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1.
Hydrogen production by rumen holotrich protozoa: effects of oxygen and implications for metabolic control by in situ conditions 总被引:1,自引:0,他引:1
Experiments with washed suspensions of holotrich protozoa (Isotricha spp. and Dasytricha ruminantium) showed that both organisms have an efficient O2-scavenging capability (apparent Km values 2.3 and 0.3 microM, respectively). Reversible inhibition of H2 production increased almost linearly with increasing O2 up to 1.5 microM; higher levels of O2 gave irreversible inhibition. In situ determinations of H2, CH4, O2 and CO2 in ovine rumen liquor, using a membrane inlet mass spectrometer probe, indicated that O2 was present before feeding at 1-1.5 microM and decreased to undetectable levels (less than 0.25 microM) within 25 min after feeding. A transient increase in O2 concentration after feeding occurred only in defaunated animals and resulted in suppression of CH4 and CO2 production. The presence of washed holotrich protozoa decreases the O2 sensitivity of CH4 production by suspensions of a cultured methanogenic bacterium Methanosarcina barkeri. It is concluded that holotrich protozoa play a role in ruminal O2 utilization as well as in the production of fermentation end products (especially short-chain volatile fatty acids) utilized by the ruminant and H2 utilized by methanogenic bacteria. These hydrogenosome-containing protozoa thus both control patterns of fermentation by influencing O2 levels, and are themselves regulated by the low ambient O2 concentrations they experience in the rumen. 相似文献
2.
Infection by pathogenic fungi involves breaching the outer layer of the host by either mechanical or enzymatic means. Subtilisin-like proteinases are considered to be important in the infection process of entomopathogenic, nematophagous, and mycoparasitic fungi. Little is known regarding the expression of such proteinases by plant pathogenic fungi. Magnaporthe poae, a fungal pathogen of Kentucky bluegrass, expressed a subtilisin-like proteinase, proteinase Mp1, in the infected roots. Antibody was produced against the purified enzyme. From immunoblot analysis, expression of the proteinase in infected roots correlated with increasing severity of disease symptoms. Sequence analysis of a genomic clone indicated proteinase Mp1 was homologous to other fungal subtilisin-like proteinases. DNA gel blot analysis indicated proteinase Mp1 was encoded by a small gene family. 相似文献
3.
Three high molecular weight protease inhibitors of rat plasma. Isolation, characterization, and acute phase changes 总被引:1,自引:0,他引:1
K Lonberg-Holm D L Reed R C Roberts R R Hebert M C Hillman R M Kutney 《The Journal of biological chemistry》1987,262(1):438-445
Rat blood plasma contains three high molecular weight thiol ester-containing proteinase inhibitors, alpha 1-macroglobulin (alpha 1M), alpha 1-inhibitor III (alpha 1I3), and alpha 2-macroglobulin (alpha 2M). Rat serums have been analyzed using a two-dimensional gel electrophoretic technique which optimizes recovery of high molecular weight proteins. alpha 1M, and (alpha beta)4-tetramer in native solution, separated in the second sodium dodecyl sulfate-containing electrophoretic dimension as a disulfide-linked (alpha beta)2-dimer with an approximate Mr of 360 kDa. alpha 1I3 separated in the gels as a single 190-kDa polypeptide. It is also a monomer in native solution by ultracentrifugation criteria. Native rat alpha 2M is a tetramer, but it separates in the gels as a disulfide-linked dimer with an Mr of approximately 360 kDa. The kinetics of changes in concentration of these proteins during the induction of polyarthritis was also measured by quantitative immunoelectrophoresis. In rats with adjuvant-induced polyarthritis, the concentration of alpha 1I3 dramatically decreases and alpha 2M appears and continues to increase in a biphasic manner for 2 weeks. The alpha 1M concentration remains relatively constant. All three macroglobulins were purified utilizing modern rapid chromatographic techniques, and parallel comparisons of their native physicochemical properties were carried out. The N-terminal sequence of the alpha-chain of rat alpha 1M was also shown to share sequence homology with that of alpha 2M. In agreement, Esnard et al. (Esnard, F., Gutman, N., El Moujahed, A., and Gauthier, F. (1985) FEBS Lett. 182, 125-129) recently reported that alpha 1I3 also contains a thiol ester bond, as do alpha 1M and alpha 2M, since it reacts covalently with [14C]methylamine and is cleaved autolytically at 80 degrees C. We have examined negatively stained preparations of native, trypsin-treated, and methylamine-treated human alpha 2M, rat alpha 2M, and rat alpha 1M in the electron microscope. Trypsin appears to convert globular ring-shaped native molecules to rectangular box-like structures, in agreement with the conclusions of a recent report on human alpha 2M (Tapon-Bretaudiere, J., Bros, A., Couture-Tosi, E., and Delain, E. (1985) EMBO J. 4, 85-89). 相似文献
4.
5.
Spermatozoa from sterile t6/tw32 and control fertile +/+, T/tw32, T/t6 mice were compared for their abilities to hydrolyse protein matrices and for their levels of acrosin activity. The data show that the immature and mature gametes from both the experimental and control males hydrolyse protein matrices. The quantitative acrosin assays show, however, that the mature gametes from the intercomplement males have significantly less total acrosin activity than any of the control groups of gametes. These findings suggest that this reduced acrosin activity is an additional phenotypic expression of the intercomplement genotype which results in male sterility. 相似文献
6.
The recessive tw5-haplotype, a complete haplotype, is transmitted by heterozygous male mice at very high frequencies (greater than 0.90) in normal matings. The present studies were undertaken to determine the effects of delayed matings and in vitro fertilizations on this phenotypic expression. Males carrying the tw5-haplotype (+/tw5) were first tested for their frequencies of transmission of the mutant 17th chromosome in both normal and delayed matings. Spermatozoa obtained from these same males were then used to fertilize eggs in vitro. The in vivo and in vitro transmission frequencies were found to be statistically equivalent in all types of inseminations. An in vitro fertilization time course study showed that the same percentages of eggs are fertilized by tw5-bearing spermatozoa when the gametes are coincubated for either 2 or 6 h. The data lead to the conclusion that the transmission frequency of the tw5-haplotype is not affected either by the length of time elapsing between insemination and fertilization or by the environment in which fertilization occurs. 相似文献
7.
The th2-haplotype is transmitted at low frequencies (less than 0.30) by +/th2 males in normal matings. In the studies described here, the transmission frequency of the th2-haplotype from Rb7/th2 males was determined for normal and delayed matings and in vitro inseminations. The data show the transmission frequency from the two in vivo inseminations to be less than 0.30 and to be statistically equivalent. However, the in vitro transmission frequency (0.80) is significantly greater than either of the in vivo frequencies. The results show that the environment in which fertilization occurs affects the transmission frequency of this specific t-haplotype significantly. 相似文献
8.
Jacquelyn A. Hank Gilda Weil-Hillman Jean E. Surfus Jeffrey A. Sosman Paul M. Sondel 《Cancer immunology, immunotherapy : CII》1990,31(1):53-59
Summary Thein vivo administration of repetitive weekly cycles of interleukin-2 (IL-2) to patients with cancer enhances the ability of freshly obtained peripheral blood lymphocytes (PBL) to lyse both the natural-killer(NK)-susceptible K562 and the NK-resistant Daudi targets. Lysis of both targets is significantly augmented by inclusion of IL-2 in the medium during the cytotoxicity assay. This boost is much greater for cells obtained following thein vivo IL-2 therapy than for cells obtained prior to the initiation of therapy or for cells from healthy control donors. In addition to direct lytic activity, the PBL obtained followingin vivo IL-2 show a rapid increase in lymphokine-activated killer (LAK) activity with more prolongedin vitro IL-2 exposure, indicating that LAK effectors primedin vivo respond with secondary-like kinetics to subsequent IL-2in vitro. Lymphocytes from healthy control individuals, cultured in IL-2 under conditions attempting to simulate thein vivo IL-2 exposure, function similarly to PBL obtained from patients following IL-2, in that low-level LAK activity was significantly boosted by inclusion of IL-2 during the cytotoxic assay and the cells also responded with secondary-like kinetics to subsequent IL-2in vitro. The augmentation of the LAK effect was also dependent on the dose of IL-2 added during the 4-h51Cr-release cytotoxicity assay, with higher doses of IL-2 having a more pronounced effect. While continuous infusion of IL-2 induces a greater cytotoxic potential per milliliter of blood obtained from patients, the peak serum IL-2 levels attained are greater with bolus IL-2 infusions. These pharmacokinetic results, together with the IL-2 dose dependence of LAK activity generatedin vivo shown in this report, suggest that a combination of treatment with bolus IL-2 infusions superimposed on continuous IL-2 infusion may transiently expose IL-2 dependent LAK cells, activatedin vivo, to higher concentrations of IL-2, facilitating theirin vivo cytotoxic potential.This work was supported by NIH contract NO1 CM-47669-02, NIH grants CA-32685, RR-031086, NO1 CM-47669-03, and American Cancer Society grant CH-237 相似文献
9.
Interspecies hydrogen transfer between rumen holotrich ciliate protoza and methanogenic bacteria has been demonstrated. As a result of the metabolic interaction with Methanosarcina barkeri , the metabolite profile of Isotricha spp. was altered and the production of butyrate and lactate was suppressed in the presence of the methanogen.
Use of membrane-inlet mass spectrometry confirmed that the presence of rumen holotrich ciliates reduced the apparent sensitivity of methanogenesis to the inhibitory effects of oxygen; a gas phase concentration of 7·4 kPa oxygen was required to inhibit methanogenesis in the presence of protozoa, while in pure cultures of M. barkeri , methanogenesis was inhibited by a gas phase oxygen concentration of 1·0 kPa. 相似文献
Use of membrane-inlet mass spectrometry confirmed that the presence of rumen holotrich ciliates reduced the apparent sensitivity of methanogenesis to the inhibitory effects of oxygen; a gas phase concentration of 7·4 kPa oxygen was required to inhibit methanogenesis in the presence of protozoa, while in pure cultures of M. barkeri , methanogenesis was inhibited by a gas phase oxygen concentration of 1·0 kPa. 相似文献
10.
Biophysical evidence that light adaptation in Limulus photoreceptors is due to a negative feedback. 总被引:3,自引:1,他引:2
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The steady-state stimulus-response curve of the Limulus ventral photoreceptor comprises a linear portion followed by a less-than-unity power law dependence, which is maintained over at least 4 decades of intensity. This progressive desensitization corresponds to light adaptation. For flash stimulation of dark-adapted cells, the stimulus-response curve again has an initial linear portion, but this is followed by a region of supralinearity before the curve saturates. In a previous article, we showed that the distribution of time integrals of the single-photon responses is consistent with a model of a single chain of first-order reactions. Starting with such a model, we have looked at relevant elementary nonlinear biochemical mechanisms to determine which of them can modulate the enzymatic amplifications of the chain in such a way as to lead to these behaviors. We assume that each of the two phenomena, adaptation and supralinearity, derives from a single mechanism that acts on a single enzymatic stage. We then conclude that the adaptation must be a cooperative negative feedback, in which an accessory material activated by a late stage of the transduction chain acts cooperatively to inhibit an earlier enzymatic amplification. In Limulus, the number of molecules that cooperate is between 3 and 5. We were not able to discard any of the mechanisms tested for the supralinearity, except to say that they must act at a stage of the chain later than that on which the adaptive material acts. If we assume the conclusions of a previous work which shows that the supralinearity mechanism is active during the steady state, we can also conclude that the supralinearity stage must precede the stage that is the source of the adaptive material. 相似文献