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Identification and estimation algorithm for stochastic neural system. II.

The algorithm for identifying the stochastic neural system and estimating the system process which reflects the dynamics of the neural network are presented in this paper. The analogous algorithm has been proposed in our preceding paper (Nakao et al., 1984), which was based on the randomly missed observations of a system process only. Since the previous algorithm mentioned above was subject to an unfavorable effect of consecutively missed observations, to reduce such an effect the algorithm proposed here is designed additionally to observe an intensity process in a neural spike train as the information for the estimation. The algorithm is constructed with the extended Kalman filters because it is naturally expected that a nonlinear and time variant structure is necessary for the filters to realize the observation of an intensity process by means of mapping from a system process to an intensity process. The performance of the algorithm is examined by applying it to some artificial neural systems and also to cat's visual nervous systems. The results in these applications are thought to prove the effectiveness of the algorithm proposed here and its superiority to the algorithm proposed previously.

Animals↗

Molecular and pharmacological identity of the alpha 2D-adrenergic receptor subtype in bovine retina and its photoreceptors.

The rat cA2-47 gene encodes the pharmacologically defined alpha 2D-adrenergic receptor (alpha 2D-AR) subtype. Previously, the expression of its mRNA was shown in bovine retina by amplification through the reverse transcription-polymerase chain reaction (RT-PCR) of a region corresponding to the rat alpha 2D-AR, amino acid (aa) residues 382-439, indicating the presence of this subtype in this neural tissue. In the present study, the structure of this gene has been probed and the encoded receptor subtype has been characterized in bovine retina and its photoreceptor cells. The deduced aa sequence of the two bovine gene fragments, aa residues 290-375 and aa residues 392-434, demonstrates 77% overall identity with the rat alpha 2D-AR subtype and 80% overall identity with the mouse alpha 2D-AR. The receptor encoded by the bovine gene was expressed in the retina and its photoreceptors with the typical pharmacological characteristics established for the rat alpha 2D-AR subtype: The receptor bound rauwolscine with a KD of 14 nM in the retina and with that of 19 nM in the photoreceptor cells; the binding association rate constant, k+1, for the ligand was 0.012 min-1, the dissociation rate constant, k-1, was 0.14 min-1 and the half-time for dissociation was 5 min. Oxymetazoline displaced the bound [3H]-rauwolscine with an EC50 value of 85 nM, while SK & F 104078, and prazosin displaced the bound [3H]-rauwolscine with the respective IC50 values of 900 nM and 3000 nM. The other alpha 2-AR subtypes -alpha 2A-AR, alpha 2B-AR, alpha 2C-AR-were not detected in the retina and its photoreceptors. Thus, this study shows that the bovine alpha 2D-AR gene is a structural variant of the rat and mouse genes, that the bovine gene encodes the typical pharmacologically defined alpha 2D-AR subtype, that this subtype is present in its exclusive form in the bovine retina and its photoreceptors, where it may be presynaptic in nature.

Adrenergic alpha-Agonists↗

Malignant versus benign paravertebral widening in children.

Dorso-medial pleural reflection line anomalies occur in malignant tumours, neuroblastoma being the commonest one, in benign tumours, spinal inflammatory disorders and in cases of pseudotumours of different origin. The latter group comprises normal structure variants, malformations and acquired disease but also a small number of transitory or permanent lesions of uncertain or obscure origin. The radiological manifestations encountered are displacement, obliteration or blurring of the paraspinal contour, sometimes a frank paravertebral mass. Based on a case material consisting of 65 patients the importance of recognizing this feature is stressed. An approach to clinical and radiological diagnosis is discussed.

Adolescent↗

Complementation analysis of locus for hypoxanthine guanine phosphoribosyltransferase in Chinese hamster cells.

The study deals with intragenic complementation between clones of Chinese hamster cells carrying mutations in the HPRT gene. All clones were of independent origin, selected in media containing one of three purine bases: 8-azaguanine (8 AG), 6-mercaptopurine (6MP), or 6-thioguanine (6TG). Some of the clones were spontaneous, others were induced by various mutagens. To make the study less time-consuming, an experimental set-up was proposed for simultaneous complementation testing of up to 10 clones. As a result, about 400 combinations of clones have been analyzed. Twelve pairs of complementating mutants have been identified in HAT medium. A linear complementation map has been constructed for the HPRT locus, showing five complementation groups. The changes in kinetic and other characteristics observed for mutant HPRT show that all the mutants studied carry structural gene mutations. Analysis of the biochemical characteristics of HPRT has revealed considerable differences between mutant enzymes in clones belonging to different complementation groups (three groups were examined). At the same time, the four mutant clones of complementation group II show similar HPRT characteristics, suggesting a relative similarity of their structural variants of the enzyme. The hybrid nature of HPRT in clones resulting from the fusion of mutant cells confirms the intragenic nature of complementation.

Animals↗

Further chromosomal studies on Ellobius lutescens: heteromorphism of chromosome No. 1 is not associated with sex determination.

Twelve animals of the species Ellobius lutescens from two generations were studied with various chromosomal banding techniques. This species carries 17 chromosomes in both sexes. In preceding studies chromosomal sex determination was assigned to different structural variants of chromosome No. 1. In the present study, no definite chromosomal basis for sex determination was found.

Animals↗

Diversity of coexisting Planktothrix (cyanobacteria) chemotypes deduced by mass spectral analysis of microystins and other oligopeptides.

Cyanobacteria are reported to produce secondary metabolites of which toxic and bioactive peptides are of scientific and public interest. Many peptides are synthesized by the non-ribosomal peptide synthesis pathway and their presence is a stable feature of individual clones. We isolated 18 clonal strains of Planktothrix from a single water sample from lake Maxsee near Berlin and analyzed them by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, HPLC, and PCR for their production of peptides and the presence of microcystin synthetase genes. Microcystins could be detected in seven of the strains with considerable variability of contents and numbers of structural variants. Other known peptides like anabaenopeptins B and E/F, microviridin I, and prenylagaramide B and new variants of known peptide classes like aeruginosins and cyanopeptolins were detected in some strains while lacking in others. The 18 strains represented 15 chemotypes with respect to their peptide patterns. In contrast, all strains were morphologically very similar with respect to cell dimensions and pigmentation. Given the diversity of chemotypes among the randomly selected isolates, an immense diversity of chemotypes in the entire population can be assumed.

Bacterial Proteins↗

Structure and origin of a novel dimeric retroposon B1-diD.

Here we describe a new short retroposon family of rodents. Like the primate Alu element consisting of two similar monomers, it is dimeric, but the left and right monomers are different and descend from B1 and ID short retroposons, respectively. Such elements (B1-dID) were found in the genomes of Gliridae, Sciuridae, Castoridae, Caviidae, and Hystricidae. Nucleotide sequences of this retroposon can be assigned to several structural variants. Phylogenetic analysis of B1-dID and related sequences suggests a possible scenario of B1-dID evolution in the context of rodent evolution.

Animals↗

High-resolution mapping of sperm function defects in the t complex fourth inversion.

Structural variants of the mouse Chr 17-specific t complex, known as t haplotypes, express factors that alter the ability of sperm to carry out their roles in the normal fertilization process. In previous studies of males carrying heterospecific combinations of the t complex, we discovered a unique M. spretus/t haplotype phenotype of male sterility. In additional studies with mice carrying a series of M. spretus-M. m. domesticus recombinant Chr 17 homologs and a complete t haplotype (S-+/t), we monitored physiological aspects of sperm function to map a locus (Hst6) responsible for expression of the t-specific "curlicue" sperm flagellar curvature phenotype to 1 cM within the fourth inversion of the t complex. In the present report, we quantitatively analyze the in vitro capability of sperm from mice with similar S-+/t Chr 17 genotypes to fertilize zona pellucida-free mouse eggs. The results identify a locus, Stop1, mapping distal to Pim1, with acute effects on the ability of sperm to penetrate the oolemma. The data suggest that Stop1 is a complex locus consisting of at least two genetic elements, a proximal one overlapping the Hst6 locus, and another, distal to the Hst6 locus. Further quantitative analyses of the "curlicue" phenotype produced by sperm derived from these same animals indicate that expression of this chronic flagellar curvature phenotype also derives from at least two elements, both mapping within the Hst6 locus. Thus, these studies provide higher resolution mapping of the molecular basis of t haplotype-specific sperm dysfunction emanating from In(17)4.

Animals↗

Complex alterations of the ribosomal gene spacers in mutant sc8 of Drosophila melanogaster.

Extreme changes in the proportions of the rDNA intergenic spacers (IGSs) have previously been demonstrated by us in the X-chromosomal rDNA array of the Drosophila melanogaster mutant sc8, where nearly all IGSs were found to be reduced in size. We have cloned different structural variants of the Xsc8 ribosomal units and mapped their spacers by restriction endonuclease analysis. Most of the IGSs exhibited complex rearrangements within the subrepeated region at their 5' terminus. The sequence data revealed the presence of an additional 100 bp subrepeat. In addition, extended deletions/substitutions down to the 18S gene were also found. Examination of the Ysc8 IGS polymorphism indicates that some of the X-chromosomal IGS variants are probably the result of X-Y interchanges. The rarity of alternative recombination products implies that the overabundant deleted spacers in sc8 are generated by nonreciprocal recombination. Our results demonstrate nonrandom distribution of deletional breakpoints within the "1900" region in sc8 and show that the deletions do not truncate the internal IGS subregions at either breakpoint but eliminate them as discrete blocks.

Animals↗

Distribution of pectic epitopes in cell walls of the sugar beet root.

Immunolabelling techniques with antibodies specific to partially methyl-esterified homogalacturonan (JIM5: unesterified residues flanked by methylesterified residues. JIM7: methyl-esterified residues flanked by unesterified residues), a blockwise de-esterified homogalacturonan (2F4), 1,4-galactan (LM5) and 1,5-arabinan (LM6) were used to map the distribution of pectin motifs in cell walls of sugar beet root (Beta vulgaris). PME and alkali treatments of sections were used in conjunction with JIM5-7 and 2F4. The JIM7 epitope was abundant and equally distributed in all cells. In storage parenchyma, the JIM5 epitope was restricted to some cell junctions and the lining of intercellular spaces while in vascular tissues it occurred at cell junctions in some phloem walls and in xylem derivatives. After secondary wall formation, the JIM5 epitope was restricted to inner cell wall regions between secondary thickenings. The 2F4 epitope was not detected without de-esterification treatment. PME treatments prior to the use of 2F4 indicated that HG at cell corners was not acetylated. The LM5 epitope was mainly present in the cambial zone and when present in storage parenchyma, it was restricted to the wall region closest to the plasma membrane. The LM6 epitope was widely distributed throughout primary walls but was more abundant in bundles than in medullar ray tissue and storage parenchyma. These data show that the occurrence of oligosaccharide motifs of pectic polysaccharides are spatially regulated in sugar beet root cell walls and that the spatial patterns vary between cell types suggesting that structural variants of pectic polymers are involved in the modulation of cell wall properties.

Antibodies, Monoclonal↗

Genetic diversity and spread of Bovine leukaemia virus isolates in Argentine dairy cattle.

Effective tools for use in control programmes against bovine leukaemia virus (BLV) infections require insight into the relationship between the variant structure of the bovine leukaemia virus and the spatial-temporal interaction of isolates and hosts. Our study showed the presence of two types of BLV isolates - Australian and Argentine - in dairy herds from various parts of Central Argentina; these isolates were characterised by RFLP on PCR amplicons, and some of them were confirmed by sequencing. One genotype (Argentine) was present in all herds, and the Australian genotype was found in two herds. Phylogenetic analysis indicated four clusters. The first cluster was composed of the Argentine isolates and one from Brazil; the second was composed of several isolates found in European countries and one from Brazil; the third cluster was composed of BLV isolates found in Japan and Germany; the fourth cluster included American and Australian isolates and those from other countries. The comparison of a number of synonymous and non-synonymous nucleotide substitutions using various BLV genes revealed purifying selection, suggesting that molecular evolution occurred under some functional constraint.

Amino Acid Sequence↗

Metolachlor-mediated selection of a microalgal strain producing novel polyunsaturated fatty acids.

Long-chain (> or = C(20)) polyunsaturated fatty acids, such as docosahexaenoic acid and eicosapentaenoic acid, are nutritionally important and provide protection against cardiovascular disease, stroke, and cancer. Structural variants of these compounds may have the potential to be used as pharmaceuticals. Marine microalgae are the key producers of long-chain polyunsaturated fatty acids in the global food web. Assuming vast biological and biochemical diversity, we devised a screen to identify microalgae that produce novel fatty acids. The herbicide metolachlor, an inhibitor of long-chain fatty acid biosynthesis, was used in microcosms containing field-collected microalgae to identify naturally resistant strains. We show that one diatom, Melosira cf. moniliformis, is naturally resistant to concentrations of metolachlor, which were cytostatic or lethal to all the other microalgae. Gas chromatography and gas chromatography-mass spectrometry revealed three fatty acids that have not previously been described-18:4 (Delta5,8,11,14), 18:4 (Delta5,9,12,15), and 18:5 (Delta5,8,11,14,17). We propose that this type of screen may be generally applicable to the search of novel compounds produced by marine microorganisms.

Acetamides↗

Balanced translocation karyotypes in patients with repetitive abortion. Case study and literature review.

Cytogenetic studies were conducted upon 100 consecutive couples with abortions. Eight balanced carrier translocation karyotypes were discovered (8%): three cases of Robertsonian translocations and five reciprocal translocations. Two structural variant karyotypes and a poly-X mosaic were also found. A review of the literature on repetitive abortion revealed 82 balanced translocations in 1,331 couples, a rate of 6.2%. Cytogenetic studied should be routine for patients with repetitive abortion. In the pooled series, 3.7% of couples with translocation had wastage, including some with normal offspring; 9.2% had malformed offspring; 62% of the carrier couples lacked the malformation history. Seventy-four percent of the translocations were reciprocal; risk rates for imbalanced progeny were undefined for 90% of the carrier couples. Only 11 imbalanced conceptuses were demonstrated cytogenetically in 262 pregnancies of the carrier group.

Abortion, Habitual↗

Anticonvulsant activity of deaminated analogues of glutamic acid diethyl ester (GDEE).

GDEE, a specific but low-potency antagonist of the quisqualate or 'Type 2' excitatory amino acid receptor, blocks seizures induced by homocysteine and quisqualic acid. Deaminated analogues of GDEE were examined for anticonvulsant activity in mice, for the purpose of determining the structural properties of GDEE required for anticonvulsant activity. The deaminated derivative of GDEE, diethyl glutarate (5 carbon chain) inhibited homocysteine thiolactone (HTL)-induced seizures with an ED50 of 533 mg/kg. A similar compound with carbon chain length increased by two (diethyl pimelate; 7 carbons) was less effective. Decreases or further increases in carbon chain length resulted in a nearly complete loss of activity. Dimethyl glutarate (5 carbons) and dimethyl adipate (6 carbons) were similar to diethyl glutarate in potency, blocking HTL-induced seizures with ED50s of about 625 and 540 mg/kg, respectively. Diethyl ethylmalonate, diethyl maleate, and diethyl fumarate were much less effective. Diethyl glutarate, but not diethyl succinate (4 carbons), blocked seizures induced by intracerebral quisqualic acid. None of the agents tested blocked pentylenetetrazole-induced seizures. Thus a number of deaminated structural variants of GDEE have anticonvulsant activity equal to that of GDEE. The amino group of GDEE appears therefore to be irrelevant for its anticonvulsant effect.

Animals↗

Dose-response studies of penicillamine and related compounds in reducing skin--tensile strength of rats.

Penicillamine (I) and certain related compounds are known to reduce the skin tensile strength (sts) of rat dorsal skin when they are given in the diet. This effect seems significant to studies of the biochemistry of collagen and of diseases of collagen, perhaps including the arthritides. The reduction of sts appears to be caused by diminution of collagen crosslinking. The effects of several such compounds were studied after intraperitoneal (ip) injection, in order to determine structure-activity relationships divorced from possible anorexic or gastrointestinal complications seen after oral dosing, and to examine the relation of ip dose to response. A cyclopentyl analog (II) of I was at least as active as I, showing that structural variants of I can be active when given ip. A dimethylthiazolidine (V) and a zinc chelate (III, rather toxic) of I were nearly as active as I, showing that probable in vivo precursors of I can be obtained that will be active when given ip. The disulfide of I and a zinc chelate of cysteine were inactive. Maximum response for several compounds seemed to occur at intermediate dose levels, with larger or smaller doses affording less sts reduction.

Animals↗

Alterations in nuclear anatomy by chemical modification of proteins in isolated rat liver nuclei.

Whole rat liver nuclei were treated with citraconic anhydride, a reagent specific for primary amines. Dramatic changes were observed in nuclear morphology and light scattering properties. An analysis for DNA and RNA content suggested that DNA was released from the nuclei with a short half-time, approximately 2-4s demonstrating a biphasic release profile. RNA was similarly released but with a monophasic profile. Analysis of SDS-PAGE gels of modified nuclei demonstrated a progressive enrichment of nuclear matrix (lamins) polypeptides with extent of modification. H1 histone was quantitatively lost as a function of modification reagent concentration, while approx. 50% of the nucleosomal histones cosedimented with DNA- and RNA-free nuclei. Modification in the presence of 2 mM EGTA released all the DNA and RNA [less than or equal to 1% remaining) while retaining structures characteristic of nuclear matrix, nucleoli, and ribonucleoprotein (predominantly hnRNA group A and B). These nucleic acid-deficient structures have been termed nuclear fossils to differentiate them from high salt detergent-prepared empty nuclear sacks, nuclear remnants, or nuclear scaffolds. Modification in the presence of 2% Triton X-100 results in structures similar to the nuclear fossils (EGTA treatment), but missing the double bilayer and a 51K polypeptide that is a major component of the other structures. The use of chemical modification on the nucleus provides an experimental approach for examining the role of ionic interactions in controlling nuclear structure. Citraconylation may thus serve two functions: (a) as a protein-specific perturbant of nuclei capable of simply and rapidly preparing a range of structural variants for the analysis of nuclear interactions; (b) offer a paradigm for control of nucleic acid-polypeptide interactions based on post-translational alterations in protein charge.

Animals↗

Variable base pairing in a helix of eubacterial 5 S ribosomal RNA points to the existence of a conformational switch.

A survey of 160 published sequences of eubacterial 5 S rRNAs shows that there exists structural variability in one of the helices of the generally accepted secondary structure model. Four structural variants are found, which differ with respect to the position and the number of bulges present. Most eubacterial 5 S RNAs fit into at least two of these conformations. A reaction scheme connecting the four observed conformations by changes in the base pairing scheme is proposed. Each of the known 5 S RNA sequences fits into conformations interconvertible by the proposed reactions.

Base Composition↗

Site-directed mutagenesis of alanine-382 of human antithrombin III.

Antithrombin III Hamilton is a structural variant of antithrombin III (AT-III) with normal heparin affinity but impaired serine protease inhibitory activity. The molecular defect of AT-III-Hamilton is a substitution of threonine for alanine at amino acid residue 382. Recently it has been shown that both plasma-derived and cell-free-derived AT-III-Hamilton polypeptides act as substrates rather than inhibitors of thrombin and factor Xa. In the present study, the cell-free expression phagemid vector pGEM-3Zf(+)-AT-III1-432 was mutated at amino acid residue 382 of AT-III to generate 7 cell-free-derived variants. All these cell-free-derived AT-III variants were able to bind heparin as effectively as cell-free-derived normal AT-III. In terms of alpha-thrombin inhibitory activity each variant reacted differently. Variants could be grouped into 3 categories with respect to thrombin-AT-III complex formation: (1) near normal activity (glycine, isoleucine, leucine, valine); (2) low activity (threonine, glutamine); (3) no detectable activity (lysine). These data suggest that mutations at position 382 of AT-III may have a variable effect on protease inhibitory activity, depending on either the stability of the P12-P9 region of the exposed loop of AT-III, or the inability of the amino acid residue at position 382 to interact with a conserved hydrophobic pocket consisting of phenylalanine (at positions 77, 221 and 422) and isoleucine (position 412) residues.

Alanine↗