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11.
12.
Determination of viability within serotypes of a soil population of Rhizobium leguminosarum bv. trifolii. 总被引:4,自引:3,他引:1 下载免费PDF全文
Concern has been raised about the percentage of viable cells within soil rhizobia populations measured by the immunofluorescence direct count method. The purpose of this study was to evaluate a direct viable count technique which is based on the fact that viable bacteria in natural populations undergo cell elongation when they are exposed to a combination of substrate and the inhibitor of DNA gyrase, nalidixic acid. A soil extraction procedure was developed to recover a high proportion of soil bacteria (ca. 10(9)/g of soil) in suspensions with an optical clarity suitable for accurate microscopic enumeration. After incubation for 16 to 20 h at 27 degrees C in the presence of yeast extract (200 mg/liter) and nalidixic acid (10 mg/liter), between 65 and 74% of the bacteria in soil suspension became significantly elongated (greater than or equal to 4.2 microns). In contrast, less than or equal to 0.5% of the same population could be cultured, regardless of the medium composition, nutrient concentration, or incubation conditions. The direct viable count method was combined with immunofluorescence to compare the percent viability and kinetics of appearance of elongated cells within serotypes of a soil population of Rhizobium leguminosarum bv. trifolii. Although the majority of these organisms were viable, as observed by immunofluorescence, we obtained evidence that subpopulations within the soil rhizobia community were in different states of competence to respond to substrate. A consistently low percentage (less than or equal to 30%) of the population of serotype 23 was elongated even after 24 h of incubation and regardless of when the soil was sampled.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
13.
J F Grillo P J Bottomley C Van Baalen F R Tabita 《Biochemical and biophysical research communications》1979,89(2):685-693
Studies on the O2 protection mechanism for nitrogenase in a mutant (PM10) of sp. CA indicated that the ability to protect nitrogenase from O2 was functionally impaired. Growth rates of PM10 were substantially improved when cells were cultured under microaerobic conditions. Nitrogenase activity was totally inhibited by exposure to O2 for 30 min; partial restoration of activity was attained when cell suspensions were subsequently made microaerobic. Experiments in which induction of nitrogenase activity was followed indicated that the synthesis of the O2 protection mechanism was temporally separated from synthesis of heterocysts and nitrogenase. 相似文献
14.
Synthesis of nitrogenase and heterocysts by Anabaena sp. CA in the presence of high levels of ammonia. 下载免费PDF全文
Anabaena sp. CA fails to synthesize heterocysts and nitrogenase when grown with KNO3 as the nitrogen source. By contrast, both heterocysts and proheterocysts are synthesized in NH4Cl-containing media to a level nearly commensurate with cells grown in the absence of combined nitrogen. The growth rate of the organism in NH4Cl-containing media was similar to that obtained with KNO3 as the nitrogen source and was independent of the presence of N2 in the atmosphere. Thus, our results indicate that the organism assimilated nitrate and ammonium nitrogen equally well to meet the nitrogen requirements for growth. Moreover, in contrast to previous studies with other cyanobacteria, the repressor singal for heterocyst differentiation in Anabaena sp. CA is not derived from the metabolism of ammonia but appears to be involved with nitrate metabolism. Nitrogenase activity was partially expressed in NH4Cl-grown cultures. Increasing the level of nitrogenase activity to a value representative of a N2-grown culture required both the inhibition of ammonia assimilation and de novo protein synthesis. An increase in the number of mature heterocysts was not required. The fact that high levels of exogenous ammonia only partially repress the synthesis of proteins required for the maximum expression of nitrogenase activity in Anabaena sp. CA has important implications. 相似文献
15.
16.
Nicky HGM Peters Inne HM Borel Rinkes Willem PTM Mali Maurice AAJ van den Bosch Remmert K Storm Peter W Plaisier Erwin de Boer Adriaan J van Overbeeke Petra HM Peeters 《Trials》2007,8(1):1-7
Background
In orthodontic treatment, anchorage control is a fundamental aspect. Usually conventional mechanism for orthodontic anchorage control can be either extraoral or intraoral that is headgear or intermaxillary elastics. Their use are combined with various side effects such as tipping of occlusal plane or undesirable movements of teeth. Especially in cases, where key-teeth are missing, conventional anchorage defined as tooth-borne anchorage will meet limitations. Therefore, the use of endosseous implants for anchorage purposes are increasingly used to achieve positional stability and maximum anchorage.Methods/Design
The intended study is designed as a prospective, multicenter randomized controlled trial (RCT), comparing and contrasting the effect of early loading of palatal implant therapy versus implant loading after 12 weeks post implantation using the new ortho-implant type II anchor system device (Orthosystem Straumann, Basel, Switzerland). 124 participants, mainly adult males or females, whose diagnoses require temporary stationary implant-based anchorage treatment will be randomized 1:1 to one of two treatment groups: group 1 will receive a loading of implant standard therapy after a healing period of 12 week (gold standard), whereas group 2 will receive an early loading of orthodontic implants within 1 week after implant insertion. Participants will be at least followed for 12 months after implant placement. The primary endpoint is to investigate the behavior of early loaded palatal implants in order to find out if shorter healing periods might be justified to accelerate active orthodontic treatment. Secondary outcomes will focus e.g. on achievement of orthodontic treatment goals and quantity of direct implant-bone interface of removed bone specimens. As tertiary objective, a histologic and microtomography evaluation of all retrieved implants will be performed to obtain data on the performance of the SLA surface in human bone evaluation of all retrieved implants. Additionally, resonance frequency analysis (RFA, Osstell? mentor) will be used at different times for clinically monitoring the implant stability and for histological comparison in order to measure the reliability of the resonance frequency measuring device.Trial registration
Current Controlled Trials ISRCTN97142521. 相似文献17.
Intrinsic fluorescence changes and rapid kinetics of proteinase deformation during serpin inhibition
The X-ray crystal structure of the serpin-proteinase complex suggested that the serpin deformed the proteinase thereby inactivating the molecule. Using a variant of alpha(1)-antitrypsin in which both tryptophan residues have been replaced by phenylalanine, we have shown that the proteinase becomes partially unfolded during serpin inhibition. The tryptophan free variant, alpha(1)-antitrypsin((FF)), is fully active as an inhibitor of thrombin. Thrombin has a fluorescence emission maximum of 340 nm which blue shifts to 346 nm, concomitant with a 40% increase in intensity, upon formation of the serpin-proteinase complex indicative of substantial conformational change within the proteinase. Stopped-flow analysis of the fluorescence changes within the proteinase indicated a two-step mechanism. A fast bimolecular reaction with a rate constant of 2.8x10(6) M(-1) s(-1) is followed by a slow unimolecular process with a rate of 0.26 s(-1) that is independent of concentration. We propose that the first rate is formation of an initial complex which is then followed by a slower process involving the partial unfolding of the proteinase during its translocation to the opposite pole of the serpin. 相似文献
18.
Chinni C de Niese MR Tew DJ Jenkins AL Bottomley SP Mackie EJ 《The Journal of biological chemistry》1999,274(14):9169-9174
Three members of the family of protease-activated receptors (PARs), PARs-1, -3 and -4, have been identified as thrombin receptors. PAR-1 is expressed by primary myoblast cultures, and expression is repressed once myoblasts fuse to form myotubes. The current study was undertaken to investigate the hypothesis that thrombin inhibits myoblast fusion. Primary rodent myoblast cultures were deprived of serum to promote myoblast fusion and then cultured in the presence or absence of thrombin. Thrombin inhibited myoblast fusion, but another notable effect was observed; 50% of control cells were apoptotic within 24 h of serum deprivation, whereas less than 15% of thrombin-treated cells showed signs of apoptosis. Proteolysis was required for the effect of thrombin, but no other serine protease tested mimicked the action of thrombin. Neither a PAR-1- nor a PAR-4-activating peptide inhibited apoptosis or fusion, and myoblast cultures were negative for PAR-3 expression. Myoblasts exposed to thrombin for 1 h and then changed to medium without thrombin accumulated apoptosis inhibitory activity in their medium over the subsequent 20 h. Thus the protective action of thrombin appears to be effected through cleavage of an unidentified thrombin receptor, leading to secretion of a downstream apoptosis inhibitory factor. These results demonstrate that thrombin functions as a survival factor for myoblasts and is likely to play an important role in muscle development and repair. 相似文献
19.
James EL Whisstock JC Gore MG Bottomley SP 《The Journal of biological chemistry》1999,274(14):9482-9488
Protein misfolding plays a role in the pathogenesis of many diseases. alpha1-Antitrypsin misfolding leads to the accumulation of long chain polymers within the hepatocyte, reducing its plasma concentration and predisposing the patient to emphysema and liver disease. In order to understand the misfolding process, it is necessary to examine the folding of alpha1-antitrypsin through the different structures involved in this process. In this study we have used a novel technique in which unique cysteine residues were introduced at various positions into alpha1-antitrypsin and fluorescently labeled with N, N'-dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)ethylenediamine. The fluorescence properties of each protein were studied in the native state and as a function of guanidine hydrochloride-mediated unfolding. The studies found that alpha1-antitrypsin unfolded through a series of intermediate structures. From the position of the fluorescence probes, the fluorescence quenching data, and the molecular modeling, we show that unfolding of alpha1-antitrypsin occurs via disruption of the A and C beta-sheets followed by the B beta-sheet. The implications of these data on both alpha1-antitrypsin function and polymerization are discussed. 相似文献
20.
The ability of proteins to fold into complex three-dimensional shapes is truly amazing. Given the difficulty of the reaction it is perhaps unsurprising that many proteins in vivo are unable to fold correctly. These misfolded proteins are generally recognized by the cell's quality control machinery and dealt with through degradation. However in an increasing number of diseases, such as Huntington's, Alzheimer's and alpha1-antitrypsin deficiency, misfolded protein accumulates both within and outside the cell. This aggregated protein is able to evade the normal cellular responses and in some cases even disable it. In this review we present an overview of protein misfolding and examine recent data which is beginning to reveal the mechanisms by which protein aggregates are toxic to cells. 相似文献