渗透压对硝化作用的影响

在气升式内循环硝化反应器中研究了渗透压对硝化作用的影响。保持进水氨氮浓度420mg·L-1,将进水渗透压逐渐从4.3×105Pa提高到18.8×105Pa,硝化反应器的氨氮转化率稳定在93%～100%。将进水渗透压进一步提高到19.2×105Pa,氨氮转化率降至69.2%。渗透压对硝化作用的影响具有突发性,临界值在18.8×105～19.2×105Pa之间。受高渗透压胁迫时,活性污泥中硝化细菌的形态趋向单一,个体变小,内膜数量减少,并产生许多不明成分的颗粒状内含物。解除渗透压胁迫后,细胞结构恢复。添加钾离子能够缓解高渗压对硝化作用的影响。高渗透压胁迫以及解除渗透压胁迫可增强污泥硝化活性,比污泥氨氮转化率(污泥以SS计)分别从0.083kg·kg-1·d-1升至0.509kg·kg-1·d-1和2.569kg·kg-1·d-1,同比提高5.1倍和30.0倍。     

Physicochemical properties and antitumor activities of water-soluble native and sulfated hyperbranched mushroom polysaccharides

A water-soluble hyperbranched beta-glucan, coded as TM3b, extracted from sclerotia of an edible fungus (Pleurotus tuber-regium) was fractioned into eight fractions coded as F1-F8 by a nonsolvent addition method. Five fractions were treated with chlorosulfonic acid at 35 degrees C to synthesize successfully sulfated derivatives coded as S-F2, S-F3, S-F4, S-F5, and S-F8 with degree of substitution of 0.28-0.54. The 13C NMR results of these sulfated beta-glucans indicated that while the C-6 position was fully substituted, C-2, C-3, and C-4 were only partially substituted by the sulfate groups. The weight-average molecular weights (Mw) and intrinsic viscosities ([eta]) of the native and sulfated TM3b fractions were determined using multi-angle laser light scattering and viscometry in 0.15M aq NaCl at 25 degrees C, respectively. The dependences of [eta] on Mw for TM3b and sulfated TM3b were found to be [eta]=0.18Mw(0.28+/-0.03) (Mw range from 3.30 x 10(4) to 3.90 x 10(7)) and [eta]=2.24 x 10(-2)Mw(0.52+/-0.06) (Mw range from 3.24 x 10(4) to 3.15 x 10(5)) in 0.15M aq NaCl at 25 degrees C, respectively. It revealed that both the native TM3b and its sulfated derivatives exist in a spherical chain conformation in 0.15M aq NaCl. Furthermore, the native and sulfated TM3b fractions showed potent antitumor activities in vivo and in vitro. The sulfated derivatives exhibited relatively higher in vitro antitumor activity against human hepatic cancer cell line HepG2 than the native TM3b. Water solubility and introduction of sulfate groups were the main factors in enhancing the antitumor activities.     

An enzymically catalysed reaction between d-glucose and the protein cyst coat of the ciliate Colpoda steinii

1. The isolated protein cyst coat of Colpoda steinii reacted with [(14)C]glucose to bind (14)C label in a reaction that was not an artifact of bacterial contamination or of adsorption. 2. The reaction was enzymically catalysed, had optimum pH7.0-7.4 and a temperature optimum of 36-38 degrees C, was non-competitively and reversibly inhibited by 10mum-p-hydroxymercuribenzoate and was specific for d-glucose. It had K(m)8.2x10(-6)m. 3. With the exception of C-1, all the carbon atoms of glucose were bound to the same extent and, measured relative to C-2 or C-6, the binding of C-1 varied between 0.45 and 0.82. The lost C-1 was not accounted for as carbon dioxide. 4. On prolonged incubation a coat preparation reacted with 3.6% of its own weight of glucose. 5. The label was tightly bound, but after acid treatment a variable proportion was recovered as glucose and there was no evidence for the release of any other (14)C-containing compound. 6. Even after dissolution of the coat protein, bound label was not removed by treatment with periodate or lead tetra-acetate.     

Volume regulation by flounder red blood cells in anisotonic media

The nucleated high K, low Na red blood cells of the winter flounder demonstrated a volume regulatory response subsequent to osmotic swelling or shrinkage. During volume regulation the net water flow was secondary to net inorganic cation flux. Volume regulation the net water flow was secondary to net inorganic cation flux. Volume regulation after osmotic swelling is referred to as regulatory volume decrease (RVD) and was characterized by net K and water loss. Since the electrochemical gradient for K is directed out of the cell there is no need to invoke active processes to explain RVD. When osmotically shrunken, the flounder erythrocyte demonstrated a regulatory volume increase (RVI) back toward control cell volume. The water movements characteristic of RVI were a consequence of net cellular NaCl and KCl uptake with Na accounting for 75 percent of the increase in intracellular cation content. Since the Na electrochemical gradient is directed into the cell, net Na uptake was the result of Na flux via dissipative pathways. The addition of 10(-4)M ouabain to suspensions of flounder erythrocytes was without effect upon net water movements during volume regulation. The presence of ouabain did however lead to a decreased ration of intracellular K:Na. Analysis of net Na and K fluxes in the presence and absence of ouabain led to the conclusion that Na and K fluxes via both conservative and dissipative pathways are increased in response to osmotic swelling or shrinkage. In addition, the Na and K flux rate through both pump and leak pathways decreased in a parallel fashion as cell volume was regulated. Taken as a whole, the Na and K movements through the flounder erythrocyte membrane demonstrated a functional dependence during volume regulation.     

Pressure regulation of the electrical properties of growing Arabidopsis thaliana L. root hairs.

Actively growing Arabidopsis thaliana L. (Columbia wild type) root hairs were used to examine the interplay between cell turgor pressure and electrical properties of the cell: membrane potential, conductance, cell-to-cell coupling, and input resistance. Pressure was directly modulated using a pressure probe or indirectly by changing the extracellular osmolarity. Direct modulation of pressure in the range of 0 to about 15 x 10(5) Pa (normal turgor pressure was 6.8 +/- 2.0 x 10(5) Pa, n = 29) did not affect the membrane potential, conductance, coupling, or input resistance. Indirect modulation of turgor pressure by adding (hyperosmotic) or removing (hypo-osmotic) 200 mM mannitol/sorbitol affected the potential and conductance but not cell-to-cell coupling. Hypo-osmotic treatment depolarized the potential about 40 mV from an initial potential of about -190 mV and increased membrane conductance, consistent with an increase in anion efflux from the cell. Hyperosmotic treatment hyperpolarized the cell about 25 mV from the same initial potential and decreased conductance, consistent with a decline in cation influx. The results are likely due to the presence of an "osmo-sensor," rather than a "turgor-sensor," regulating the cell's response to osmotic stress.     

Thermal denaturation of the erythrocyte cytoskeleton alters the morphological changes associated with osmotic swelling

1. 1. The shape changes during osmotic swelling of human erythrocytes in a hypotonic medium at room temperature, at 45°C and at the denaturation temperature (49.5°C) of the cytoskeletal protein, spectrin, have been monitored by video microscopy.

2. 2. At room temperature the great majority of cells (which were discoid prior to injection of hypotonic medium) swelled to a spherical shape through an intermediate ellipsoidal form.

3. 3.At 49.5°C (where cells had cupped shapes prior to injection) the transition to the spherical form often involved a stomatocytic rather than ellipsoidal intermediate shape.

4. 4. The cupped form of the cells prior to injection did not account for the high incidence of cells swelling through a stomatocytic intermediate shape at 49.5°C.

5. 5. A theoretical treatment by Svetina and Zeks (1983) attributes the nature of the osmotic swelling transition shape to the difference in area between the outer and inner faces of the membrane. Our results would be consistent with the theoretical predictions if it is assumed that an increase in the area of the inner face of the membrane follows thermal denaturation of the cytoskeleton of cells in hypotonic medium.

Differential effect of NaCl and polyethylene glycol on the ultrastructure of chloroplasts in rice seedlings

Ionic and osmotic effects of salinity on the ultrastructure of chloroplasts in salt-treated rice seedlings were investigated. After rice seedlings were grown in hydroponic culture for three weeks, they were treated with NaCl and polyethylene glycol (PEG) 4000 both at a water potential of -1.0 MPa for 3 days. The most notable difference in ultrastructural change between NaCl and PEG treatment was observed in the damage in chloroplast membranes. NaCl induced swelling of thylakoids and caused only a slight destruction of the chloroplast envelope. PEG caused severe destruction of the chloroplast envelope compared with NaCl, however thylakoids did not swell. Our observations suggested that in salt-treated rice plants, the ionic effects induced swelling of thylakoids and the osmotic effects caused the destruction of chloroplast envelope.     

Renal blood flow, early distal sodium, and plasma renin concentrations during osmotic diuresis

Inconsistencies in previous reports regarding changes in early distal NaCl concentration (ED(NaCl)) and renin secretion during osmotic diuresis motivated our reinvestigation. After intravenous infusion of 10% mannitol, ED(NaCl) fell from 42.6 to 34.2 mM. Proximal tubular pressure increased by 12.6 mmHg. Urine flow increased 10-fold, and sodium excretion increased by 177%. Plasma renin concentration (PRC) increased by 58%. Renal blood flow and glomerular filtration rate decreased, however end-proximal flow remained unchanged. After a similar volume of hypotonic glucose (152 mM), ED(NaCl) increased by 3.6 mM, (P < 0.01) without changes in renal hemodynamics, urine flow, sodium excretion rate, or PRC. Infusion of 300 micromol NaCl in a smaller volume caused ED(NaCl) to increase by 6.4 mM without significant changes in PRC. Urine flow and sodium excretion increased significantly. There was a significant inverse relationship between superficial nephron ED(NaCl) and PRC. We conclude that ED(Na) decreases during osmotic diuresis, suggesting that the increase in PRC was mediated by the macula densa. The results suggest that the natriuresis during osmotic diuresis is a result of impaired sodium reabsorption in distal tubules and collecting ducts.     

Effects of Salinity and Relative Humidity on Two Melon Cultivars Differing in Salt Tolerance

The effects of increasing relative humidity on the growth and salt tolerance of two melon (Cucumis melo L.) cultivars, Revigal C-8 (salt sensitive) and Galia (salt tolerant) was investigated. One month after germination, the plants were exposed for 15 d to 0 (control) and 80 mM NaCl, under relative humidity (RH), 30 and 70 %. The growth of the whole plant, leaf, stem and root of cv. Revigal C-8 was increased with increasing RH. On the other hand, cv. Galia showed an increase in root growth with increasing RH only under the NaCl treatment. Under salinity, most of the Na⁺ was withheld in the stems. An increase in RH in the NaCl treatment significantly decreased Na⁺ and Cl^– concentrations in leaves of cv. Revigal C-8, while it had no effect on their concentrations in cv. Galia. In both cultivars, increasing RH under NaCl condition significantly decreased water contents in leaves and stems, and increased osmotic potential in roots. The amount of the root exudate of cv. Galia was significantly decreased with increasing RH, while it was not affected in cv. Revigal C-8. Under the NaCl treatment, cv. Galia had significantly higher leaf osmotic potential than cv. Revigal C-8 at both relative humidities and higher amount of root exudate at 30 % RH.     

Matric Water Potential of Leaf Tissue--Measurement and Significance

A thermocouple psychrometer method, previously described foruse in determining tissue and sap water potentials, has beenadapted for determining matric potentials. Matric values ranging from approximately zero to –10 x10⁵ Pa were observed with wilting wheat leaves. Mean valuesof total tissue water potential and of its solute (osmotic),matric, and cell wall pressure components were –16.6,–18.3, –30 and +4.7 Pa x 10⁵, respectively. Matric potentials are often ignored in investigations of plantwater relations. This practice is shown to result in underestimationof wall pressures (in this case by a mean of 64 per cent) andsometimes in spuriously negative values.     

Glibenclamide interferes with mitochondrial bioenergetics by inducing changes on membrane ion permeability

The interference of glibenclamide, an antidiabetic sulfonylurea, with mitochondrial bioenergetics was assessed on mitochondrial ion fluxes (H+, K+, and Cl-) by passive osmotic swelling of rat liver mitochondria in K-acetate, KNO3, and KCl media, by O2 consumption, and by mitochondrial transmembrane potential (Deltapsi). Glibenclamide did not permeabilize the inner mitochondrial membrane to H+, but induced permeabilization to Cl- by opening the inner mitochondrial anion channel (IMAC). Cl- influx induced by glibenclamide facilitates K+ entry into mitochondria, thus promoting a net Cl-/K+ cotransport, Deltapsi dissipation, and stimulation of state 4 respiration rate. It was concluded that glibenclamide interferes with mitochondrial bioenergetics of rat liver by permeabilizing the inner mitochondrial membrane to Cl- and promoting a net Cl-/K+ cotransport inside mitochondria, without significant changes on membrane permeabilization to H+.     

盐胁迫下小麦苗叶片吡咯-5-羧酸还原酶活性和游离脯氨酸积累

受NaCl、KCl或MsCl_2胁迫的小麦幼苗,当外部溶液的渗透势由—160 kPa下降到—900 kPa时,叶片吡咯—5—羧酸还原酶(PSC)活性增高;渗透势由—900 kPa降低到—1500kPa,酶活性下降;胁迫 1d和 2d的幼苗,还原酶活性显著增加;3～6d酶活性无大变化。游离脯氨酸含量随溶液渗透势下降和培养时间的延长而提高。胁迫解除后酶活性和脯氨酸含量均降低。受NcCl胁迫义在ABA影响下的幼苗,P5C还原酶活性和脯氨酸含量高于仅受NaCl胁迫的幼苗。     

Biochemical characterisation of mitochondrial and lysosomal particles in foetal and neonatal rats

1. The sedimentation coefficients of the mitochondria are larger in the foetal as compared to the adult rats. For cytochrome oxidase, the values are respectively 18.700 x 10(3) and 13,550 x 10(3) S. 2. Lysosomal sedimentation coefficients are smaller in the foetal as compared to the adults. For beta galactosidase, the values are respectively 5.090 x 10(3) and 10.430 x 10(3) S. 3. It is concluded that the mitochondria are larger and the lysosomes smaller in size in the foetus as compared to the adults. 4. The external membrane of neonatal mitochondria and the membrane of foetal and neonatal lysosomes are more resistant to osmotic swelling.     

CO2-stimulated NaCl absorption in the mouse renal cortical thick ascending limb of Henle. Evidence for synchronous Na +/H+ and Cl-/HCO3- exchange in apical plasma membranes

These experiments evaluated salt transport processes in isolated cortical thick limbs of Henle (cTALH) obtained from mouse kidney. When the external solutions consisted of Krebs-Ringer bicarbonate (KRB), pH 7.4, and a 95% O2-5% CO2 gas phase, the spontaneous transepithelial voltage (Ve, mV, lumen-to-bath) was approximately mV; the net rate of Cl- absorption (JnetCl) was approximately 3,600 pmols s-1 cm-2; the net rate of osmotic solute absorption Jnetosm was twice JnetCl; and the net rate of total CO2 transport (JnetCO2) was indistinguishable from zero. Thus, net Cl- absorption was accompanied by the net absorption of a monovalent cation, presumably Na+, and net HCO3- absorption was negligible. This salt transport process was stimulated by (CO2 + HCO3- ): omission of CO2 from the gas phase and HCO3- from external solutions reduced JnetCl, Jnetosm, and Ve by 50%. Furthermore, 10(-4) M luminal furosemide abolished JnetCl and Ve entirely. The lipophilic carbonic anhydrase inhibitor ethoxzolamide (10(-4) M, either luminal or peritubular) inhibited (CO2 + HCO3-)-stimulated JnetCl, Jnetosm, and Ve by approximately 50%; however, when the combination (CO2 + HCO3-) was absent, ethoxzolamide had no detectable effect on salt transport. Ve was reduced or abolished entirely by omission of either Na+ or Cl- from external solutions, by peritubular K+ removal, by 10(-3) M peritubular ouabain, and by 10(-4) M luminal SITS. However, Ve was unaffected by 10(-3) M peritubular SITS, or by the hydrophilic carbonic anhydrase inhibitor acetazolamide (2.2 x 10(-4) M, lumen plus bath). We interpret these data to indicate that (CO2 + HCO3-)-stimulated NaCl absorption in the cTALH involved two synchronous apical membrane antiport processes: one exchanging luminal Na+ for cellular H+; and the other exchanging luminal Cl- for cellular HCO3- or OH-, operating in parallel with a (CO2+ HCO3-)-independent apical membrane NaCl cotransport mechanism.     

Effect of water stress and gibberellic acid on germination of flax,sesame and onion seeds

The effects of different osmotic stresses (from 0 to –8× 10⁵ Pa) obtained with NaC or polyethylene glycol 6000 solutions on the germination of flax, sesame and onion seeds were investigated. The effect of presoaking with gibberellic acid (GA³) on the germination of the above mentioned seeds was also studied. It was found that the rate of seed germination and the final germination percentages as well as the amount of water absorbed by the seeds were considerably lowered with the rise of osmotic stress levels whatever the stress agent used, more considerable reduction was obtained under polyethylene glycol 6000 than under NaCl. Presoaking with gibberellic acid increased the rate and the final germination percentage of osmotically stressed flax and sesame seeds, while those of stressed onion seeds were slightly retarded.     

Mechanics of microtubule bundles in pillar cells from the inner ear.

The mechanical properties of cross-linked microtubule bundles were measured from outer pillar cells isolated from the mammalian inner ear. Measurements were made using a three-point bending test and were incorporated into a mathematical model designed to distinguish between the stiffness contributions from microtubules and their cross-linking proteins. Outer pillar cells were composed of 1000-3000 parallel bundled microtubules in a square array that was interdigitated and cross-linked with actin filaments. The average midpoint bending stiffness of intact cells was 7 x 10(-4) N/m. After removal of both the actin filaments and cross-links with detergent in the presence of DNase I, the square array was disrupted and the stiffness decreased by a factor of 4, to 1.7 x 10(-4) N/m. The bending modulus for individual microtubules was calculated to be 7 x 10(-23) Nm2, and the Young's modulus for these 15 protofilament microtubules was 2 x 10(9) Pa. The shear modulus between microtubules in intact cells was calculated to be 10(3) Pa. It was concluded that cross-linking proteins provided shear resistance between microtubules, which resulted in a fourfold increase in stiffness. The model can be used to estimate the mechanical properties of cross-linked microtubule bundles in cells from which direct measurements are not available.     

Stimulation of potassium cycling in mitochondria by long-chain fatty acids

Nonesterified long-chain fatty acids (myristic, palmitic, oleic and arachidonic), added at low amounts (around 20 nmol/mg protein) to rat liver mitochondria, energized by respiratory substrates and suspended in isotonic solutions of KCl, NaCl, RbCl or CsCl, adjusted to pH 8.0, induce a large-scale swelling followed by a spontaneous contraction. Such swelling does not occur in alkaline solutions of choline chloride or potassium gluconate or sucrose. These changes in the matrix volume reflect a net uptake, followed by net extrusion, of KCl (or another alkali metal chloride) and are characterized by the following features: (1) Lowering of medium pH from 8.0 to 7.2 results in a disappearance of the swelling-contraction reaction. (2) The contraction phase disappears when the respiration is blocked by antimycin A. (3) Quinine, an inhibitor of the K(+)/H(+) antiporter, does not affect swelling but suppresses the contraction phase. (4) The swelling phase is accompanied by a decrease of the transmembrane potential and an increase of respiration, whereas the contraction is followed by an increase of the membrane potential and a decrease of oxygen uptake. (5) Nigericin, a catalyst of the K(+)/H(+) exchange, prevents or partly reverses the swelling and partly restores the depressed membrane potential. These results indicate that long-chain fatty acids activate in liver mitochondria suspended in alkaline saline media the uniporter of monovalent alkali metal cations, the K(+)/H(+) antiporter and the inner membrane anion channel. These effects are presumably related to depletion of mitochondrial Mg(2+), as reported previously [Arch. Biochem. Biophys. 403 (2002) 16], and are responsible for the energy-dissipating K(+) cycling. The uniporter and the K(+)/H(+) antiporter are in different ways activated by membrane stretching and/or unfolding, resulting in swelling followed by contraction.     

Ultrastructural transformations in bean inner mitochondrial membranes

The ultrastructure of inner membrane-matrix mitochondria isolated from bean (Phaseolus vulgaris) shoots was examined in different metabolic states. Gross ultrastructural transformations analogous to the condensed-to-orthodox configurational changes reported in mammalian mitochondria are observed on transistion from nonrespiring to respiring metabolism. With the induction of oxidative phosphorylation, the particles remain in the orthodox configurational state. The reverse orthodox-to-condensed configurational changes observed in mammalian preparations does not occur. Optically monitored absorbancy studies with bean particles show a substrate-supported P_i-induced swelling under the same conditions that induce the condensed-to-orthodox ultrastructural transformation. The swelling is associated with the net uptake of K⁺ and P_i as well as a small P_i-induced respiratory stimulation. When phosphorylation is initiated with these swollen particles, the optically monitored volume remains unchanged. Thus a positive correlation exists between the ultrastructural configuration and the osmotic volume changes, which supports the conclusion that configurational changes reflect internal osmotic adjustments.     

Structure and chain conformation of five water-soluble derivatives of a beta-D-glucan isolated from Ganoderma lucidum

A linear water-insoluble (1-->3)-beta-D-glucan, coded as GL-IV-I, was isolated from the fruit body of Ganoderma lucidum by extracting with NaOH solution. Its derivatives were prepared by using sulfation, carboxymethylation, hydroxyethylation, hydroxypropylation, and methylation, respectively, and these were labeled as S-GL, CM-GL, HE-GL, HP-GL and M-GL. Five derivatives exhibited good water solubility. Their structures and chain conformations were investigated with infrared spectroscopy, elemental analysis (EA), one- and two-dimensional NMR spectroscopy, laser light scattering (LLS), and size-exclusion chromatography combined with LLS (SEC-LLS). The reactivity of the hydroxyl group of GL-IV-I was ordered as C-6>C-4>C-2 for the five derivatives. The degree of substitution (DS) of the derivatives was calculated from EA and NMR spectroscopy to be from 0.32 to 1.18. The weight-average molecular mass (M(w)) of GL-IV-I, S-GL, CM-GL, HE-GL, HP-GL, and M-GL was 13.3 x 10(4), 10.1 x 10(4), 6.3 x 10(4), 7.2 x 10(4), 5.1 x 10(4), and 14.1 x 10(4), respectively. The conformation analysis studies revealed that GL-IV-I exists as a compact coil in dimethyl sulfoxide, whereas the five derivatives are slightly expanded flexible chains in 0.9% aqueous NaCl solution.     

Osmotic stress regulates the anticoagulant efficiency of dermatan sulfate.

Glycosaminoglycans (GAGs) in pericellular and interstitial spaces help to maintain local water homeostasis and blood coagulation balance. This study explored whether dehydrating microenvironment conditions influence dermatan sulfate's (DS) anticoagulant activity. Water transfer during antithrombin activation by dermatan sulfate was measured using osmotic stress techniques. Anticoagulant activity was determined from the change in the rate of coagulation factor Xa (fXa) inhibition. Osmotic stress accelerated reaction rates, indicating water transfer from reactants to bulk. The net volume transferred, measured using osmotic probes similar in size to the reacting proteins, was approximately 2500 mol of water per mole of fXa inhibited. The reaction efficiency, V(sat)/K 1/2 (rate at saturation/concentration resulting in half-maximal rates), determined in titrations with monosulfated dermatan sulfate and disulfated dermatan sulfate (DDS), were 4x10(4) and 2x10(5) M-1 s-1 under osmotic stress and in the presence of calcium, corresponding to 34- and 81-fold increases over efficiency measured under standard conditions. These results indicate that dermatan sulfate can contribute significantly to antithrombin activation, and that in dehydrating environments and depending of ionic conditions, its anticoagulant efficiency can exceed that of heparan sulfate (HS).