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151.
Saxiphilin is a 91 kDa saxitoxin-binding protein that is homologous to members of the transferrin family of Fe(3+)-binding proteins noted for pH-dependent release of Fe3+. The mechanism of toxin binding to purified native saxiphilin from the bullfrog (Rana catesbeiana) was studied using [3H]saxitoxin. At pH 7.4 and 0 degrees C [3H]saxitoxin binds to a single site on saxiphilin with a KD of approximately 0.2 nM. The pH dependence of [3H]saxitoxin binding follows a one-site titration curve in the range of pH 9-4 with maximal binding from pH 9 to 7 and half-inhibition at pH 5.7. Inhibition of toxin binding at low pH is the combined result of a decrease in the rate of toxin association and an increase in the rate of toxin dissociation. The dependence of the apparent rate constants for [3H]saxitoxin association and dissociation on [H+] can be accounted for by a four-state model of allosteric interaction between the toxin-binding site and a single titratable residue of saxiphilin with a pKa of 7.2 in the toxin-free form and 4.3 in the toxin-bound form. From 0 to 25 degrees C, the temperature dependence of [3H]saxitoxin binding to saxiphilin is characterized by delta H degrees = -8.3 kcal mol-1, delta S degrees = 13.8 cal mol-1 K-1, and activation energies of 22.5 kcal mol-1 for dissociation and 11.1 kcal mol-1 for association. Binding of [3H]saxitoxin to saxiphilin is competitively inhibited with low affinity by a variety of divalent metal and lanthanide cations. Inhibition of toxin binding by the carboxyl-methylating reagent trimethyloxonium is prevented by pre-equilibration with [3H]saxitoxin, implicating the presence of one or more carboxyl groups in the binding site. Functional similarities suggest that the saxitoxin-binding site of saxiphilin is located in an interdomain cleft analogous to the location of one of the two homologous Fe(3+)-binding sites of transferrins. On the basis of residue substitutions between saxiphilin and transferrins, it is proposed that the saxitoxin-binding site is located in the carboxy terminal lobe of saxiphilin and that binding is modulated by protonation of a conserved histidine residue. 相似文献
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153.
LF Cardoso M Grinberg MA Ratti LP de Magalh?es I Antelmi CC Medeiros F Tarasoutchi EG Rossi WS Avila G Bellotti 《Canadian Metallurgical Quarterly》1993,61(2):87-91
PURPOSE: To compare immediate and long term results balloon mitral valvuloplasty (BMV) using double balloon or bifoil balloon. METHODS: One hundred and thirteen consecutive cases of BMV used aleatory double balloon (group DB--55 cases) or bifoil balloon (group BF--16 cases). Patients were similar regarding to age, sex, valvopaty etiology, functional class and echocardiographic score. Seventy one (63%) patients achieved 12 months follow-up. RESULTS: In group DB there were 2 (4%) insuccess, 2 (4%) cardiac tamponade and 2 (4%) deaths, 91% patients had immediate criteria of success. Mitral valve area (MVA) increased from 0.8 to 1.69cm2 and mitral gradient (G) by echodopplercardiographic (ECHO) decreased from 17.9 to 4.8mmHg. Three (5%) patients developed severe mitral regurgitation (MR) and needed surgical intervention. At follow-up 2 (4%) developed mitral restenosis. MVA estimated by ECHO study after one year follow-up was inferior to 1.15cm2 in 15 (32%) cases, between 1.5 and 2.0cm2 in 17 (37%) and superior to 2.0cm2 in 14 (31%). In group BF there were 2 (12.5%) insuccess, 4 (25%) developed severe MR occurring 1 death immediate post-operative mitral valve replacement. Among 14 (87%) success cases, MVA increased from 0.8 to 1.89cm2 and G decreased from 18 to 6.4mmHg. Lately 2 (12.5%) needed surgical intervention because significative MR. At 12th month follow-up the ECHO study showed that in one (10%) case MVA was < 1.5cm2, and in 3 (27%) cases was between 1.5 and 2.0cm2. CONCLUSION: There were similar good results in both groups, however group DB had more restenosis, cardiac tamponade and vascular complications and group BF had more severe MR. 相似文献
154.
OBJECTIVES: Nitric oxide (NO), a highly reactive species produced by the activity of NO synthases (NOS), is normally present in the exhaled air of humans and animals. Exhaled NO concentration increases significantly in humans with sepsis and animals, but neither the source nor NOS isoforms responsible for this rise in pulmonary NO production are known. The main objective of this study is to determine the sites and the mechanisms of enhanced NO production in the exhaled air of endotoxemic pigs. DESIGN: Randomized, controlled, animal study. SETTING: University-based animal research facility. SUBJECTS: Thirteen pathogen-free adult female pigs (22 to 27 kg). INTERVENTIONS: Anesthetized pigs were divided into two groups: control and lipopolysaccharides (LPS) (septic) groups. In both groups, extrathoracic (upper airways, nasal, and paranasal) and intrathoracic (bronchi, bronchioles, and alveoli) compartments were ventilated equally with two separate ventilators connected to two tracheal tubes. The LPS group received slow infusion (over 2 h) of Escherichia coli endotoxin (10 microg/kg/h), whereas saline solution was infused into the control group. Expired air of the two compartments was collected throughout the 2-h observation period. The animals were then killed and the lungs were quickly excised and frozen. MEASUREMENTS: Hemodynamic variables were measured in both groups. NO concentration in the exhaled air of both compartments was measured with a chemiluminescence analyser. Pulmonary NOS activity was evaluated by measuring the conversion of L-[2,3H]-arginine to L-[2,3H]-citrulline, and pulmonary expression of NOS was evaluated by immunoblotting. RESULTS: Baseline NO concentration in both groups was significantly higher in the extrathoracic vs intrathoracic compartment (average of 5.2 vs 3.4 parts per billion). Endotoxin infusion elicited a significant and early (after 45 min) rise in exhaled NO concentration in the extrathoracic compartment. Exhaled NO in the intrathoracic compartment also rose significantly but after 90 min of endotoxin infusion. Measurement of lung NOS activity showed a substantial rise in Ca++/calmodulin-dependent activity in the LPS group with no rise in Ca++/calmodulin-independent activity. Immunoblotting of lung tissue samples indicated the absence of the inducible isoform in both groups of animals. Moreover, LPS injection elicited no significant alterations in the pulmonary expression of the endothelial and the neuronal isoforms. CONCLUSIONS: Both extrathoracic and intrathoracic compartments contribute to the rise in exhaled NO production in experimental septic shock. The rise in exhaled NO production is due to increased activity of constitutive NOS isoforms as a result of increased cofactor availability and/or downregulation of the endogenous inhibitors of NOS. 相似文献
155.
The effects of enhanced HSP27 expression or expression of a nonphosphorylatable form of HSP27 on the migration of bovine arterial endothelial cells was assessed. Expression of the wild-type protein enhanced migration by twofold compared to control transfectants, whereas expression of the mutant protein retarded migration by 40%. Since homologs of the small heat shock protein inhibit F-actin polymerization in vitro and may alter basolateral F-actin content in vivo, it was postulated that the 27 kDa heat shock protein affects microfilament extension essential for cell motility. Expression of the wild-type protein promoted the generation of long cellular extensions, whereas expression of the dominant negative mutant protein resulted in a marked reduction of lamellipodia and generated aberrant microfilament morphology at the wound edge. Immunofluorescence combined with phalloidin staining demonstrated the colocalization of the HSP27 gene products with lamellipodial microfilament structures. These data suggest that the 27 kDa heat shock protein regulates migration by affecting the generation lamellipodia microfilaments. 相似文献
156.
Little is known about the role of the N-linked oligosaccharides in the function of the epidermal growth factor (EGF) receptor (EGF-R). In a human glioma cell line, U373 MG, EGF-Rs contain the bisecting N-linked oligosaccharide sequence recognized by erythroagglutinating phytohemagglutinin lectin from Phaseolus vulgaris (E-PHA). Incubation of E-PHA with cultured U373 MG cells results in inhibition of EGF binding to its receptor and consequently inhibition of EGF-induced autophosphorylation of the receptor. Consistent with the inhibitory effects on the EGF-R, phenotypic events that depend on EGF-R signaling, such as cell spreading and proliferation, were also found to be modified. The effect of this lectin seems to be specific because leukoagglutinating phytohemagglutinin lectin from P. vulgaris (L-PHA), an isolectin of E-PHA, had no effect on EGF-R activity or the biological functions of these cells even though L-PHA was able to bind to the EGF-R. These findings suggest the presence of an important bisecting N-linked oligosaccharide structure in close proximity to the EGF binding site on the receptor. Furthermore, these results suggest the possibility that E-PHA lectin binding may provide an additional approach to blocking EGF-dependent glioma cell growth. 相似文献
157.
BJ Kroesen J Nieken DT Sleijfer G Molema EG de Vries HJ Groen W Helfrich TH The NH Mulder L de Leij 《Canadian Metallurgical Quarterly》1997,45(3-4):203-206
OBJECTIVES: To evaluate the clinical importance of the interaction between carbamazepine (CBZ) and dextropropoxyphene in elderly patients. METHODS: All patients (n = 7263) in Gothenburg, Sweden, who were part of a drug-dispensing programme, were included in the study. Eight per cent of the patients took CBZ and 18% took dextropropoxyphene, continuously. Patients who used a combination of these drugs were compared with patients who took only CBZ or dextropropoxyphene or neither of the two drugs. These four groups of patients were matched to each other with reference to gender, age and concomitant medication, which finally resulted in 21 patients in each group. A questionnaire with 30 symptoms of well-being, including symptoms typical of adverse effects of CBZ, were answered by the patients with the help of a registered nurse. Venous blood samples were drawn from the patients for the analysis of CBZ, its metabolite CBZ 10,11-epoxide (CBZ-E) and dextropropoxyphene. RESULTS: The doses of CBZ and dextropropoxyphene were lower among patients who used the combination of the two drugs than among those who only used one of the drugs. The mean level of CBZ in serum (S-CBZ) was, however, significantly higher and the level of CBZ-E in serum (S-CBZ-E) significantly lower among the patients who used the combination of CBZ and dextropropoxyphene, thus indicating an inhibition of the metabolism of CBZ. The prevalence of symptoms indicating side effects of CBZ was significantly higher in the group of patients who used both drugs. CONCLUSION: This study has shown that the combination of CBZ and dextropropoxyphene is hazardous in elderly patients and should be used with caution. 相似文献
158.
159.
M Bakker J Renes A Groenhuijzen P Visser H Timmer-Bosscha M Müller HJ Groen EF Smit EG de Vries 《Canadian Metallurgical Quarterly》1997,73(3):362-366
While a large body of literature depicting relationships between depression or stress and immunity exists, few such studies have dealt with children, and none investigated myeloid cell-derived immunity. We investigated both phagocytosis and bactericidal activity against Staphylococcus aureus in children with major depressive disorder (MDD). We found that both MDD and stress influence the bactericidal but not the phagocytic activity of polymorphonuclear leukocytes. The data support the existence of psychobiologic effects in children and suggest possible mechanisms by which depression and stress may affect health. 相似文献
160.