Dependence of radiostrontium uptake by pea and lupin on the content of calcium in the nutrient solution

In lupin (Lupinus albus L.) and pea (Pisum sativum L., cv. Raman) it was shown that the uptake of⁸⁹Sr from Knop's nutrient solution is significantly increased from a solution with decreased calcium content (one tenth of the normal content) and is slightly decreased from a solution with higher calcium content (150% of the normal content). The calciphile pea absorbs approximately 50% more calcium than the calciphobe lupin, and accordingly 50% radiostrontium less. The pea plant more strongly blocks the translocation of radiostrontium from roots to overground parts, as is proved by the higher discrimination factor of pea (i.e. by the ratio of specific activities of mCi⁸⁹ Sr/g Ca of roots to overground parts). The presence of chlorine in the nutrient solution decreases the content of radiostrontium per gram of dry matter, both in pea and lupin. Radiostrontium is absorbed quickly by both species of plants and is autoradiographically detectable as early as 2 hours after the introduction of radiostrontium to the nutrient solution. *** DIRECT SUPPORT *** A01GP049 00004     

Haematological studies on90Sr-90Y-toxicity

Summary Erythropoietic changes were observed, measured by⁵⁹Fe-uptake into red blood cells, and on radioiron turnover from blood plasma, at different time intervals (2–64 days) after treating adult female mice with varying activities of⁹⁰Sr-⁹⁰Y. Activities of 2.5 or 5.0 µCi radiostrontium per animal lead to a depression at time intervals of two and four days, at longer periods there was an overshoot. With activities of 0.5 or 1.0 µCi radiostrontium disturbances in the radioiron uptake are still observed, although these effects are not as pronounced as in experiments with higher burdens. In comparison with results obtained in experiments in which the plasma⁵⁹Fe-turnover was applied, even with an activity of 5 µCi radiostrontium per mouse, no deviation as against the untreated controls was detected.Dedicated to Prof. Dr. Hermann Muth, Homburg/Saar, on the occasion of his 65th birthday     

Regulation of the in vitro anamnestic antibody response by cyclic AMP: I. Antigen-induced uptake of nucleotides and nucleosides by lymphocytes

Addition of N⁶, O²′-dibutyryl cAMP (DbcAMP) to keyhole limpet hemocyanin (KLH)-primed rabbit lymph node cells for 1 hr, followed by its removal and the addition of KLH, had no effect on the subsequent antibody response, whereas addition of KLH for 1 hr followed by DbcAMP resulted in a 100% enhancement of antibody synthesis. Addition of cholera enterotoxin (CT), which rapidly and irreversibly binds to lymphocytes and activates adenylate cyclase, either before or after the addition of antigen, elevated the antibody response by 100%. These results suggested that some antigen-induced event(s) may be required for DbcAMP to exert its enhancing effects on the antibody response. The effect of KLH on the uptake of DbcAMP by KLH-primed lymph node cells was investigated. One and one hundred micrograms of KLH, which induce optimal and supraoptimal antibody synthesis, respectively, promoted maximal uptake of DbcAMP. This induced uptake was first detectable about 12 hr after addition of KLH, and it peaked during 24–48 hr of culture. DbcAMP uptake induced by a brief exposure of KLH (0–1 hr) was equivalent to that observed with long-term KLH addition (0–24 hr). KLH-induced DbcAMP uptake required KLH-reactive lymphocytes and represented active transport. Antibody to rabbit T lymphocytes inhibited this antigen-induced uptake. The mitogens concanavalin A (Con A) (T cells) and goat anti-rabbit Fab' (anti-Fab') (B cells) also stimulated DbcAMP uptake, as did human serum albumin (HSA) and myoglobulin (Mb) when added to homologously primed cells, indicating the generality of the phenomenon. [³H]DbcAMP entered the cells as di- or monobutyryl cAMP with about 40% metabolized to 5′AMP. This uptake could be competitively inhibited by other adenine or guanine nucleotides and nucleosides.     

Enhanced uptake of calcium by transforming lymphocytes

Phytohemagglutinin caused a rapid increase in calcium accumulation by lymphocytes. The enhanced uptake was observed within 1 hr of initiation of transformation in both human lymphocyte and mouse spleen cell cultures. Increased uptake was also found in mixed lymphocyte cultures although not until late in the response. The rate of calcium uptake increased with time after stimulation and depended upon the PHA concentration. The lowtemperature coefficient (Q₁₀) for calcium permeability in unstimulated cells was indicative of a passive diffusion process, but the Q₁₀ was slightly greater for PHA-stimulated cells. Various chemical agents which alter membrane properties and/or cellular metabolism inhibited uptake to a greater extent in stimulated cultures than in control cultures. Ouabain did not affect the calcium permeability of controls or stimulated cells within 1 hr after PHA addition, but it partially inhibited calcium uptake 12 hr after PHA treatment. Cyclic AMP, dibutyryl cyclic AMP, and theophylline also altered calcium transport providing evidence for an effect of cyclic AMP on an early event in the transformation process.     

Megalodiscus temperatus: absorption and incorporation of tritiated tyrosine, thymidine, and adenosine

Autoradiographic and liquid scintillation counting studies indicate adults of Megalodiscus temperatus can absorb large amounts of thymidine and adenosine, but use little of the thymidine in in vitro exposures of up to 72 hr. Labeling of worms exposed to ³H-adenosine in periods as short as 3 hr was achieved especially over young eggs, testes, and the ovary. Large amounts of tyrosine are found in the free amino acid pool of this worm but little is incorporated into tissues for exposure periods of up to 24 hr. Ligation experiments and autoradiograms of frozen-dried worms show thymidine and adenosine can enter via the tegument. Adenosine added to the exposure medium did not inhibit absorption of tyrosine but did markedly reduce the uptake of ³H-thymidine in 8-hr in vitro exposures. Bacteria adhering to the glycocalyx of M. temperatus were identified as Escherichia coli, but their role in uptake phenomena is unknown. The association of thymidine with the lymph vessels indicates a circulatory or excretory role for this system in M. temperatus. Adenosine was considered the best compound of the three to use for labeling cells for studies on the reproductive system.     

Sodium balance in adult atlantic salmon (Salmo salar L.) during migration into neutral and acid fresh water

• 1.1. In sea-water, adult salmon (S. salar) exchange an average of 12.6% of total body sodium/hr.
• 2.2. Following transfer to fresh water sodium uptake follows Michaelis-Menton kinetics. F_max = 2.40 mmol Na/1 ECF/hr, K_m = 0.26 mmol Na/1. The uptake system is fully activated immediately following transfer to fresh water.
• 3.3. Post smolts adapted to sea-water for 3 months take up sodium at only one third of the rate of adult fish following return to fresh water.
• 4.4. The concentration of prolactin in the plasma is low in sea-water adapted fish and does not rise during the first 8 hr in fresh water.
• 5.5. At pH 5 sodium uptake is reduced by almost 90%, even in the absence of aluminium, but recovers immediately on return to neutral water.
• 6.6. At pH 5 and 20 μmol Al/1 there is little further effect on sodium uptake but after 6 hr in aluminium the inhibition of sodium uptake continues after return to neutral aluminium fresh water and uptake is only 50% of normal 24 hr later.

     

Studies on Inhibition of Intestinal Absorption of Radioactive Strontium: I. Prevention of Absorption from Ligated Intestinal Segments

A method is reported which permits selective suppression of absorption of radioactive strontium from ingested food material, permitting the calcium to be available to the body. Studies were carried out in vivo by injection of Sr⁸⁹ and Ca⁴⁵ in the presence of inert carrier into ligated intestinal segments in rats, and the amount of absorption was measured by standard monitoring techniques. The pattern of absorption of both ions is very similar but the rate of absorption is different. It was found that the polyelectrolyte, sodium alginate, obtained from brown algae (Phaeophyceae), injected simultaneously with radiostrontium effectively reduces the absortion of Sr⁸⁹ from all segments of the intestine by as much as 50-80% of the control values. No significant reduction in absorption of Ca⁴⁵ was observed in equivalent concentrations. The reduction in blood levels of Sr⁸⁹ and in bone uptake corresponded to the absorption pattern. The difference in the effect on strontium and calcium absorption may be due to differences in the binding capacity of sodium alginate from the two metal ions under the conditions present in vivo.     

Influence of Cold on Radiostrontium Excretion

IN most work on the problems of releasing radiostrontium which has been incorporated into the vertebrate skeleton the effects of various parenterally administered chemical agents on the excretion rate of radiostrontium have been studied. Someriecrease in skeletal retention was obtained when inactive salts of strontium^1–4 or zirconium⁵ were given almost simultaneously with radiostrontium; so far all other efforts have been practically useless.     

Rumen Bacterial and Protozoal Responses to Insecticide Substrates

Insecticides containing organophosphate, chlorinated hydrocarbon, and carbamate were tested with bovine ruminal ingesta fractions. Rumen bacteria exposed to insecticide levels of 0 to 500 ppm in rumen fluid for 4 hr were inoculated into rumen fluid-starch feed extract medium. No apparent significant bacterial count inhibitions were noted. Also, when insecticides were used as carbon sources at concentrations of 500 ppm in carbohydrate-limited media, no increases in bacterial counts were indicated. Warburg manometric data showed that paraffin oil-Triton X-155 preparations of dimethoate, Diazinon, lindane, Thiodan and Sevin stimulated gas production in holotrich protozoa. Entodinium simplex, an oligotrich, produced less gas with insecticide substrates per unit of dry weight than did an Isotricha sp. Rumen bacteria and plant debris fractions from ruminal ingesta provided with insecticides did not give increased manometric responses over the endogenous control vessels. Washed suspensions of I. intestinalis produced volatile fatty acids in excess of the endogenous suspensions when provided insecticide substrates. Thiodan dissimilation by I. intestinalis was followed colorimetrically with 15% loss in substrate in 1 hr of incubation at 39 C. Diazinon-C¹⁴ substrate uptake was demonstrated with suspensions of E. simplex and I. intestinalis. Rumen ciliates are suggested as a possible means for screening out useful insecticides susceptible to microbial dissimilation for use on forage and other cattle-feed crops.     

Dynamics of vitellogenin uptake in Aedes aegypti as demonstrated by trypan blue

Trypan blue has been shown to be a reliable indicator of the micropinocytotic uptake of vitellogenin by developing oöcytes. Trypan blue was injected into the mosquito Aedes aegypti to determine at what times after the blood meal vitellogenin was taken up. Histological sections examined by light microscopy showed that trypan blue began to be sequestered from 2 to 5 hr after the blood meal. Any association between dye and ovariole ended from 39 to 42 hr after the blood meal, in which period no dye was incorporated into spheres of yolk protein. Of the times investigated in this experiment, the greatest amount of dye was seen in the oöcyte at 24 hr after the blood meal. The onset and conclusion of trypan blue uptake correspond with the related events in the synthesis of vitellogenin by the fat body. The presence of trypan blue in occasional interfollicular spaces suggests that the route of entry of vitellogenin in Aedes aegypti is indeed an interfollicular one.     

Studies of Inhibition of Intestinal Absorption of Radioactive Strontium: IV. Estimation of the Suppressant Effect of Sodium Alginate

A method is reported which permits selective suppression of absorption of radioactive strontium from ingested food material, allowing calcium to be available to the body. Studies were carried out on the inhibitory effect of various amounts of sodium alginate and the dose-response relationship of Sr⁸⁹ and bone uptake. The results obtained indicated that under laboratory conditions sodium alginate effectively reduces Sr⁸⁹ uptake in a constant proportion. This effect was observed at the three levels of administration of 1.4%, 12% and 24% of sodium alginate. The linear relationship between the dosage of the radioisotope and the bone uptake in the presence of sodium alginate suggests that the same proportion is maintained at the lower levels of intake of radioactive strontium.Previous studies with small constant doses of sodium alginate were extended in rats to a period corresponding approximately to three years of human life span. Low doses were sufficient to reduce appreciably bone uptake of radiostrontium.     

Increased muscle glucose uptake in response to chronic glyburide treatment is not related to changes in glucose transporter (GLUT4) protein

• 1.1. The mechanism of action of glyburide (a sulfonylurea) on muscle has been investigated by measuring glucose uptake and glucose transporter (GLUT4) protein levels after chronic glyburide treatment.
• 2.2. A dietary induced insulin resistant rat model (4 wk of high-fat, high-sucrose feeding) was given glyburide (2mg/kg/day) for 10 days and glucose uptake was measured in a perfused hindquarter preparation.
• 3.3. Protein levels of the GLUT4 glucose transporter were determined by Western analysis.
• 4.4. After 7 days of treatment, rats fed glyburide had lower blood glucose concentrations 2 hr (72 ± 5 vs 103 ± 12 mg/dl) and 24 hr (97 ± 7 vs 123 ± 7 mg/dl) after glyburide administration with no difference in serum insulin levels compared to vehicle treated animals.
• 5.5. Glucose uptake was approx doubled in basal state (0 insulin) in response to glyburide (2.8 + 0.4 vs 1.7 ± 0.2μ mol/g per hr).
• 6.6. Maximal insulin (100 nM) stimulated glucose uptake tended to be higher in the glyburide treated group, but did not reach statistical significance (8.0 ± 0.7 vs 7.0 ± 0.6 μmol/g per hr).
• 7.7. Western analysis revealed no significant effect of glyburide on the GLUT4 protein level in skeletal muscle.
• 8.8. These results suggest that glyburide alters glucose uptake through some mechanism other than alterations in the level of the GLUT4 glucose transporter protein.

     

Trypanosoma brucei: An evaluation of salicylhydroxamic acid as a trypanocidal drug

The chemotherapeutic potential of salicylhydroxamic acid (SHAM) was studied in adult rats infected with a strain of Trypanosoma brucei that kills the rats in about 100 hr. The median lethal dose, administered intraperitoneally in a carboxymethyl-cellulose suspension, is approximately 820 mg/kg body weight for male and 850 mg/kg for female rats. The apparent cause of death is severe depression of the central nervous system.Half-maximal inhibition of O₂ uptake by trypanosomes in vitro requires 15 μM SHAM, whereas 100 μM inhibits over 90%. This inhibitory effect on trypanosome respiration was used as a biological assay for the effective SHAM concentration in rat plasma. After administration of a sublethal SHAM dose to rats, the effective plasma SHAM concentration rose rapidly to about 500 μM and then fell to about 10 μM at 4 hr. Nevertheless, this dose did not significantly affect the survival time of rats infected with T. brucei. Even if, by repeated SHAM administration, the plasma SHAM concentration was kept at around 100 μM for more than 4 hr, no therapeutic effect was observed.These results show that O₂ uptake is not essential for the survival of trypanosomes in rats and they support the idea that bloodstream trypanosomes have an alternative pathway for glycolysis, allowing energy production in the absence of respiration.The possibility that SHAM or other inhibitors of trypanosome respiration could stilll be trypanocidal if used in conjunction with another inhibitor of glycolysis is discussed.     

Plasmodium berghei: uptake of clindamycin and its metabolites by mouse erythrocytes with clindamycin-sensitive and clindamycin-resistant parasites.

Tritiated Clindamycin was used to compare the uptake of Clindamycin in plasma and red cells of mice infected with clindamycin-sensitive or clindamycin-resistant Plasmodium berghei and in uninfected mice. Red cells infected with either sensitive or resistant parasites have a higher concentration of [³H]clindamycin and its active metabolites 1 hr after drug administration than uninfected red blood cells. There was no significant difference in uptake of Clindamycin by red blood cells parasitized by sensitive or resistant parasites. Levels of Clindamycin and its metabolites were consistently higher in red cells than in plasma, both in infected and uninfected mice, but the drug was readily removed by washing red cells with phosphate buffered saline in either case. It is concluded that resistance to Clindamycin is not due to an impaired uptake of the drug by the parasitized red cell as has been shown for chloroquine resistance in P. falciparum and P. berghei.     

Effects of Estrogen on Glucose Uptake by Rat Muscle

The isolated rat diaphragm was used to study the effects of 17β-estradiol on basal and insulin-mediated glucose uptake. Rats were injected with estradiol for 2 wk in daily doses of 10 μg/100 g of body weight and were compared to untreated control animals. Estrogen treatment resulted in a 16% decrease in basal glucose uptake by diaphragm muscle as compared to controls. In contrast, in the presence of insulin, glucose uptake by muscle increased 103% above basal in estradiol-treated animals as compared to a 38% rise in the control group. The absolute rate of glucose uptake induced by insulin in the estradiol treated animals (5.8 mg/g/hr) was 22% higher than in controls. These findings were not accompanied by changes in weight gain, plasma glucose and plasma immunoreactive insulin concentrations in the treated animals. In vitro incubation of diaphragm muscle with estradiol did not have an effect on basal or insulin-mediated glucose uptake.     

A neighboring-inhibition model for stomate patterning

The patternization of stomate distribution was investigated in the first leaf of the sporophyte of the fern Dryopteris thelypteris and in the leaves of the jade plant Crassula argentae. In the fern leaf, stomates arise over a period of 1.1 days (26.4 hr) and attain a frequency of 0.186 of the epidermal cells while in the jade plant the formative period is over only 0.8 days (19.2 hr) and stomates reach a frequency of 0.090 of the epidermal cells. A computer model was devised to simulate the appearance of stomates by an induction process for a new stomate which then inhibits contiguous cells from becoming stomates. The validity of the model was demonstrated in that it gave values for stomate and stomate cluster frequencies upon calculating the frequency of free cells. For the fern leaf, the model required 57 iteration intervals, six iterations for the time from stomate induction until the adjacent cells were inhibited (a period of induction plus inhibitions), and an induction rate of 0.02 cells per time interval. From these theoretical values and the measured period of 26.4 hr when stomates arise during leaf development, a period of induction/inhibitions is calculated as 2.7 hr. In the model for the jade plant, 81 iteration intervals are required with six iterations per period of induction/inhibitions along with an induction rate of 0.002 cells per interation interval. These values give the duration for a period of induction/inhibitions in jade of 1.4 hr. This study describes the patterning process of stomate formation by an explicit, mathematical algorithm, and, from measurements of actual leaves, the various periods in the model can be assigned real time values.     

Exploring the pattern of phenotypic and genetic polymorphism in the arsenic hyperaccumulator Pteris vittata L. (Chinese brake fern)

Background and aims

The trace element uptake process of plants is a key factor in assessing the risks of trace element build-up in agricultural soils. Scarce information exists on the trace element dynamic uptake of plants grown in the field, especially on those potentially hazardous. In this study, the uptake process of As, Cd, Cu, and Zn in maize plants was quantified and characterized throughout the entire season.

Methods

Along two seasons, the uptake dynamics of field-grown maize plants in absorbing the soil borne trace elements was examined. Biomass production and the concentration of the elements in plant and soil solution samples were determined. A kinetic model was employed to characterize the uptake by plants.

Results

The kinetic parameters of the uptake process, maximum cumulative uptake rate, U _max , time to reach 50 % of U _max , t _U50 , and reciprocal of the uptake rate, b _U when followed throughout the season in terms of the plant’s growing degree days remained constant between seasons and were element specific. In spite of the large amount biomass produced, maize plants extracted minute quantities of Cd and As. Increasing cumulative uptake rates of As, Cd, Cu, and Zn from the soil took place primarily in the early half of the growing season when the biomass accumulation was still less than 50 % of the maximum harvested biomass. The element-specific plant uptake factor (PUF), which denote the partition of trace elements between the soil solution and plant phases, decreased following a first-order kinetics along the growing period, did not show any significant difference between seasons, and, at maturity stage, followed the sequence Cd≥Zn>Cu≥As.

Conclusions

The uptake process of the elements was adequately described by the kinetic model, showing similar patterns but different magnitude and distribution in the plant. The extraction of Cd and As by plants is low in comparison to common inputs through fertilizer applications into maize production systems, indicating potential risk of trace element accumulation in soils. The PUF may be estimated according to the kinetics parameters of the uptake process. On a per-unit-soil solution element basis, Cd and Zn would be more susceptible to the soil-to-plant transfer than As and Cu.     

Pharmacokinetics of chlorogenic acid and rutin in larvae of Heliothis zea

Studies using [³H]chlorogenic acid and [³H]rutin demonstrated that the kinetics of uptake of these plant phenolics into the haemolymph of 5th-instar Heliothis zea (Boddie) following actue oral administration is a first-order process. The total quantity of either phenolic present in the haemolymph within 1 hr amounts to 5% or less of the total ingested dose. Based on TLC analyses, 80% or more of the radioactivity in the haemolymph occurs as the parent phenolic. Retention of [³H]-chlorogenic acid or [³H]-rutin in H. zea following chronic feeding from 1st to 3rd-instar larvae is also linearly related to dietary dose. Chlorogenic acid and rutin are both equitoxic and equivalent in bioavailability to H. zea.Loss of [³H]-rutin from the haemolymph of 5th-instar larvae following injection is biphasic. One half of the injected dose is excreted in the frass in the first 6 hr after injection; the other half is thereafter eliminated at 1/20th of the initial rate. Analyses of extracts of frass by thin-layer chromatography indicate that after either chronic or acute feeding 90% of the ingested phenolic is excreted unchanged. Possible sites and modes of action of phenolics in insects are discussed in light of these findings.     

Rhizosphere colonization and arsenic translocation in sunflower (Helianthus annuus L.) by arsenate reducing Alcaligenes sp. strain Dhal-L

In the present study, six arsenic-resistant strains previously isolated were tested for their plant growth promoting characteristics and heavy metal resistance, in order to choose one model strain as an inoculum for sunflower plants in pot experiments. The aim was to investigate the effect of arsenic-resistant strain on sunflower growth and on arsenic uptake from arsenic contaminated soil. Based on plant growth promoting characteristics and heavy metal resistance, Alcaligenes sp. strain Dhal-L was chosen as an inoculum. Beside the ability to reduce arsenate to arsenite via an Ars operon, the strain exhibited 1-amino-cyclopropane-1-carboxylic acid deaminase activity and it was also able to produce siderophore and indole acetic acid. Pot experiments were conducted with an agricultural soil contaminated with arsenic (214 mg kg^?1). A real time PCR method was set up based on the quantification of ACR3(2) type of arsenite efflux pump carried by Alcaligenes sp. strain Dhal-L, in order to monitor presence and colonisation of the strain in the bulk and rhizospheric soil. As a result of strain inoculation, arsenic uptake by plants was increased by 53 %, whereas ACR3(2) gene copy number in rhizospheric soil was 100 times higher in inoculated than in control pots, indicating the colonisation of strain. The results indicated that the presence of arsenate reducing strains in the rhizosphere of sunflower influences arsenic mobilization and promotes arsenic uptake by plant.