Purification from human plasma of endogenous sodium transport inhibitor(s)

Na+,K+-ATPase inhibitors extracted from plasma of healthy human subjects displaced 3H-ouabain binding to human erythrocytes and inhibited the Na+ efflux catalyzed by the Na+,K+-pump and unexpectedly the Na+,K+-cotransport system without alteration of the Na+,Na+-exchange or the Na+ passive permeability. This suggests the presence in healthy human plasma of endogenous factors with ouabain-like and furosemide-like activities.     

Sr2+ can become incorporated into an agonist-sensitive,cytoplasmic Ca2+ store in a cell line derived from the equine sweat gland epithelium

We have explored the properties of a Ca²⁺-dependent cell-signalling pathway that becomes active when cultured equine sweat gland cells are stimulated with ATP. The ATP-regulated, Ca²⁺-influx pathway allowed Sr²⁺ to enter the cytoplasm but permitted only a minimal influx of Ba²⁺. Experiments in which cells were repeatedly stimulated with ATP suggested that Sr²⁺, but not Ba²⁺, could become incorporated into the agonist-sensitive, cytoplasmic Ca²⁺ store. Further evidence for this was provided by experiments using ionomycin, a Ca²⁺ ionophore which has no affinity for Sr²⁺.     

Studies on the metal-complex of acetyl salicylic acid (aspirin).

The present communication deals with the isolation of acetyl salicylic acid (aspirin complexes with Bi+3 Zn+2 and UO2+2. The characterization of 1:2 complexes have been carried out with the help of conductometric, pH met;ric, elemental analysis and IR spectral studies. Spectrophotometric studies in case of UO2+2 (the only colored complex) in range of 4.2 to 5.5 pH show absorption at 490 nm and complex obey Beers Law at the concentration range of 0.01 M to 0.1 M.     

Blockade by zinc ions of K+-induced contraction and calcium in guinea pigTaenia coli

Preincubation with 0.3 mM Zn²⁺ markedly inhibited both the tonic response and Ca²⁺ binding at low affinity sites induced by K⁺ (60 mM), with smaller effects on the phasic response and the high affinity Ca²⁺ sites, inTaenia coli. However, when the muscle was kept in Zn²⁺-containing medium following the first stimulation with the K⁺, the phasic response and the high affinity Ca²⁺ sites were more severely inhibited during the second stimulation with K⁺. This probably indicates that Zn²⁺ reduced the tonic tension response to K⁺ mainly by inhibiting Ca²⁺ influx at the cell membranes ofTaenia coli. However, when Zn²⁺ is continuously present, Ca²⁺ is not supplied at the storage sites and is not available for the phasic response to a second stimulation with K⁺.     

Calcium signalling: a historical account, recent developments and future perspectives

Ca2+ is a uniquely important messenger that penetrates into cells through gated channels to transmit signals to a large number of enzymes. The evolutionary choice of Ca2+ was dictated by its unusual chemical properties, which permit its reversible complexation by specific proteins in the presence of much larger amounts of other potentially competing cations. The decoding of the Ca2+ signal consists in two conformational changes of the complexing proteins, of which calmodulin is the most important. The first occurs when Ca2+ is bound, the second (a collapse of the elongated protein) when interaction with the targeted enzymes occurs. Soluble proteins such as calmodulin contribute to the buffering of cell Ca2+, but membrane intrinsic transporting proteins are more important. Ca2+ is transported across the plasma membrane (channel, a pump, a Na+/Ca2+ exchanger) and across the membrane of the organelles. The endoplasmic reticulum is the most dynamic store: it accumulates Ca2+ by a pump, and releases it via channels gated by either inositol 1,4,5-trisphosphate (IP3) and cyclic adenosine diphosphate ribose (cADPr). The mitochondrion is more sluggish, but it is closed-connected with the reticulum, and senses microdomains of high Ca2+ close to IP3 or cADPr release channels. The regulation of Ca2+ in the nucleus, where important Ca(2+)-sensitive processes reside, is a debated issue. Finally, if the control of cellular Ca2+ homeostasis somehow fails (excess penetration), mitochondria 'buy time' by precipitating inside Ca2+ and phosphate. If injury persists, Ca2(+)-death eventually ensues.     

Ca2+ release-activated Ca2+ (CRAC) current, structure, and function

Store-operated Ca²⁺ entry describes the phenomenon that connects a depletion of internal Ca²⁺ stores to an activation of plasma membrane-located Ca²⁺ selective ion channels. Tremendous progress towards the underlying molecular mechanism came with the discovery of the two respective limiting components, STIM and Orai. STIM1 represents the ER-located Ca²⁺ sensor and transmits the signal of store depletion to the plasma membrane. Here it couples to and activates Orai, the highly Ca²⁺-selective pore-forming subunit of Ca²⁺ release-activated Ca²⁺ channels. In this review, we focus on the molecular steps that these two proteins undergo from store-depletion to their coupling, the activation, and regulation of Ca²⁺ currents.     

Sulfhydryl oxidation induces calcium release from fragmented sarcoplasmic reticulum even in the presence of glutathione

Alcian blue and plumbagin induced transient Ca²⁺ release from fragmented sarcoplasmic reticulum. Dithiothreitol (DTT) and glutathione (GSH) partially blocked Ca²⁺ release induced by these oxidizing compounds. Pretreatment of alcian blue and plumbagin with DTT or GSH for more than 1 min was required to abolish the ability of the oxidizing compounds to release Ca²⁺. Mg²⁺ and ruthenium red completely blocked alcian blue-and plumbagin-induced Ca²⁺ release. These results suggest that oxidation of sulfhydryls on Ca²⁺ release channels induces Ca²⁺ release even in the presence of GSH in situ.     

Net ATP synthesis by running the red cell calcium pump backwards.

Ca2+ loaded inside-out vesicles from human red blood cells, yielding C2+ into a Ca2+ free medium with 4 mM EGTA, 2 mM ADP and 10 mM phosphate, produced an excess of 14.9 pmoles . min-1 . (mg protein)-1 of ATP compared to controls in which the transmembrane Ca2+ gradient was abolished by the ionophore A 23 187.     

Tetrasomy and quadruple trisomy in pea

Summary Two off-type plants, morphologically distinguishable from each other and from their respective sister euploid, were isolated in the M₃ generation of pea interchange heterozygotes. Pollen sterility was very high, ranging from 63.0 to 90.0%. Cytologically one of them was tetrasomic (2n+2=16) and the other one was quadruple trisomic (2n+1+1+1+1=18). In the tetrasomic plant 1IV+6II was the most frequent (46.7%) chromosome configuration, while cells with 4III+3II were predominant (40.0% cells) in the quadruple trisomic plant.Financial assistance of C.S.I.R., New Delhi, India is acknowledged.     

Influence of medium amino acids on the ouabain sensitive and insensitive 86Rb+-fluxes in HeLa cells

Components of the 86Rb+-influx in HeLa cells were investigated in Joklik minimal essential medium, or in Earle's balanced salt solution with and without medium amino acids. The presence of amino acids led to the stimulation of the ouabain sensitive 86Rb+-uptake and inhibition of the diuretic-sensitive and residual 86Rb+-fluxes. These results show that the presence of amino acids is an important regulator of the K+/Rb+-fluxes under normal conditions in growth medium.     

Complex modulation of Cav3.1 T-type calcium channel by nickel

Nickel is considered to be a selective blocker of low-voltage-activated T-type calcium channel. Recently, the Ni²⁺-binding site with critical histidine-191 (H191) within the extracellular IS3–IS4 domain of the most Ni²⁺-sensitive Ca_v3.2 T-channel isoform has been identified. All calcium channels are postulated to also have intrapore-binding site limiting maximal current carried by permeating divalent cations (PDC) and determining the blockade by non-permeating ones. However, the contribution of the two sites to the overall Ni²⁺ effect and its dependence on PDC remain uncertain. Here we compared Ni²⁺ action on the wild-type “Ni²⁺-insensitive” Ca_v3.1_w/t channel and Ca_v3.1_Q172H mutant having glutamine (Q) equivalent to H191 of Ca_v3.2 replaced by histidine. Each channel was expressed in Xenopus oocytes, and Ni²⁺ blockade of Ca²⁺, Sr²⁺, or Ba²⁺ currents was assessed by electrophysiology. Inhibition of Ca_v3.1_w/t by Ni²⁺ conformed to two sites binding. Ni²⁺ binding with high-affinity site (IC₅₀ = 0.03–3 μM depending on PDC) produced maximal inhibition of 20–30 % and was voltage-dependent, consistent with its location within the channel’s pore. Most of the inhibition (70–80 %) was produced by Ni²⁺ binding with low-affinity site (IC₅₀ = 240–700 μM). Q172H-mutation mainly affected low-affinity binding (IC₅₀ = 120–160 μM). The IC₅₀ of Ni²⁺ binding with both sites in the Ca_v3.1_w/t and Ca_v3.1_Q172H was differentially modulated by PDC, suggesting a varying degree of competition of Ca²⁺, Sr²⁺, or Ba²⁺ with Ni²⁺. We conclude that differential Ni²⁺-sensitivity of T-channel subtypes is determined only by H-containing external binding sites, which, in the absence of Ni²⁺, may be occupied by PDC, influencing in turn the channel’s permeation.     

Effects of barium on separately recorded fast and slow PIII responses in bullfrog retina

Investigation of Ba2+ effects on fast and slow PIII responses in isolated bullfrog retina revealed that Ba2+ suppressed slow PIII completely with little effect on fast PIII. A light-induced [K+]0 decrease in the photoreceptor layer was observed in spite of Ba2+ perfusion, indicating the suppressive action of Ba2+ on the K+ conductance of the Müller cell membrane.     

Insulin-like effect of dichloroacetic acid on hexose transport in Swiss 3T3 cells

Hexose transport in Swiss 3T3 cells was increased by treatment with dichloroacetic acid as well as by treatment with insulin. Neither extra- nor intracellular Ca2+ was found to be involved in their stimulatory action. On the other hand, the removal of intracellular Mg2+ resulted in a loss of the stimulation. These results suggest that dichloroacetic acid stimulates the hexose transport in Mg2+-dependent manner, similar to that of insulin.     

Hyper-tetraploids in pea

Summary Cytomorphologically, five hypertetraploids; three hexasomic (4n+2=30), one double pentasomic (4n+1+1=30) and one multiple aneuploid tetraploid (4n+2+1+1+1=33), where isolated in the C₃ generation of pea autotetraploids. Plants with 30 chromosomes were morphologically very distinct from their euploids and characterized by variation in plant height and morphology of leaves. Hexasomic and double pentasomic tetraploids were characterized by the presence of a hexavalent and two pentavalents, respectively. The multiple aneuploid tetraploid showed very vigorous growth and varying frequencies of hexavalents and pentavalents. All the five aneuploids showed high Anaphase-I anomalies; pollen sterility ranged from 50% to 81%.Financial assistance of C.S.I.R., New Delhi, India is acknowledged.     

Non-enzymatic active transport in vitro. Conversion of chemical to osmotic energy]

The active transport of ammonia gas was obtained with an asymmetric composite membrane-model using H+ as carrier and the formation of water as chemical energy source. The experimental molecular pump externally limited by two silicone films permeable to gasses but not to liquids or ions is separated into two liquid layers by a central cation-exchange membrane. The first acid layer produces the aspiration of NH3, due to NH4+ formation, through the inlet silicone film; NH4+ is transferred to the second alkaline layer through the ion exchange membrane, by interdiffusion with K+; there NH4+ reacts with OH-, water is formed and NH3 is expelled throught the oulet silicone film. This new pump enlarges the applicability of in vitro active transport to non-enzymatic systems and energy sources different from ATP.     

Extracellular Mg2+ regulates intracellular Mg2+ and its subcellular compartmentation in fission yeast, Schizosaccharomyces pombe

Effects of extracellular magnesium ions ([Mg²⁺]_o ) on intracellular free Mg²⁺ ([Mg²⁺]_i ) and its subcellular distribution in single fission yeast cells, Schizosaccharomyces pombe, were studied with digital-imaging microscopy and an Mg²⁺ fluorescent probe (mag-fura-2). Using 0.44 mM [Mg²⁺]_o , [Mg²⁺]_i in yeast cells was 0.91±0.08 mM. Elevation of [Mg²⁺]_o to 1.97 mM induced rapid (within 5 min) increments in [Mg²⁺]_i (2.18±0.11 mM). Lowering [Mg²⁺]_o to 0.06 mM, however, exerted no significant effects on [Mg²⁺]_i (0.93±0.14 mM), at least for periods of up to 30 min. Irrespective of the [Mg²⁺]_o used, the subcellular distribution of [Mg²⁺]_i remained hetero geneous, i.e. where the sub-plasma membrane region >cytoplasm >nucleus. [Mg²⁺] in all three subcellular compartments increased significantly, two- to threefold, concomitant with [Mg²⁺]_i when placed in 1.97 mM [Mg²⁺]_o . We conclude that [Mg²⁺]_i in fission yeast is maintained at a physiologic level when [Mg²⁺]_o is low, but intracellular free Mg²⁺ rapidly rises when [Mg²⁺]_o is elevated. Like most eukaryotic cells, yeast may have a Mg²⁺ transport system(s) which functions to maintain gradients of Mg²⁺ from the outside to inside the cell and among its subcellular compartments. Received 18 April 1996; received after revision 4 July 1996; accepted 26 July 1996     

红叶石楠(Photinia X Fraseri)愈伤组织诱导研究

利用红叶石楠叶片和茎段作为外植体进行愈伤组织试验,研究了TDZ和AgNO_3及其他植物生长调节刑对红叶石楠愈伤组织诱导的影响效果.结果表明:TDZ和AgNO_3对红叶石楠愈伤组织诱导具有一定的促进作用,但需NAA配合.方能形成愈伤组织.本试验中,培养基1/2MS+AgNO_32.5 m/l+TDZ0.5 mg/l+NAA2.Omg/l对叶片有较好诱导效果,1/2MS+AgNO_32.5 mg/l+TDZ0.5 mg/l+NAA1.0 mg/l对茎段有较好诱导效果.此外,1.5 mg/l的TDZ与0.5 mg/l的2,4-D配比有较好诱导效果.光培养条件下,叶片的诱导率高于茎段;在暗培养下,茎段的诱导率高于叶片.     

Action and interactions at microtubule ends

Microtubule dynamic instability is fundamentally important to the way cells respond to their environment and segregate their genetic material. A disparate class of proteins defined by their localisation to growing microtubule plus ends ('+TIPS') play a key role in controlling microtubule dynamics and organisation. They directly impact upon the behaviour of the microtubule tip and link this structure to interfaces that include kinetochores and the cortex of the cell. Surprisingly, some +TIPs also have important functions at the microtubule minus end. These properties contribute to the important roles played by +TIPs in processes such as mitosis and cell migration. This review examines how recent advances have impacted our understanding of +TIP function in mammalian cells, with emphasis on the emergence of the EB1 family as a core component of +TIP activities. An overview of the use of +TIP imaging as a tool for the cell biologist is also presented.     

Evidence of abnormalities in net sodium and potassium fluxes in erythrocytes of patients with essential hypertension]

A new and simple laboratory test for measuring net Na+ and K+ fluxes in Na+-loaded/K+-depleted human erythrocytes was developed and applied to hypertension. Moderate essential hypertension was characterized by a constant increase in net K+ influx; more severe cases showed a drop in net Na+ efflux. Na+ and K+ erythrocyte fluxes were found to be normal in hypertension of renal origin.