Steroid hormones: modulators of Na+ absorption and Cl- secretion in cultured tracheal epithelia.

Cultures of tracheal epithelial cells from adult nonpregnant female rabbits maintained a different balance of Na+ absorption and Cl- secretion from cells of pregnant animals and fetal cultures. Cells of nonpregnant rabbits had a large Na+ absorptive current (11 microA/cm2 or 53% of baseline short circuit current) and a small Cl- secretory current (2.4 microA/cm2 or 11%). In contrast, fetal cells had greater Cl- secretion (15.1 microA/cm2 or 61%) and much less Na+ absorption (1.6 microA/cm2 or less than 10%). Cells of pregnant animals had amounts of Cl- secretion (11.4 microA/cm2 or 41%) and Na+ absorption (6.7 microA/cm2 or 22%) that were in between that of fetal and adult nonpregnant animals. In addition to these striking differences in baseline ion transport among the three groups, epinephrine was found to stimulate Cl- secretion in cells from pregnant rabbits, both Na+ absorption and Cl- secretion in cells from nonpregnant animals, and neither in fetal cells. Steroid hormones of pregnancy, progesterone and estradiol, when added to cells cultured from nonpregnant rabbits, altered the magnitudes of the individual transport pathways to mimic cells cultured from pregnant animals. In addition to changing baseline transport, these steroids modified the epinephrine-stimulated currents to resemble those in the fetus and in pregnancy. We conclude that steroid hormones regulate the balance of Na+ absorption and Cl- secretion in tracheal epithelia in utero and in adulthood.     

The loss of the chloride channel, ClC-5, delays apical iodide efflux and induces a euthyroid goiter in the mouse thyroid gland

Genetic inactivation of ClC-5, a voltage-gated chloride channel prominently expressed in the kidney, leads to proteinuria because of defective apical endocytosis in proximal tubular cells. Because thyroid hormone secretion depends on apical endocytosis of thyroglobulin (Tg), we investigated whether ClC-5 is expressed in the thyroid and affects its function, using Clcn5-deficient knockout (KO) mice. We found that ClC-5 is highly expressed in wild-type mouse thyroid ( approximately 40% of mRNA kidney level). The protein was immunolocalized at the apical pole of thyrocytes. In Percoll gradients, ClC-5 overlapped with plasma membrane and early endosome markers, but best codistributed with the late endosomal marker, Rab7. ClC-5 KO mice were euthyroid (normal T4 and TSH serum levels) but developed a goiter with parallel iodine and Tg accumulation (i.e. normal Tg iodination level). When comparing ClC-5 KO with wild-type mice, thyroid 125I uptake after 1 h was doubled, incorporation into Tg was decreased by approximately 2-fold, so that trichloroacetic acid-soluble 125I increased approximately 4-fold. Enhanced 125I- efflux upon perchlorate and presence of 125I-Tg as autoradiographic rings at follicle periphery demonstrated delayed iodide organification. Endocytic trafficking of 125I-Tg toward lysosomes was not inhibited. Expression of pendrin, an I-/Cl- exchanger involved in apical iodide efflux, was selectively decreased by 60% in KO mice at mRNA and protein levels. Thus, ClC-5 is well expressed in the thyroid but is not critical for apical endocytosis, contrary to the kidney. Instead, the goiter associated with ClC-5 KO results from impaired rate of apical iodide efflux by down-regulation of pendrin expression.     

Long-term results of surgical treatment for pulmonary valve stenosis

109 children who survived surgical treatment for isolated pulmonary valve stenosis were followed for up to 17 years. In all the postoperative status was assessed as satisfactory. Cardiac catheterization repeated in 43 gave a resting valve gradient below 40 mmHg. The 22 children whose pulmonary valves had been excised were as healthy as the 87 who had undergone pulmonary valvotomy. Consideration was given to the desirable length of postoperative review. Except for the few children with symptoms before operation, a postoperative increase in exercise tolerance was not a feature.     

Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney

BACKGROUND: The mRNAs of several types of calcium channels have been identified in intact rat kidney, and L-type calcium channels cause changes in intracellular calcium in primary cultures of distal tubule cells. The aim of this study was to evaluate the tubular and cellular distribution of the alpha1C subunit of the L-type calcium channel in intact kidney. METHODS: RT-PCR and Northern blot analysis were used to assess the regional abundance of the mRNA of this channel. Immunocytochemistry combined with confocal microscopy and surface biotinylation were applied to determine the tubular and cellular localization of the protein. RESULTS: Northern blot and RT-PCR analysis indicated that the mRNA of the alpha1C subunit of the cardiac L-type calcium channel was present in whole rat kidney, kidney tubules and kidney cell lines. Western blot of lysates from whole kidney, kidney tubules or cell lines revealed bands of approximately 190 kD for the alpha1C subunit and approximately 60 kD for the beta3 subunit. Confocal immunohistochemistry indicated that the alpha1C subunit of this channel was co-expressed in cells of the distal tubule that express calbindin-D28K, but not in intercalated cells. The alpha1C subunit was also highly expressed in both outer and inner medullary collecting ducts. Serial confocal microscopic images or surface biotinylation experiments determined that the channel was predominantly on the basolateral membrane but had some distribution on the apical membrane. CONCLUSIONS: The distribution and cellular localization of the alpha1C subunit of cardiac L-type calcium channel suggest it is probably involved in intracellular and membrane calcium signaling.     

(-)-Epigallocatechin-3-gallate inhibition of ultraviolet B-induced AP-1 activity

Green tea polyphenols have been shown to inhibit cancer in a variety of tumor models, including ultraviolet B (UVB)-induced non-melanoma skin cancer. In green tea extracts, the major dry mass constituent is the family of catechins, of which (-)-epigallocatechin-(3)-gallate (EGCG) is considered to be important for the chemopreventive activity. EGCG has been shown to have antioxidant properties, but there has been little progress toward identifying the specific targets and mechanisms of its action. Using cultured human keratinocytes, we show that UVB- induced AP-1 activity is inhibited by EGCG in a dose range of 5.45 nM to 54.5 microM. EGCG is effective at inhibiting AP-1 activity when applied before, after or both before and after UVB irradiation. EGCG also inhibits AP-1 activity in the epidermis of a transgenic mouse model. This work begins to define a mechanism by which EGCG could be acting to inhibit UVB-induced tumor formation.      

Potassium transport in the early distal tubule ofAmphiuma kidney

Studies were performed to investigate potassium transport in early distal tubule of the doubly-perfused kidney ofAmphiuma under control conditions and following K-adaptation. Double barreled K-sensitive microelectrodes were used in stationary microperfusion experiments. Net K-flux was evaluated along with measurements of both cell membrane potential and cell K activity. Net K flux and electrochemical driving forces of K were described over a wide range of peritubular K concentrations. Whereas in control animals, at normal and low peritubular K concentrations K reabsorption occurs, K secretion is induced by elevating peritubular K. In contrast, net K secretion is seen at all peritubular K levels in the K-adapted kidney. Net K secretion approaches saturation at high peritubular K concentrations. Intracellular K activities also approach plateau values which are shifted upward in the state of K-adaptation. In control animals at zero net flux conditions intracellular K is maintained above electrochemical equilibrium across both the peritubular and the luminal cell membrane. After K-adaptation, however, K approaches electrochemical equilibrium across the luminal cell membrane. The results indicate that in control conditions, K is taken up actively into the cell across the peritubularand across the luminal cell barrier. It is likely that both luminal and peritubular transport components (increased luminal K conductance, diminished luminal K cotransport, stimulation of peritubular K-uptake) are responsible for increased K secretion during K adaptation.     

Glucocorticoid receptor and heat shock protein 90 in peripheral blood mononuclear cells from asthmatics

目的探讨糖皮质激素受体(GR)和热休克蛋白90(HSP90)mRNA在糖皮质激素敏感型(SS)、依赖型(SD) 和抵抗型(SR)哮喘中的表达及其在SR哮喘发病中的作用。 方法采用反转录-聚合酶链(RT-PCR)的方法分别测定正常人(10例)、SS哮喘(10例)、SD哮喘(5例)和 SR哮喘(6例)的外周血单个核细胞(PBMC)中GR mRNA和HSP90 mRNA的表达,并在体外用IL-2、IL-4分 别、联合刺激上述细胞观察其受刺激后GR mRNA和HSP90 mRNA表达的改变情况。 结果 SR哮喘的GR和HSP90 mRNA表达水平最高(分别为0.730±0.171和1.122±0.165),SS哮喘次之 (分别为0.359±0.350和0.885±0.250),SD哮喘最低(分别为0.017±0.008和0.078±0.039)。正常人有 一定表达(分别为0.052±0.013和0.362±0.101)。GR和HSP90 mRNA的表达各组间相比P＜0.05。正 常人、SS、SD和SR哮喘HSP90/GR的比值分别为7.15±1.84、8.39±7.95、5.51±3.30、1.57±0.18,SR哮喘 HSP90/GR比值明显低于前三组(P＜0.05)。IL-2和IL-4单独刺激对SS、SD和SR哮喘的GR、HSP90 mRNA表达无明显影响,二者联合刺激可使SS、SD和SR哮喘GR mRNA表达以及SS、SD哮喘HSP90 mRNA 表达增强,但不能使SR哮喘HSP90 mRNA表达增强。 结论 SR哮喘中HSP90 mRNA表达相对不足造成HSP90/GR比值降低可能是SR哮喘形成的原因之一, IL-2+IL-4对GR和HSP90 mRNA表达的不同调节作用可能是形成SR哮喘HSP90/GR比值降低的原因之一。     

Children of alcoholics: Attention, information processing and event-related brain potentials

Recent studies on biological markers and risk factors for alcoholism have distinguished between nonalcoholic individuals with a family history of alcoholism and those without such a family history on measures of event-related brain potentials. The main finding of these "high-risk" studies is a smaller amplitude of the P300 component in males with a history of paternal alcoholism. This relationship between P300 amplitude and a family history of paternal alcoholism has been observed in adults and children. Consequently, several authors have suggested that a reduced P300 amplitude could serve as a vulnerability marker for alcoholism. We address several conceptual and methodological issues involved in the study of event-related potentials in children at high risk for alcoholism. Subsequently, the ongoing high-risk study of the Amsterdam Institute for Addiction Research is described briefly.     

Psychological mechanisms of enhanced risk of addiction in children of alcoholics: a dual pathway?

The background and rationale of a recently started project of the Amsterdam Institute for Addiction Research are outlined. This project is aimed at the psychological mechanisms underlying an enhanced risk of (later) addiction in children of alcoholics and the relationship with childhood psychopathology. A dual pathway mechanism is proposed, in which the type of alcoholism of the parent plays a major role. The child of a multigenerational primary alcoholic parent may suffer from an inherited mild dysfunction of the prefrontal cortex, expressed in neuropsychological and personality characteristics similar to those of the alcoholic parent. These are impulsive, aggressive and reward-seeking behaviour, response perseveration and, in some children, related psychopathology such as conduct disorders. For a child of a secondary alcoholic parent, another mechanism is proposed. In these children, stress and social learning may lead to negative affectivity and repressive coping style, with emotional problems at a later age, and the risk of falling into the "circle of secondary alcoholism". In both pathways, alcohol-related expectancies are suggested to constitute a "final common pathway" between different risk factors and later alcohol abuse. Specific expectancies might be related to different pathways and to gender differences in later drinking patterns.