coordinate regulation between the gene coding for phosphoribosylglycineamide synthetase [5-phosphoribosylamine:glycine ligase (ADP-forming), EC 6.3.4.13], defective in our Ade-C mutants, and the gene coding for phoshoribosylaminoimidazole synthetase [5'-phosphoribosylformylglycinamidine cyclo-ligase (ADP-forming), EC 6.3.3.1], defective in our Ade-G mutants. Moreover, both genes can be assigned to human chromosome 21. Because at least two genes for purine biosynthesis have now been assigned to chromosome 21, and because patients with trisomy 21 (Down syndrome) show increased levels of serum purines, it may be that cells of these patients overproduce purines and that this overproduction may be relevant to the pathology of the syndrome.
Primary structure of the Fc region of human immunoglobulin D: implications for evolutionary origin and biological function.
Authors: Lin L C , Putnam F W
Published: May 2006

We have determined the complete amino acid sequence of a tryptic Fc delta fragment generated from an intact human IgD (WAH); it is 226 residues long and includes the second (C delta 2) and the third (C delta 3) constant domains of the delta chain. Comparison of the homology of the Fc sequence of the five human immunoglobulin classes suggests that either the delta-chain gene evolved from the alpha-chain gene soon after the divergence of a mu-alpha common ancestor or it evolved from an ancestral gene distinct from both the mu-alpha and the gamma-epsilon common ancestors. Comparative study using a spatial model of the Fc region indicates that the structure of the C delta 3 domain differs extensively from that of the carboxy-terminal domains of other heavy chain classes; this, together with the unique hinge region structure, probably reflects the biological role of IgD as a receptor molecule on the B-lymphocyte surface.
Cytoplasmic polyadenylate processing events accompany the transfer of mRNA from the free mRNP particles to the polysomes in Physarum.
Authors: Adams D S , Noonan D , Jeffery W R
Published: May 2006

The relationship between the mRNA in the polysomes and the free cytoplasmic messenger ribonucleoprotein of Physarum polycephalum was studied by microinjection techniques. Labeled free cytoplasmic ribonucleoprotein, prepared from donor plasmodia, was microinjected into unlabeled host plasmodia, and its fat was followed in the host ribonucleoprotein particles. Approximately one-half of the poly(A)-containing RNA [poly(A)+RNA] that originated from the microinjected particles was incorporated into the host polysomes by normal translational processes within 1 hr. Very short poly(A) sequences (approximately 15 nucleotide residues) were found in these poly(A)+RNA molecules. These short poly(A) sequences were sensitive to digestion with micrococcal nuclease, suggesting that they were not associated with protein. Because the poly(A)+RNA molecules of the microinjected free cytoplasmic mRNP had originally contained poly(A) sequences 50-65 nucleotides long and were associated with protein extensive poly(A) degradation and poly(A).protein complex dissociation must have occurred during their incorporation into the polysomes or during their translation. These results demonstrate a precursor-product relationship between free cytoplasmic mRNP and polysomal mRNA and suggest that the incorporation process in Physarum is accompanied by structural modifications in the poly(A) region of mRNA. They also imply that the polysome is a site for disruption of the poly(A).protein complex and poly(A) degradation.
Evolution of DNA sequence organization in mitochondrial genomes of
Zea
Authors: Sederoff R. R. , Levings C. S. , Timothy D. H. , Hu W. W. L.
Published: May 2006

Molecular mechanisms underlying the evolution of mtDNA in maize and three species of teosinte have been investigated. By using DNA transfer techniques and cloned fragments of maize mtDNA, changes in the position of homologous sequences in restriction digests were analyzed in related taxa. The resulting patterns indicate general conservation of sequence homology in all four species. One-third of the cloned fragments showed sequences conserved in homology and position of
Bam
HI restriction fragments. Other fragments produced patterns indicating that extensive rearrangement of DNA sequences has taken place. In two cases, hybridization patterns revealed major changes in relative abundance of specific sequences that we believe are related to the molecular heterogeneity phenomenon of maize mtDNA. The evolution of mtDNA in these plants appears dramatically different from that in animal mtDNA, where base substitutions may account for most of the observed changes in restriction patterns.
Mechanisms of Kirsten murine sarcoma virus transformation-induced changes in the collagen phenotype and synthetic rate of BALB 3T3 cells.
Authors: Bateman J F , Peterkofsky B
Published: May 2006

Specific viral transformation rather than cell selection can explain the previously observed increase in the proportion of type III procollagen compared to type I procollagen in BALB 3T3 cells transformed by Kirsten murine sarcoma virus (Ki-MSV). Two subclones of BALB 3T3 A31 were productively infected with with a temperature-sensitive Ki-MSV in the presence of helper murine leukemia virus (MLV), resulting in virtually complete transformation of cultures and eliminating selection of transformed foci. Analysis of radioactive collagen, derived from procollagen by pepsin treatment, showed that both of the tsKi-MSV/MLV-transformed subclones contained a 4-fold greater proportion of type III procollagen than did control MLV-infected cultures. A nonproducer derivative exhibited an even greater change (10-fold), indicating that viral replication was irrelevant. After 48 hr at a nonpermissive temperature, tsKi-MSV-transformed cells retained a high proportion of type III procollagen, suggesting that either this change is not induced by src protein or else there is a slowly reversible or irreversible step involved. Alternatively, type III procollagen mRNA may be long lived. In contrast, the relative rate of procollagen synthesis in transformed cells was clearly regulated by src protein. Translation of mRNA from cells preincubated at permissive or nonpermissive temperatures revealed that the decreased relative rate can be explained by a simultaneous small decrease in the level of procollagen mRNA and a large increase in mRNA for noncollagen proteins.
Two periplasmic transport proteins which interact with a common membrane receptor show extensive homology: complete nucleotide sequences.
Authors: Higgins C F , Ames G F
Published: May 2006

The hisJ and argT genes of Salmonella typhimurium encode two periplasmic binding proteins, J and LAO, which are involved in histidine and arginine transport, respectively, and which interact with a common membrane-bound component, the P protein. The complete nucleotide sequences of these two genes have been determined. The two genes show extensive homology (70%) and presumably arose by tandem duplication of a single ancestral gene. The two encoded proteins now perform distinct functions but still retain sufficient homology to permit interaction with the same site on the membrane-bound P protein. Three lines of evidence have allowed both the amino acid-binding site and the site involved in the interaction with the P protein to be assigned to specific regions of each binding protein: (i) the distribution of amino acid differences between the two proteins; (ii) the properties of a functional chimeric protein, produced by a deletion mutant in which the first half of the argT gene is fused to the second half of the hisJ gene; (iii) the sequence change in a mutant J protein unable to interact with P.
Membrane potential depolarization and increased intracellular pH accompany the acrosome reaction of sea urchin sperm.
Authors: Schackmann R W , Christen R , Shapiro B M
Published: May 2006

The intracellular pH and membrane potential in sperm of the sea urchin Strongylocentrotus purpuratus were investigated by using fluorescent and radiolabeled probes. The weak bases [14C]methylamine, [14C]diethylamine, and 9-aminoacridine were concentrated within sperm 5-fold or greater. The weak acid [14C]dimethyloxazolidine-2,4-dione (DMO) was excluded from sperm. These data suggested that the apparent intracellular pH is acidic with respect to seawater (pH 8.0). Induction of the acrosome reaction caused efflux of the amines and uptake of DMO, consistent with an increase in apparent intracellular pH of 0.1-0.2 pH unit. The presence of an internally negative membrane potential was indicated by estimating the distribution of [3H]tetraphenylphosphonium (Ph4P+) and [14C]SCN-. From SCN- exclusion we estimated a value of about -30 mV for the nonmitochondrial membrane potential, whereas from Ph4P+ accumulation an apparent potential of -90 to -150 mV was demonstrated. The membrane potentials obtained with Ph4P+ and SCN- were dependent upon the external K+ concentration, with increasing K+ leading to depolarization. Induction of the acrosome reaction led to efflux of Ph4P+ and uptake of SCN- for an approximate depolarization of about 30 mV, primarily due to the collapse of the plasma membrane potential.
Uninfected cell polymerase efficiently transcribes early but not late herpes simplex virus type 1 mRNA.
Authors: Frink R J , Draper K G , Wagner E K
Published: May 2006

The sequences of the DNAs encoding the 5' ends of one early and one late herpes simplex virus type 1 mRNA were analyzed, and the 5' ends of these mRNA species were precisely located. Neither mRNA species is spliced and the noncoding strand of the DNA contains recognizable T-A-T-A and C-A-T boxes upstream from their respective 5' ends. The early mRNA was efficiently transcribed by a commercially available uninfected cell lysate system, but the late mRNA was not. This difference between early and late mRNAs appears to be general in this virus.
Synthesis of parvovirus H-1 replicative form from viral DNA by DNA polymerase gamma.
Authors: Kollek R , Goulian M
Published: May 2006

The initial event in the replication cycle of parvovirus H-1 is conversion of the single-stranded linear viral DNA to the double-stranded linear replicative form. We describe here detection of an activity in uninfected cell extracts that carries out this reaction. The activity was purified and identified as DNA polymerase gamma.
Induction of plasminogen activator by UV light in normal and xeroderma pigmentosum fibroblasts.
Authors: Miskin R , Ben-Ishai R
Published: May 2006

Normal and DNA repair-deficient human fibroblasts have been used to study induction of plasminogen activator (PA) by DNA damage. UV light induced the synthesis of PA in skin fibroblasts of all types of xeroderma pigmentosum (XP) in XP heterozygotes and in human amniotic cells. Enzyme induction was, however, not observed in fibroblasts of normal adults. In classical XP, which are deficient in excision repair, PA synthesis occurred in a narrow range of low-UV fluences. In such strains, the level of enzyme produced was correlated with the extent of repair deficiency. UV fluences required for PA induction in XP variants and XP heterozygotes were at least 10 times those inducing enzyme synthesis in excision-deficient XP. Maximum enzyme induction occurred 48 hr after irradiation, and the highest levels of enzyme produced were 15-20 times those of PA baseline levels. Electrophoretic analysis showed that UV irradiation enhances the synthesis of the Mr 60,000 human urokinase-type PA, which is present in low amounts in untreated cells. Our results suggest that PA induction in human cells is caused by unrepaired DNA damage and represents a eukaryotic SOS-like function. In addition, PA induction may provide a sensitive assay for detection of cellular DNA repair deficiencies and identification of XP heterozygotes.
pH homeostasis in Escherichia coli: measurement by 31P nuclear magnetic resonance of methylphosphonate and phosphate.
Authors: Slonczewski J L , Rosen B P , Alger J R , Macnab R M
Published: May 2006

The intracellular pH of Escherichia coli cells, respiring on endogenous energy sources, was monitored continuously by 31P NMR over an extracellular pH range between 5.5 and 9. pH homeostasis was found to be good over the entire range, with the data conforming to the simple relationship intracellular pH = 7.6 + 0.1(external pH - 7.6) so that the extreme values observed for intracellular pH were 7.4 and 7.8 at external pH 5.5 and 9, respectively. As well as inorganic phosphate, we employed the pH-sensitive NMR probe methylphosphonate, which was taken up by glycerol-grown cells and was nontoxic; its pKa of 7.65 made it an ideal probe for measurement of cytoplasmic pH and alkaline external pH.
Mouse alpha-globin genes and alpha-globin-like pseudogenes are not syntenic.
Authors: Popp R A , Lalley P A , Whitney J B , Anderson W F
Published: May 2006

A genetic polymorphism for a Bgl I endonuclease site near the alpha-globin-like pseudogene alpha-4 of C57BL/6 and C3H/HeN mice was used to show that alpha-4 was not affected by three independent mutations in which the adult globin genes alpha-1 and alpha-2 were deleted. These results indicated that alpha-4 might not be located adjacent to the adult alpha-globin genes on chromosome 11. Restriction endonuclease analysis of DNA of a primary clone of a Chinese hamster--mouse somatic cell hybrid that had lost mouse chromosomes 11 and 18 showed that this clone lacked the adult murine globin genes alpha-1 and alpha-2 but it did contain the alpha-globin-like pseudogenes alpha-3 and alpha-4. These results indicated that the adult alpha-globin genes and alpha-globin-like pseudogenes are not located on the same chromosome. Similar analyses of several other Chinese hamster--mouse somatic cell hybrids that had segregated other mouse chromosomes indicated that the alpha-globin-like pseudogenes alpha-3 and alpha-4 are located on mouse chromosomes 15 and 17, respectively. These data explain why alpha-3 and alpha-4 were not affected by the three independently induced deletion-type mutations that cause alpha-thalassemia in the mouse.
Transcription of the simian virus 40 genome in DNA-transformed murine teratocarcinoma stem cells.
Authors: Linnenbach A , Huebner K , Croce C M
Published: May 2006

To study the molecular basis for lack of expression of the simian virus 40 (SV40) early region genes in murine teratocarcinoma-derived stem cells, we introduced a recombinant plasmid consisting of pBR322 linked to the herpes simplex virus type 1 thymidine kinase gene and SV40 genome into thymidine kinase-deficient F9 stem cells. The resulting stem cell clone, 12-1, and a retinoic acid-induced differentiated daughter cell clone, 12-1a, each contain one copy per cell of the entire recombinant plasmid integrated into the cellular genome through a site on the pBR322 genome. Restriction endonuclease analyses indicate that there is no difference in integration site or organization of the three component parts of the plasmid genome within cellular DNA of stem and differentiated cells; yet the differentiated cells, 12-1a, express SV40 large tumor antigen whereas the stem cells, 12-1, do not. Both stem and differentiated cells produce two size classes of polyadenylylated RNA, 2900 and 2600 bases in length, homologous to the early region of the SV40 genome, detectable by RNA blotting analysis. S1 nuclease analysis of the SV40 transcripts present in stem and differentiated cells indicate that the SV40 mRNAs were identically spliced in the two cell types, in a manner consistent with that observed for spliced large and small tumor antigen mRNAs in SV40-infected monkey kidney cells. Thus, the failure of 12-1 teratocarcinoma stem cells, containing an integrated SV40 genome, to express SV40 tumor antigen is not due to a lack of transcription of the SV40 early region or to an inability to splice primary transcripts.
Protein variations associated with Lesch-Nyhan syndrome.
Authors: Merril C R , Goldman D , Ebert M
Published: May 2006

Patients having Lesch--Nyhan syndrome were studied by using enzymatic, immunologic, and two-dimensional electrophoretic techniques. Four hundred proteins were analyzed on each two-dimensional electrophoretogram for positional or quantitative variation. In autoradiograms of lymphocytes stimulated with phytohemagglutinin, there were 11 quantitative differences found in all patients that were significant at the 2P less than 0.01 level. A significant quantitative difference was also found in an analysis of silver-stained gels of unstimulated lymphocytes. Patients had trace amounts of erythrocyte hypoxanthine phosphoribosyl transferase (HPRT) activity and trace or no immunoprecipitable HPRT. However, HPRT was observed in silver-stained erythrocyte electrophoretograms and in autoradiograms from phytohemagglutinin-stimulated lymphocytes. Unstimulated lymphocytes contained 65% of the control HPRT concentration. Currently, the technology of two-dimensional electrophoresis detects a fraction of the total cellular proteins and defective proteins may not show electrophoretic alterations. However, specific secondary changes in other polypeptides may be observed and, when catalogued, will serve as an aid in the diagnosis and understanding of the pathophysiology of metabolic diseases.
Manifold expression of new cellular genes in human lymphoid neoplasia.
Authors: Hanania N , Shaool D , Poncy C , Harel J
Published: May 2006

Previous studies performed in our laboratory have shown that a minor single-stranded DNA component (ssDNA) isolated from the bulk of double-stranded nuclear DNA (nDNA) of human, murine, or avian cells consisted mainly of active transcription sites. In the present work, ssDNA isolated from human lymphoid cells, either malignant or not, amounted to 1.2-1.4% of the total nDNA. This was labeled with 125I and utilized as a probe in RNA-driven iterative hybridization and cross-hybridization assays. In all cases of neoplasia studied, the same subfraction of ssDNA (equivalent to 10-12% of the total ssDNA) from malignant cells could be hybridized with cytoplasmic RNA from malignant cells, whatever their origin: acute lymphoid leukemia cells collected from blood of leukemic patients, local lymphosarcomas, or cells from two established strains grown in suspension, one Epstein-Barr virus positive (Raji strain), the other Epstein--Barr virus negative (Ramos strain). The majority of these tumor-specific DNA sequences could be reassociated with unique-copy DNA from both normal and malignant cells and were not expressed in normal lymphoid cells, including those of an established strain immortalized by Epstein--Barr virus and multiplied in culture. It is concluded that a great number of new transcription units, in the range of 2500-3000 per cell genome, were activated after spontaneous malignant transformation of lymphoid cells.
Inhibition of prostate tumor growth in two rat models by chronic administration of D-Trp6 analogue of luteinizing hormone-releasing hormone.
Authors: Redding T W , Schally A V
Published: May 2006

We have investigated the effect of the D-Trp6 analogue of luteinizing hormone-releasing hormone (LH-RH), a superactive analogue of LH-RH, on the growth of two different models of prostate tumors in rats. Chronic administration of D-Trp6-LH-RH in a dose of 25 micrograms/day for 14-21 days significantly inhibited the growth of the chemically induced squamous cell carcinoma 11095 in Fisher 344 male rats. The weights of the ventral prostate and testes were also significantly reduced by treatment with this analogue. After 21 days of treatment, the animals no longer showed increases in serum luteinizing hormone and follicle-stimulating hormone levels in response to D-Trp6-LH-RH. Treatment of male Copenhagen F-1 rats bearing the Dunning 3327 prostate adenocarcinoma with 25 micrograms of D-Trp6-LH-RH per day for 42 days decreased the weights of both the ventral prostate and testes but had no effect on the weight of the anterior pituitary gland. The percentage increase in tumor volume was decreased to one-third and the actual tumor weight was decreased by 58% compared to untreated controls. The tumor doubling time was more than 4 times longer in rats receiving D-Trp6-LH-RH than in controls. Serum levels of luteinizing hormone and follicle-stimulating hormone were significantly decreased in rats receiving this analogue. In both Fisher 344 and Copenhagen F-1 rats, serum prolactin and testosterone levels were significantly decreased after treatment with D-Trp6-LH-RH, whereas progesterone levels were increased.
Immunocytochemical localization of aspartate aminotransferase immunoreactivity in cochlear nucleus of the guinea pig.
Authors: Altschuler R A , Neises G R , Harmison G G , Wenthold R J , Fex J
Published: May 2006

There is substantial evidence supporting the role of aspartate or glutamate as the neurotransmitter of the auditory nerve. The concentration of aspartate aminotransferase (L-aspartate:2-oxoglutarate aminotransferase, EC 2.6.1.1), an enzyme associated with the metabolism of these amino acids, is high in axons and terminals of the auditory nerve. Antibodies were raised against aspartate aminotransferase and used in immunocytochemical studies to determine its localization in the cochlear nucleus of the guinea pig. Indirect immunofluorescence techniques were used for light microscopic localization of aspartate aminotransferase-like immunoreactivity in normal guinea pigs and guinea pigs with auditory nerve lesions. Fluorescent rings of aspartate aminotransferase-like immunoreactivity were seen around spherical cells in the anteroventral cochlear nucleus. In animals with auditory nerve lesions, rings were no longer seen in the ipsilateral cochlear nucleus. Immunoreactivity was also seen on cells in the posteroventral cochlear nucleus and in auditory nerve fibers. Ultrastructural studies were done in the rostral anteroventral cochlear nucleus, using the peroxidase-antiperoxidase technique. Aspartate aminotransferase-like immunoreactivity was seen at axosomatic synapses on large spherical cells in terminals with the morphological characteristics of auditory nerve terminals. Other classes of terminals on the soma of large spherical cells showed no immunoreactivity. It was concluded that aspartate aminotransferase-like immunoreactivity is present in axons and terminals of the auditory nerve. These findings indicate that aspartate aminotransferase-like immunoreactivity may serve as a marker at terminals where aspartate or glutamate is a neurotransmitter.
Synthesis and determination of configuration of natural 25-hydroxyvitamin D
3
26,23-lactone
Authors: Eguchi Tadashi , Takatsuto Suguru , Ishiguro Masaji , Ikekawa Nobuo , Tanaka Yoko , DeLuca Hector F.
Published: May 2006

The four stereoisomers of 25-hydroxyvitamin D
3
26,23-lactone were synthesized by a stereoselective lactonization method. Natural 25-hydroxyvitamin D
3
26,23-lactone was produced from 25-hydroxy-[3α-
3
H]vitamin D
3
by
in vitro
incubation of kidney homogenate prepared from vitamin D-supplemented chickens or was isolated from the serum of rats given 1,25-dihydroxyvitamin D
3
and 25-hydroxy-[3α-
3
H]vitamin D
3
. The four synthetic isomers and the naturally produced 25-hydroxyvitamin D
3
26,23-lactone were subjected to high-performance liquid chromatography in a system capable of separating the four isomers. The natural lactone comigrated with synthetic (23
S
,25
R
)-25-hydroxyvitamin D
3
26,23-lactone, establishing it as the natural vitamin D
3
metabolite.
Purine metabolism in the protozoan parasite Eimeria tenella.
Authors: Wang C C , Simashkevich P M
Published: May 2006

Crude extracts of the oocysts of Eimeria tenella, a protozoan parasite of the coccidium family that develops inside the caecal epithelial cells of infected chickens, do not incorporate glycine or formate into purine nucleotides; this suggests lack of capability for de novo purine synthesis by the parasite. The extracts, however, contain high levels of activity of the purine salvage enzymes: hypoxanthine, guanine, xanthine, and adenine phosphoribosyltransferases and adenosine kinase. The absence of AMP deaminase from the parasite indicates that E. tenella cannot convert AMP to GMP; the latter thus has to be supplied by the hypoxanthine, xanthine, or guanine phosphoribosyltransferase of the parasite. These three activities are associated with one enzyme (HXGPRTase), which has been purified to near homogeneity in high yield (71-80%) in a single step by GMP-agarose affinity column chromatography. The size of the enzyme subunit is estimated to be 23,000 daltons by NaDodSO4 gel electrophoresis. Kinetic studies suggest differences in purine substrate specificity between E. tenella HXGPRTase and chicken liver HGPRTase. Allopurinol preferentially inhibits the parasite enzyme by competing with hypoxanthine; a Ki approximately 22 microM.
Purified glucocorticoid receptors bind selectively in vitro to a cloned DNA fragment whose transcription is regulated by glucocorticoids in vivo.
Authors: Payvar F , Wrange O , Carlstedt-Duke J , Okret S , Gustafsson J A , Yamamoto K R
Published: May 2006

Activated glucocorticoid receptor protein, purified to 40-60% homogeneity from rat liver extracts, binds selectively in vitro to a cloned fragment of murine mammary tumor virus (MTV) DNA. The DNA fragment tested contains about half of the sequences present in intact MTV DNA, and its rate of transcription, like that of the intact viral element, is strongly stimulated by glucocorticoids when it is introduced into the genome of a receptor-containing cell. In contrast, the receptor fails to bind selectively to DNA restriction fragments from E. coli plasmids pBR322 and RSF2124 or from bacteriophages lambda and T4. Preliminary experiments to localize regions within MTV DNA responsible for selective binding have revealed thus far one subfragment that fails to bind the receptor and one selectively bound subfragment that maps far downstream from the 5' terminus of the normal RNA transcript. These studies are consistent with the notion that steroid receptors may modulate rates of transcription by recognizing specific DNA sequences within or near the regulated genes.
Internal control regions for transcription of eukaryotic tRNA genes.
Authors: Sharp S , DeFranco D , Dingermann T , Farrell P , Söll D
Published: May 2006

We have identified the region within a eukaryotic tRNA gene required for initiation of transcription. These results were obtained by systematically constructing deletions extending from the 5' or the 3' flanking regions into a cloned Drosophila tRNAArg gene by using nuclease BAL 31. The ability of the newly generated deletion clones to direct the in vitro synthesis of tRNA precursors was measured in transcription systems from Xenopus laevis oocytes, Drosophila Kc cells, and HeLa cells. Two control regions within the coding sequence were identified. The first was essential for transcription and was contained between nucleotides 8 and 25 of the mature tRNA sequence. Genes devoid of the second control region, which was contained between nucleotides 50 and 58 of the mature tRNA sequence, could be transcribed but with reduced efficiency. Thus, the promoter regions within a tRNA gene encode the tRNA sequences of the D stem and D loop, the invariant uridine at position 8, and the semi-invariant G-T-psi-C sequence.
Molecular cloning of equine herpesvirus type 1 DNA: analysis of standard and defective viral genomes and viral sequences in oncogenically transformed cells.
Authors: Robinson R A , Tucker P W , Dauenhauer S A , O'Callaghan D J
Published: May 2006

Genomic DNA sequences of equine herpesvirus type 1 (EHV-1) have been cloned as BamHI and EcoRI restriction fragments into the plasmid pBR322 and propagated in Escherichia coli. With the exception of two EcoRI restriction fragments that reside in the S region of the viral genome, all of the cloned fragments demonstrated the same electrophoretic mobilities, restriction cleavage sites, and blot-hybridization patterns as did the parent fragments produced by BamHI or EcoRI digestion of virion DNA. The EcoRI J fragment and the BamHI E fragment of the L-region terminus were cloned after the addition of appropriate linker oligonucleotides. Fragments originating from each of the two isomeric forms of EHV-1 DNA were contained in this library of clones. Supramolar DNA fragments present only in the DNA of defective interfering (DI) particles of EHV-1 were generated from Bgl II digestion of DNA preparations enriched for EHV-1 DI particles and were cloned as Bgl II and EcoRI fragments into the plasmid vector. The cloned viral sequences represented in this defective genome mapped to the S region of EHV-1 DNA. Blot-hybridization analysis of EHV-1 transformed and tumor cell DNAs with the cloned BamHI B fragment confirmed that subgenomic viral sequences are present and indicated that those sequences map to the viral genome between 0.32 and 0.43 map unit.
Human beta-endorphin: specific binding in neuroblastoma N18TG2 cells.
Authors: Hammonds R G , Li C H
Published: May 2006

The mouse neuroblastoma N18TG2 has specific, saturable binding sites for human beta-endorphin (beta h-EP). The affinity and number of sites are 1.1 nM and 280,000 per cell, respectively, beta h-EP binding is not inhibited by [Leu]enkephalin or morphine at concentrations up to 0.1 mM; beta h-EP-(6--31) is a potent inhibitor of binding, and camel beta-EP is much less potent. The data suggest the importance of the nonenkephalin segment of the beta h-EP molecule for interaction with the binding site in N18TG2 cells.
Modulation of nucleosome structure by histone subtypes in sea urchin embryos.
Authors: Simpson R T
Published: May 2006

Switches of the types of histones synthesized and incorporated into chromatin occur during sea urchin embryogenesis. In an attempt to define the possible effects of these variant histones on chromatin structure, I have isolated and characterized nucleosome core particles from Strongylocentrotus purpuratus blastula (nearly 100% early histones) and pluteus (75% late histones). Both particles contain 146-base-pair lengths of DNA wrapped around an octamer of H2A, H2B, H3, and H4. Although sharing these similarities with the canonical core particle, the nucleosome structures have certain features that differ from those of typical adult tissues. Both the reversible and the irreversible conformational transitions occurring on heating core particles are destabilized in the embryonic particles vs. "typical" core particles. The blastula core particle unfolds more easily than pluteus (or other) nucleosomes under the stress of low ionic strength. The rate of DNase I digestion of pluteus core particles is about half that of particles from blastula; certain cutting sites differ in their susceptibility between the two embryonic particles and between these two and the canonical core particle. The data demonstrate that the variant histones synthesized during early embryogenesis have demonstrable effects on chromatin structure, even at this basic level.
Bridging of IgE receptors activates phospholipid methylation and adenylate cyclase in mast cell plasma membranes.
Authors: Ishizaka T , Hirata F , Sterk A R , Ishizaka K , Axelrod J A
Published: May 2006

Bridging of IgE receptors on normal rat mast cells by divalent anti-receptor antibodies induced phospholipid methylation and an increase in intracellular cyclic AMP within 15 sec after the receptor bridging. These biochemical events were followed by Ca2+ influx and histamine release. When IgE receptors on isolated plasma membranes were bridged by the antibody, both the increase in the incorporation of [3H]methyl into lipid fraction and the synthesis of cyclic AMP were demonstrated. The synthesis of cyclic AMP in this system was enhanced in the presence of GTP. The results indicated that the bridged IgE receptors are linked to both methyltransferases and adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] in the plasma membrane. An increase in cyclic AMP prior to receptor bridging suppressed phospholipid methylation in the plasma membrane, Ca2+ uptake, and subsequent histamine release. On the other hand, inhibition of phospholipid methylation by (S)-isobutyryl-3-deazaadenosine resulted in the suppression of cyclic AMP synthesis in the plasma membrane. These findings suggest that the activation of phospholipid methylation and the activation of adenylate cyclase are e, and subsequent histamine release. On the other hand, inhibition of phospholipid methylation by (S)-isobutyryl-3-deazadenosine resulted in the suppression of cyclic AMP synthesis in the plasma membrane. These findings suggest that the activation of phospholipid methylation and the activation of adenylate cyclase are e, and subsequent histamine release. On the other hand, inhibition of phospholipid methylation by (S)-isobutyryl-3-deazadenosine resulted in the suppression of cyclic AMP synthesis in the plasma membrane. These findings suggest that the activation of phospholipid methylation and the activation of adenylate cyclase are mutually regulated.
Somatomedin-like peptide(s) isolated from fetal bovine cartilage (cartilage-derived factor): isolation and some properties.
Authors: Kato Y , Nomura Y , Tsuji M , Kinoshita M , Ohmae H , Suzuki F
Published: May 2006

Fetal bovine cartilage contains a polypeptide(s) that has somatomedin-like effects on rat and rabbit costal chondrocytes in culture. This factor, named the cartilage-derived factor, was extracted from fetal bovine cartilage, fractionated with acetone, and purified by gel filtration on Toyopearl HW 55-F in 4 M guanidine hydrochloride, preparative isoelectric focusing, and subsequent gel filtration on Toyopearl HW 55-F in 1 M formic acid. The resulting preparation, which focused in the neutral pH region and eluted from a Toyopearl column in a fraction with apparent Mr 10,000--11,000, appeared homogenous by NaDodSO4 gel electrophoresis. The purified preparation markedly enhanced not only proteoglycan synthesis but also DNA synthesis in rabbit costal chondrocytes and, on a protein basis, it was 1000 times more active than insulin and 1,000,000 times more active than fetal calf serum in stimulating proteoglycan synthesis.
Amino acid sequences of bacterial cytochromes c' and c-556.
Authors: Ambler R P , Bartsch R G , Daniel M , Kamen M D , McLellan L , Meyer T E , Van Beeumen J
Published: May 2006

The cytochrome c' are electron transport proteins widely distributed in photosynthetic and aerobic bacteria. We report the amino acid sequences of the proteins from 12 different bacterial species, and we show by sequences that the cytochromes c-556 from 2 different bacteria are structurally related to the cytochromes c'. Unlike the mitochondrial cytochromes c, the heme binding site in the cytochromes c' and c-556 is near the COOH terminus. The cytochromes c-556 probably have a methionine sixth heme ligand located near the NH2 terminus, whereas the cytochromes c' may be pentacoordinate. Quantitative comparison of cytochrome c' and c-556 sequences indicates a relatively low 28% average identity.
Uric acid provides an antioxidant defense in humans against oxidant- and radical-caused aging and cancer: a hypothesis.
Authors: Ames B N , Cathcart R , Schwiers E , Hochstein P
Published: May 2006

During primate evolution, a major factor in lengthening life-span and decreasing age-specific cancer rates may have been improved protective mechanisms against oxygen radicals. We propose that one of these protective systems is plasma uric acid, the level of which increased markedly during primate evolution as a consequence of a series of mutations. Uric acid is a powerful antioxidant and is a scavenger of singlet oxygen and radicals. We show that, at physiological concentrations, urate reduces the oxo-heme oxidant formed by peroxide reaction with hemoglobin, protects erythrocyte ghosts against lipid peroxidation, and protects erythrocytes from peroxidative damage leading to lysis. Urate is about as effective an antioxidant as ascorbate in these experiments. Urate is much more easily oxidized than deoxynucleosides by singlet oxygen and is destroyed by hydroxyl radicals at a comparable rate. The plasma urate levels in humans (about 300 microM) is considerably higher than the ascorbate level, making it one of the major antioxidants in humans. Previous work on urate reported in the literature supports our experiments and interpretations, although the findings were not discussed in a physiological context.
Cholesteryl myristate conformation in liquid crystalline mesophases determined by neutron scattering.
Authors: Burks C , Engelman D M
Published: May 2006

The possible involvement of cholesteryl ester states in the development and persistence of atherosclerosis and the transport and storage of cholesteryl esters has led to questions concerning the organization and conformation of cholesteryl ester molecules in both pure phases and membranes. The experiments we report here were designed to measure the distance between the center of mass of the fatty acyl terminal methyl group and the center of mass of the three-carbon branched terminus of the cholesterol moiety at the opposite end of the molecule. The distance obtained is thus a gauge of cholesteryl ester conformation through the conformational range from a completely extended conformation to a U-shaped conformation. Neutron scattering experiments on partially deuterated samples of pure cholesteryl myristate in the crystalline, smectic, cholesteric, and isotropic phases indicate that the molecule is extended in each of these states. A discussion of specific molecular models consistent with these results and extension of these conclusions to other cholesteryl esters is included.
Computer-enhanced video microscopy: digitally processed microscope images can be produced in real time.
Authors: Walter R J , Berns M W
Published: May 2006

Digital processing techniques can be used to greatly enhance the available information in an optical image. Although this technology has been routinely used in many fields for a number of years, little application of digital image-processing techniques have been made toward analysis and enhancement of the types of images seen most often by the research biologist. We describe here a computer-based video microscope system that is capable of performing extensive manipulation and enhancement of microscope images in real time. The types of manipulations possible with these techniques greatly surpass the enhancement capabilities of photographic or video techniques alone. The speed and flexibility of this system enables experimental manipulation of the microscopic specimen based on its live processed image. These features greatly extend the power and versatility of the light microscope.
Nuclear DNA from uninfected or potato spindle tuber viroid-infected tomato plants contains no detectable sequences complementary to cloned double-stranded viroid cDNA.
Authors: Hadidi A , Cress D E , Diener T O
Published: May 2006

High molecular weight tomato nuclear DNA was isolated from uninfected and potato spindle tuber viroid (PSTV)-infected tomato leaves. Restriction digests were fractionated on agarose gels, denatured and transferred to diazobenzyloxymethylpaper, and hybridized to 32P-labeled cloned double-stranded PSTV cDNA. No hybridization to DNA from either uninfected or infected tissue could be detected under conditions that permitted detection of cloned double-stranded PSTV cDNA at a concentration equivalent to one-fifth copy of PSTV-related DNA per haploid tomato genome. Hybridization of tomato DNA to 32P-labeled cloned soybean 18S and 28S ribosomal DNA sequences showed that the restricted nuclear DNA was suitable for hybridization to probes containing homologous sequences. Our results indicate that neither PSTV nor its complementary strand is transcribed from nuclear DNA but do not rule out the possibility of sequence homology between host DNA and a small portion of PSTV or its complement.
Electrical stimulation of sympathetic nerves increases the concentration of cyclic AMP in rat pineal gland.
Authors: Heydorn W E , Frazer A , Weiss B
Published: May 2006

Electrical stimulation of the superior cervical ganglia causes a rapid increase in the concentration of cyclic AMP in the pineal gland of rats. This effect is dependent upon the frequency, voltage, and duration of the stimulus and is markedly potentiated by pretreating the animals with desmethylimipramine. The increase in cyclic AMP is blocked by prior treatment of the rats with reserpine, bretylium, or propanolol but not with phentolamine. These results provide direct evidence that electrical stimulation of sympathetic nerves increases cyclic AMP in a target organ through the release of norepinephrine from presynaptic terminals acting on postsynaptic beta-adrenergic receptors.