A method for the accurate determination of anti-hapten cytotoxic T lymphocyte precursors: correction for apparent 'anti-self' reactivity

A method is described for accurately determining the frequency of precursors of hapten specific cytotoxic T cells. The method is based on a standard Poisson analysis of limit dilution cultures, but makes a correction of 'anti-self' reacting clones and for spontaneously arising clones that recognise modified self. These corrections are shown to be especially important when low hapten densities are used, where there may be more than a 10-fold difference between the corrected and uncorrected frequency estimates. Determined levels of antigen specificity and of H-2 restriction are significantly enhanced by application of this method.     

Pitfalls in acid/base experiments with conscious dogs

Arterial pH and blood gases were measured at intervals in conscious dogs after their first human contact of the day. Blood was sampled through an indwelling catheter in the aorta without disturbing the animals. It appeared that in the first 90 min arterial PO₂, oxygen saturation and haemoglobin concentration significantly declined. PCO₂ and pH changed less consistently when the acid/base status of the dogs was normal, but when a non-respiratory acidosis was present there was a significant decrease in pH and a significant increase in PCO₂. Arterial pH and blood gases were also measured before and after feeding the animals. It appeared that an appreciable metabolic alkalosis developed within 2 h after a meal. The alkaline tide was accompanied by a trend to higher values for PCO₂. It is concluded that, after a period of seclusion, renewed human contact causes behavioural changes in a dog, which may result in appreciable transitory changes in arterial pH and blood gas values. Blood sampling from conscious dogs should therefore take place after a proper period of habituation; preferably, a few samples should be taken at intervals to check that a steady state has been reached. If possible, blood should be collected before feeding; in any case the relationship in time of blood sampling to feeding should be constant throughout.     

Masked excitatory action of noradrenaline on rat islet β-cells via activation of phospholipase C

The effect of noradrenaline (NE) on rat islet -cells was examined. NE reduced insulin secretion from rat islets exposed to extracellular solutions containing glucose at 5.5 or 16.6 mM. In islets treated with pertussis toxin (PTX), however, NE increased insulin secretion. The NE-induced augmentation of insulin secretion was inhibited by prazosin. In intact islets, NE increased phospholipase C (PLC) activity, an effect that was prevented by treatment of islets with U-73122. NE elevated intracellular [Ca²⁺] ([Ca²⁺]_i) in isolated -cells independently of PTX. Although this NE effect was inhibited by prazosin, phenylephrine did not mimic it. The [Ca²⁺]_i response to NE was also prevented by the treatment of cells with U-73122. NE produced depolarization of -cells followed by nifedipine-sensitive action potentials. NE reduced the whole-cell membrane currents through ATP-sensitive K⁺ channels (K_ATP), responsible for the depolarization. This NE effect was prevented by treatment of -cells with U-73122 or BAPTA/AM. Although at least some of our results imply the presence of ₁-adrenoceptors, -cells were not stained by a polyclonal IgG antibody recognizing all adrenergic ₁-receptor subtypes so far identified. These results suggest that an interaction of NE with an unknown type of receptor activates rat islet -cells via a PLC-dependent signal pathway. This effect is, however, masked by the inhibitory action via a PTX-sensitive pathway also activated by NE.     

gamma-Aminobutyric acid and taurine release in the striatum of the rat during hypoglycemic coma, studied by microdialysis

Extracellular levels of striatal gamma-aminobutyric acid (GABA) and taurine were monitored during insulin-induced hypoglycemia using microdialysis. At the onset of isoelectricity in the electroencephalogram (EEG), a transient 5-fold increase in the levels of GABA occurred. Taurine levels increased 5 min following the onset of isoelectricity and continued to increase during the entire isoelectric period. The results demonstrate that events associated with the onset of isoelectricity during hypoglycemia trigger an increase in extracellular concentrations of GABA and taurine. The discrepancy in time-course of these changes may reflect differences in compartmentation, function and metabolism of the two amino acids.     

On the interaction between phenytoin and digoxin

Summary An open, randomized, single-blind cross over trial to investigate phenytoin-digoxin interactions at steady state was performed in 6 healthy male volunteers. Coadministration of phenytoin caused a significant reduction in the elimination half-life of digoxin from 33.9 to 23.7 h and a diminution in AUC_0–48 from 31.6 to 24.4 ng · ml^–1 · h. Renal digoxin clearance was not significantly altered from 135.7 to 120.3 ml · min^–1. Assuming no change in -acetyldigoxin absorption, the in decrease time-course the serum digoxin concentration was due to a significantly increased total digoxin clearance from 258.6 to 328.3 ml · min^–1. An insignificant reduction in the digoxin distribution volume from 749.4 to 668.0 l was also observed. No relevant change in the pharmacokinetic parameters (elimination half-life, area under the serum concentration time-curve, protein binding) of phenytoin was observed when phenytoin and digoxin were co-administered. The data suggest that with this drug combination the serum digoxin concentration should be carefully monitored and, if necessary, the daily digoxin dose should be increased.     

Perioperative Cefamandolprophylaxe bei aortocoronaren Bypass-Operationen Serumkonzentrationsverlauf während der extrakorporalen Zirkulation

Zusammenfassung Eine perioperative antibiotische Kurzprophylaxe mit 2 g Cefamandol intravenös bei Narkoseeinleitung wurde bei 12 Patienten während coronarchirurgischer Eingriffe unter Verwendung der Herz-Lungen-Maschine durchgeführt. Bei Beginn der extrakorporalen Zirkulation (= EKZ) kam es infolge Hämodilution zu einem Absinken der Serumkonzentrationen von 110,96 ± 40,29 mcg/ml auf 70,89 ± 34,65 g/ml innerhalb von 10 min. Im weiteren Verlauf der EKZ war der Abfall der Serumspiegel gleich schnell wie davor und danach. Nach 240 min fanden sich noch Serumspiegel von 16,80 ± 9,32 g/ml. Als Ursache für das Versagen einer antibiotischen Prophylaxe kommt bei einer Operationsdauer von mehr als 4 h das Absinken der Serumspiegel unter die minimale Hemmkonzentration der entsprechenden Keime in Frage.

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Perioperative cefamandole prophylaxis in aortocoronary bypass operations: Course of serum concentration during extracorporeal circulation

Summary Antibiotic prophylaxis with 2 g Cefamandole at induction of anaesthesia was performed in 12 male patients undergoing aortocoronary bypass surgery. Caused by hemodilution, there was a marked decrease of serum concentration at the beginning of extracorporeal circulation, from 110.96 ± 40.29 mcg/ml to 70.89 ± 34.65 mcg/ml within 10 min. During extracorporeal circulation, elimination was as fast as before and after perfusion. 240 min after application, mean serum concentrations of 16.80 ± 9.32 mcg/ml were measured. Failure of antibiotic prophylaxis in operations exceeding 4 h might be due to unadaequate antibiotic concentrations, beyond the minimal inhibitory concentration for the pathogens, reported to cause infections after cardiac operations.

     

活性氧对红细胞膜流动性影响的研究

目的 :研究活性氧诱导红细胞发生氧化溶血的机制。方法 :用不同浓度的 H2 O2 作用于离体正常红细胞 ,分别用比色法、硫代巴比妥酸荧光法、荧光偏振法测定红细胞溶血度、红细胞膜脂质过氧化物 (L PO)、红细胞膜微粘度。结果 :5 0 ,10 0 ,12 0 mm ol/ L 浓度的 H2 O2 作用于红细胞后 ,均引起红细胞发生氧化溶血 ,红细胞溶血度、红细胞膜 L PO、红细胞膜微粘度均明显增加 ;H2 O2 在 5 0～ 12 0 mm ol· L- 1浓度范围内 ,红细胞溶血度、红细胞膜 L PO、红细胞膜微粘度的增加与 H2 O2 有明显的量效关系 ;红细胞膜微粘度与红细胞溶血度呈明显正相关 ,红细胞膜 L PO与红细胞膜微粘度呈明显正关。结论 :活性氧引发红细胞膜脂质过氧化导致红细胞膜流动性降低在活性氧诱导红细胞发生氧化溶血的机制中起一定的作用     

高脂血症大鼠红细胞扫描电镜观察

目的 :探讨高脂血症红细胞形态变化和临床意义。方法 :Wistar大鼠分为正常对照组 ( NC)和高脂血症组 ( HL,喂饲高脂饲料 ) ,利用扫描电镜观察红细胞形态改变并计数分类。结果 :与 NC组比较 ,HL组的正常红细胞百分比由 76.4 %减少到 4 5.7%、棘型红细胞、球口型红细胞、靶型红细胞分别由4 %、2 .2 %、1.5%增加到 17.3%、10 .4 %、5.8% ,嵴型和其它异型红细胞也略有增加。结论 :高脂血症的异型红细胞明显增加。     

原发性高血压患者红细胞[Ca2+]i浓度变化及影响因素

目的：观察原发性高血压患者红细胞[Ca^2 ]i,多巴胺β羟化酶及ATP含量变化并分析其结果。方法：测定35例高血压患者的红细[Ca^2 ]i、ATP、血清多巴胺β羟化酶活性,血糖及血浆胰岛素含量,并以30例健康成年人为对照。结果：高血压患者的红细胞[Ca^2 ]i、ATP、多巴胺β羟化酶均明显高于对照组（P＜0．05,P＜0．01）,但血糖与胰岛素未见明显变化。结论：高血压患者血清多巴胺β羟化酶活性增强伴随ATP与[Ca^2 ]i升高。