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81.
The application of electron-nuclear double resonance (ENDOR) spectroscopy for the investigation of photosynthetic systems is reviewed. The basic principles of continuous wave and pulse ENDOR are presented. Selected examples of the application of the ENDOR technique for studying stable and transient paramagnetic species, including cofactor radical ions, radical pairs, triplet states, and the oxygen-evolving complex in plant Photosystem II (PSII) are discussed. Limitations and perspectives of ENDOR spectroscopy are outlined.
相似文献82.
Reini W Bretveld Chris MG Thomas Paul TJ Scheepers Gerhard A Zielhuis Nel Roeleveld 《Reproductive biology and endocrinology : RB&E》2006,4(1):30
Some pesticides may interfere with the female hormonal function, which may lead to negative effects on the reproductive system
through disruption of the hormonal balance necessary for proper functioning. Previous studies primarily focused on interference
with the estrogen and/or androgen receptor, but the hormonal function may be disrupted in many more ways through pesticide
exposure. The aim of this review is to give an overview of the various ways in which pesticides may disrupt the hormonal function
of the female reproductive system and in particular the ovarian cycle. Disruption can occur in all stages of hormonal regulation:
1. hormone synthesis; 2. hormone release and storage; 3. hormone transport and clearance; 4. hormone receptor recognition
and binding; 5. hormone postreceptor activation; 6. the thyroid function; and 7. the central nervous system. These mechanisms
are described for effects of pesticide exposure in vitro and on experimental animals in vivo. For the latter, potential effects of endocrine disrupting pesticides on the female reproductive system, i.e. modulation
of hormone concentrations, ovarian cycle irregularities, and impaired fertility, are also reviewed. In epidemiological studies,
exposure to pesticides has been associated with menstrual cycle disturbances, reduced fertility, prolonged time-to-pregnancy,
spontaneous abortion, stillbirths, and developmental defects, which may or may not be due to disruption of the female hormonal
function. Because pesticides comprise a large number of distinct substances with dissimilar structures and diverse toxicity,
it is most likely that several of the above-mentioned mechanisms are involved in the pathophysiological pathways explaining
the role of pesticide exposure in ovarian cycle disturbances, ultimately leading to fertility problems and other reproductive
effects. In future research, information on the ways in which pesticides may disrupt the hormonal function as described in
this review, can be used to generate specific hypotheses for studies on the effects of pesticides on the ovarian cycle, both
in toxicological and epidemiological settings. 相似文献
83.
84.
Goldiner I Overmars H Groen AK Kulik W 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2003,795(1):35-40
Fatty acid-bile acid conjugates and especially arachidyl amido cholic acid are synthetic molecules that were shown to prevent cholesterol gallstone formation in mice and hamsters as well as to dissolve pre-existing gallstones in mice. To measure these novel compounds we developed a liquid chromatography electrospray tandem mass spectrometry method based on the analysis of 100 microL of plasma with stearyl amido cholic acid (stamchol, 1.5 microM/L) added as internal standard. Repeatable calibrations between 0 and 50 microM/L exhibited consistent linearity and reproducibility. Inter- and intraassay C.V.s were 5.3-11.4% and 2.6-6.4%, respectively, at targeted concentrations of 0.1, 2.3 and 50 microM/L. 相似文献
85.
R S English J S Lampel TJ Vanden Boom 《Journal of industrial microbiology & biotechnology》1998,21(4-5):219-224
The introduction of plasmid DNA into germinating spores of an industrially improved strain of Saccharopolyspora erythraea was accomplished by electroporation. Various parameters affecting the efficiency of electroporation were examined. The most
critical factor was the extent of spore germination. Electrocompetence was limited to a 4-h period following the initial emergence
of the germ tube. Electroporation efficiencies as high as 2 × 105 CFU μg−1 of plasmid DNA were obtained using electrocompetent germlings. The optimal field strength was 12–14 kV cm−1 with a pulse duration of 15–20 ms. Electrocompetent germlings were stored at −80°C without a significant decrease in transformation
efficiency. The utility of this protocol was demonstrated by isolating a propionyl-CoA carboxylase mutant through targeted
gene disruption and replacement.
Received 3 April 1998/ Accepted in revised form 28 September 1998 相似文献
86.
Sequence variations in small-subunit ribosomal RNAs of Hartmannella vermiformis and their phylogenetic implications 总被引:1,自引:0,他引:1
Evidence of associations between free-living amoebas and human disease has
been increasing in recent years. Knowledge about phylogenetic relationships
that may be important for the understanding of pathogenicity in the genera
involved is very limited at present. Consequently, we have begun to study
these relationships and report here on the phylogeny of Hartmannella
vermiformis, a free-living amoeba that can harbor the etiologic agent of
Legionnaires' disease. Our analysis is based on studies of small-subunit
ribosomal RNA genes (srDNA). Nucleotide sequences were determined for
nuclear srDNA from three strains of H. vermiformis isolated from the United
Kingdom, Germany, and the United States. These sequences then were compared
with a sequence previously obtained for a North American isolate by J. H.
Gunderson and M. L. Sogin. The four genes are 1,840 bp long, with an
average GC content of 49.6%. Sequence differences among the strains range
are 0.38%-0.76%. Variation occurs at 19 positions and includes 2
single-base indels plus 14 monotypic and 3 ditypic single-base
substitutions. Variation is limited to eight helix/loop structures
according to a current model for srRNA secondary structure. Parsimony,
distance, and bootstrap analyses used to examine phylogenetic relationships
between the srDNA sequences of H. vermiformis and other eukaryotes
indicated that Hartmannella sequences were most closely related to those of
Acanthamoeba and the alga Cryptomonas. All ditypic sites were consistent
with a separation between European and North American strains of
Hartmannella, but results of other tests of this relationship were
statistically inconclusive.
相似文献
87.
Ouabain-insensitive salt and water movements in duck red cells. I. Kinetics of cation transport under hypertonic conditions 总被引:10,自引:6,他引:4 下载免费PDF全文
Duck red cells in hypertonic media experience rapid osmotic shrinkage followed by gradual reswelling back toward their original volume. This uptake of salt and water is self limiting and demands a specific ionic composition of the external solution. Although ouabain (10(-4)M) alters the pattern of cation accumulation from predominantly potassium to sodium, it does not affect the rate of the reaction, or the total amount of salt or water taken up. To study the response without the complications of active Na-K transport, ouabain was added to most incubations. All water accumulated by the cells can be accounted for by net salt uptake. Specific external cation requirements for reswelling include: sufficient sodium (more than 23 mM), and elevated potassium (more than 7 mM). In the absence of external potassium cells lose potassium without gaining sodium and continue to shrink instead of reswelling. Adding rubidium to the potassium- free solution promotes an even greater loss of cell potassium, yet causes swelling due to a net uptake of sodium and rubidium followed by chloride. The diuretic furosemide (10(-3)M) inhibits net sodium uptake which depends on potassium (or rubidium), as well as inhibits net sodium uptake which depends on sodium. As a result, cell volume is stabilized in the presence of this drug by inhibition of shrinkage, at low, and of swelling at high external potassium. The response has a high apparent energy of activation (15-20 kcal/mol). We propose that net salt and water movements in hypertonic solutions containing ouabain are mediated by direct coupling or cis-interaction, between sodium and potassium so that the uphill movement of one is driven by the downhill movement of the other in the same direction. 相似文献
88.
The temperature dependence of the electron spin-lattice relaxation time T1 was measured for the S0 state of the oxygen-evolving complex (OEC) in photosystem II and for two dinuclear manganese model complexes by pulse EPR using the inversion-recovery method. For [Mn(III)Mn(IV)(mu-O)2 bipy4]ClO4, the Raman relaxation process dominates at temperatures below 50 K. In contrast, Orbach type relaxation was found for [Mn(II)Mn(III)(mu-OH)(mu-piv)2(Me3 tacn)2](ClO4)2 between 4.3 and 9 K. For the latter complex, an energy separation of 24.7-28.0 cm(-1) between the ground and the first excited electronic state was determined. In the S0 state of photosystem II, the T1 relaxation times were measured in the range of 4.3-6.5 K. A comparison with the relaxation data (rate and pre-exponential factor) of the two model complexes and of the S2 state of photosystem II indicates that the Orbach relaxation process is dominant for the S0 state and that its first excited state lies 21.7 +/- 0.4 cm(-1) above its ground state. The results are discussed with respect to the structure of the OEC in photosystem II. 相似文献
89.
Birrell L Kulik L Morgan BP Holers VM Marchbank KJ 《Journal of immunology (Baltimore, Md. : 1950)》2005,174(11):6974-6982
Complement receptor type 2 (CR2/CD21), in association with CD19, plays an important role in enhancing mature B cell responses to opsonized Ags. We have shown that mice expressing a human CR2/CD21 (hCR2/CD21) transgene during the CD43(+)/CD25(-) late pro-B cell stage of B cell development demonstrate marked changes in subsequent B cell ontogeny. In the present study, we show that the humoral immune response to the T cell-dependent Ag, sheep RBC, is muted severely in a manner inversely proportional to B cell expression level of hCR2. Individual Ag-specific IgG isotypes vary in the degree to which they are affected but all are reduced while IgM titers are normal. A substantial reduction in germinal centers, both in size and frequency, in the spleens of immunized hCR2 transgenic mice demonstrates a failure to maintain germinal center reaction. However, both IgM expression levels and LPS-proliferative responses appear fully intact in B cells from hCR2-positive mice, suggesting that this alteration in B cell phenotype is different qualitatively from that of specific Ag-defined anergy models. These data suggest that the unresponsiveness to T-dependent Ags displayed by hCR2-positive B cells is linked to an increase in the level of stimulus required to propel the B cell into a fully activated state and thus a normal humoral immune response to Ags. We conclude that this phenotype and these mice may offer an additional means to dissect mechanisms underlying B cell tolerance and Ag responsiveness both in bone marrow and periphery. 相似文献
90.
Pulse EPR, 55Mn-ENDOR and ELDOR-detected NMR experiments were performed on the S2-state of the oxygen-evolving complex from spinach Photosystem II. The novel technique of random acquisition in ENDOR was used to suppress heating artefacts. Our data unambiguously shows that four Mn ions have significant hyperfine coupling constants. Numerical simulation of the 55Mn-ENDOR spectrum allowed the determination of the principal values of the hyperfine interaction tensors for all four Mn ions of the oxygen-evolving complex. The results of our 55Mn-ENDOR experiments are in good agreement with previously published data [Peloquin JM et al. (2000) J Am Chem Soc 122: 10926–10942]. For the first time ELDOR-detected NMR was applied to the S2-state and revealed a broad peak that can be simulated numerically with the same parameters that were used for the simulation of the 55Mn-ENDOR spectrum. This provides strong independent support for the assigned hyperfine parameters. 相似文献