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1.
Hu  Qiuhui  Pan  Genxing  Zhu  Jianchun 《Plant and Soil》2002,238(1):91-95
The present study examined the effect of fertilization with sodium selenite on the selenium content of tea and the nutritional function of Se-enriched tea. Selenium content of tea leaves was increased up to 0.36 g g–1 by the application of sodium selenite to soil at 0.5 and 1.0 kg Se ha–1. Application by a Se-enriched organic manure at a rate of 0.5 kg Se ha–1 provided a higher biological availability of selenium for plant uptake compared with a similar amount of sodium selenite. Foliar spray of sodium selenite at 50–100 g Se ha–1 increased the selenium content to 0.32–1.45 g g–1 in tea leaves sampled at the 8–26 days after spraying. Selenium content in the blood and liver, glutathione peroxidase activity in blood of rats were significantly enhanced by feeding of an extracted solution of Se-enriched tea leaves and sodium selenite. Glutathione peroxidase activity in liver of rats fed with Se-enriched tea was higher than that fed with sodium selenite, indicating that the selenium in Se-enriched tea leaves is a more effective Se source than sodium selenite. Increasing the Se level in food products through the application of a selenium fertilizer is a safe, effective and feasible means of increasing the selenium intake of human and animals in low selenium areas of China.  相似文献   

2.
Following acute liver injury, hepatocytes divide to facilitate regeneration. However, during chronic injury, hepatocyte proliferation is typically blocked and repair is mediated through liver progenitor (oval) cells. Signalling of the p55 tumour necrosis factor (TNF) receptor is central to these processes. Two ligands for p55 are known: TNF and lymphotoxin-alpha (LT). However, one study suggests that another exists that mediates liver injury following viral challenge. We have therefore investigated whether ligands other than TNF and LT are required for liver regeneration following either acute or chronic injury. Wild-type and double TNF/LT knockout (TNF–/–LT–/–) mice were subjected to either partial hepatectomy (PHx) or a choline-deficient ethionine-supplemented (CDE) diet. Proliferating hepatocytes, oval cells and inflammatory cells were identified and quantified in liver sections by immunohistochemistry. Liver inflammatory cells were characterised by cell surface antigen expression. Liver damage and mortality were monitored. Both hepatocyte and oval cell proliferation was reduced in TNF–/–LT–/– mice. Lymphocyte clusters were evident in all TNF–/–LT–/– livers and were heterogeneous, comprising B and T lymphocytes. PHx evoked liver inflammation in TNF–/–LT–/– but not wild-type mice, whereas no difference was apparent between genotypes in CDE experiments. Thus, TNF/LT signalling mediates liver regeneration involving both hepatocytes and progenitor cells. The hyper-inflammatory response following PHx in TNF–/–LT–/– animals, which is absent following CDE-induced injury, demonstrates that the two forms of liver injury evoke discrete inflammatory responses and provides a model in which such differences can be examined further.  相似文献   

3.
Nitrification and denitrification rates were estimated simultaneously in soil-floodwater columns of a Crowley silt loam (Typic Albaqualfs) rice soil by an15N isotopic dilution technique. Labeled NO 3 was added to the floodwater of soil-water columns, half were treated with urea fertilizer. The (NO 3 +NO 2 )–N and (NO 3 +NO 2 )–N concentrations in the floodwater were measured over time and production and reduction rates for NO 3 calculated. Nitrate reduction in the urea amended columns averaged 515 mol N m–2h–1 and nitrification averaged 395 mol N m–2h–1 over the 35–153 d incubation. The nitrification rate for 4–19 d sampling period (1,560 mol N m–2h–1) in the urea amended columns was almost 9 times greater than the reduction rate (175 mol N m–2h–1) over the same period. Without the addition of urea the NO 3 production rate averaged 32 mol N m–2h–1 and reduction 101 mol N m–2h–1.  相似文献   

4.
Phosphorus and nitrogen excretion rates by zooplankton communities from two eutrophic and shallow Dutch lakes were measured in laboratory. The variations in excretion rates in the lakes (May–October) were caused mainly by fluctuation in zooplankton biomass. Mean summer excretion rates (June–September) were 2.4 and 0.9 µg PO4P·1–1·d–1 in Lake Loosdercht and Lake Breukeleveen, respectively. This difference between the lakes was caused mainly by the lower zooplankton biomass in Lake Breukeleveen. The excretion of 2.4 µg PO4P·1–1·d compared with the calculated P-demand of phytoplankton of 8.0 µg PO4P·1–1·d–1 is substantial in the summer (June–September) and far more important than the external P-supply of 0.4 µg P·1–1·d–1 and sediment release of 0.5 µg P·1–1·d–1. Both temperature and composition of zooplankton affected the weight specific excretion rates of the zooplankton community. The weight specific community excretion rates of P and N increased with temperature (exponential model); 1–8 g PO4P·mg–1 zooplankton-C·d–1 and 5–42 µg NH3N·mg–1 zooplankton-C·d–1 (10°C–20°C).  相似文献   

5.
Summary The kinetics ofBordetella pertussis growth was studied in a glutamate-limited continuous culture. Growth kinetics corresponded to Monod's model. The saturation constant and maximum specific growth rate were estimated as well as the energetic parameters, theoretical yield of cells and maintenance coefficient. Release of pertussis toxin (PT) and lipopolysaccharide (LPS) were growth-associated. In addition, they showed a linear relationship between them. Growth rate affected neither outer membrane proteins nor the cell-bound LPS pattern.Nomenclature X cell concentration (g L–1) - specific growth rate (h–1) - m maximum specific growth rate (h–1) - D dilution rate (h–1) - S concentration of growth rate-limiting nutrient (glutamate) (mmol L–1 or g L–1) - Ks substrate saturation constant (mol L–1) - ms maintenance coefficient (g g–1 h–1) - Yx/s theoretical yield of cells from glutamate (g g–1) - Yx/s yield of cells from glutamate (g g–1) - YPT/s yield of soluble PT from glutamate (mg g–1) - YKDO/s yield of cell-free KDO from glutamate (g g–1) - YPT/x specific yield of soluble PT (mg g–1) - YKDO/x specific yield of cell-free KDO (g g–1) - qPT specific soluble PT production rate (mg g–1 h–1) - qKDO specific cell-free KDO production rate (g g–1 h–1)  相似文献   

6.
The effect of medium composition on the growth form of Aspergillus terreus NRRL 1960 in relation to itaconic acid fermentation has been studied. Four types of mycelial pellets were obtained under the conditions used and may be classified as (a) frayed and loose with 0.1–0.5 mm diameter (b) compact with 0.1–0.5 mm diameter (c) loose with 0.5–2.0 mm diameter and (d) compact with 0.5–2.0 mm diameter. Their respective maximum specific rates of formation and yields of itaconic acid, based on 100 g sucrose supplied, were (a) 1.25 mol mg–1h–1 and 55–59 g, (b) 0.27–0.43 mol mg–1 h–1 and 26–38 g, (c) 0.75–0.90 mol mg–1 h–1 and 45–51 g and (d) 0.12 mol mg–1 h–1 and 10 g. The presence of Ca2+, Zn2+ and Fe2+ in the basal medium at concentrations of 23.3 mg/100 ml, 0.01 mg/100 ml and 0.006 mg/100 ml respectively were found to be adequate and crucial in obtaining the desired outgrowth for both high production rates and consistent yields of itaconic acid. The further addition of either commercial plaster of Paris or analytical-reagent-grade CaSO4, especially when activated by heating to 530°C and present in excess of solubility, results in small and frayed pellets, which lead to itaconic acid yields of 55–59 g acid/100 g sugar supplied.  相似文献   

7.
This study examines the influence of current velocity in the toxiceffect of copper in diatom-dominated biofilms grown in artificial channels.Effects on community structure, algal biomass and photosynthesis (carbonincorporation) caused by 15 g L–1 of copperwere tested at contrasting (1 and 15 cm s–1)velocities. Moreover, a possible threshold on the effect of copper on algalbiomass and photosynthesis related to current velocity was examined by usingprogressively increasing current velocity (1 to 50 cms–1) at 15 g L–1 Cu.Chlorophyll-a decreased ca. 50% as a result of addition of15 g L–1 Cu. Chlorophyll decrease occurredearlier at 15 cm s–1 than at 1 cms–1 when adding 15 g L–1Cu. Copper also caused a remarkable decrease in carbon incorporation(from 30 to ca. 50%), which was produced earlier at 15 cms–1 (three days) than at 1 cms–1 (seven days). Some taxa were affected by thecombination of copper and current velocity. Both Achnanthesminutissima and Stigeoclonium tenue becomedominant at 15 cm s–1 in the presence of copper.Significant inhibition of algal growth in 15 g L–1Cu occurred at low (1 cm s–1) and highvelocities (50 cm s–1), but not at intermediatevelocity (20 cm s–1). The experiments indicatethat current velocity triggers the effect that copper has on diatom-dominatedbiofilms, and that the effect is more remarkable at low and high than atintermediate current velocities.  相似文献   

8.
Summary The transfer of a Chl element, causing resistance to chloramphenicol in Streptomyces coelicolor A3(2), was studied in NF x SCP1 superfertile crosses. When the Chl element is on the donor side (NF) its transfer to the recombinant cells was virtually total as if the element acted as a second concomitant transfer origin. When the Chl element was on the recipient side (SCP1) it was never displaced by the immigrant chromosome even when the region facing chl + was selected for. A fraction of the original Chl mutants presented a requirement for arginine (ArgB). A Chl mutant gave rise spontaneously to ArgB derivatives at high frequency. The same ArgB requirement come out at high frequency among Chl derivatives from a cross NFChl x SCP1Chl+ in which neither parent required arginine or produced spontaneously arginineless derivatives. It is suggested that the Chl element is a transposable element (Tn) presumably associated with insertion sequences (IS). The insertional inactivation of the Chl element may be accompanied or followed by a deletion in the adjacent ArgB gene.  相似文献   

9.
Graphic presentation of weekly rates of change of algal biomass (expressed as chlorophyll a) and nutrient and dissolved oxygen concentrations can be regarded as harmonic oscillation motion. Maximum amplitudes of these oscillations provide a useful tool to assess the degree of stability of aquatic ecosystems in relation to their trophic state. Data sets from seven different lakes ranging from hypereutrophic to oligo-mesotrophic were processed using a computerized method. The high values of oscillation amplitudes of approximately 150 g l–1 wk–1 chlorophyll a, 500 g l–1 wk–1 ammonia nitrogen, 50 g l–1 wk–1 soluble reactive phosphorus and 10 mg l–1 wk–1 dissolved oxygen, indicated strong ecosystem instability, while low values of less than 10 g l–1 wk–1 of chlorophyll a, 20 g l–1 wk–1 ammonia nitrogen, 2 g l–1 wk–1 soluble reactive phosphorus, and 3 mg l–1 wk–1 dissolved oxygen represented a stable system. Oscillation amplitudes of the chlorophyll a values were found to be the most representative indicator of ecosystem stability.  相似文献   

10.
A method for callus induction, adventitious bud regeneration, shoot multiplication and rooting of in vitro formed shoots of Helianthus annuus L. var. Argentario is described. Hypocotyl and cotyledon explants formed callus on medium containing 2 mgl–1 naphthalene acetic acid and 0.5 mgl–1 benzyladenine. Adventitious buds were formed on hypocotyl segments on medium containing 0.5–2 mgl–1 benzyladenine. The optimal level of sucrose concentration for shoot regeneration from hypocotyls was 1.5%. Multiplication from shoot apices was promoted by kinetin (2 mgl–1) plus gibberellic acid (5 mgl–1), benzyladenine (2 mgl–1) plus gibberellic acid (10 mgl–1) or at lower frequency by benzyladenine (1 mgl–1). A general feature of the plantlets formed in vitro was the precocious flowering.  相似文献   

11.
The effects of eight microelements (I, BO3 3–, MoO4 2–, Co2+, Cu2+, Mn2+, Fe2+, Zn2+) on the biosynthesis of camptothecin and the growth of suspension cultures of Camptotheca acuminata were studied. The increase of I to 25 M l–1, Cu2+ to 1 M l–1, Co2+ to 2 M l–1 and MoO4 2– to 10 M l–1 in Murashige and Skoog (MS) medium resulted in 1.66, 2.84, 2.53 and 2.04 times higher of camptothecin yield than that in standard MS medium respectively. Combined treatment of I (25 M l–1), Cu2+ (1 M l–1), Co2+ (2 M l–1) and MoO4 2– (10 M l–1) lead to improve cell dry weight, camptothecin content, and camptothecin yield to 30.56 g l–1, 0.0299%, and 9.15 mg l–1, respectively, which were 20.2, 208.9 and 273.8% increment respectively when compared with those of control.  相似文献   

12.
Isolation of bacteroids from effective (Fix+) and ineffective (Fix) pea nodules, inoculated withRhizobium leguminosarum K, were performed by a density gradient centrifugation method using silica sol (Percoll). Only one zone (=1.064–1.072; n-zone) was recognized in the Fix+ nodule which contained typical Y-shaped bacteroids while two zones (n-zone and =1.125–1.145; n'-zone) were obtained from the Fix nodule. The cells in the n'-zone, which are long rods differed morphologically from free-living cells at any growth phase (=1.108–1.125; f-zone and =1.074–1.078; f'-zone), and differed from Y-shaped bacteroids by cell density. The esterase isozyme pattern of bacteroids in the n-and n'-zones also showed clear differences from that of f-and f'-zone of free-living cells.  相似文献   

13.
Fallisia biporcati n. sp. parasitises thrombocytes and lymphocytes of Anolis biporcatus and A. lionotus in Panama. Round or oval schizonts average 13.3 × 11.5 (10.5–16 × 9–13) m, with LW 153.8 (94–208) m2, and produce 38.3 (28–60) merozoites. Gametocytes are variably shaped, from round or oval to nearly triangular or rectangular, and average 12.6 × 9.0 (10–15 × 6–12) m, with LW 113.1 (82–150) m2 and L/W ratio 1.43 (1.0–2.2). Thrombocytes and lymphocytes of A. poecilopus in Panama are parasitised by F. poecilopi n. sp. Schizonts, oval to elongate in shape, 7.7 × 4.7 (5.5–9 × 3–6) m, with LW 36.5 (22–54) m2, are filled with 31.0 (20–51) tiny nuclei or merozoites. Gametocytes are 10.1 × 8.0 (7.5–14 × 6–11) m, with LW 82.0 (45–132) m2, round to elongate with L/W ratio 1.27 (1.0–1.6). F. thecadactyli n. sp. parasitises thrombocytes and lymphocytes of Thecadactylus rapicaudus in Panama and Venezuela. Oval, oblong, or triangular schizonts average 10.3 × 8.0 (7–13 × 5–12) m, with LW 86.6 (37–156) m2, and produce 40.2 (26–61) merozoites. Gametocytes are round, oval, triangular or elongate, 10.4 × 7.0 (7–15 × 5–11) m, with LW 74.8 (40–154) m2 and L/W ratio 1.51 (1.1–2.2). F. dominicensis n. sp. parasitises thrombocytes of A. cybotes on Hispaniola. Schizonts, 6.0 × 4.8 (4–8 × 3–7) m, with LW 29.1 (12–56) m2, round, oval, elongate, oblong or lentiform in shape, produce 12.4 (8–22) merozoites. Gametocytes are 6.6 × 5.0 (5–9 × 4–7) m , with LW 33.8 (20–56) m2, round, oval or elongate, and L/W ratio of 1.34 (1.1–2.0).  相似文献   

14.
Predictive models for phosphorus retention in wetlands   总被引:1,自引:0,他引:1  
The potential of wetlands to efficiently remove (i.e., act as a nutrient sink) or to transform nutrients like phosphorus under high nutrient loading has resulted in their consideration as a cost-effective means of treating wastewater on the landscape. Few predictive models exist which can accurately assess P retention capacity. An analysis of the north American data base (NADB) allowed us to develop a mass loading model that can be used to predict P storage and effluent concentrations from wetlands. Phosphorus storage in wetlands is proportional to P loadings but the output total phosphorus (TP) concentrations increase exponentially after a P loading threshold is reached. The threshold P assimilative capacity based on the NADB and a test site in the Everglades is approximately 1 g m–2 yr–1. We hypothesize that once loadings exceed 1 g m–2 yr–1 and short-term mechanisms are saturated, that the mechanisms controlling the uptake and storage of P in wetlands are exceeded and effluent concentrations of TP rise exponentially. We propose a One Gram Rule for freshwater wetlands and contend that this loading is near the assimilative capacity of wetlands. Our analysis further suggests that P loadings must be reduced to 1 g m–2 yr–1 or lower within the wetland if maintaining long-term low P output concentrations from the wetlands is the central goal. A carbon based phosphorus retention model developed for peatlands and tested in the Everglades of Florida provided further evidence of the proposed One Gram Rule for wetlands. This model is based on data from the Everglades areas impacted by agricultural runoff during the past 30 years. Preliminary estimates indicate that these wetlands store P primarily as humic organic-P, insoluble P, and Ca bound P at 0.44 g m–2 yr–1 on average. Areas loaded with 4.0 g m–2 yr–1 (at water concentrations>150 g·L–1 TP) stored 0.8 to 0.6 g m–2 yr–1 P, areas loaded with 3.3 g m–2 yr–1 P retained 0.6 to 0.4 g m–2 yr–1 P, and areas receiving 0.6 g m–2 yr–1 P retained 0.3 to 0.2 g m–2 yr–1. The TP water concentrations in the wetland did not drop below 50 g·L–1 until loadings were below 1 g m2 yr–1 P.  相似文献   

15.
Mercury budget of an upland-peatland watershed   总被引:6,自引:2,他引:6  
Inputs, outputs, and pool sizes oftotal mercury (Hg) were measured in a forested 10 hawatershed consisting of a 7 ha hardwood-dominatedupland surrounding a 3 ha conifer-dominatedpeatland. Hydrologic inputs via throughfall andstemflow, 13±0.4 g m–2 yr–1over the entire watershed, were about doubleprecipitation inputs in the open and weresignificantly higher in the peatland than in theupland (19.6 vs. 9.8 g m–2 yr–1). Inputs of Hg via litterfall were 12.3±0.7g m–2 yr–1, not different in thepeatland and upland (11.7 vs. 12.5 g m–2yr–1). Hydrologic outputs via streamflow were2.8±0.3 g m–2 yr–1 and thecontribution from the peatland was higher despiteits smaller area. The sum of Hg inputs were lessthan that in the overstory trees, 33±3 gm–2 above-ground, and much less than eitherthat in the upland soil, 5250±520 gm–2, or in the peat, 3900±100 gm–2 in the upper 50 cm. The annual flux of Hgmeasured in streamflow and the calculated annualaccumulation in the peatland are consistent withvalues reported by others. A sink for Hg of about20 g m–2 yr–1 apparently exists inthe upland, and could be due to either or bothstorage in the soil or volatilization.  相似文献   

16.
The reproductibility of tank experiments concerning unicellular marine algal development was analyzed by means of parallel experiments with cultures ofThalassiosira rotula andSkeletonema costatum, using large flexible plastic tanks under semi-natural conditions. The tanks (3–4 m3, 4–5 m deep) were exposed in the German Bight at a station in the outer harbour of helgoland. The water was obtained from the open North Sea in towable tanks; it was filtered (plate filter), enriched with nitrate (20–30 gat dm–3), phosphate (1.3–2.3 gat dm–3) and silicate (15–23 gat dm–3)-nearly natural springtime concentrations in this area-and inoculated with 103–105 cells dm–3. The water was mixed with non-metal stirring equipment. Within 5 days, concentrations of 106–107 cells dm–3 in an exponential growth phase were obtained. In experiments withT. rotula a parallel development was achieved in spite of some contamination by surrounding water. This is the case for nearly all parameters analyzed (nutrient salts, phytoplankton, bacteria, C, N and particulate carbohydrates). The heterotrophic bacteria, which were determined by means of the plate method, reached concentrations of up to 106 (T. rotula) and 105 (S. costatum) CFU cm–3, respectively. They showed a consistent retrograde development at diatom concentrations above a certain level. The crop did not increase again until the diatoms had reached the stationary phase. During exponential growth ofT. rotula (G=8.9–11.7 h) a partially synchronous cell division was observed. There were also rhythms with respect to cell size (pervalvar axes) and chain length (number of cells). For the experiments withS. costatum (G=10–11.4 h) diurnal variations of cell size and chain length occurred. The present results indicate acceptable reproducibility of algal development and related phenomena in enclosed water bodies.  相似文献   

17.
Hansen  J.P.  Vinther  F.P. 《Plant and Soil》2001,237(2):257-266
Aiming at estimating the spatial variability in N2 fixation, and to evaluate the appropriateness of the 15N isotope dilution (ID) method and the natural15N abundance (NA) method in reflecting spatial variability under the influence of cattle grazing, the symbiotic N2 fixation in grass–white clover mixture was studied. At the Foulum site, where the ID method was used, differences in the climatic conditions between the two years of investigations caused a considerable difference in plant growth rates and proportion of clover. Consequently, the total N2 fixation in ungrazed reference plots was significantly less in 1998 than in 1997, being 5.9 and 12.5 g N m–2, respectively. In both years there was a wide range in concentration of inorganic N in the soil with coefficients of variance of approximately 60–190% for ammonium and 70–340% for nitrate. Significant negative correlations between pNdfa, determined by the ID method, and the log-transformed values of inorganic N and total N in grass were found. The NA method was applied on three nearby commercial dairy farms. They also showed high coefficients of variation. The coefficient of variance for NO3 -N ranged from 37 to 282% and for NH4 +-N from 29 to 237%. Average estimates of pNdfa values, which in the NA method were calculated using apparent B values ranging from –2.10 to –2.59, were generally lower (0.7–0.87) for these farms than for the Foulum site (0.89–0.95) using the ID method. For the NA method the 15N values, i.e. deviation in 15N concentration from atmospheric N2, ranged from –7.0 to 5.7 for the grass N, which in several cases was lower than for clover N. Due to this high variability of the 15N values, probably caused by deposition and plant assimilation of 15N depleted urinary N in the pastures, the NA method was marginal for accurate determination of pNdfa. Consequently no significant correlation between the pNdfa determined by this method, and the log-transformed values of inorganic N in soil or total N in grass were found.  相似文献   

18.
A method has been suggested for the synthesis of conjugates of oligodeoxyribonucleotides with chemical constructs mimicking the ribonuclease A active center for directed fragmentation of RNA. The method is based on sequential addition of a linker group, 9-(methylamino)anthracene, to the 5"- or 3"-terminal phosphate of oligonucleotide, and then an imidazole-containing construct by cycloaddition. The conjugates of oligonucleotides complementary to regions 44–61 (2B–R) and 60–76 (1C–R) of yeast phenylalanine tRNA proved able to cleave tRNAPhe under physiological conditions preferentially at the sole phosphodiester bond (C63–A64 for 2B–R and C56–G57 for 1C–R, respectively). The half-time of tRNAPhe hydrolysis in the presence of 2B–R conjugate was 30 min at a 2B–R concentration of 10 M and several minutes at conjugate concentration of 50 M.  相似文献   

19.
Finfish culture is a growing industry, and it causes a nutrient loading problem. To investigate the feasibility of an integrated culture of kelp and salmon, 15-cm long kelp (Laminaria saccharina) was grown in salmon culture effluent. The objectives were to test the effects of flow rate and kelp density on dissolved inorganic nitrogen removal (DIN), and DIN uptake and growth by the kelp. NH 4 + , NO 3 and DIN (NH 4 + + NO 3 ) loadings were in the ranges 6.2–25.4, 12.9–40.0, 19.7–52.7 mol 1–1, respectively, over the experimental period.Surplus uptake of nitrogen was not evident, because the C:N ratio (10–11) was constant in all experiments. During light periods, the kelp removed from 170–339 mol 1–1 h–1, and approximately 26–40% of the incoming DIN. The DIN uptake rate, based on daylight sampling periods, ranged between 6.1–22.5 mol g–1 dry mass h–1. The highest-flow rate, lowest-density tank had the highest DIN uptake rate. Debris from the fish effluent settling on the kelp thalli in the low-flow rate tanks affected uptake. Mean DIN uptake rate based on 3 days of growth for all flow-density combinations ranged between 5.4–8.3 mol g–1 dry mass h–1. The kelp utilized NH 4 + and NO 3 equally.The growth ranged between 6.5–9% d–1. The biomass production ranged from 1–2 g per sampling period. The highest growth rate and biomass production were achieved by kelp in the highest-flow rate, lowest-density tank. Lower DIN concentrations due to higher DIN removal rates in the other tanks and light limitation due to self-shading in the high-density tanks were probably responsible for the reduced growth rate in these tanks.Author for correspondence  相似文献   

20.
Summary The basolateral membrane of the thick ascending loop of Henle (TALH) of the mammalian kidney is highly enriched in Na+/K+ ATPase and has been shown by electrophysiological methods to be highly conductive to Cl. In order to study the Cl conductive pathways, membrane vesicles were isolated from the TALH-containing region of the porcine kidney, the red outer medulla, and Cl channel activity was determined by a36Cl uptake assay where the uptake of the radioactive tracer is driven by the membrane potential (positive inside) generated by an outward Cl gradient. The accumulation of36Cl inside the vesicles was found to be dependent on the intravesicular Cl concentration and was abolished by clamping the membrane potential with valinomycin. The latter finding indicated the involvement of conductive pathways. Cl channel activity was also observed using a fluorescent potential-sensitive carbocyanine dye, which detected a diffusion potential induced by an imposed inward Cl gradient. The anion selectivity of the channels was Cl>NO 3 =I gluconate. Among the Cl transport inhibitors tested, 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPAB), 4,4-diisothiocyano-stilbene-2,2-disulfonate (DIDS), and diphenylamine-2-carboxylate (DPC) showed IC50 of 110, 200 and 550 m, respectively. Inhibition of36Cl uptake by NPPAB and two other structural analogues was fully reversible, whereas that by DIDS was not. The nonreactive analogue of DIDS, 4,4-dinitrostilbene-2,2-disulfonate (DNDS), was considerably less inhibitory than DIDS (25% inhibition at 200 m). The irreversible inhibition by DIDS was prevented by NPPAB, whereas DPC was ineffective, consistent with its low inhibitory potency. It is proposed that NPPAB and DIDS bind to the same or functionally related site on the Cl channel protein.  相似文献   

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