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41.
P W Jungblut E Kallweit W Sierralta A J Truitt R K Wagner 《Hoppe-Seyler's Zeitschrift für physiologische Chemie》1978,359(10):1259-1268
New techniques for the adrenalectomy of pigs and for the isolation of pig uterus nuclei are described. The isolated nuclei were analysed for their content of estradiol and of estradiol receptor. The concentration of the latter exceeded that of the hormone in extracts of uterine nuclei from ovariectomized pigs by ratios of 1.8--10.3. Substantial amounts of both monomer and "activated" dimer receptor but no estradiol at all were extracted from uterine nuclei of ovariectomized/adrenalectomized pigs. The mechanism of action of steroid hormones is discussed on the basis of these results. 相似文献
42.
G A Truitt D K Dennison R R Rich S S Rich 《Journal of immunology (Baltimore, Md. : 1950)》1979,123(2):745-750
Generation of cytotoxic T lymphocytes (CTL) in mixed leukocyte cultures was suppressed by a factor elaborated by alloantigen-activated T cells. This suppressor factor, CTL-TsF, in contrast to a factor that suppresses proliferative responses in mixed leukocyte reactions (MLR-TsF), was effective only when added during the first 24 hr of a 6-day-culture period. Moreover, removal of CTL-TsF 24 hr after culture initiation failed to restore CTL responses. CTL activity could be rescued from suppressed cultures, however, by addition of 2-mercaptoethanol on days 3 or 4. Similarly, transfer of nonadherent cells at 3 or 4 days from cultures treated with CTL-TsF to cultures of adherent cells initiated in control factor restored CTL responses. Mixing experiments with cells pulsed with CTL-TsF for 4 hr at culture initiation identified a target of CTL-TsF as a Thy-1 negative cell that was adherent to plastic and to Sephadex G-10. Suppression was not due to interference with physiologic accessory cell function, but more likely was accomplished via a negative signal from CTL-TsF-pulsed cells. The results thus suggest that CTL-TsF acts early, but reversibly, in the CTL differentiative process via a second suppressor effector cell, possibly a macrophage. 相似文献
43.
Barley aleurone cells contain two types of vacuoles. Characterization Of lytic organelles by use of fluorescent probes 总被引:10,自引:1,他引:9 下载免费PDF全文
Light microscopy was used to study the structure and function of vacuoles in living protoplasts of barley (Hordeum vulgare cv Himalaya) aleurone. Light microscopy showed that aleurone protoplasts contain two distinct types of vacuole: the protein storage vacuole and a lysosome-like organelle, which we have called the secondary vacuole. Fluorescence microscopy using pH-sensitive fluorescent probes and a fluorogenic substrate for cysteine proteases showed that both protein storage vacuoles and secondary vacuoles are acidic, lytic organelles. Ratio imaging showed that the pH of secondary vacuoles was lower in aleurone protoplasts incubated in gibberellic acid than in those incubated in abscisic acid. Uptake of fluorescent probes into intact, isolated protein storage vacuoles and secondary vacuoles required ATP and occurred via at least two types of vanadate-sensitive, ATP-dependent tonoplast transporters. One transporter catalyzed the accumulation of glutathione-conjugated probes, and another transported probes not conjugated to glutathione. 相似文献
44.
W Sierralta A J Truitt P W Jungblut 《Hoppe-Seyler's Zeitschrift für physiologische Chemie》1978,359(4):517-528
Pig endometrium cells, collected by curettage and homogenized in an all-glass Potter Elvehjem homogenizer, gave a considerably higher yield of intact mitochondria and lysosomes than homogenates of whole uterus obtained with the Ultraturrax or the Parr bomb. After homogenization of the cells and subfractionation in the presence of Mg2, mitochondria and lysosomes equilibrated at the same modal density in isopycnic centrifugation. Homogenization and subfractionation in buffers devoid of divalent cations and containing EDTA resulted in a decrease in the buoyant density of mitochondria, allowing for a separation from lysosomes. The pH optima and the specific activities of two mitochondrial enzymes and eight hydrolyases used as marker enzymes were determined. The morphological characteristics of fractions were established by electron microscopy. Preliminary results indicate an involvement of lysosomes in steroid metabolism rather than in steroid and receptor translocation into the nucleus. 相似文献
45.
46.
Suppression of in vitro cytotoxic lymphocyte (CL) responses was mediated by soluble factor(s) produced when in vivo alloantigen-activated suppressor cells were re-exposed to alloantigen in vitro. Elaboration of suppressor factor (SF) was T cell dependent and was optimal 7 days after alloantigen injection. Suppressor factor failed to inhibit CL generation when alloantigen-primed cells rather than normal spleen cells were used as responders. Moreover, SF added at day 3 of incubation rather than at culture initiation was also ineffective, suggesting that suppression probably occurs during antigen induction or early differentiation. Additionally, suppression was abrogated by the presence of 2-mercaptoethanol. Studies combining SF and CL responder cells from a variety of H-2 disparate mouse sdrains revealed that suppression of CL responses: 1) was not alloantigen specific; 2) did not require H-2 homology between responder and suppressor strains; and 3) could not be demonstrated with CBA/J mice. Although CBA/J CL responses were not suppressed by any SF preparation, allo-sensitized CBA/J spleen cells did elaborate SF that inhibited BALB/c CL responses. 相似文献
47.
48.
Cheng Liao Jonathan Hsu Yong Kim Dong-Qing Hu Daigen Xu Jun Zhang Achal Pashine John Menke Toni Whittard Natasha Romero Theresa Truitt Michelle Slade Christine Lukacs Johannes Hermann Mingyan Zhou Matthew Lucas Satwant Narula Julie DeMartino Seng-Lai Tan 《Arthritis research & therapy》2013,15(5):R146
Introduction
Spleen tyrosine kinase (SYK) is a key integrator of intracellular signals triggered by activated immunoreceptors, including Bcell receptors (BCR) and Fc receptors, which are important for the development and function of lymphoid cells. Given the clinical efficacy of Bcell depletion in the treatment of rheumatoid arthritis and multiple sclerosis, pharmacological modulation of Bcells using orally active small molecules that selectively target SYK presents an attractive alternative therapeutic strategy.Methods
A SYK inhibitor was developed and assayed in various in vitro systems and in the mouse model of collagen-induced arthritis (mCIA).Results
A novel ATP-competitive inhibitor of SYK, 6-[(1R,2S)-2-Amino-cyclohexylamino]-4-(5,6-dimethyl-pyridin-2-ylamino)-pyridazine-3-carboxylic acid amide, designated RO9021, with an adequate kinase selectivity profile and oral bioavailability, was developed. In addition to suppression of BCR signaling in human peripheral blood mononuclear cells (PBMC) and whole blood, FcγR signaling in human monocytes, and FcϵR signaling in human mast cells, RO9021 blocked osteoclastogenesis from mouse bone marrow macrophages in vitro. Interestingly, Toll-like Receptor (TLR) 9 signaling in human Bcells was inhibited by RO9021, resulting in decreased levels of plasmablasts, immunoglobulin (Ig) M and IgG upon B-cell differentiation. RO9021 also potently inhibited type I interferon production by human plasmacytoid dendritic cells (pDC) upon TLR9 activation. This effect is specific to TLR9 as RO9021 did not inhibit TLR4- or JAK-STAT-mediated signaling. Finally, oral administration of RO9021 inhibited arthritis progression in the mCIA model, with observable pharmacokinetics (PK)-pharmacodynamic (PD) correlation.Conclusions
Inhibition of SYK kinase activity impinges on various innate and adaptive immune responses. RO9021 could serve as a starting point for the development of selective SYK inhibitors for the treatment of inflammation-related and autoimmune-related disorders. 相似文献49.
KH Richau RL Kudahettige P Pujic NP Kudahettige A Sellstedt 《Journal of biosciences》2013,38(4):703-712
The actinorhizal bacterium Frankia expresses nitrogenase and can therefore convert molecular nitrogen into ammonia and the by-product hydrogen. However, nitrogenase is inhibited by oxygen. Consequently, Frankia and its actinorhizal hosts have developed various mechanisms for excluding oxygen from their nitrogen-containing compartments. These include the expression of oxygen-scavenging uptake hydrogenases, the formation of hopanoid-rich vesicles, enclosed by multi-layered hopanoid structures, the lignification of hyphal cell walls, and the production of haemoglobins in the symbiotic nodule. In this work, we analysed the expression and structure of the so-called uptake hydrogenase (Hup), which catalyses the in vivo dissociation of hydrogen to recycle the energy locked up in this ‘waste’ product. Two uptake hydrogenase syntons have been identified in Frankia: synton 1 is expressed under free-living conditions while synton 2 is expressed during symbiosis. We used qPCR to determine synton 1 hup gene expression in two Frankia strains under aerobic and anaerobic conditions. We also predicted the 3D structures of the Hup protein subunits based on multiple sequence alignments and remote homology modelling. Finally, we performed BLAST searches of genome and protein databases to identify genes that may contribute to the protection of nitrogenase against oxygen in the two Frankia strains. Our results show that in Frankia strain ACN14a, the expression patterns of the large (HupL1) and small (HupS1) uptake hydrogenase subunits depend on the abundance of oxygen in the external environment. Structural models of the membrane-bound hydrogenase subunits of ACN14a showed that both subunits resemble the structures of known [NiFe] hydrogenases (Volbeda et al. 1995), but contain fewer cysteine residues than the uptake hydrogenase of the Frankia DC12 and Eu1c strains. Moreover, we show that all of the investigated Frankia strains have two squalene hopane cyclase genes (shc1 and shc2). The only exceptions were CcI3 and the symbiont of Datisca glomerata, which possess shc1 but not shc2. Four truncated haemoglobin genes were identified in Frankia ACN14a and Eu1f, three in CcI3, two in EANpec1 and one in the Datisca glomerata symbiont (Dg). 相似文献
50.
Mark A. Marsili Michael K. Robinson Gary A. Truitt E. Frederick Wheelock 《Cancer immunology, immunotherapy : CII》1999,16(1):59-64
Summary Previous experiments have demonstrated a temporal relationship between the decline of cytotoxic T lymphocyte (CTL) activity in the peritoneal cavity of DBA/2 mice harboring L5178Y cells in a tumor-dormant state and the appearance of ascitic tumors. Some tumor-dormant mice remain clinically normal for many weeks after the decline of CTL activity, and this activity can be rapidly restimulated by an IP inoculation of irradiated L5178Y cells. We report here that the peritoneal cells from many tumor-dormant mice can be stimulated to cytolytic activity in vitro when cultured for 4 days either with or without the addition of irradiated L5178Y cells. Peritoneal cell populations which cannot be stimulated in vitro can suppress the generation of CTL in those populations which can be stimulated. The tumor-dormant state may terminate when suppressor cells in the peritoneal cavity of tumor-dormant mice inhibit the generation of CTL activity and permit tumor cells to produce an ascitic tumor.
Abbreviations used in this paper: C, complement; CTL, cytotoxic thymus-derived lymphocyte; PC, peritoneal cells; DPC, days post challange; NAD, nonadherent; SC, subcutaneous; IP, intraperitoneal 相似文献