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31.
Catherine Digonnet-Kerhoas Gilles Gay Jean Claude Duplan Christian Dumas 《Planta》1989,179(2):165-170
During ageing of the short-lived pollen grains of Cucurbita pepo L., water loss was examined in relation to viability using biophysical (1H-nuclear magnetic resonance, NMR) and cytological methods (fluorochromatic reaction test, freezefracture and scanning electron microscopy). A semi-logarithmic representation of the pollen weight loss demonstrated the complexity of the dehydration process. A the study of proton loss using 1H-NMR indicated that two major releases water of had taken place, each with different flux rates. Pulse 1H-NMR experiments showed the occurrene of non-exponential signal decay as a function of time, indicating the existence of different fractions of water in a pollen grain sample. These fractions leave the pollen grain at different times during pollen dehydration, and one of them (that of the so-called vital water) can be related to pollen viability. The quantity of protons giving a signal during pulse 1H-NMR experiments was very low when the pollen grains were judged to be dead according to the fluorochromatic test. Freeze-fracture replicas of these dead pollen grains (less than 25% water content) showed that the plasma membrane had become detached from the intine surface; this ultrastructural feature might therefore be involved in the loss of pollen viability.Abbreviations A
initial amplitude of the NMR signal
- A2
quantity of water charcterized by T2-2
- A5
quantity of water characterized by T2–5
- FCR
fluorochromatic reaction
- NMR
nuclear magnetic resonance
- T2
transverse relaxation time
- T2-2
T2 measured with 2 ms between each pulse of radiofrequency
- T2–5
T2 measured with 5 ms between each pulse of radiofrequency 相似文献
32.
Valérie Toulon Hervé Sentenac Jean-Baptiste Thibaud André Soler David Clarkson Claude Grignon 《Planta》1989,179(2):235-241
The effect of HCO
3
-
on ion absorption by young corn roots was studied in conditions allowing the independent control of both the pH of uptake solution and the CO2 partial pressure in air bubbled through the solution. The surface pH shift in the vicinity of the outer surface of the plasmalemma induced by active H+ excretion was estimated using the initial uptake rate of acetic acid as a pH probe (Sentenac and Grignon (1987) Plant Physiol. 84, 1367). Acetic acid and orthophosphate uptake rates and NO
3
-
accumulation were slowed down, while 86Rb+ uptake and K+ accumulation rates were increased by HCO
3
-
. These effects were similar to those induced by 4-(2-hydroxyethyl)-1-piperazineethane sulfonic acid/2-amino-2-(hydroxymethyl)-1,3-propanediol (Hepes-Tris). They were more pronounced when the H+ excretion was strong, were rapidly reversible and were not additive to those of Hepes-Tris. The hypothesis is advanced that the buffering system CO2/H2CO3/HCO
3
-
accelerated the diffusion of equivalent H+ inside the cell wall towards the medium. This attenuated the surface pH shift in the vicinity the plasma membrane and affected the coupling between the proton pump and cotransport systems.Abbreviations FW
fresh weight
- Hepes
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- Jaa
acetic acid influx
- JK
+
K+ influx
- JPi
orthophosphate influx
- Mes
2-(N-morpholino)ethanesulfonic acid
- pCO2
CO2 partial pressure
- Tris
2-amino-2-(hydroxymethyl)-1,3-propanediol 相似文献
33.
Catherine Manin Jean Noël Barbotin Daniel Thomas Jean Claude Lazzaroni 《Applied microbiology and biotechnology》1989,32(2):143-147
Summary In continuous cultures, alkaline phosphatase was synthesised and excreted for more than 250 h by immobilized growing cells in contrast to free cells for which the excretion decreased after 150 h of culture. This observed increase in alkaline phosphatase synthesis and excretion by immobilized cells may have resulted from growing conditions within the gel beads.Offprint requests to: C. Manin 相似文献
34.
Neuromedin U-immunoreactivity in the nervous system of the small intestine of the pig and its coexistence with substance P and CGRP 总被引:2,自引:0,他引:2
Jean-Pierre Timmermans Prof. Dietrich W. Scheuermann Werner Stach Dirk Adriaensen Marie H. A. De Groodt-Lasseel Julia M. Polak 《Cell and tissue research》1989,258(2):331-337
Summary In the small intestine of the pig, neuromedin U (NMU)-immunoreactivity was mainly confined to the nerve plexus of the inner submucosal and mucosal regions. After colchicine treatment, a high number of immunoreactive nerve cell bodies was observed in the plexus submucosus internus (Meissner), whereas only a low number was found in the plexus submucosus externus (Schabadasch). The plexus myentericus as well as the aganglionic nerve meshworks in the circular and longitudinal smooth muscle layers almost completely lacked NMU-immunoreactivity. Double-labeling experiments demonstrated the occurrence of distinct NMU-containing neuron populations in the plexus submucosus internus: (1) relatively large type-II neurons revealing immunoreactivity for NMU and calcitonin gene-related peptide (CGRP) and/or substance P (SP); (2) a group of small NMU- and SP-immunoreactive neurons; (3) a relatively low number of small neurons displaying immunoreactivity for NMU but not for SP. Based on its distributional pattern, it is concluded that NMU plays an important role in the regulation and control of mucosal functions. 相似文献
35.
Ricardo Benavente Marie Christine Dabauvalle Ulrich Scheer Nathalie Chaly 《Chromosoma》1989,98(4):233-241
Many nuclear proteins are released into the cytoplasm at prometaphase and are transported back into the daughter nuclei at the end of mitosis. To determine the role of this reentry in nuclear remodelling during early interphase, we experimentally manipulated nuclear protein uptake in dividing cells. Recently we and others have shown that signal-dependent, pore complex-mediated uptake of nuclear protein is blocked in living cells on microinjection of the lectin wheat germ agglutinin (WGA), or of antibodies such as PI1 that are directed against WGA-binding pore complex glycoproteins. In the present study, we microinjected mitotic PtK2 cells with WGA or antibody PI1 and followed nuclear reorganization of the daughter cells by immunofluorescence and electron microscopy. The inhibitory effect on nuclear protein uptake was monitored by co-injection of the karyophilic protein nucleoplasmin. When injected by itself early in mitosis, nucleoplasmin became sequestered into the daughter nuclei as they entered telophase. In contrast, nucleoplasmin was excluded from the daughter nuclei in the presence of WGA or antibody PI1. Although PtK2 cells with blocked nuclear protein uptake completed cytokinesis, their nuclei showed a telophaselike organization characterized by highly condensed chromatin surrounded by a nuclear envelope containing a few pore complexes. These findings suggest that pore complexes become functional as early as telophase, in close coincidence with nuclear envelope reformation. They further indicate that the extensive structural rearrangement of the nucleus during the telophase-G1 transition is dependent on the influx of karyophilic proteins from the cytoplasm through the pore complexes, and is not due solely to chromosome-associated components.Abbreviations
WGA
wheat germ agglutinin
-
GlcNAc
N-acetylglucosamine 相似文献
36.
Philippe Frachet Georges Uzan Dominique Thevenon Eric Denarier Marie Hélène Prandini Gérard Marguerie 《Molecular biology reports》1990,14(1):27-33
Platelet GPIIbIIIa is only synthesized in megakaryocyte or in cell lines with megakaryocytic features. The sequence for GPIIb and GPIIIa have recently been derived from cDNAs obtained from HEL cells. The sequence of these proteins produced by the megakaryocyte, has however, not been determined yet. This study describes full length cDNAs for GPIIb and GPIIIa isolated from megakaryocyte cDNA libraries. The cDNA sequences indicate the presence of nucleotide differences, between the sequence of the GPIIIa cDNAs from HEL cells, endothelial cells and megakaryocytes. One difference was also observed between HEL and megakaryocyte GPIIb at position 633 where a cystein in the megakaryocyte GPIIb, is replaced by a serine in the HEL sequence. The mRNA species for GPIIb (3.4kb) and GPIIIa (6.1 kb) were of the same size in HEL cells and human megakaryocytes. 相似文献
37.
Cystic fibrosis typing with DNA probes and screening for ΔF508 deletion in families from Southern France 总被引:1,自引:1,他引:0
Mireille Claustres Marie Desgeorges Paule Kjellherg Hélène Bellet Jacques Demaille Michelle Ramsay 《Human genetics》1990,85(4):398-399
Summary A sample of 235 individuals from 49 French cystic fibrosis (CF) families with at least one living affected child was typed
with probes for restriction fragment length polymorphisms (RFLPs) known to be linked to the CF gene, and was screened for
the ΔF508 mutation. Using a combination of six probes, 44 out of the 49 families were sufficiently informative to enable prenatal
diagnosis or carrier determination. As in many other populations, linkage disequilibrium was found between the CF locus and
the haplotype B (XV2c: allele 1; KM19: allele 2), which accounts for about 78% of CF chromosomes in our families. The ΔF508
deletion was present in 64.3% of CF chromosomes. 相似文献
38.
Marie-Hélène Chaput Darasinh Sihachakr Georges Ducreux Dominique Marie Nasrine Barghi 《Plant cell reports》1990,9(8):411-414
In order to regenerate somatic hybrids, mesophyll protoplasts from a dihaploid potato, BF15 (H1), were electrofused with those from two other dihaploid clones, Aminca (H6) and Cardinal (H3). Determination of the ploidy level by flow cytometry showed that 10% of plants regenerated from the fusion experiment with BF15 + Aminca were diploids, 14% triploids, 63% tetraploids and very few were mixoploids or had a higher ploidy level. Using morphological markers and vigour in plant growth, we were able to recover a total of 24 somatic hybrid plants, respectively 20 and 4 hybrids (accounting for 12% and 13% of regenerants) from the fusions BF15 + Aminca and BF15 + Cardinal. Most of the somatic hybrids were at the expected tetraploid level (2n=4x=48). The hybrid nature was confirmed by examining isoenzyme patterns for malate dehydrogenase (MDH) and isocitrate dehydrogenase (ICD). 相似文献
39.
A Barakat J C Marie G Rosselin 《Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie》1990,310(5):189-194
We demonstrate here that membranes prepared from beta cells which release insulin contain specific binding sites for calcitonin gene-related peptide (CGRP). The binding of 125I(His) human CGRP to beta cell membranes was protein concentration, time, temperature and pH dependent. Scatchard analysis of the data revealed a single class of binding sites with an apparent dissociation constant (Kd) of 1.5 nM and a maximal binding capacity of 19 fmol/mg of protein. Chicken CGRP inhibited the label binding whereas salmon calcitonin had only a weak effect. These results suggest that the effect of CGRP on insulin secretion is due to a direct action on beta cells. 相似文献
40.
A new recombinant procoagulant protein derived from the cDNA encoding human factor VIII 总被引:1,自引:0,他引:1
P Meulien T Faure F Mischler H Harrer P Ulrich B Bouderbala K Dott M Sainte Marie C Mazurier M L Wiesel 《Protein engineering》1988,2(4):301-306
We have constructed new B domain deletion derivatives of human factor VIII (FVIII) by manipulating the cDNA using recombinant DNA techniques. One of these new derivatives, FVIII delta II, in which amino acids 771(pro)-1666(asp) have been deleted, no longer contains the protease cleavage site at amino acid position 1648(arg)-1649(glu) known to be involved in the initial step of FVIII processing. We have expressed this molecule in both baby hamster kidney (BHK) 21 cells using the vaccinia virus (VV) expression system and have established Chinese hamster ovary (CHO) derived permanent cell lines expressing either recombinant (r)FVIII or FVIII delta II. The characteristics of FVIII delta II have been compared to those of rFVIII and/or plasma derived (pd) FVIII. FVIII delta II has the following properties: (i) it exhibits FVIII procoagulant activity; (ii) it is expressed at 5-fold higher levels than is the complete molecule in comparable systems; (iii) it migrates for the most part as a single major band on SDS-PAGE, in contrast to the complete molecule; (iv) it is activated to a greater extent by thrombin than is either rFVIII or pdFVIII; and (v) it retains the ability to bind von Willebrand factor (vWf). 相似文献