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71.
The effector domain mutants of oncogenic Ras, V12S35 Ras, V12G37 Ras, and V12C40 Ras were tested for their abilities to mediate tumorigenic and metastatic phenotypes in athymic nude mice when expressed in NIH 3T3 fibroblasts. All mutants displayed comparable tumorigenic properties, but only the mutant that activates the Raf-mitogen-activated protein kinase kinase (MEK)-extracellular regulated kinase (ERK) 1/2 pathway, V12S35 Ras, induced tumors in the experimental metastasis assay. Furthermore, direct activation of the MEK-ERK1/2 pathway in NIH 3T3 cells by mos or a constitutively active form of MEK was sufficient to induce metastasis whereas R-Ras, which fails to activate the ERK1/2 pathway, is tumorigenic but nonmetastatic. The subcutaneous tumors and lung metastases derived from V12S35 Ras-transformed NIH 3T3 cells expressed higher levels of activated ERK1/2 in culture when compared with the parental cellular pool before injection, indicating that selection for cells with higher levels of activated ERK1/2 occurred during tumor growth and metastasis. By contrast, cells explanted from V12G37-Ras or V12C40-Ras-induced tumors did not show changes in the level of ERK1/2 activation when compared with the parental cells. When tumor-explanted cell lines derived from each of the effector domain mutants were passaged one additional time in vivo, all mediated rapid tumor growth, but, again, only cells derived from V12S35 Ras-tumors formed numerous metastatic lesions within the lung. These results show that the metastatic properties of the Ras effector domain mutants segregate, and that, whereas Ras-mediated tumorigenicity can arise independently of ERK1/2 activation, experimental metastasis appears to require constitutive activation of the ERK1/2 pathway. 相似文献
72.
73.
Serotonin [5-hydroxytryptamine(5-HT)] in the paraventricular nucleus (PVN) of rats has a suppressive effect on feeding behavior and causes a selective decrease in carbohydrate ingestion, specifically at the onset of the natural (dark) feeding period. Studies conducted here provide further evidence for this phenomena, showing a similar dose-related decrease in carbohydrate ingestion at dark onset after PVN injection of 5-HT or of the agonists, d-norfenfluramine or fluoxetine, which act through endogenous 5-HT. To further characterize the effects of this indoleamine on the macrostructure of feeding, a computer-automated data acquisition system was used to analyze macronutrient feeding patterns in freely feeding animals maintained on the pure diets of protein, carbohydrate, and fat. Results indicate that PVN administration of 5-HT at dark onset decreases intake of the carbohydrate nutrient by decreasing meal size, feeding time, and feeding rate for this nutrient and increasing the satiating effect of carbohydrate. These effects, which occur specifically during the first meal after injection, are opposite those seen after peripheral administration of the 5-HT receptor antagonist, metergoline. This drug stimulates feeding through a selective increase in carbohydrate intake, characterized by an increase in meal size, percent composition, and feeding time for this nutrient and a decrease in the satiety ratio for carbohydrate. These results implicate the serotonergic system in the termination of carbohydrate-rich meals that are prevalent during the early hours of the natural feeding cycle. 相似文献
74.
Mice persistently infected with lymphocytic choriomeningitis virus (LCMV) exhibit impaired learning ability. In this report, we determined whether clearance of the virus was associated with restoration of behavioral function. Neonatal Balb/cByJ mice were persistently infected with LCMV and tested as adults in a nonconditional spatial discrimination task. The presence of viral proteins in neurons was confirmed immunohistochemically and infectious virus was quantified in the blood by plaque assay. LCMV-infected adult mice made more errors in a Y-maze avoidance task compared to sham-inoculated controls. After the initial behavioral analysis, infected and control mice received a dose of cytotoxic T-lymphocytes sufficient to clear virus from these mice. Following complete clearance of the virus, mice were re-tested in the behavioral task, 5 months after the original test. No reversal of the learning deficit was seen following viral clearance; mice that had been cleared of the virus and those that remained persistently infected behaved similarly. These data indicate that persistent LCMV infection of the CNS lasting up to 7 months results in discriminated learning impairments that are not reversed by subsequent anti-viral immunocytotherapy. 相似文献
75.
76.
M Mori T Shiraishi S Tanaka M Yamagata K Mafune Y Tanaka H Ueo GF Barnard K Sugimachi 《Canadian Metallurgical Quarterly》1999,79(2):211-213
The cell-wall characteristics of Gardnerella vaginalis grown in conventional and biofilm systems were studied by electron microscopy. The gram-positive nature of the cell wall was confirmed. Novel cell-wall particles which appeared to be associated with cell division were also identified, particularly in organisms of biofilm origin. 相似文献
77.
To confirm the role of peptides as principal precursors of acrylamide formation in sterilized table olives, peptides from olive water were fractionated. After their partial fractionation by solid phase extraction (SPE) and ultrafiltration (<10,000 Da), respectively, small peptides from olive water were isolated by size-exclusion chromatography (SEC). In the fractions collected, peptides and polyphenolic compounds were determined colorimetrically, and acrylamide was quantitated by LC–MS/MS after heating of the samples. Subsequently, peptides were characterized by matrix-assisted laser desorption/ionization-tandem time of flight mass spectrometry (MALDI-TOF/TOF-MS), and polyphenols were analyzed by LC–MS in the respective fractions. Finally, peptides containing fractions were purified on a polymeric resin (Amberlite XAD 16HP) to remove unbound phenolic compounds by adsorption. The results of the different experiments performed in complete absence of free asparagine and reducing sugars strongly support small peptides bound to polyphenols to be the principal precursors of acrylamide in sterilized table olives. 相似文献
78.
BACKGROUND: So far only a few cases of carcinoma in situ of the fallopian tube have been reported, but its detailed clinical and pathologic findings, including cytology, have not been fully described. CASE: A 70-year-old female was admitted to our hospital because of irregular genital bleeding. Endometrial smear revealed a small number of atypical cells with a clear background. Hysterectomy, bilateral salpingo-oophorectomy and omentectomy were performed. Grossly, a grayish white papillary tumor, measuring 1.5 x 1.0 cm, was observed within the lumen of the left fallopian tube. Microscopically, the diagnosis of papillary adenocarcinoma in situ of the left fallopian tube was made according to 1992 International Federation of Gynecologists and Obstetricians fallopian tube staging. CONCLUSION: Although endometrial brush cytology is not sensitive enough to detect a primary carcinoma of the fallopian tube, our case indicates that it may contribute useful information on extrauterine diseases and can detect a stage 0 cancer of the fallopian tube. Clinicians, as well as pathologists, should consider the possibility of fallopian tube cancer if cervical or endometrial cytology shows atypical cells with papillary patterns with a clear background but endometrial curettage cannot prove malignancy. 相似文献
79.
GF Joyce 《Canadian Metallurgical Quarterly》1998,95(11):5845-5847
80.
Equilibrium dialysis and Scatchard plots were used to establish that human and rabbit paraoxonases both have two calcium binding sites. Independent-site and stepwise constant analyses were used to calculate a higher affinity site (Kd1) of 3.6 +/- 0.9 x 10(-7) M for human A paraoxonase, and 1.4 +/- 0.5 x 10(-8) M for rabbit paraoxonase, and a lower affinity site (Kd2) of 6.6 +/- 1.2 x 10(-6) M for human A paraoxonase, and 5.3 +/- 0.94 x 10(-6) M for rabbit paraoxonase. In both species, the higher affinity sites were found to be essential to maintain hydrolytic activity; complete removal of calcium led to irreversible inactivation. The lower affinity sites were required for catalytic activity, and their binding of calcium was reversible. Experimentally estimated values of Kd2 based on the concentration of calcium required to obtain half the maximum enzymatic activity were 3 microM for human A and B paraoxonases, and also in the order of 3 microM for rabbit paraoxonase, using three different substrates. Calcium was the only metal found that protects against denaturation and also confers hydrolytic activity with these two mammalian paraoxonases. 相似文献