Inhibition of the Na+/K+ cotransport system by cyclic AMP and intracellular Ca2+ in human red cells

Human erythrocytes are able to incorporate cyclic AMP (cAMP) in amounts larger than those required to saturate cAMP-dependent protein kinase. In contrast to previous observations in avian red blood cells in which cAMP stimulates the Na⁺/K⁺ cotransport system, we demonstrate that cAMP inhibits this system in human erythrocytes. The cotransport inhibition is enhanced by addition of phosphodiesterase inhibitor 1-methyl-3-isobutylxanthine to the incubation medium. The cAMP concentration giving half-maximal cotransport inhibition showed a wide variation among different individuals (from 0.1 to 5 mM external cAMP concentration). In contrast to cAMP, cyclic GMP showed little effect on the cotransport system. Ca²⁺ introduced into the cell interior was an inhibitor of the Na⁺/K⁺ cotransport system. These results suggest that in human cells in which endogeneous levels of cAMP and Ca²⁺ are modulated by hormones, the Na⁺/K⁺ cotransport system may be under hormonal regulation.     

Conformation Changes,N-terminal Involvement,and cGMP Signal Relay in the Phosphodiesterase-5 GAF Domain

The activity of phosphodiesterase-5 (PDE5) is specific for cGMP and is regulated by cGMP binding to GAF-A in its regulatory domain. To better understand the regulatory mechanism, x-ray crystallographic and biochemical studies were performed on constructs of human PDE5A1 containing the N-terminal phosphorylation segment, GAF-A, and GAF-B. Superposition of this unliganded GAF-A with the previously reported NMR structure of cGMP-bound PDE5 revealed dramatic conformational differences and suggested that helix H4 and strand B3 probably serve as two lids to gate the cGMP-binding pocket in GAF-A. The structure also identified an interfacial region among GAF-A, GAF-B, and the N-terminal loop, which may serve as a relay of the cGMP signal from GAF-A to GAF-B. N-terminal loop 98–147 was physically associated with GAF-B domains of the dimer. Biochemical analyses showed an inhibitory effect of this loop on cGMP binding and its involvement in the cGMP-induced conformation changes.     

Regulation of phospholipid synthesis inMycobacterium smegmatis by cyclic adenosine monophosphate

Forskolin, an adenylate cyclase activator and a cyclic AMP analogue, dibutyryl cyclic AMP have been used to examine the relationship between intracellular levels of cyclic AMP and lipid synthesis inMycobacterium smegmatis. Total phospholipid content was found to be increased in forskolin grown cells as a result of increased cyclic AMP levels caused by activation of adenylate cyclase. Increased phospholipid content was supported by increased [¹⁴C] acetate incorporation as well as increased activity of glycerol-3-phosphate acyltransferase. Pretreatment of cells with dibutyryl cyclic AMP had similar effects on lipid synthesis. Taking all these observations together it is suggested that lipid synthesis is being controlled by cyclic AMP in mycobacteria.     

整治区植烟土壤养分空间变异及肥力适宜性等级评价

采用地理信息系统(GIS)技术和地统计学方法,研究了湖北省恩施市“清江源”现代烟草农业科技园区、恩施城郊和利川柏杨基地单元新整治区域的土壤属性和速效养分空间变异特征,并采用模糊数学法对其土壤肥力适宜性指数(SFI)进行评价.结果表明: 土地整治对土壤速效磷含量的变异和空间分布的影响较大,这种影响程度可能与地形地貌特征和整治程度有关;土地整治对土壤pH值的影响较小,但是柏杨土壤酸化严重(pH<5.5).77.6%的城郊土壤具有较低的SFI,而柏杨和清江源分别仅有17.1%和31.4%的土壤具有较低的SFI.柏杨需要重点解决土壤酸化问题,城郊需要重点解决整个区域土壤培肥和肥力均衡化问题,而清江源则需解决SFI较低区域的培肥问题.     

Sea level and faunal changes during the latest Llandovery and earliest Ludlow (Silurian)

It is now recognized that the late Telychian and early Gorstian sea level changes are, like many others in the Silurian, of world‐wide extent. The 30–50 m deepening events at these times were between 1 and 2 Ma in duration, so melting continental ice caps appear to be the most probable cause. The faunal changes associated with the two events are respectively very close to the base and the top of the Wenlock Series and thus segregate many of the faunas diagnostic of the Llandovery, Wenlock, and Ludlow series. Permanent extinctions (often followed by radiations) are more prevalent in the graptolites, conodonts, and acritarchs, while benthic faunas are more affected by regional shifts in community distributions. This means that, in the benthos, slowly evolving lineages are the only reliable guides to time. Such phyletic evolution, however, appears to have been unaffected by sea‐level events.     

Regulation of gap junction coupling in the developing neocortex

In the developing mammalian, neocortex gap junctions represent a transient, metabolic, and electrical communication system. These gap junctions may play a crucial role during the formation and refinement of neocortical synaptic circuitries. This article focuses on two major points. First, the influence of gap junctions on electrotonic cell properties will be considered. Both the time-course and the amplitude of synaptic potentials depend,inter alia, on the integration capabilities of the postsynaptic neurons. These capabilities are, to a considerable extent, determined by the electrotonic characteristics of the postsynaptic cell. As a consequence, the efficacy of chemical synaptic inputs may be crucially affected by the presence of gap junctions. The second major topic is the regulation of gap junctional communication by neurotransmitters via second messenger pathways. The monoaminergic neuromodulators dopamine, nordrenaline, and serotonin reduce gap junction coupling via activation of two different intracellular signaling cascades—the cAMP/protein kinase A pathway and the IP₃/Ca²⁺/protein kinase C pathway, 013 respectively. In addition, gap junctional communication seems to be modulated by the nitric oxide (NO)/cGMP system. Since NO production can be stimulated by glutamate-induced calcium influx, the NO/cGMP-dependent modulation of gap junctions might represent a functional link between developing glutamatergic synaptic transmission and the gap junctional network. Thus, it might be of particular importance in view of a role of gap junctions during the process of circuit formation.     

Characterization of K+ Channels in the Basolateral Membrane of Rat Tracheal Epithelia

To study K⁺ channels in the basolateral membrane of chloride-secreting epithelia, rat tracheal epithelial monolayers were cultured on permeable filters and mounted into an Ussing chamber system. The mucosal membrane was permeabilized with nystatin (180 μg/ml) in the symmetrical high K⁺ (145 mm) Ringer solution. During measurement of the macroscopic K⁺ conductance properties of the basolateral membrane under a transepithelial voltage clamp, we detected at least two types of K⁺ currents: one is an inwardly rectifying K⁺ current and the other is a slowly activating outwardly rectifying K⁺ current. The inwardly rectifying K⁺ current is inhibited by Ba²⁺. The slowly activating K⁺ current was potentiated by cAMP and inhibited by clofilium, phorbol 12-myristae 13-acetate (PMA) and lowering temperature. This is consistent with the biophysical characteristics of I _SK channel. RT-PCR analysis revealed the presence of I _SK cDNA in the rat trachea epithelia. Although 0.1 mm Ba²⁺ only had minimal affect on short-circuit current (I _sc) induced by cAMP in intact epithelia, 0.1 mm clofilium strongly inhibited it. These results indicate that I _SK might be important for maintaining cAMP-induced chloride secretion in the rat trachea epithelia. Received: 1 March 1996/Revised: 5 August 1996     

Rapid regulation of electrolyte absorption in sweat duct

Even though the same Cl channel (CFTR) is common to certain fluid transport functions that are oppositely directed, i.e., secretion and absorption, only fluid secretion has clearly been shown to be acutely regulated. It is now clear that fluid secretion activated by -adrenergic stimulation is controlled by cAMP-mediated opening and closing of CFTR-Cl channels. Since the conductance of the human sweat duct is almost wholly due to CFTR-Cl conductance (CFTR-G_Cl), we sought to determine whether salt absorption via CFTR-Cl channels could also be subject to acute regulation in this purely absorptive epithelium. After -toxin permeabilization, we found that addition of cAMP resulted in a large increase in Cl diffusion potentials across the apical membrane and a more than twofold increase in the average membrane conductance. Since the cAMP effects were dependent on Cl alone, not on Na, and since apical Cl conductance appears to be almost exclusively comprised of CFTR-G_Cl, we surmise that this form of electrolyte absorption like secretion is also subject to acute control through CFTR-G_Cl. Acute regulation of absorption involves both activation by phosphorylation (PKA) and inactivation by dephosphorylation (unknown endogenous phosphatase) of CFTR. Phosphorylation of CFTR was shown by the facts that CFTR-G_Cl could be activated by cAMP and inhibited by the kinase antagonist staurosporine, or by removal of either substrate ATP or Mg²⁺ cofactor. Inactivation of CFTR-G_Cl by endogenous phosphatase(s) was indicated by a spontaneous but reversible loss of CFTR-G_Cl upon removal of cAMP. Such loss of CFTR-G_Cl activity could be prevented either by application of phosphatase inhibitors or by using phosphatase-resistant ATP--S as substrate to phosphorylate CFTR. We surmise that absorptive function is subject to rapid regulation which can be switched on and off acutely by a control system that is common to both absorptive and secretory processes and that this control is crucial to switching between conductive and nonconductive transport mechanisms during salt absorption.The authors are grateful to Mr. Kirk Taylor for expert technical assistance, and to numerous volunteer human subjects for participating in the experiments with informed consent. Supported by grants from the National Institutes of Health, DK-41329-04 and the National Cystic Fibrosis Foundation, Z 439.     

Response by macrozoobenthos biomass to water level regulation in some Finnish lake littoral zones

The relationship of the macrozoobenthos biomass in the littoral area to the yearly fluctuation in water level and the characteristics of the area or lake are studied using data collected from sheltered bays in regulated and natural waters. Most of the lakes were clear and oligotrophic. The benthos biomass at all depths in the littoral decreased with increased water level fluctuation, provided that the transparency of the water was uniform.The macrozoobenthos biomass in the 0–3 m depth zone could be predicted fromlog macrozoobenthos biomass (mg ODW) m^-2=4.25-1.33 (log Biomass Index) in which the Biomass Index is calculated as% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaaeOqaiaabM% gacaqGVbGaaeyBaiaabggacaqGZbGaae4CaiaabccacaqGjbGaaeOB% aiaabsgacaqGLbGaaeiEaiaab2dacaqGGaGaaeiiaiaabccacaqGGa% GaaeiiaiaabccacaqGGaGaaeiiaiaabccacaqGGaGaaeiiaiaabcca% caqGGaGaaeiiaiaabccacaqGGaGaaeiiaiaabccacaqGGaGaaeiiai% aabccadaWcaaabaeqabaGaae4DaiaabggacaqG0bGaaeyzaiaabkha% caqGGaGaaeiBaiaabwgacaqG2bGaaeyzaiaabYgacaqGGaGaaeOzai% aabYgacaqG1bGaae4yaiaabshacaqG1bGaaeyyaiaabshacaqGPbGa% ae4Baiaab6gacaqGGaGaaeyAaiaab6gacaqGGaGaaeiDaiaabIgaca% qGLbGaaeiiaiaabchacaqGYbGaaeyzaiaabAhacaqGPbGaae4Baiaa% bwhacaqGZbGaaeiiaiaabMhacaqGLbGaaeyyaiaabkhaaeaacaqGOa% GaaeyBaiaabUdacaqGGaGaae4yaiaabggacaqGSbGaae4yaiaabwha% caqGSbGaaeyyaiaabshacaqGLbGaaeizaiaabccacaqGMbGaaeOCai% aab+gacaqGTbGaaeiiaiaab2gacaqGVbGaaeOBaiaabshacaqGObGa% aeiBaiaabMhacaqGGaGaaeyBaiaabwgacaqGHbGaaeOBaiaabccaca% qG2bGaaeyyaiaabYgacaqG1bGaaeyzaiaabohacaqGPaaaaeaacaqG% tbGaaeyzaiaabogacaqGJbGaaeiAaiaabMgacaqGGaGaaeizaiaabM% gacaqGZbGaae4AaiaabccacaqG2bGaaeyyaiaabYgacaqG1bGaaeyz% aiaabccacaqGPbGaaeOBaiaabccacaqG0bGaaeiAaiaabwgacaqGGa% Gaae4CaiaabggacaqGTbGaaeyzaiaabccacaqGVbGaaeiCaiaabwga% caqGUbGaaeiiaiaabEhacaqGHbGaaeiDaiaabwgacaqGYbGaaeiiai% aabohacaqGLbGaaeyyaiaabohacaqGVbGaaeOBaiaabccacaqGOaGa% aeyBaiaabMcaaaaccaGae8hiaaIaaKiEaiab-bcaGiaaigdacaaIWa% GaaGimaiaac6caaaa!CBD8!\[{\text{Biomass Index = }}\frac{\begin{gathered} {\text{water level fluctuation in the previous year}} \hfill \\ {\text{(m; calculated from monthly mean values)}} \hfill \\ \end{gathered} }{{{\text{Secchi disk value in the same open water season (m)}}}} \user1{x} 100.\]The whole illuminated littoral shifts due to water level fluctuation, which disturbs the zonation of the benthos. Such an increase or decrease in benthic biomass has been observed after one year of disturbance due to water level fluctuation. It need, however, a study based on the carefully planned and collected data, in which it can be taken account by a multivariate statistical analysis also the interactions between the important factors affected the littoral benthos.