Volatile anesthetics gate a chloride current in postnatal rat hippocampal neurons.

A volatile anesthetic-gated current was characterized in patch-clamped cultured postnatal rat hippocampal neurons. In this preparation, the major volatile anesthetics, isoflurane, halothane, and enflurane, open an anion-selective conductance. This volatile anesthetic-gated current exhibits anion selectivity with a chloride-to-acetate permeability ratio of 15, shows outward rectification well described by the constant field equation, and is activated in a dose-dependent fashion with half-maximal response to isoflurane at 0.8 mM (0.032 atm). The current persists in the absence of external Ca2+ and is not blocked by strychnine, a glycine antagonist. However, the gamma-aminobutyric acidA (GABAA) antagonists, bicuculline and picrotoxinin, and the nonspecific anion channel blocker, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), completely block the response. These observations suggest that volatile anesthetics, like several other general anesthetics such as barbiturates, steroids, and etomidate, have a GABA-mimetic effect on vertebrate central neurons in culture. It is not clear whether this GABAA-gating property is a prerequisite for all general anesthetics. However, under normal physiological conditions of low intracellular Cl-, it is likely that drugs with both direct GABA agonist and GABA modulatory properties will produce overall depression of the central nervous system by increasing the normal inhibitory synaptic influence and by directly hyperpolarizing neurons.     

Studies on the Analysis of Fluorescence Decay Data by the Method of Moments

The method of moments, as presented by Isenberg and Dyson (1969; Biophys. J. 9:1337) has been shown to be a reliable way of obtaining the amplitudes and time constants of several simultaneously emitting species, even in the presence of an overlapping excitation. Recent improvements in the method include (a) a component incrementation test for determining the number of relaxations, (b) a procedure, which we call exponential depression, for dramatically improving convergence, and (c) a new algorithm for implementing the method of moments on a digital computer with a high degree of flexibility and efficiency. These improvements, as well as new general theory, are described and tested using both synthetic and real experimental data. Component incrementation consists of examining models with increasing numbers of exponential terms. Given adequate precision, we find that an analysis for N + 1 components, of data that are actually represented by N components, provides the correct amplitudes and time constants plus an N + 1 term with an insignificant amplitude. Exponential depression is a transformation in which the original excitation and fluorescence, E(t) and F(t), are multiplied by exp (-λt), where λ is an arbitrary parameter. While the convolution is invariant to this transformation, the proper choice of λ greatly reduces the number of iterations necessary to obtain the amplitudes and time constants and may even improve their accuracy. In addition, an appendix by John P. Mullooly presents a statistical analysis of the effect of counting error on the method of moments estimates of fluorescence decay parameters, applicable when data are obtained by the monophoton technique. Formulas are derived that give the approximate precision of the decay parameters for the general case of N exponential components, with calculational details for one and two component systems.     

Intensity of photosynthesis and chlorophyll content as related to leaf age inNicotiana Sanderae hort

U r?zně starých list? v listové r??ici 90 a? 110 denních rostlin Nicotiana sanderae hort. byly sledovány rozdály v intensitě ?isté fotosynthesy a v obsahu chlorofylu (a + b). Ke stanovení intensity fotosynthesy bylo pou?ito dvou odli?ných metod, a to váhového stanovení p?ír?stku su?iny podle Barto?e, KubÍna a ?et-lÍka (1960) a gazometrického stanovení infra?erveným analyzátorem CO₂. Nejvy??í intensitu fotosynthesy i nejvy??í obsah chlorofylu (vzhledem k plo?e listové) mají mladé, ale ji? dob?e rozvinuté listy, tj. t?etí a? ?tvrté od vrcholu (prvním listem se rozumí list o plo?e asi 20 cm²). Tyto listy nazýváme ?fotosyntheticky dospělými“. Listy nejmlad?í a zejména pak listy star?í mají intensitu fotosynthesy i obsah chlorofylu ni??í; u nejstar?ích list? je intensita fotosynthesy prakticky nulová. Intensita fotosynthesy i obsah chlorofylu se během vývoje mění: jejich momentální rozdíly u list? v genetické spirále jsou z?ejmě shodné s jejich změnami v ontogenesi listu. Pokles intensity fotosynthesy p?i stárnutí list? je rychlej?í ne? pokles obsahu chlorofylu. P?i ur?itém obsahu chlorofylu (tj. asi 2,25 a? 2,45 mg/dm²) klesá intensita ?isté fotosynthesy k nule. Intensita fotosynthesy je v lineárním vztahu k mno?ství chlorofylu (p?i p?epo?tu na plo?nou jednotku), a to nezávisle na poloze listu v genetické spirále. Obě pou?ité metody ke stanovení intensity fotosynthesy poskytly obdobné výsledky.     

Structure of Major Surface Determinants and DNA Diagnosis of Pneumocystis carinii

Pneumocystis carinii is a pathogen which, causes fatal pneumonia in patients with the acquired immune deficiency syndrome (AIDS). To facilitate the basic study of P. carinii , we have analyzed its major surface proteins by both immunochemical and biochemical methods. The major protein components of both cysts and trophozoites are a group of proteins called "P115" with apparent masses of 105–120 kd. It includes 6 isoelcclric variants. A monoclonal antibody raised against cysts recognizes all 6 variants and reacts with epitopes located in the cell wall indicating that P115 is an immunorcactive surface component. The isoelectric variants contain identical or closely related protein components and they are mannose-rich glycoproteins. The isoelectric variation may be due primarily to differences in glycosylation. The majority of sera from humans with diagnosed pneumocystosis that were tested reacted strongly with the P115 proteins. To develop probes for DNA diagnosis and to facilitate molecular studies, a genomic DNA library of P. carinii has been constructed. Some of these clones were used for DNA hybridization analysis of rat and human lungs.     

镉引起小麦苗逆境乙烯的产生及其和镉的吸收分布的关系

溶液培养小麦幼苗转移至含Cd~(2 )的营养液中,根系乙烯产生较快地增加,约在12h达高峰,然后下降;ACC含量亦呈先升后降的趋势。未和Cd~(2 )溶液直接接触的地上部乙烯亦增加,至36h达高峰,此后急剧下降,而ACG和 MAGC含量持续上升。地上部乙烯的增加,主要是由通过根系运往地上部的镉直接作用的结果,不是根部合成ACG运往地上部后再产生的。电镜观察表明,地上部乙烯产生和ACC含量变化的时间进程,可以与镉进入叶细胞内的部位及其对细胞膜和细胞器的影响相联系。     

Immunogold-surface replica study of ADP-induced ligand binding and fibrinogen receptor clustering in human platelets

Platelet cohesion requires the binding of fibrinogen to its receptor, a heterodimer consisting of the plasma-membrane glycoproteins GPIIb and GPIIIa. Although the GPIIb-IIIa complex is present on the surface of unstimulated platelets, it binds fibrinogen only after platelet activation. We have used an immunogold-surface replica technique to study the distribution of GPIIb-IIIa and bound fibrinogen over broad expanses of surface membranes in unstimulated and ADP-activated human platelets. We found that the gold prove was monodispersed over the surface of unstimulated platelets, although the cell surface lacked immunoreactive fibrinogen. To ascertain whether the receptors clustered prior to ligand binding or as a consequence thereof, we studied the surface distribution of GPIIb-IIIa after stimulation with ADP, which causes activation of the fibrinogen receptor function of GPIIb-IIIa without inducing the secretion of fibrinogen. In the absence of added fibrinogen, the unoccupied, yet binding-competent receptors on ADP-stimulated platelets were monodispersed. The addition of fibrinogen caused the GPIIb-IIIa molecules to cluster on the cell surface. Clustering was also induced by the addition of the GPIIb-IIIa binding domains of fibrinogen--namely, the tetrapeptide Arg-Gly-Asp-Ser on the alpha-chain or the gamma-chain decapeptide gamma 402-411. These results show that receptor occupancy causes clustering of GPIIb-IIIa in activated platelets.     

Probing actin and liposome interaction of talin and talin-vinculin complexes: a kinetic, thermodynamic and lipid labeling study.

Talin purified from human platelets and chicken gizzard smooth muscle is an actin and lipid binding protein. Here, we have investigated the effect of vinculin on (a) talin-nucleated actin polymerization and (b) insertion of talin into lipid bilayers. Calorimetric data show ternary complex formation between talin, vinculin, and actin. Actin-talin, actin-vinculin and actin-(talin-vinculin) binding and rate constants as well as actin polymerization rates for all three protein species have been determined by steady state titration, stopped-flow, and fluorescence assay. In contrast to an increase of the polymerization rate by a factor of less than 2 for actin-talin and actin-(talin-vinculin) when lowering the temperature, we measured a decrease in rates for actin alone and actin-vinculin. The overall equilibrium constants (Keq) in the van't Hoff plot proved linear and were of one-step reactions. Thermodynamic data exhibited signs of van der Waal's binding forces. Using the photoactivatable lipid analogue [3H]PTPC/11, which selectively labels membrane-embedded hydrophobic domains of proteins, we also show that talin partially inserts into the hydrophobic bilayer of liposomes. This insertion occurs in a similar manner irrespective of preincubation with vinculin.     

固定化虫荧光素酶光纤传感器

固定化虫荧光素酶光纤传感器蔡谨,王顺光,杨歧生,吉鑫松（浙江大学化工系生化教研室,杭州３１００２７;中国科学院上海生物化学研究所,２０００３１）关键词虫荧光素酶,ＡＴＰ,固定化酶,光纤生物传感器ＡＴＰ是生物体内极为重要的能量物质。如何准确快速地定量Ａ...