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31.
32.
Barbara Huisamen Elna Ellis Magdalena van Dyk Amanda Lochner 《Molecular and cellular biochemistry》1996,162(1):1-9
Although it is well-accepted that the phosphatidylinositol signalling transduction pathway, producing inositol-1,4,5-P3 (InsP3) and inositol-1,3,4,5-P4 (InsP4) as second messengers, functions in heart muscle, virtually nothing is known about the roles of the higher inositol polyphosphates such as inositolhexakisphosphate (InsP6). This study demonstrates that InSP6 has the ability to bind intracellularly, with different binding characteristics, to different myocardial membranes. Binding to purified sarcoplasmic reticulum (SR) membranes, purified sarcolemmal (SL) membranes as well as to viable mitochondria were characterized. Binding to all these membranes display high as well as low affinity binding sites, with differing affinities. Kd values of binding to SR were 32 and 383 nM, to SL 61 and 1312 nM, while those of mitochondrial binding were 230 and 2200 nM respectively.InsP4 binding was also investigated and displayed the following characteristics: to SR, one low affinity binding site (Kd = 203 nM) and to SL, a high as well as a low affinity binding site with Kd values of 41 and 2075 nM respectively. Presence of InsP3, the second messenger for SR calcium release, at concentrations of 1 nM, elevated the binding of InsP4 to SR and SL by a mean of 30% and 20% respectively.Fractionation of SR and SL membranes on sucrose density gradients, after solubilization with CHAPS, indicated that InsP6 bound to two separate protein peaks in both these membranes, while InsP4 bound to only one. In SR membranes, InsP4 bound preferentially to a protein separating at high sucrose density while it bound to a protein separating at low sucrose density in SL membranes. 相似文献
33.
Evidence for natural horizontal transfer of tetQ between bacteria that normally colonize humans and bacteria that normally colonize livestock. 总被引:8,自引:1,他引:7
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Though numerous studies have shown that gene transfer occurs between distantly related bacterial genera under laboratory conditions, the frequency and breadth of horizontal transfer events in nature remain unknown. Previous evidence for natural intergeneric transfers came from studies of genes in human pathogens, bacteria that colonize the same host. We present evidence that natural transfer of a tetracycline resistance gene, tetQ, has occurred between bacterial genera that normally colonize different hosts. A DNA sequence comparative approach was taken to examine the extent of horizontal tetQ dissemination between species of Bacteroides, the predominant genus of the human colonic microflora, and between species of Bacteroides and of the distantly related genus Prevotella, a predominant genus of the microflora of the rumens and intestinal tracts of farm animals. Virtually identical tetQ sequences were found in a number of isolate pairs differing in taxonomy and geographic origin, indicating that extensive natural gene transmission has occurred. Among the exchange events indicated by the evidence was the very recent transfer of an allele of tetQ usually found in Prevotella spp. to a Bacteroides fragilis strain. 相似文献
34.
David J. Bunyan David O. Robinson Amanda L. Collins Annette E. Cockwell Hilary M. S. Bullman Paul A. Whittaker 《Human genetics》1994,93(5):541-544
The family of a male with Duchenne muscular dystrophy (DMD) and a deletion within the dystrophin gene has been studied. Polymerase chain reaction analysis of ectopic mRNA from peripheral blood T+B lymphocytes and the use of (CA)
n
repeat polymorphisms in and around the deleted region showed the proband's mother to be both a germline mosaic and a somatic mosaic for the deletion seen in her son. The mutation therefore occurred as a mitotic event early in embryogenesis. 相似文献
35.
The mercury-silver (Hg-Ag) argentaffin technique, known to stain specifically proteins in the lateral components of triads/diads in striated muscle cells, was applied to the central nervous system of adult rats. Following fixation in glutaraldehyde, axons in white and gray matter were selectively stained, but not perikarya or their proximal axon and dendrites. Neural tissues were postfixed 24 hr in 5% (w/v) mercuric acetate in 2% (v/v) acetic acid in distilled water, stained for 12-24 hr in darkness at 37-43 C with ammoniacal silver nitrate solution, freshly prepared by adding concentrated ammonia to 60% (w/v) silver nitrate solution until a small amount of silver oxide precipitate remained undissolved. Samples were then washed with freshly prepared 5% (w/v) sodium sulfite and distilled water. All steps were carried out using dark-colored glass flasks. Samples were dehydrated with ethanol and embedded in Paraplast or Poly Bed. Electron microscopy showed the silver-reducing protein inside the axons. Methylation abolished Hg-Ag axonal reactivity indicating that carboxyl groups were necessary for silver staining. Proteins with solubility properties characteristic of neurofilament proteins were involved in Hg-Ag staining. In the cerebellum the plexus of parallel fibers in the molecular layer were not stained, while basket cell axonal processes reacted intensely. The method appears to distinguish neuronal protein variants related to cytotypic differences in cytoskeletal neurofilaments. 相似文献
36.
D. P. Chora L. Reddy S. K. Gupta L. Wan P. A. Mathieu R. L. Shoemaker J. S. Rhim 《In vitro cellular & developmental biology. Animal》1994,30(8):539-546
Summary Cystic fibrosis (CF) involves abnormalities in mucus production and secretion of the airway. Studies of the regulation of
airway mucin production and secretion has been difficult due to the lack of in vitro models of the airway epithelial cells
which express functional differentiation. Because the majority of the mucin in the airway is apparently produced by the submucosal
glands, we have focused our attention on the development of cell culture models of human airway submucosal glands. This report
describes the propagation of CF airway submucosal gland epithelial cells which continue to express mucin production. The CF
bronchus was obtained from a 31-yr-old patient who received a double lung transplant. The glands were dissected out and primary
cultures prepared by the explant/outgrowth procedure. The cells were immortalized by infection with Adl2-SV40 hybrid virus.
The cultures are maintained in serum-free keratinocyte basal medium supplemented with insulin (5μg/ml), hydrocortisone (0.5μg/ml), epidermal growth factor (10 ng/ml), bovine pituitary extract (25μg/ml), and antibiotics. Cultures were passaged using 0.125% trypsin in Ca+2 and Mg+2-free Hanks’, balanced salt solution. Polymerase chain reaction (PCR) analysis demonstrated that the cells were homozygous
for the ΔF508 mutation. Morphologic observations showed that the cells were epithelial and were interconnected by sparsely
distributed desmosomes. Their cytoplasm contained secretory-type structures including abundant Golgi, rough endoplasmic reticulum,
and secretory vesicles. Immunofluorescent studies determined that all cells were positive for cytokeratins, mucin glycoconjugates,
and cystic fibrosis transmembrane conductance regulator. The cultures secreted substantial amounts of mucin glycoproteins
and expressed the MUC-2 mucin gene. Patch clamp experiments revealed that the cells expressed defective Cl− channels which were not activated by Forskolin. 相似文献
37.
Induction of B-A transitions of deoxyoligonucleotides by multivalent cations in dilute aqueous solution. 总被引:4,自引:1,他引:3
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Circular dichroism (CD) spectra of d(CCCCGGGG) in the presence of Co(NH3)6(3+) are very similar to spectra of r(CCCCGGGG). In contrast, B-form characteristics are observed for d(CCCCGGGG) in the presence of Na+ and Mg2+, even at high salt concentrations. Spermidine induces modest changes of the CD of d(CCCCGGGG). The NMR chemical shifts of the nonexchangeable protons of d(CCCCGGGG) in the absence and presence of Co(NH3)6(3+) were assigned by proton two-dimensional (2D) NOESY and COSY measurements. The chemical shifts of the GH8 protons of d(CCCCGGGG) move upfield upon titration with Co(NH3)6Cl3. The sums of the sugar H1' coupling constants decrease with added Co(NH3)6Cl3. Cross peak intensities in the 2D proton NOESY spectra show a transformation from B-DNA to A-DNA characteristics upon the addition of Co(NH3)6Cl3. The temperature-dependent 59Co transverse and longitudinal relaxation rates demonstrate that Co(NH3)6(3+) is site-bound to the oligomer. Such localization is not a general feature of Co(NH3)6(3+) binding to oligonucleotides. 59Co NMR relaxation and CD measurements demonstrate chiral discrimination by d(CCCCGGGG) for the two stereoisomers of Co(en)3(3+). Both stereoisomers bind tightly as judged by 59Co NMR, and both cause large (but nonequivalent) changes in the CD of this oligomer. 相似文献
38.
M. S. Hawbaker W. R. Fehr L. M. Mansur R. C. Shoemaker R. G. Palmer 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,87(1-2):49-53
Tissue culture may generate useful genetic variation for quantitative traits. The objective of this study was to analyze genetic variation for ten quantitative traits of soybean [Glycine max (L.) Merr.] among lines derived from the tissue culture of three cultivars. The three cultivars used to obtain R0 plants from tissue culture were BSR 101, Hodgson 78, and Jilin 3. A total of 63 R0-derived lines of BSR 101, eight of Hodgson 78, and 42 of Jilin 3 was planted with the untreated controls in row plots in a randomized complete-block design with three replications at two locations for each of 2 years. The traits evaluated were days to beginning bloom (R1), beginning seed (R5), beginning maturity (R7), full maturity (R8), height, lodging, seed yield, seed weight, protein content, and oil content. Significant (P < 0.05) variation was observed among lines for each of the ten quantitative traits. There was 57.1% of the BSR 101 lines, 87.5% of the Hodgson 78 lines, and 76.2% of the Jilin 3 lines that were significantly different from the controls for at least one trait. The percentages of lines that were significantly different from the control for an individual trait ranged from 2.7% for oil content to 25.7% for R7. The magnitude of the changes was relatively small. Although this genetic variation may be useful for cultivar development, greater variability at less expense would be expected with conventional artificial hybridization.Journal Paper No. J-14958 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IOWA, USA Project No. 2475. 相似文献
39.
David W. Kikuchi William L. Allen Kevin Arbuckle Thomas G. Aubier Emmanuelle S. Briolat Emily R. Burdfield-Steel Karen L. Cheney Klára Daňková Marianne Elias Liisa Hämäläinen Marie E. Herberstein Thomas J. Hossie Mathieu Joron Krushnamegh Kunte Brian C. Leavell Carita Lindstedt Ugo Lorioux-Chevalier Melanie McClure Callum F. McLellan Iliana Medina Viraj Nawge Erika Páez Arka Pal Stano Pekár Olivier Penacchio Jan Raška Tom Reader Bibiana Rojas Katja H. Rönkä Daniela C. Rößler Candy Rowe Hannah M. Rowland Arlety Roy Kaitlin A. Schaal Thomas N. Sherratt John Skelhorn Hannah R. Smart Ted Stankowich Amanda M. Stefan Kyle Summers Christopher H. Taylor Rose Thorogood Kate Umbers Anne E. Winters Justin Yeager Alice Exnerová 《Journal of evolutionary biology》2023,36(7):975-991
Prey seldom rely on a single type of antipredator defence, often using multiple defences to avoid predation. In many cases, selection in different contexts may favour the evolution of multiple defences in a prey. However, a prey may use multiple defences to protect itself during a single predator encounter. Such “defence portfolios” that defend prey against a single instance of predation are distributed across and within successive stages of the predation sequence (encounter, detection, identification, approach (attack), subjugation and consumption). We contend that at present, our understanding of defence portfolio evolution is incomplete, and seen from the fragmentary perspective of specific sensory systems (e.g., visual) or specific types of defences (especially aposematism). In this review, we aim to build a comprehensive framework for conceptualizing the evolution of multiple prey defences, beginning with hypotheses for the evolution of multiple defences in general, and defence portfolios in particular. We then examine idealized models of resource trade-offs and functional interactions between traits, along with evidence supporting them. We find that defence portfolios are constrained by resource allocation to other aspects of life history, as well as functional incompatibilities between different defences. We also find that selection is likely to favour combinations of defences that have synergistic effects on predator behaviour and prey survival. Next, we examine specific aspects of prey ecology, genetics and development, and predator cognition that modify the predictions of current hypotheses or introduce competing hypotheses. We outline schema for gathering data on the distribution of prey defences across species and geography, determining how multiple defences are produced, and testing the proximate mechanisms by which multiple prey defences impact predator behaviour. Adopting these approaches will strengthen our understanding of multiple defensive strategies. 相似文献
40.
Amanda dos Santos Marina Tenório Botelho Willian Reina Joviano Vicente Gomes José Roberto Machado Cunha da Silva Gisela de Aragão Umbuzeiro 《Invertebrate Biology》2023,142(1):e12394
Hemocytes are circulating blood cells that play a crucial function in amphipods and other crustacean immune systems. The hemocytes of the marine tropical amphipod Parhyale hawaiensis have been used for the evaluation of DNA damage and micronuclei, but they have not been characterized in the scientific literature. The aim of this study was to describe the hemolymph cells of P. hawaiensis and study their phagocytotic activity. Basic dyes were used to differentiate the cell types and the presence of lipids. The total hemocyte counts (THCs) and the proportion and sizes of the hemocyte types were determined. Hemolymph was exposed to Escherichia coli for verification of the presence of phagocytosis. Three cell types, all containing lipids, were identified in P. hawaiensis: granulocytes (oval shape, 13.4 × 7.6 μm), semi-granulocytes (oval shape, 14.1 × 7.2 μm), and hyalinocytes (round shape, 9.6 × 7.2 μm). Those three cell types were found in different percentages in males (64.8%, 31.1%, and 4.2%) and females (70.1%, 28.2%, and 1.7%). THCs for males were 9007 ± 3800 cells per individual and 4695 ± 1892 cells per individual for females. The cells of E. coli were phagocytized by the hemocytes. Our findings increased the knowledge of hemocytes in P. hawaiensis and is a step forward in using hemocyte-based immune responses as an endpoint in ecotoxicology. 相似文献