Drug-induced intrahepatic cholestasis: characterization of different pathomechanisms

The pathogenesis of intrahepatic cholestasis in rats was studied using isolated perfused livers as an experimental model. Three basic mechanisms were differentiated: 1. Permeabilization of the bilio-sinusoidal barrier associated with electron microscopic alterations of the tight junctional complexes was found in livers of rats treated with -naphthylisothiocyanate (ANIT, 250 mg/kg body weight). Consequences of these alterations were: reflux of bile constituents such as taurocholate and sulfobromophthalein and increased access to the biliary space of paracellular markers such as inulin and sucrose. The clear-cut mechanism of ANIT cholestasis was used to distinguish other mechanisms of intrahepatic cholestasis. 2. Inhibition of the basic process of fluid secretion was found to be the primary event in the development of cholestasis induced by estrogens. After 5 days of treating rats with ethinyl estradiol (5 mg/kg/day), bile flow was diminished in isolated livers while the permeability of the biliary tract to sucrose and inulin was not affected. Accordingly, the maximal concentration of taurocholate in bile was increased, indicating that its secretion was sustained. The same effect was observed after 1 week of treatment with the depot estrogen estradiol valerate (1 mg/kg/week). After 3 weeks of treatment, however, the taurocholate concentration in bile was lowered and the clearance of sucrose was increased. Bile flow remained at the same cholestatic level for 20 weeks. These results suggest that estrogens have the potency to increase tight junctional permeability only in a second step in the development of cholestasis, following the inhibition of bile flow. 3. An additional mode of secretory inhibition was induced by lowering the concentration of Ca²⁺ in the perfusate of isolated liver. Using ANIT-pretreated livers, i. e., livers with very low capacity to secrete foreign dyes, a high rate of efflux of sulfobromophthalein into the perfusate of preloaded livers suggests stimulation of the efflux of cholephilic solutes across the sinusoidal membrane of liver cells.The results demonstrate that the term intrahepatic cholestasis comprises a number of different sites of interference with the complex process of bile secretion.Dedicated to Professor Dr. med. Herbert Remmer on the occasion of his 65th birthday     

Pharmacological modification of tremor and enhanced acoustic startle by chlordecone and p,p′-DDT

Pretreatment of rats with phenoxybenzamine (5 mg/kg; SC), an alpha adrenergic antagonist, decreased the peak tremor power and startle magnitude of rats subsequently given DDT (75 mg/kg; PO) or chlordecone (60 mg/kg; IP), without having a significant effect on control animals. Pretreatment with an intracerebroventricular injection of calcium (3.75 M in 5 l NaCl) decreased the peak tremor power due to subsequently administered DDT, while increasing the tremor response in rats later dosed with chlordecone. The effects of phenoxybenzamine are postulated to be due to a blockade of an excitatory influence of the adrenergic system. Calcium may decrease DDT-induced tremor by acting as a neuronal stabilizer. Potentiation of the tremorigenic effect of chlordecone by calcium may be due to increased levels of intracellular calcium, resulting in augmented release of neurotransmitters in chlordecone-exposed animals.     

Crystalluria in marathon runners

Summary Epidemiological evidence suggests that marathon runners have a higher incidence of renal stone formation than occurs in the general population. Since crystalluria and stone disease are thought to be related, we subjected urine samples from a group of marathon runners to particle counting and sizing in a Coulter Counter equipped with a population accessory unit. The volume-size distribution curves so obtained were bimodal with one peak occurring in the 2–5 m diameter range and a second in the 15–32 m diameter range—a pattern that is remarkably similar to the distributions reported for recurrent idiopathic stone formers and distinctly different to those recorded for control subjects. Analyses by scanning electron microscopy and X-ray powder diffraction revealed other features which are regarded as typical of stone formers' crystalluria. These physicochemical data indicate that marathon runners may be at increased risk of urinary stone formation.     

Inhibitory effects of procaine on the contractile responses of the guinea-pig Taenia caecum to acetylcholine,substance P and potassium chloride

Summary The effect of procaine on the contractile responses to acetylcholine, substance P and KCl was investigated using the isolated guinea-pig taenia caecum. In normal Tyrode solution (37°C), procaine (10–100 mol/l) caused a parallel shift to the right of only the dose-response curve of acetylcholine (pA₂ value, 5.11). The pA₂ value of procaine against acetylcholine was not significantly affected by increasing the Ca concentration in the bathing solution from 0.9 to 7.2 mmol/l. On the other hand, a high concentration of procaine (10 mmol/l) caused a transient contraction of the taenia caecum, but completely suppressed contractions to all concentrations of the agonists. In K-depolarized preparations, procaine (1–10 mmol/l) shifted the dose-response curve for the CaCl₂-induced contraction to the right. Substance P (3 mol/l)-induced contraction of the taenia caecum incubated with Ca-free EGTA (0.1 mmol/l) solution (20°C) was markedly reduced by procaine (10 mmol/l). Using the single sucrose-gap technique, it was found that procaine (10 mmol/l) produced a membrane depolarization and increases in both amplitude and frequency of spontaneous spike discharge. These potential changes were still observed even after the procaine-induced contraction had disappeared. The spike discharges and contraction caused by procaine were abolished in the presence of a Ca-entry blocker, verapamil (10 mol/l). From these observations, it is concluded that at low concentrations procaine acts as a competitive antagonist of muscarinic receptors in the guinea-pig taenia caecum while high concentrations of procaine may depress the contractile responses to acetylcholine, substance P and KCl by inhibiting the Ca-induced Ca release from the intracellular store site or by reducing the transmembrane Ca influx during depolarization.     

Interactions between the putative calcium entry promotor Bay k 8644 and pressor responses produced by α-1 and α2-adrenocepter agonists in the pithed normotensive rat

Summary Interactions between the putative calcium entry promotor Bay k 8644 and both -₁ and ₁-adrenocepter mediated increases in diastolic pressure were studied in the pithed normotensive rat. The ₁-adrenocepter mediated pressor responses elicited by B-HT920, TL-99, DP-6,7-ADTN and B-HT958 were potentiated by Bay k 8644, reflected by a leftward shift and an increase in the maximum of the log dose-pressor respinse curves. The -₁-adrenocepter effects elicited by cirazoline, methoxamine, (–)-amidephrine, St 587, (–)-phenylephrine and Sgd 101/75 were less enhanced by Bay k 8644. Only a leftward shift of the dose-response curves was observed, which was most pronounced for (–)-phenylephrine and Sgd 101/75. The -₁ and ₂-adrenocepter pressor components of (–)-noradrenaline were similarly distinguished by Bay k 8644 as observed for the selective -₁ or ₂-adrenocepter agonists.Effects of Bay k 8644 on the increase in diastolic pressure mediated by B-HT 920, St 587 and cirazoline were also studied after pretreatment with the calcium entry blocker nifedipine. After additional pretreatment with nifedipine the potentiation by Bay k 8644 observed for B-HT 920 and St 587 was more pronounced. The presence of nifedipine had no effect on the interaction between Bay k 8644 and cirazoline.It is concluded that Bay k 8644 behaves as a mirror image of nifedipine. The observation that Bay k 8644 enhances ₂-adrenocepter mediated pressor effects more effectively than ₁-adrenocepter increases in diastolic pressure is in accordance with the hypothesis of the more pronounced calcium dependency of ₂-adrenocepter mediated pressor responses. The data obtained for ceptor mediated pressor responses. The data obtained for St 587 and (–)-phenylephrine are in apparent contradiction to the finding that the pressor responses to the former drug are more markedly inhibited by calcium entry blockade than those of the latter. It is suggested that St 587 employs calcium channels which are already maximally modulated and that (–)-phenylephrine makes use of calcium channels which are in a rather inactive state. The hypothesis is put forward that the intrinsic activity of ₂-adrenocepter agonists reflects their ability to bring calcium channels in an active state.     

复合纳米人工骨的注射、凝固及其机械性能研究

目的 构建半水硫酸钙和纳米羟基磷灰石为主的复合人工骨材料并对其注射性能、凝固性能和机械强度的影响因素进行考察.方法 测试不同液固比条件下复合材料的注射特性,25℃和37℃时分别测试不同液固比、不同二水硫酸钙促凝剂条件下的材料初、终凝时间和压缩强度,均与纯硫酸钙作对比.结果 液固比0.50以上时注射性能满意.无论何种液/固比,复合材料的凝固时间均较硫酸钙延长,37℃下的凝固时间较25℃下延长.一定范围内促凝剂用量过大或过小均使凝固时间延长.液固比越大或促凝剂用量越高,材料压缩强度越低.纳米磷灰石含量增大则材料强度降低.结论 合理掌握纳米磷灰石的比例,液固比和促凝剂的用量,是开发可注射纳米人工骨的关键.     

Single channel activity associated with the calcium dependent outward current inHelix pomatia

A recently improved version of the extracellular patch clamp technique (9, 13) was used to record currents from microscopic membrane areas of Helix neurons with predominant Ca²⁺ dependent outward currents. Current fluctuations in the patches consisted mainly of frequently interrupted, one-sided steps indicating discrete open-closed state changes of single channels with an ohmic conductance of approximately 19 pS. Frequency of occurrence of the elementary events compares with amplitudes of macroscopic currents during depolarizing voltage steps of varied amplitude. Average delays in appearance of the events vary in line with delayed time courses of the cell's outward current.     

羟苯磺酸钙对血液流变及血栓形成作用的实验研究

目的研究国产抗糖尿病微血管病变药物羟苯磺酸钙对血小板聚集、血栓形成、血液粘滞度、红细胞脆性以及出血时间等影响 ,为临床应用提供理论依据。方法与结果羟苯磺酸钙0.15、0.21和0.30g/kg,灌胃 ,每日1次 ,连续3d ,可明显抑制ADP和胶原引起的大鼠血小板聚集 ,其抑制率与对照组比较有显著性差异(P<0.01) ;明显降低大鼠血液粘滞度及改善大鼠红细胞中间脆性 ,其作用与对照组比较均有显著性差异(P<0.01)。羟苯磺酸钙0.15、0.30和0.60g/kg灌胃 ,一次给药 ,可明显抑制大鼠颈动静脉旁路血栓形成 ,其作用呈剂量依赖性 ,与对照组比较有显著性差异(P<0.01 ,P<0.05) ;对于小鼠 ,羟苯磺酸钙0.15、0.30和0.60g/kg 灌胃 ,一次给药 ,可明显对抗胶原诱导的血栓形成 ,其降低血栓引起的死亡和偏瘫率呈剂量依赖性 ,与对照组比较有显著性差异(P<0.01 ,P<0.05) ;能明显延长小鼠出血时间 ,与对照组比较有显著性差异(P<0.01)。结论羟苯磺酸钙对于糖尿病微血管病变相关的血液学指标有改善作用。     

The action of caffeine on inward barium current through voltage-dependent calcium channels in single rabbit ear artery cells

The effect of caffeine on inward current carried by barium ions through voltage-dependent calcium channels has been investigated in single rabbit ear artery cells using whole-cell voltage-clamp techniques. Caffeine (1 –30 mM) caused a rapid and reversible concentration-dependent blockade of barium current and a related compound, 3-isobutyl-1-methylxanthine (IBMX), was a more potent inhibitor of barium current. Caffeine-induced inhibition of barium current showed no voltage- or usedependence and caffeine did not alter the steady-state inactivation of barium current. The effect of caffeine was not blocked by extracellular or by intracellular ryanodine or inclusion of both 5 mM 1,2-bis(2-aminophenoxy)-ethane N,N,N,N,-tetraacetic acid (BAPTA) and 2 mM ethylene glycol-bis(-amino ethyl ether) N,N,N,N,-tetraacetic acid (EGTA) in the intracellular solution. Rolipram and M&B 22984, non-xanthine inhibitors of phosphodiesterase, did not diminish inward barium current. The data indicate that caffeine and IBMX block voltage-operated calcium channels and it is suggested that this is due to a direct interaction of methylxanthines with the calcium channel.     

Studies on subcutaneous fat necrosis of the newborn

Summary Biopsy specimens from the skin and subcutaneous fat tissue of four cases with neonatal subcutaneous fat necrosis were made and investigated by light and electron microscopy at 2, 4, and 6 weeks, and 5 months (Case 2) from the onset of the disease. Three stages of ultrastructural change of fat cells were observed. The evolution of crystal formation in the fat cells was seen and phagocytosis of crystals and fat droplets by macrophages and foreign-body giant cells was also noted. In the light microscope accumulation of calcium concretions in the spaces between and inside the fat cells was found. In the electron microscope we detected foci of highly electrondense granules, which were similar in distribution and structure to calcium salts stained with the von Kossa method. Changes in small and medium size blood vessels were observed.This work was written during a stay supported by Max-Planck-Gesellschaft from Dec. 1. 1973 to March 31. 1975 in the Max-Planck-Institut für Klinische und Physiologische Forschung (W.G. Kerckhoff-Institut), Bad Nauheim, West Germany (Director: Prof. Dr. Wolfgang Schaper)