Amiloride-sensitive Na+ transport across cultured renal (A6) epithelium: evidence for large currents and high NaK selectivity

Electrical techniques were used to determine the NaK selectivity of the amiloride-sensitive pathway and to characterize cellular and paracellular properties of A6 epithelium. Under control conditions, the mean transepithelial voltage (V _T) was –57±5 mV, the short-circuit current (I _sc) averaged 23±2 A/cm² and the transepithelial resistance (R _T) was 2.8±0.3 kcm² (n=13). V _T and I _sc were larger than reported in previous studies and were increased by aldosterone. The conductance of the amiloride-sensitive pathway (G _amil) was assessed before and after replacement of Na⁺ in the mucosal bath by K⁺, using two independent measurements: (1) the slope conductance (G _T), determined from current-voltage (I-V) relationships for control and amiloride-treated tissues and (2) the maximum amiloride-sensitive conductance (G _max) calculated from the amiloride dose-response relationship. The ratio of G _amil in mucosal Na⁺ solutions to G _amil for mucosal K⁺ solutions was 22±6 for G _T measurements and 15±2 for G _max data. Serosal ion replacements in tissues treated with mucosal nystatin indicated a potassium conductance in the basolateral membrane. Equivalent circuit analyses of nystatin and amiloride data were used to resolve the cellular (E _c) and paracellular (R _j) resistances (5 kcm² and 8–9 kcm², respectively). Analysis I-V relationships for tissues depolarized with serosal K⁺ solutions revealed that the amiloride-sensitive pathway could be described as a Na⁺ conductance with a permeability coefficient (P _Na)=1.5±0.2× 10^–6 cm/s and the intracellular Na⁺ concentration (Na_i)=5±1 mM (n=5), similar to values from other tight epithelia. We conclude that A6 epithelia are capable of expressing large amiloride-sensitive currents which are highly Na⁺ selective.     

Delayed onset and slow time course of the non-M-type muscarinic current in bullfrog sympathetic neurons

The onset and time course of the muscarinic currents induced by brief applications of acetylcholine (ACh) were examined in voltage-clamped neurons of bullfrog sympathetic ganglia bathed in a solution containing d-tubocurarine. At a potential of –40 mV, the ACh-induced current (I _ACh) appeared within 1.2 s and rapidly increased to its peak with a half-activation time of 2.2 s. This initial current was termed the fastI _ACh and was blocked by 4 mM Ba²⁺. At a potential more negative than –60 mV, the fastI _ACh disappeared and the remainingI _ACh activated with a delay of 3.9 s and slowly increased to its peak with a half-activation time of 8.2 s. This delayed current was termed the slowI _ACh and is thought to be associated with inhibition of a K⁺ current, orI _M, as well as activation of an inward current through non-M-type muscarinic cation channels. The slowI _ACh was not inhibited by Ba²⁺, but its amplitude was reduced with depolarization (the extra-polated reversal potential was +3 mV). In Na⁺-free solution, the amplitude of the slowI _ACh reduced, but its polarity did not reverse in the voltage region examined (-30 to –100 mV). The slow excitatory postsynaptic current was also recorded, and was shown to have a similar delay in onset and slow time course. The results demonstrate that ACh activates the non-M-type muscarinic current three times more slowly than it inhibitsI _M.     

Analysis of Electromagnetic Exposure in Wireless Power Transfer System for Endoscopic Micro-robot

A wireless power transfer system for endoscopic micro-robot operating at 36 kHz is presented in this paper. The issue of patient＇ s health and safety regarding exposure to the electromagnetic field is addressed. The specific absorption rate and current density can be used to investigate the electromagnetic influences on the biological tissues surrounded by the wireless power launching coil. In view of this purpose, the limited close-ound solenoid electromagnetic model is built, the relationship between the electric intensity and the specific absorption rate and current density is deduced, and the simulation experiments are done. Experimental results show that the values of SAR and current density related to different tissue catalogs are all very small and do not exceed their own limits respectively when the resonance frequency of operation is 36 kHz.     

Membrane attack complex of complement and 20 kDa homologous restriction factor (CD59) in myocardial infarction

In order to investigate the mechanism of deposition of the complement membrane attack complex (MAC) in cardiomyocytes in areas of human myocardial infarction, the 20 kDA homologous restriction factor of complement (HRF20; CD59) and complement components (C1q, C3d and MAC) were analysed immunohistochemically using specific antibodies. Myocardial tissues obtained at autopsy from nine patients who died of acute myocardial infarction were fixed in acetone and embedded in paraffin. The ages of the infarcts ranged from about 3.5 h to 12 days. In cases of myocardial infarction of 20 h or less, MAC deposition was shown in the infarcted cardiomyocytes without loss of HRF20. Where the duration was 4 days or more, the cardiomyocytes with MAC deposition in the infarcted areas also showed complete loss of HRF20. Outside the infarcts, HRF20 in the cardiomyocytes was well preserved without MAC deposition. The present study suggests that the initial MAC deposition in dead cardiomyocytes can occur as a result of degradation of plasma-membrane by a mechanism independent of complement-mediated injury to the membrane. Loss of HRF20 from dead cardiomyocytes may not be the initial cause of MAC deposition, but may accelerate the deposition process of MAC in later stages of infarction.     

A model of electrical activity and cytosolic calcium dynamics in vascular endothelial cells in response to fluid shear stress

A mathematical model is proposed to describe the intracellularCa ²⁺ (Ca _i) transient and electrical activity of vascular endothelial cells (VEC) elicited by fluid shear stress (τ). The intracellularCa ²⁺ store of the model VEC is comprised of aCa _i-sensitive (sc) and an inositol (1,4,5)-trisphosphate (IP ₃)-sensitive compartment (dc). The dc [Ca ²⁺] is refilled by the sc whose [Ca ²⁺] is the same as extracellular [Ca ²⁺].IP ₃ produced by the τ-deformed mechanoreceptors discharges the dcCa ²⁺ into the cytosol. The increase of cytosolic[Ca ²⁺] inducesCa ²⁺ release (CICR) from the sc. The raisedCa _i activates aCa _i-activatedK ⁺ current (I _{K, Ca}) and inhibitsIP ₃ production. The cell membrane potential is determined byI _{K, Ca}, voltage-dependentNa ⁺ andK ⁺ currents. Steady τ>0.1 dyne/cm² elicits aCa _i varies sigmoidally withLog ₁₀(τ) with a maximal peakCa _i of 150 nM at τ=4 dynes/cm². Step increases of τ fail to elicit aCa ²⁺ response in cells previously stimulated by a lower shear. TheCa ²⁺ response gradually decreases with repetitive τ stimuli. Pulsatile shear elicits two to three times higherCa _i and hyperpolarizes the cell more than steady shear of the same magnitude. The simulatedCa ²⁺ responses to τ are quantitatively and qualitatively similar to those observed in cultured VEC. The model provides a possible explanation of why the vasodilating stimulus is greater for pulsatile flow than for nonpulsatile flow.     

Microelectrode amplifier with improved method of input-capacitance neutralisation

A circuit is described which allows the input capacitance of an f.e.t. input integrated circuit to be used both as the feedback capacitance to neutralise the total input capacitance and to inject current pulses into the input. Compared with the conventional method of adding discrete capacitors to perform these functions, this design results in a lower total capacitance at the input, which reduces the high-frequency noise generated by the amplifier and facilitates the achievement of a low effective capacitance. A modified version having an ultralow (0·1pA) input current, for use with ion-sensitive microelectrodes, is also described.     

Inhibition of the slow inward current and the time-dependent outward current of mammalian ventricular muscle by gentamicin

The aminoglycoside antibiotic, gentamicin (GM), depressed the plateau phase and shortened the duration of the action potential in guinea pig papillary muscle. Its effect on the membrane currents was studied by a single sucrose gap voltage clamp method. The slow inward current (i_s) was remarkably diminished by GM with little change in its time course, in the voltage-dependency of the steady-state inactivation and activation or in its reversal potential. The maximal amplitude of i_s, obtained by subtracting the Co²⁺-resistant current, was reduced to 57% by 0.1 mmol/l GM and almost reduced to zero by 1 mmol/l GM. The efficacy of GM in inhibiting i_s was reduced by increasing the external Ca²⁺ concentration from 1.8 to 5.4 or 10.8 mmol/l, but not by the application of adrenaline. The time-dependent outward current (i_K) was also decreased by GM but only at higher concentrations. It is proposed that the depressant action of GM on i_s was due to a blockade of slow channels, whereby GM may have dislocated Ca from the binding sites at slow channels on the external surface of the membrane.     

Calcium-dependent slow outward current in visceral primary afferent neurones of the rabbit

Slow outward currents were recorded from voltage-clamped neurones in nodose ganglia excised from rabbits. In the majority of Type C neurones, a short depolarizing command pulse evoked a slow outward tail current (I _SAH) with a decay time constant ranging from 0.5 to 2 s. TheI _SAH was due to an increase in membrane conductance to K⁺ because its reversal potential was approximately equal to the Nernst potential for K⁺. TheI _SAH was reversibly blocked by removal of external Ca²⁺ or by Ca²⁺ antagonists. A Ca²⁺ ionophore, A23187, produced an outward current which was similar to theI _SAH. TheI _SAH was resistant to tetraethylammonium and depressed by Ba²⁺, whereas it was not affected by Cs⁺ and 4-aminopyridine. TheI _SAH was initially augmented and subsequently depressed by apamin (1–10 nM) and (+)-tubocurarine (100–600 M). It is concluded that theI _SAH in visceral primary neurones may be due to a long-lasting increase in K⁺ conductance caused by an increase in the concentration of intracellular Ca²⁺, resulting from Ca²⁺ entry during the depolarizing command pulse.     

温敏型智能化靶向式药物载体研究（I）——具有温敏开关的多孔膜

研究了膜孔内接枝聚异丙基丙烯酰胺 (PNIPAM)“开关”的温敏型智能膜的制备 ,并对其进行了温度感应开关性能实验。实验中采用等离子体接枝填孔聚合法将 PNIPAM接枝在多孔平板膜的膜孔中 ,结果表明 ,这种接枝了 PNIPAM“开关”的多孔膜具有温度感应特性 ,其利用膜孔内 PNIPAM接枝链的膨胀 -收缩特性实现了感温性开关性能。当环境温度低于 PNIPAM的低临界溶解温度 (L CST)时 ,膜孔内 PNIPAM分子链膨胀而使膜孔呈“关闭”状态 ;而当环境温度高于 L CST时 ,PNIPAM分子链变为收缩状态而使膜孔“开启”。温敏开关的 L CST可通过添加丙烯酰胺 (AAM)与异丙基丙烯酰胺 (NIPAM)共聚来调节 ,AAM与 NIPAM共聚开关的 L CST随 AAM添加量的增加而单调上升。     

隔膜条件下骨再生成骨细胞来源的实验观察

目的:研究在隔膜条件下引导性骨再生过程中成骨细胞的来源,进一步认识引导性骨再生的机制.方法:以成年新西兰兔为研究对象,在双侧挠骨中段制作标准骨缺损不愈合模型,用硅胶膜成管状包裹一侧骨缺损,另一侧无特殊处理为对照.术后分别进行X线检查、常规HE染色以及SP方法BMP、BGP抗体的免疫组化染色.结果:硅胶膜在骨缺损处形成隔离密闭的腔室,将周围组织阻挡于骨缺损之外.早期骨端骨内膜、骨髓基质细胞大量增殖,形成肉芽组织占据骨缺损.骨再生过程中表现出明显的组织学特征:骨痂表面为2～3层成骨细胞,骨缺损中央为肉芽组织,两者之间为数层细胞形成的移行区,细胞排列疏松.早期骨端骨内膜、骨髓基质细胞BMP、BGP呈强阳性染色,骨痂生长过程中,移行区部分细胞呈阳性染色.结论:结果表明在隔膜条件下骨再生的成骨细胞在早期来源于髓内的骨内膜和骨髓基质细胞,骨痂形成后,成骨细胞则来源于骨内膜、骨髓、骨膜增殖细胞共同形成的肉芽组织中的间质细胞或成纤维细胞.