EFFECTS OF PHENOLIC INHIBITORS ON GROWTH AND METABOLISM OF GLUCOSE-UL-14C IN PAUL'S SCARLET ROSE CELL-SUSPENSION CULTURES

ABSTRACT Paul's Scarlet rose cell-suspension cultures were incubated in varying concentrations of the following phenolic inhibitors; chlorogenic acid, cinnamic acid, p-coumaric acid, ferulic acid, and scopoletin. All test compounds except chlorogenic acid were completely inhibitory at a 10⁻³m concentration, resulting in death of the cells prior to completion of the growth cycle. To assess the cellular effects of two commonly named plant inhibitors, ferulic and cinnamic acids, these compounds were provided to cultures during incubation of cells with glucose-UL-¹⁴C. Incubation of cells with glucose-UL-¹⁴C in the presence of 10⁻⁴m ferulic acid resulted in increased incorporation of ¹⁴C into the soluble lipid fraction along with decreased incorporation of ¹⁴C into protein, organic acids, and soluble amino acids. Treatment of the cells with 10⁻⁵m cinnamic acid during the incubation period resulted in a significant decrease in incorporation of ¹⁴C into protein. These alterations in the flow of carbon into cellular constituents when cells are treated with cinnamic and ferulic acids explain, at least in part, why these compounds inhibit growth, seed germination, and seedling development.     

FORMATION AND DISSOCIATION OF CELL AGGREGATES IN SUSPENSION CULTURES OF PAUL'S SCARLET ROSE

In liquid culture stem tissue of Paul's Scarlet rose produces a suspension containing cell aggregates of extremely variable dimensions. There is, however, a definite pattern of change in the degree of cell aggregation over time. During the period of most rapid cell division large aggregates form as the result of a minimal separation of the proliferating cells. As the rate of cell division slows, the average number of cells per aggregate decreases. The dissociation of cell aggregates continues at a uniform rate after cell division has stopped. Cell separation is inhibited at low (0.1 mg/1) auxin (NAA) concentrations and by substitution of sucrose for glucose in the culture medium. Cell separation is delayed (but not greatly inhibited) by kinetin. The presence of casein hydrolysate prevents the formation of the large cell aggregates normally occurring in the early stages of the culture cycle. A variant strain which shows a much higher degree of cell separation has been isolated from stock callus tissue grown on solid medium.     

The Uptake of Growth Substances: III. INFLUENCE OF INDOLEACETIC ACID AND OTHER AUXINS ON THE UPTAKE OF 2,4-DICHLOROPHENOXY-ACETIC ACID BY CHLORELLA

When 2,4-dichlorophenoxyacetic acid, labelled with ¹⁴C, is accumulatedby Chlorella pyrenoidosa, an appreciable fraction, which increaseswith time, is held within the cell in a form which is not removedwhen the cells are placed in a solution containing the unlabelledcompound. The accumulation of this fraction is dependent uponthe supply of metabolic energy since it is affected by temperature,light, and the addition of inhibitors or citrate to the externalmedium. The uptake of this metabolically accumulated 2,4-D follows apattern which indicates that a single enzymic reaction is apredominant component of the uptake process. In the presenceof the other auxins, including indoleacetic acid, 2,4,5-trichlorophenoxyaceticacid and 4-chlorophenoxyacetic acid, the uptake of 2,4-D isdepressed and the changes suggest that there is competitionat the site of this reaction. The nature of the competitionis seemingly of the type where each compound serves as a substratefor the enzyme. It may be significant that the extent to which the individualauxins compete with 2,4-D is in the same order as their generaleffectiveness as growth regulators for other species.     

NUTRIENT CONTROLS ON NITROGEN UPTAKE AND METABOLISM BY NATURAL POPULATIONS AND CULTURES OF TRICHODESMIUM (CYANOBACTERIA)

The effects of inorganic nutrient (ammonium [NH₄ ⁺ ] and nitrate [NO₃ ⁻ ]) and amino acid (glutamate [glu] and glutamine [gln]) additions on rates of N₂ fixation, N uptake, glutamine synthetase (GS) activity, and concentrations of intracellular pools of gln and glu were examined in natural and cultured populations of Trichodesmium. Additions of 1 μM glu, gln, NO₃ ⁻ , or NH₄ ⁺ did not affect short-term rates of N₂ fixation. This may be an important factor that allows for continued N₂ fixation in oligotrophic areas where recycling processes are active. N₂ fixation rates decreased when nutrients were supplied at higher concentrations (e.g. 10 μM). Uptake of combined N (NH₄ ⁺ , NO₃ ⁻ , and amino acids) by Trichodesmium was stimulated by increased concentrations. For NO₃ ⁻ , proportional increases in NO₃ ⁻ uptake and decreases in N₂ fixation were observed when additions were made to cultures before the onset of the light period. GS activity did not change much in response to the addition of NH₄ ⁺ , NO₃ ⁻ , glu, or gln. GS is necessary for N metabolism, and the bulk of this enzyme pool may be conserved. Intracellular pools of glu and gln varied in response to 10 μM additions of NH₄ ⁺ , glu, or gln. Cells incubated with NH₄ ⁺ became depleted in intracellular glu and enriched with intracellular gln. The increase in the gln/glu ratio corresponded to a decrease in the rate of N₂ fixation. Although the gln/glu ratio decreased in cells exposed to the amino acids, there was only a corresponding decrease in N₂ fixation after the gln addition. The results presented here suggest that combined N concentrations on the order of 1 μM do not affect rates of N₂ fixation and metabolism, although higher concentrations (e.g. 10 μM) can. Moreover, these effects are exerted through products of NH₄ ⁺ assimilation rather than exogenous N, as has been suggested for other species. These results may help explain how cultures of Trichodesmium are able to simultaneously fix N₂ and take up NH₄ ⁺ and how natural populations continue to fix N₂ once combined N concentrations increase within a bloom.     

UPTAKE AND RELEASE OF NIPECOTIC ACID BY RAT BRAIN SLICES

—Nipecotic acid, a potent inhibitor of GABA uptake, is taken up by slices of rat cerebral cortex by a sodium-dependent, ‘high affinity’ system (K_m 11 μM), and can be released from these slices by an increased potassium ion concentration in a calcium-dependent manner. Nipecotic acid and GABA appear to be taken up by the same osmotically-sensitive structures. GABA and substances which inhibit GABA uptake also inhibit the uptake of nipecotic acid. GABA can release preloaded nipecotic acid from brain slices, and nipecotic acid can release preloaded GABA. This indicates that GABA and nipecotic acid can be counter-transported using the same mobile carrier. Nipecotic acid appears to have a higher affinity than GABA for this carrier.     

UPTAKE OF LIPOPHILIC CADMIUM COMPLEXES BY THREE GREEN ALGAE: INFLUENCE OF HUMIC ACID AND ITS pH DEPENDENCE1

Cadmium forms neutral, lipophilic CdL₂⁰ complexes with diethyldithiocarbamate (L = DDC) and with ethylxanthate (L = XANT). In a synthetic solution and in the absence of natural dissolved organic matter (DOM), for a given total Cd concentration, uptake of these complexes by unicellular algae is much faster than the uptake of the free Cd²⁺ cation. The objective of the present study was to determine how this enhanced uptake of the lipophilic CdL₂⁰ complexes was affected by the presence of natural DOM (Suwannee River humic acid, SRHA). Experiments were performed with Cd(DDC)₂⁰ and Cd(XANT)₂⁰ at two pH values (7.0 and 5.5) and with the three chlorophytes [Chlamydomonas reinhardtii P. A. Dang., Pseudokirchneriella subcapitata (Korshikov) Hindák, Chlorella fusca var. vacuolata Shihira et R. W. Krauss]. Short‐term uptake (30–40 min) of the CdL₂⁰ complexes was followed in the absence and presence of SRHA (6.5 mg C · L^?1). Acidification from pH 7.0 to 5.5 decreased CdL₂⁰ uptake by the three algae, in the presence or absence of humic acid (HA). The dominant effect of the HA was to decrease Cd uptake, due to its interaction with the CdL₂⁰ complexes in solution. However, if uptake of the free CdL₂⁰ complexes was compared in the presence and absence of HA, in four of eight cases initial uptake rate constants (k_i) were significantly higher (P < 0.05) in the presence of the HA, suggesting the operation of an interfacial effect of the HA at the algal cell membrane, favoring uptake of CdL₂⁰. Overall, the experimental results suggest that neutral metal complexes will be less bioavailable in natural waters than they are in synthetic laboratory media in the absence of natural DOM.     

EFFECT OF CYSTEINE AND METHIONINE ON SULFATE UPTAKE AND PRIMARY PRODUCTIVITY BY AXENIC ALGAL CULTURES AND LAKE MICROPLANKTON COMMUNITIES1

The presence of up to 500 μg sulfur·l^?1 of an equimolar mixture of cysteine and methionine had virtually no effect on the SO₄^2- uptake rate of Navicula pelliculosa, (Bréb.) Hilse whereas the rate of Ankistrodesmus falcatus (Corda) Ralfs was decreased by the presence of 500 μg S· l^?1 and Anabaena flos-aquae (Lyngbye) Bréb. by 50 μg S·l^?1. Primary productivity in these axenic cultures was affected (decreased) only in A. falcatus. The C:S uptake ratio was lowest in N. pelliculosa and highest in A. falcatus. Considering these species as representative of groups of naturally occurring algae, patterns of SO₄^2- uptake and primary productivity in a eutrophic and a moderately oligotrophic lake reflected the results of the algal culturing experiments: SO₄^2- uptake rates, relative to primary productivity, were higher in the presence of diatoms and bluegreen algae and lower when green algae were present; the addition of the cysteine I methionine mixture to the lake waters decreased the rate of microplankton SO₄^2- uptake in correlation with the makeup of the algal community; primary productivity decreased upon the addition of cysteine I methionine when green algae were relatively abundant. It is concluded that, in most fresh water systems, the effects of organic sulfur pollution on algal SO₄^2- uptake and primary productivity are insignificant as compared to other ecological changes that occur due to that pollution.     

UPTAKE AND METABOLISM OF 5-METHYL-TETRAHYDROFOLIC ACID BY RAT BRAIN SLICES

DNA-dependent DNA polymerases were partially purified from nuclei of cells from the occipital lobe of human brain. The purification procedure included successive DEAE-cellulose and phosphocellulose column chromatography, gel filtration and sucrose density gradient centrifugation steps. Four enzymes corresponding to DNA polymerases-α, β, γ, and terminal deoxynucleotidyl transferase were found. Brain DNA polymerases could be differentiated from one another by size, template preferences and sensitivity to sulfhydryl blocking agents.     

13C AND 15N UPTAKE BY MARINE PHYTOPLANKTON. I. INFLUENCE OF NITROGEN SOURCE AND CONCENTRATION IN LABORATORY CULTURES OF DIATOMS1

Two species of marine diatoms, Skeletonema costatum (Grev.) Cleve and Phaeodactylum tricornutum Bohlin were grown in batch and continuous cultures on four different nitrogen compounds (nitrate, nitrite, ammonium, urea). Carbon and nitrogen uptake were measured simultaneously with the stable isotopes ¹³C and ¹⁵N. Nitrogen uptake generally increased with N concentration in the medium, but no clear difference existed between the N sources. Carbon fixation was decreased for up to 5 h following the addition of the N compound. Nitrite generally had the greatest inhibitory effect on C uptake. Carbon-to-nitrogen uptake ratios decreased with increasing dissolved N concentration, becoming lower than one in nutrient-limited cultures. In contrast, batch cultures exhibited C:N uptake ratios greater than one. These effects are essentially short-term and differ from long-term influences of the N source on the cellular chemical composition.     

INFLUENCE OF THE CLAY TYPES ILLITE AND MONTMORILLONITE ON THE UPTAKE OF65 Zn BY SCENEDESMUS OBLIQUUS.

SUMMARY

The stimulatory effect of the clays illite and montmorillonite on zinc uptake by Scenedesmus obliquus is discussed. The competitive effects of hydrogen, phosphate, magnesium and copper ions on zinc uptake are illustrated. This competition is reduced in the presence of illite and montmorillonite and zinc uptake is stimulated. Ferrous ions have no effect on zinc uptake, while calcium ions antagonize zinc uptake at high concentrations. The latter effect was remarkable in the presence of montmorillonite.     

幽门螺杆菌感染对胃酸及胃液氨浓度的影响

目的 :探讨幽门螺杆菌 (Hp)感染对胃酸分泌及氨浓度的影响以及十二指肠 (DU)的关系。方法 :对DU患者 ,Hp根治前后的胃液pH ,空腹胃酸及氨浓度之间的关系进行研究。结果 :Hp阳性的UC患者其空腹胃酸、氨浓度显著高于正常对照组 (P <0 0 5 ) ,而根除Hp后 ,空腹胃酸显著下降 ,接近正常水平 (P >0 0 5 ) ,氨浓度明显下降。结论 :Hp感染使DU患者胃酸分泌增多 ,二者之间的相互作用在DU的发病中占有重要地位。     

ADSORPTION OF FULVIC ACID ON ALGAL SURFACES AND ITS EFFECT ON CARBON UPTAKE

Adsorption of Suwannee River fulvic acid (SRFA) to algal surfaces of three green algae was studied at environmentally relevant pH values (4 –7) and SRFA concentrations (5–100 mg·L ^{− 1}). The influence of adsorbed SRFA on carbon uptake of Scenedesmus subspicatus Chodat was also examined. Although no adsorption was observed at neutral pH values (pH 6 and 7), at pH 4 up to 31 mg SRFA·m ^{− 2} and at pH 5 up to 4 mg SRFA·m ^{− 2} was adsorbed to the algal surfaces. Electrophoretic mobility measurements of S. subspicatus demonstrated an increase in the negative surface charge of the alga in the presence of SRFA at pH 4. The adsorbed SRFA also influenced ¹⁴C uptake in S. subspicatus; in this case, enhanced carbon uptake could be related to the amount of adsorbed SRFA. The binding of humic substances by algal surfaces was interpreted as the result of hydrogen bonding and hydrophobic interactions.     

EFFECTS OF SINGLE AND MULTIPLE EXPOSURES OF FERULIC ACID ON THE VEGETATIVE AND REPRODUCTIVE GROWTH OF PHASEOLUS VULGARIS BBL-290

Phaseolus vulgaris BBL-290 plants were grown in growth chambers in the Southeastern Plant Environment Laboratory and exposed to either single (at seedling, flower, or podfill) or multiple (biweekly or weekly) treatments of ferulic acid (FA). In the first experiment, plants were harvested one week after FA treatment (0, 1.0, 2.0 mM) and at final harvest (56 days old). FA delayed leaf expansion during the seedling and flowering stages. The total plant leaf area and the plant dry weight of plants treated with 1.0 and 2.0 mM FA as seedlings were reduced one week after treatment by 38–48%. The total plant leaf area and the plant dry weight of plants treated at flowering with 2.0 mM FA were reduced by 25% one week after treatment. Treatment with 2.0 mM FA at podfill caused the senescence and abscission of older leaves and reduced total plant leaf area, plant dry weight and mean pod dry weight by 54, 40, and 48%, respectively, one week after treatment. The plants treated at the seedling and flowering stages recovered by final harvest. In a subsequent experiment, FA (0, 0.50, 1.0, 1.5 mM) reduced total plant leaf area at the seedling and flowering stages but not at podfill. The youngest expanding leaves were most sensitive to FA at flowering. The leaf area of these leaves was reduced by 35 and 25%, one and two weeks after treatment, respectively. Their absolute growth rates were reduced from 31 to 56% one week after treatment at flowering. Their relative growth rates were reduced by 50% one week after treatment. Growth rates then recovered within two weeks after treatment. In the final experiment, biweekly exposures of FA (0.25, 0.50, 0.75, 1.0) reduced total plant leaf area but did not affect any other growth parameters. Weekly exposures of FA (0.25, 0.50, 0.75, 1.0) reduced total plant leaf area up to 34%, absolute growth rate up to 58%, leaf number up to 31% and pod number up to 58%. As the frequency of exposure to FA increased, the concentration necessary to affect bean plant growth and development decreased.     

运动小鼠心肌和骨骼肌对支链氨基酸的摄取及其对蛋白质合成的作用

目的和方法：本实验采用昆明种小鼠游泳运动模型,及１５ＮＧｌｙｃｉｎｅ和３ＨＬｅｕｃｉｎｅ同位素示踪技术探讨了运动状态下心肌和骨骼肌对ＢＣＡＡ的摄取量及ＢＣＡＡ对蛋白质合成的作用。结果：运动时心肌和骨骼肌从血循环中摄取ＢＣＡＡ显著增加,同时血清中ＢＣＡＡ的含量降低。结论：心肌和骨骼肌的蛋白质代谢存在差异,骨骼肌的蛋白质合成率高于心肌;运动使心肌蛋白代谢加速,补充ＢＣＡＡ降低了运动对心肌蛋白质代谢的影响,有利于骨骼肌蛋白质合成或使蛋白质分解降低。     

PROPERTIES OF THE UPTAKE AND RELEASE OF GLUTAMIC ACID BY SYNAPTOSOMES FROM RAT CEREBRAL CORTEX

Abstract– (1) The uptake and release of glutamic acid by guinea-pig cerebral cortex slices and rat synaptosomal fractions were studied, comparing the naturally occurring l - and non-natural d -isomers. Negligible metabolism of d -glutamic acid was observed in the slices. (2) Whereas in the cerebral slices the accumulation of glutamic acid was almost the same for the two isomers, d -glutamic acid was accumulated into the synaptosomal fraction at a markedly lower rate than was the L-isomer. (3) The uptake systems for d -isomer into the slices and synaptosomal fraction were found to be of single component, in contrast with the two component systems, high and low affinity components, for the uptake of l -glutamic acid. The apparent K_m values for the uptake of d -glutamic acid into the slices and synaptosomal fraction were comparable with those reported for the low affinity components for l -isomer. The uptake systems for d -glutamic acid were dependent on the presence of Na⁺ ions in the medium, like those for l -glutamic acid and GABA. (4) The evoked release of radioactive preloaded d -glutamic acid was observed both from the slices and synaptosomal fraction following stimulation by high K⁺ ions in the medium. From these observations, it is evident that the evoked release of an amino acid by depolarization in vitro is not necessarily accompanied by a high affinity uptake process. (5) The uptake of l -glutamic acid, expecially into the synaptosomal fraction, was highly resistant to ouabain. On the other hand, the uptake rate of d -glutamic acid and GABA into the synaptosomal fraction was inhibited by varying concentrations of ouabain in accordance with the inhibition for brain Na-K ATPase. (6) The uptake of l -glutamic acid into subfractions of the P₂ fraction was studied in relation to the distribution of the ‘synaptosomal marker enzymes’. An attempt to correlate the activities of enzymes of glutamic acid metabolism with the uptake of l -glutamic acid into the synaptosomal fraction from various parts of brain was unsuccessful. The high affinity uptake of l -glutamic acid was found to be very active in the synaptosomal fraction from any part of brain examined.