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991.
Pafenolol is a -blocker with unusual oral absorption properties. The blood concentration–time profile exhibits two peaks, and the bioavailability is low and dose dependent because of incomplete and nonlinear intestinal uptake. We addressed the question whether the intestinal absorption of pafenolol was affected by bile depletion in the gut lumen of rats. Further, the hypothesis that variable gastric emptying accounts for double peaks in blood was tested by duodenal administration of pafenolol. Following intraduodenal administration to rats with intact bile secretion, double peaks were observed in the blood concentration–time curve. The bioavailability was 6.8 ± 0.7% for the low dose (1 µmol/kg) and increased significantly to 28 ± 10% following the high duodenal dose (25 µmol/kg). These blood concentration–time profiles exclude interrupted gastric emptying as cause of the twin peaks. In bile duct-cannulated rats the intestinal absorption of the low dose (1 µmol/kg) was still poor (F = 10.7 ± 5.5%) and the blood concentration–time profile contained two peaks. Following administration of a high duodenal dose (25 µmol/kg) to rats with an almost bile-free small intestine, the absorption rate increased and the double-peak phenomenon disappeared in five of seven rats, while the bioavailability increased significantly, to 62 ± 27%. These results suggest that the low bioavailability of pafenolol is due to a complexation between bile and pafenolol in the gut lumen, preventing intestinal uptake in the major part of the small intestine. Further, such complex formation in the intestinal lumen may be the underlying mechanism of the double peaks observed in the blood concentration–time profile. 相似文献
992.
Nika Adham Laurence A. Borden Lee E. Schechter Eric L. Gustafson Tamara L. Cochran Pierrre J.-J. Vaysse Richard L. Weinshank Theresa A. Branchek 《Naunyn-Schmiedeberg's archives of pharmacology》1993,348(6):566-575
We recently described the cloning of a fifth member of the 5-hydroxytryptamine (5-HT)1 (serotonin1) receptor class that inhibits adenylyl cyclase, namely the human 5-HT1F receptor (Adham et al. 1993 a). In the present study we have examined in greater detail the functional coupling of the 5-HT1F receptor in two different cell lines, NIH-3T3 and LM(tk–) fibroblasts (receptor densities of 1.7 and 4.4 pmol/mg protein, respectively). The maximal inhibitory response elicited by 5-HT was significantly greater in NIH-3T3 as compared to LM(tk–) cells, whereas the EC50 values were comparable.To investigate the relationship between receptor occupancy and inhibition of cAMP accumulation mediated by 5-HT1F receptors in NIH-3T3 cells (and hence the degree of receptor reserve), we used the irreversible receptor antagonist N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). The half-maximal response required only about 10% receptor occupancy, consistent with a receptor reserve of 90% (88±2.1%, n = 4) for 5-HT-induced inhibition of FSCA. Despite the presence of such a high degree of receptor reserve, a range of intrinsic activities was displayed by structurally diverse classes of compounds. For example, sumatriptan and lysergol were as efficacious as 5-HT itself and thus acted as full agonists, whereas metergoline and 1-NP behaved as partial agonists and as shown previously (Adham et al. 1993a), methiothepin was a silent antagonist (Kb = 438 nM).We have also investigated activation of additional signal transduction pathways by the 5-HT1F receptor and found that the responses differ in the two cell lines with respect to stimulation of phospholipase C. For example, in NIH-3T3 cells no elevation of inositol phosphates (IP) of [Ca2+]i was observed even at very high agonist concentrations (100 M). In contrast, in LM(tk–) cells concentrations of 5-HT as low as 10 nM induced stimulation of IP and a rapid increase of [Ca2+]i. The 5-HT1F receptor failed to alter arachidonic acid release in either cell line.The maximal increase in IP accumulation in LM(tk–) cells was modest, averaging about 100% above basal. The increases of IP and [Ca2+]i required 5-HT concentrations less than one order of magnitude greater than those inhibiting FSCA (EC50 = 17, 55 and 8 nM, respectively), and both responses were blocked by 100 M methiothepin. All three responses (cAMP, IP, and [Ca2+]i) were sensitive to pertussis toxin pre-treatment, suggesting the involvement of Gi/Go protein(s) in these signal transduction pathways. [Ca2+]i was also elevated by sumatriptan, which may provide a mechanism by which this drug causes constriction of the vasculature. In conclusion, these data indicate that the human 5-HT1F receptor can couple to multiple effectors, and that this coupling is cell-type dependent.Abbreviations FSCA
forskolin-stimulated cAMP accumulation
- [Ca2+]
intracellular free calcium concentration
- AA
arachidonic acid
- EEDQ
N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline
- CHO
chinese hamster ovary cell
- LM(tk–)
mouse fibroblast cell
- Bmax
maximal binding site density
- Ki
apparent dissociation constant obtained from competition binding studies
- G protein
guanine nucleotide-binding protein
- HBS
HEPES-buffered saline
- IP
inositol phosphates
- IP3
inositol 1,4,5 trisphosphate
- PLC
phospholipase C
- Kb
antagonist dissociation constant
- Kd
equilibrium dissociation constant
- N-1F-6
stable NIH-3T3 cells expressing the cloned 5-HT1F receptor
- L-1F-3
stable LM(tk–) cells expressing the cloned 5-HT1F receptor
- PTX
pertussis toxin
- BSA
bovine serum albumin
- METH
methiothepin
- SUMA
sumatriptan
- 5-MeO-DMT
5-methoxy-N,N-dimethyltryptamine
- 1-NP
1-(1-napthyl)piperazine
- 5-CT
5-carboxyamidotryptamine
Correspondence to: N. Adham at the above address 相似文献
993.
Masaaki Oka Hitoshi Mitsunaga Shoichi Hazama Shigefumi Yoshino Takashi Suzuki 《Surgery today》1993,23(8):669-674
The natural killer (NK) activity of peripheral blood mononuclear cells and serum immunosuppressive acidic protein (IAP) levels were examined in patients with esophageal or gastric cancer, before and after surgery. Patients with stage IV esophageal or stage IV gastric cancer had significantly lower NK activity (39.5±14.8% and 37±11.6%, respectively), and also higher serum IAP levels (778±264 g/mL and 633±156 g/mL, respectively), than the corresponding control values (50±5.6% and 375±26 g/mL, respectively). Patients with esophageal or gastric cancer who underwent curative resection had high NK activity (54.8±11.6% and 54.8±8.0%, respectively), and low IAP levels (471±116 g/mL and 490±42 g/mL, respectively), compared with those who underwent non-curative resection. Patients who underwent non-curative resection had lower NK activity and higher serum IAP levels than those who underwent curative resection, even 1 month after surgery. Mononuclear cells in the regional lymph nodes and tumor specimens showed significantly lower NK activity than those in the peripheral blood and spleen. Thus, NK activity and the IAP level reflected the immunocompetence, clinical course, and surgical curability of those patients. NK cells appeared not to have any significant antitumor activity in the regional lymph nodes or in the tumor itself, although they were still active in the peripheral blood. 相似文献
994.
Endre Sulyok Tibor Ertl Károly Adamovit Sarolta Hovanyovszky Wolfgang Rascher 《Pediatric nephrology (Berlin, Germany)》1993,7(6):881-885
The present study was undertaken to establish the developmental pattern of urinary endothelin-1 (ET-1) excretion and to define its possible role in mediating pathophysiological changes related to perinatal asphyxia/infection and dopamine treatment. Urinary ET-1 levels were measured by radioimmunoassay in 7 full-term neonates (mean gestational age 39.3 weeks) on days 1, 3 and 5, and in 9 pre-term neonates (mean gestational age 30.8 weeks) on days 1, 3, 5, 7 and weekly thereafter for 5 consecutive, weeks. The results were compared with those of three age-groups of 30 normal children (4–8 years, 9–12 years and 13–18 years); each group, consisted of 10 children. The influence of severe cardiopulmonary distress (n=16, mean gestational age 33.9 weeks, post-natal age 3.3 days) and dopamine administration in a dose of 2 g/min per kg (n=10, mean gestational and post-natal ages 32.1 weeks and 5.6 days, respectively) were also studied. In full-term infants, ET-1 concentration fell from 34.3±1.8 pmol/l on day 1 to 21.5±1.5 pmol/l on day 5 (P<0.01). In premature infants its absolute value and its post-natal fall were similar in the 1st week and no further change occurred in weeks 2–5; it stabilized at levels between 17.1±2.2 and 16.7±1.7 pmol/l. These concentrations tended to be lower than those of 25.5±1.3, 23.0±1.0 and 26.2±0.7 pmol/l measured in three groups of older children. During the 1st week, daily ET-1 excretion remained unchanged in term infants (3.1±1.0 vs. 3.7±1.5 pmol/m2 per day), but there was a significant increase from 6.5±1.0 to 12.4±0.7 pmol/m2 per day (P<0.01) in premature infants. During weeks 2–5, preterm infants excreted more ET-1 than older children (P<0.01). In response to perinatal ashphyxia/infection and dopamine therapy, urinary ET-1 excretion markedly rose and there was a significant positive correlation between urine flow rate and ET-1 excretion (P<0.001). We conclude that ET-1 concentration rather than excretion rate may have a role in mediating the changes in renal functions that occur soon after birth. The pathophysiological significance of the flow-dependent increase in urinary ET-1 excretion needs to be further studied. 相似文献
995.
G. Grimmer G. Dettbarn J. Jacob 《International archives of occupational and environmental health》1993,65(3):189-199
Summary A filter combination consisting of an impregnated glass fibre and a control filter was used for the collection of air samples in which gaseous and particulate polycyclic aromatic hydrocarbons (PAHs) were determined. To estimate the loss of lower boiling PAHs, d10-phenanthrene was applied as internal standard. A simple, well-reproducible method for the determination of 1-, 2-, 3-, 4- and 9-hydroxyphenanthrene, 1,2-, 3,4- and 9,10 dihydroxydihydrophenanthrene, 1-hydroxypyrene and 1,2-dihydroxy-1,2-dihydropyrene is described. By means of personal air samplers the exposure to PAHs of four coke plant employees working at different locations was measured over 4 days. Simultaneously the 24-h urine was collected and stored frozen until analysed. The main excretion product of pyrene is a 1-hydroxypyrene conjugate, whereas phenanthrene is excreted predominantly as dihydrodiol conjugate. As expected, workers on the battery topside were exposed the most and accordingly excreted by far the highest amounts of PAHs. Up to 34.0 g phenanthrol conjugates (total of all isomeric phenols) and 195.5 g dihydrodiol conjugates (total of all isomeric dihydrodiols) were excreted in the 24-h urine (mean of 4 days). The metabolite profiles of five isomeric phenanthrene phenols and three isomeric dihydrodiols exhibited only small percentage variations within one individual whereas significant interindividual differences were observed. These findings may indicate a genetically determined enzyme pattern responsible for the metabolic conversion of PAHs.This paper is dedicated to Professor Dr. R. Preußmann on the occasion of his 65th birthday 相似文献
996.
The binding characteristics of valproic acid (VPA) and its pharmacologically active monounsaturated metabolite, E-2-VPA, to rat plasma proteins were compared. The plasma free fraction was determined by a rapid equilibrium procedure, which minimizes the interfering effects of nonesterified fatty acids liberated by in vitro lipolysis. Nonlinear binding behavior was observed with both compounds over their respective pharmacologic concentration range. Multiple binding-site models were invoked to explain the binding isotherm. The 2-unsaturated compound has a much higher affinity for the rat plasma proteins (mainly albumin) than its saturated precursor. The equilibrium association constants for the high- and intermediate-affinity sites were more than an order of magnitude higher with E-2-VPA than with VPA (104–106 versus 103
M
–1). This difference in binding affinity was also reflected by a lower plasma free fraction for E-2-VPA compared with VPA (<<10 versus >20% at total concentrations of less than 100 µg/ml). A more pronounced dose- and concentration-dependent variation in the distribution and clearance kinetics is predicted for the 2-unsaturated analogue compared to VPA. Also, the structural dependency in plasma protein binding observed with these branched-chain fatty acids may provide insights into the mechanism of interaction between fatty acyl molecules and albumin. 相似文献
997.
目的 阐明鼻咽癌细胞中EB病毒编码的潜伏膜蛋白 1(LMP1)活化核转录因子NF κB的机制。方法 利用强力霉素Dox诱导表达LMP1的鼻咽癌细胞株Tet on LMP1 HNE2为实验模型 ,首先应用免疫印迹方法测定Dox诱导后不同时相LMP1的表达动力学以及IκBs蛋白量及功能的改变。进而用间接免疫荧光法检测NF κB的亚细胞定位。最后采用瞬间共转染及报道基因活性分析分析NF κB的活性。结果 在鼻咽癌细胞Tet on LMP1 HNE2中 ,Dox处理 15分钟后LMP1的表达迅速升高并维持与较高水平直至 12 0分钟。LMP1的诱导性表达导致IκBα的磷酸化并降解 ,但IκBα蛋白总量无改变。继IκBα的磷酸化并降解 ,NF κB(P6 5 )自胞浆易位至胞核且活性升高。IκBα的显性负性突变子抑制NF κB(P6 5 )的核易位及报道活性。LMP1的诱导性表达并未引起IκBβ蛋白水平变化。结论 在鼻咽癌细胞Tet on LMP1 HNE2中 ,EB病毒LMP1通过IκBα的磷酸化并降解激活核转录因子NF κB的活性 ,并且 ,LMP1诱导的NF κB活性能被IκBα的显性负性突变子完全抑制。IκBβ在此信号传导途径中无改变。LMP1表达前后IκBα蛋白总量维持恒定可能是由于NF κB的活化迅速启动了IκBα的重头合成这一自身调节环路所致。 相似文献
998.
Construction of a recombinant plasmid harbouring the rhoptry protein 1 gene of Toxoplasma gondii and preliminary observations on DNA immunity 总被引:9,自引:1,他引:8
Objective To observe the immune responses elicited in BALB/c mice by a DNA vaccine. A ge ne encoding rhoptry protein 1 (ROP1) from Toxoplasma gondii (T. gondii) was cloned into vector pcDNA3.Methods Amplifyied gene fragments coding for ROP1 from the genomic DNA of T.gondii ZS2 were inserted into cloning vector, pUC18, and sub-cloned into pcDNA3. Mice wer e injected at a dosage of 100 μg recombinant plasmid DNA by intramuscular inje ction and boosted after 2 weeks. pcDNA3 and normal saline were used as control . 30, 50 and 70 days after the second immunization, NK cell activity, T lympho cyte proliferation and sub-clusters and serum IgG antibody were assayed.Results The specific gene fragment coding for ROP1 was amplified and a pcROP1 recombinan t was constructed. At 30 days after immunization, the spleens of the mice were obviously enlarged evidently. NKC activity and the proliferation of spleen T ly mphocytes seen on MTT assay were higher in pcROP1 group than in the controls. T he number of CD4(+) T cells exhibited no obvious increase compared with that of the control, but CD8(+) T cells were obviously increased (P<0.05). At 90 d ays after vaccination, the titer of IgG antibody in the serum of vaccinated mice was positive (1∶100).Conclusion pcROP1 was constructed and it could elicit both cellular and humoral immune r esponses in immunized mice. 相似文献
999.
Objective To construct a eukaryotic expression system with pcDNA3-PfCSP/Hela for the Circ umsporozoite protein (CSP) gene of Plasmodium falciparum (P.falciparum), t o observe the immune responses in BALB/c mice induced by the expressed proteins .Methods The recombinant plasmid pcDNA3-PfCSP was transformed into the Hela cell line. The expressed protein was isolated and analyzed by using SDS-PAGE and used for immunization of BALB/c mice by subcutaneous, intravenous, and intraperitone al adminstration.Enzyme-linked immunosorbent assay(ELISA), Dot-ELISA, Wester n blot, T lymphocyte proliferation test, natural killer cell(NKC) activity assay , and CD4(+) and CD8(+) T cell detection were used for observation of humoral an d cellular immune responses.Results Immune sera strongly reacted with the expressed protein, antibody titer was up to 1∶6400 as detected by ELISA.Western blot analysis revealed a specific b and at 38.3 Kda.When the spleen cells of normal and immunized BALB/c mice we re specifically stimulated with expressed protein, the optical densities were 0 .12±0.03 and 0.34±0.04, respectively.The latter were significantly highe r than the former (P<0.01).We used the MTT colorimetric assay to measure NKC activity of mice spleen.The results showed that the NKC activity of immuni zed BALB/c mice was remarkably higher than that of the controls (P<0.05). CD4(+) and CD8(+) T cells were detected by using monoclonal antibody immunofluor escence methods.The results showed that the percentage of CD4(+) and CD8(+) T cells of immunized group were significantly higher than that of control group ( P<0.05).Conclusions The humoral and cell-mediated immune responses and elevated NKC activity to pr oducts made with a eukaryotic expression system could be specifically detected i n BALB/c mice.These findings indicate that the expressed protein could enhance the immune function in mice. 相似文献
1000.
目的 寻找可能存在的抗丙型肝炎病毒 (HCV)感染的中和抗体。方法 构建两个分别含HCVE1和E2抗原基因的真核表达载体 ,用瞬时表达法检测其在哺乳动物细胞中的表达。将构建的载体连同IL 2基因一起经皮下给BalB/c小鼠进行注射 ,然后用ELISA法检测特异性抗体产生情况。最后通过研究抗体和病毒间的相互作用分析基因免疫诱导产生的抗血清在体外与HCV的相互作用情况。结果 经过基因免疫的小鼠分别产生了E 1和E2抗体。其中 ,用含E2基因的质粒载体免疫的小鼠所产生的E2抗血清不仅可以免疫沉淀来源血清中的HCV病毒颗粒 ,而且可以和与来源株非相关的HCV病毒颗粒相互作用。结论 我们的研究表明基因免疫诱导小鼠产生的E2抗体可以和HCV病毒颗粒发生交叉反应 ,这使我们看到了诱导产生具有广泛针对性的E2抗体的希望。 相似文献