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Induction time measurement of barbital tolerance in mice given recurrent subhypnotic dosage of barbital.

A procedure designed to estimate the degree of development of neuronal tolerance to barbiturates is described. With this technique, it has been shown that a subhypnotic (125 mg/kg) peritoneal dose of barbital sodium, given twice daily, over the course of either five or 10 consecutive days, can produce significant elevation in tolerance over that found for analogous groups of saline control animals. Mean tolerances were considerably lower (about 20 percent) than those obtained when the recurrent barbital sodium dosages were at the hypnotic (300 mg/kg) level given once per day for 10 days. The results obtained support the proposition that the method employed is sufficiently sensitive to estimate neuronal tolerance developed as a result of recurrent administrations of subhypnotic dosages of barbiturates when given at adequate frequency and duration.

Animals↗

Associative control of tolerance to the sedative effects of a short-acting benzodiazepine.

The role of Pavlovian conditioning in tolerance to the depressant effect of a benzodiazepine (midazolam) on the ambulatory activity of rats was examined. The depression of activity by low doses (1.0 and 4.0 mg/kg, ip) of midazolam diminished quickly over repeated doses given at 48-hr intervals (Experiment 1). Equivalent tolerance was observed in groups measured at 2 min and 30 min after drug injection. When challenged with saline, however, drug-tolerant animals tested immediately after injection were hyperactive in comparison with nontolerant controls, whereas equivalent groups tested 30 min after injection were not. A second context was designed, and its discriminability from the original was established by assessing context-specific suppression of activity following exposure to mild electric shock (Experiment 2). In Experiment 3A, although tolerant animals tested in the drug-associated context remained fully tolerant, a second group demonstrated a complete loss of tolerance when given the drug in a saline-associated context. Both groups were fully tolerant when tested again in the drug-associated context after 14 drug-free days. In Experiment 3B, tolerance was significantly reduced by 14 extinction exposures to the drug-associated environment without the drug. These results are uniquely predicted by associative models of drug tolerance and may have implications for the clinical use of this class of drugs.

Animals↗

Transgenic tobacco plants overexpressing glyoxalase enzymes resist an increase in methylglyoxal and maintain higher reduced glutathione levels under salinity stress.

The mechanism behind enhanced salt tolerance conferred by the overexpression of glyoxalase pathway enzymes was studied in transgenic vis-à-vis wild-type (WT) plants. We have recently documented that salinity stress induces higher level accumulation of methylglyoxal (MG), a potent cytotoxin and primary substrate for glyoxalase pathway, in various plant species [Yadav, S.K., Singla-Pareek, S.L., Ray, M., Reddy, M.K. and Sopory, S.K. (2005) MG levels in plants under salinity stress are dependent on glyoxalase I and glutathione. Biochem. Biophys. Res. Commun. 337, 61-67]. The transgenic tobacco plants overexpressing glyoxalase pathway enzymes, resist an increase in the level of MG that increased to over 70% in WT plants under salinity stress. These plants showed enhanced basal activity of various glutathione related antioxidative enzymes that increased further upon salinity stress. These plants suffered minimal salinity stress induced oxidative damage measured in terms of the lipid peroxidation. The reduced glutathione (GSH) content was high in these transgenic plants and also maintained a higher reduced to oxidized glutathione (GSH:GSSG) ratio under salinity. Manipulation of glutathione ratio by exogenous application of GSSG retarded the growth of non-transgenic plants whereas transgenic plants sustained their growth. These results suggest that resisting an increase in MG together with maintaining higher reduced glutathione levels can be efficiently achieved by the overexpression of glyoxalase pathway enzymes towards developing salinity stress tolerant plants.

Antioxidants↗

Morphine analgesia, tolerance and addiction in the cricket Pteronemobius sp. (Orthoptera, Insecta).

The escape reaction time (ERT) of the cricket Pteronemobius sp. from the heated box begins at 48 degrees and increases with temperature until 56 degrees C, beyond which there is no further increase. The ERT (2.2 +/- 1.39 s) from the hot box at 54 degrees C is used as a model for studying the analgesic effects of opiates. Results of the present paper show that the ERT did not change after injecting the insect in the abdominal haemocoel with 0.9% saline solution, but ERT increased when 0.32, 0.52 or 0.69 mg/g of morphine is injected in the same place. The maximum ERT increase is reached at 90 min after drug injection, and the drug effect disappears 3 h after the injection. At 90 min after drug injections, the dose of 0.50 mg/g of morphine produces 50% of ERT increase, and it is referred to as the median analgesic dose (D50). 1.05 mg/g of morphine produces an ERT longer than 30 s that results in an irreversible damage to the insect. Sixty-four micrograms/g of naloxone given in addition to D50 of morphine fully blocked the effect of morphine during its 3-h action. However, more than 64 micrograms/g of naloxone alone also increase the ERT in the cricket, similar to what has been described for vertebrates. Four daily morphine injections of D50 decreased ERT in such a way that, at the fourth day, the ERT is similar to the ERT produced by saline solution; i.e., tolerance is shown. The suppression of daily morphine injections of D50 during the fifth day produced a hyperresponse to vibration (big jumps) not shown in the case of the injections of saline solution; i.e., addiction is shown.

Analgesia↗

A comparison of sputum induction methods: ultrasonic vs compressed-air nebulizer and hypertonic vs isotonic saline inhalation.

Airway inflammation can be demonstrated by the modem method of sputum induction using ultrasonic nebulizer and hypertonic saline. We studied whether compressed-air nebulizer and isotonic saline which are commonly available and cost less, are as effective in inducing sputum in normal adult subjects as the above mentioned tools. Sixteen subjects underwent weekly sputum induction in the following manner: ultrasonic nebulizer (Medix Sonix 2000, Clement Clarke, UK) using hypertonic saline, ultrasonic nebulizer using isotonic saline, compressed-air nebulizer (BestNeb, Taiwan) using hypertonic saline, and compressed-air nebulizer using isotonic saline. Overall, the use of an ultrasonic nebulizer and hypertonic saline yielded significantly higher total sputum cell counts and a higher percentage of cell viability than compressed-air nebulizers and isotonic saline. With the latter, there was a trend towards squamous cell contaminations. The proportion of various sputum cell types was not significantly different between the groups, and the reproducibility in sputum macrophages and neutrophils was high (Intraclass correlation coefficient, r [95%CI]: 0.65 [0.30-0.91] and 0.58 [0.22-0.89], p < 0.001). Overall changes in median FEV, were small and comparable between all groups. Induction using ultrasonic nebulizers together with hypertonic saline was generally less well tolerated than compressed-air nebulizers and isotonic saline. We conclude that in normal subjects, although both nebulizers and saline types can induce sputum with reproducible cellular profile, ultrasonic nebulizers and hypertonic saline are more effective but less well tolerated.

Administration, Inhalation↗

Strains of the heterotrophic flagellate Bodo designis from different environments vary considerably with respect to salinity preference and SSU rRNA gene composition.

The morpho species Bodo designis is widespread and abundant globally in highly contrasting terrestrial and aquatic ecosystems. Whether the forms of Bodo designis from contrasting environments are conspecific, i.e. largely genetically identical, or whether they merely share the external morphology is presently not known. We examined the ability of different strains of Bodo designis isolated from different environments at different geographical sites to survive and grow at a salinity range of 0.5-45%. The Bodo designis strains from marine, freshwater, and terrestrial environments showed a different ability to cope with altered physiological conditions. Most of the tested strains were only able to tolerate a small salinity range, whereas others were able to withstand all tested salinity levels. We further examined the phylogenetic relationship between the different strains by sequencing the small subunit (SSU) rRNA gene. The resulting phylogenetic analyses suggest a huge genetic variation within Bodo designis, and also imply that Dimastigella and Rhyncomonas are developed inside Bodo designis. If the biological species concept is used, the genetic differences as well as the physiological barriers between the different strains of Bodo designis, would suggest that they should be assigned to different species.

Animals↗

Crop and irrigation management strategies for saline-sodic soils and waters aimed at environmentally sustainable agriculture.

Irrigation has long played a key role in feeding the expanding world population and is expected to play a still greater role in the future. As supplies of good-quality irrigation water are expected to decrease in several regions due to increased municipal-industrial-agricultural competition, available freshwater supplies need to be used more efficiently. In addition, reliance on the use and reuse of saline and/or sodic drainage waters, generated by irrigated agriculture, seems inevitable for irrigation. The same applies to salt-affected soils, which occupy more than 20% of the irrigated lands, and warrant attention for efficient, inexpensive and environmentally acceptable management. Technologically and from a management perspective, a couple of strategies have shown the potential to improve crop production under irrigated agriculture while minimizing the adverse environmental impacts. The first strategy, vegetative bioremediation--a plant-assisted reclamation approach--relies on growing appropriate plant species that can tolerate ambient soil salinity and sodicity levels during reclamation of salt-affected soils. A variety of plant species of agricultural significance have been found to be effective in sustainable reclamation of calcareous and moderately sodic and saline-sodic soils. The second strategy fosters dedicating soils to crop production systems where saline and/or sodic waters predominate and their disposal options are limited. Production systems based on salt-tolerant plant species using drainage waters may be sustainable with the potential of transforming such waters from an environmental burden into an economic asset. Such a strategy would encourage the disposal of drainage waters within the irrigated regions where they are generated rather than exporting these waters to other regions via discharge into main irrigation canals, local streams, or rivers. Being economically and environmentally sustainable, these strategies could be the key to future agricultural and economic growth and social wealth in regions where salt-affected soils exist and/or where saline-sodic drainage waters are generated.

Agriculture↗

Evaluation of anorectal functions of children with anorectal malformations using fecoflowmetry.

BACKGROUND/PURPOSE: A simple and objective method evaluating the bowel functions of patients with anorectal anomalies is necessary. The authors applied fecoflowmetry and saline enama test for patients with anorectal anomalies to evaluate the fecodynamics and anorectal motilities in these children. METHODS: The bowel functions of 16 patients who underwent repair for anorectal malformations and 5 normal controls were evaluated by saline enema test and fecoflowmetry. The correlations between the clinical scores for the bowel functions and the parameters in fecodynamic studies were investigated. RESULTS: Seven of 16 patients exhibited periodical contractions of the rectum synchronized with relaxations of the anal canal during saline infusion as did the controls and had significantly higher clinical scores than the other patients. Two patients with severe chronic constipation lacked rectal contractions. Among the fecoflowmetric parameters, the maximum flow, average flow, and tolerable volume of saline infused into the rectum were significantly lower in the patients with low clinical scores than those of the controls. The maximal squeeze pressure and resting anal pressure were not significantly different between the patients and controls. CONCLUSION: Fecodynamic studies, such as fecoflowmetry and saline enema test, help in obtaining clinical indicators for the bowel functions of patients with anorectal anomalies.

Adolescent↗

Vasopressin secretion in the DIDMOAD (Wolfram) syndrome.

The diabetes insipidus which accompanies the DIDMOAD (Wolfram) syndrome is thought to be hypothalamic in origin, though no formal study of vasopressin secretion in the syndrome has been published, and some data in the literature suggest a renal tubular defect. We have studied vasopressin secretion in seven patients with the Wolfram/DIDMOAD syndrome during three dynamic stimuli: an osmotic stimulus (hypertonic saline infusion), hypoglycaemia (insulin tolerance test) and a baroregulatory stimulus (trimetaphan infusion). Hypertonic saline infusion demonstrated three patients to have complete and four to have partial hypothalamic diabetes insipidus; administration of (per nasal) desmopressin excluded nephrogenic diabetes insipidus in all seven patients. Insulin hypoglycaemia failed to stimulate vasopressin release, but trimetaphan-induced hypotension produced significant though subnormal rises in plasma vasopressin in three patients with partial diabetes insipidus, though it produced a negligible rise and no rise in plasma vasopressin in two patients with complete diabetes insipidus. The data suggest a much greater frequency of hypothalamic diabetes insipidus in the Wolfram/DIDMOAD syndrome than is reported, but did not identify nephrogenic diabetes insipidus. The absence of vasopressin responses to non-osmotic stimuli in patients with complete diabetes insipidus suggests global lack of vasopressin secreting neurones, rather than an isolated osmoreceptor defect or selective vasopressin secreting neuronal loss, as the lesion producing diabetes insipidus in the DIDMOAD syndrome.

Adult↗

An investigation of the negative-reinforcing properties of naloxone and cyclazocine in squirrel monkeys.

Monkeys were trained to emit 10 responses to terminate a schedule complex consisting of a light and masking noise in the presence of which brief, intravenous injections of naloxone (0.01 mg/kg per injection), dl-cyclazocine (0.003, 0.01 mg/kg per injection), or saline were scheduled to occur over blocks of consecutive daily sessions. Completion of the fixed ratio (FR 10) response requirement terminated the schedule complex, producing a time-out of 60 s in which responses were without consequence and injections did not occur. Sessions lasted for 100 min or ended after 100 injections, whichever came first. When monkeys received an intramuscular injection of morphine (10 mg/kg) 60 min preceding each session, responding to avoid injections of naloxone or cyclazocine was well maintained, though morphine-dependent monkeys tolerated injections of saline or the vehicle in which cyclazocine was dissolved. When the morphine pretreatment regimen was discontinued, monkeys tolerated injections of naloxone and cyclazocine though, at the larger cyclazocine dose (0.01 mg/kg per injection), responding was maintained for at least 30 consecutive sessions in two postdependent animals, but not in two other monkeys treated identically. The capacity of these morphine-antagonist drugs to support responding that avoids their injection depends evidently upon whether monkeys are rendered morphine dependent or not; in the case of cyclazocine, however, an increased sensitivity to the negative-reinforcing effects of the drug appears to persist in some "postdependent" monkeys.

Animals↗

Treatment of murine lupus with F(ab')2 fragments of monoclonal antibody to L3T4. Suppression of autoimmunity does not depend on T helper cell depletion.

Treatment with mAb to the L3T4 Ag on Th cells can inhibit autoimmunity in mice. However, the mechanism by which anti-L3T4 inhibits autoimmunity is not known. In these studies, lupus-prone NZB/NZW F1 (B/W) mice were treated with F(ab')2 fragments of mAb to L3T4 to determine whether Th cell depletion is required for the beneficial effects of anti-L3T4. We first showed that treatment of female B/W mice with F(ab')2 anti-L3T4 from age 5 to 9 mo significantly reduced autoantibody production without depleting L3T4+ cells. However, treatment was complicated by the development of a host immune response to the rat mAb fragments. To circumvent this problem, female B/W mice were treated with a single high-dose of intact rat mAb to L3T4 (GK1.5) at age two mo. to induce immune tolerance to the mAb. Then, after recovery of L3T4+ cells, the mice were treated from age four to 14 mo with either F(ab')2 anti-L3T4 (0.5 mg 3 times per wk), intact anti-L3T4, or saline. In mice tolerized by this regimen, neither the F(ab')2 rat mAb nor the intact rat mAb elicited a host response. The mAb fragments bound target Ag but did not deplete the Th cells, whereas intact mAb to L3T4 profoundly depleted the L3T4+ cells. Despite this difference, both therapies had the same substantial beneficial effects on autoimmunity. They significantly decreased anti-DNA Ab production, improved renal function and prolonged survival. The initial tolerizing dose, by itself, did not inhibit autoimmunity. These findings show that anti-L3T4 suppresses autoimmunity by directly altering Th cell function through the L3T4 Ag, and not solely by depleting Th cells. They also document the detrimental effects of the host immune response to therapy with anti-L3T4 mAb, and they demonstrate a new strategy by which this response may be prevented.

Animals↗

Cardiovascular effects of infused adenosine in man: potentiation by dipyridamole.

In a single blind randomized study, eight normal subjects (23-40 years) received on two separate days adenosine as a constant i.v. infusion with doses increasing from 0.005-0.1 mg kg-1 min-1 following either i.v. dipyridamole 0.4 mg kg-1 per 10 min or a corresponding volume of saline. Heart rate, blood pressure and skin temperature were measured. Following saline all subjects tolerated adenosine 0.07 mg kg-1 min-1, six also tolerated 0.09 mg kg-1 min-1. Both heart rate and skin temperature increased with adenosine in a dose-related manner. Thus, adenosine 0.09 mg kg-1 min-1 was associated with an increase in heart rate (mean +/- SD) from baseline before saline with 16 +/- 10 b.p.m. (P less than 0.01) and an increase in skin temperature with 1.3 +/- 0.8 degrees C (P less than 0.02). Dipyridamole, which inhibits the cellular uptake of adenosine was associated with a change in heart rate similar to that induced by adenosine. Furthermore, when adenosine was infused following dipyridamole the changes in heart rate and skin temperature were potentiated as compared with adenosine following saline. Thus, following dipyridamole an increase in heart rate with 15 b.p.m. above baseline was obtained with 0.005 mg kg-1 min-1 of adenosine as compared with 0.08-0.09 mg kg-1 min-1 of adenosine following saline. Blood pressure did not change in any of the studies. Exogenous adenosine in man has a dose-related positive chronotropic effect and a local dilatory effect in the skin microcirculation. Dipyridamole potentiates the cardiovascular effects of infused adenosine in man.

Adenosine↗

The role of ethanol in diluents of drugs that protect mice from hypoxia.

This study evaluates the hypothesis that ethanol alone, or in diluents for drugs used to protect hypoxic mice, is responsible in part for an increased tolerance to hypoxia (4-5% oxygen). The change in hypoxic tolerance following i.v. or i.p. administration of ethanol, diazepam, nimodipine and various diluent components was measured. Diazepam (50 mg/kg i.v.) increased hypoxic tolerance to 700 +/- 47% (n = 11) of saline control, its diluent increased hypoxic tolerance to 468 +/- 60% (n = 10) of saline control but the ethanol component of the diluent accounted for almost half of this diluent effect. Nimodipine (2 mg/kg i.p.), a calcium antagonist, increased tolerance to 180 +/- 18% of control (n = 19) and nimodipine diluent showed an even greater increase to 226 +/- 25% of control (n = 15). In this case essentially all of the protective effect of nimodipine diluent (81.3%) is accounted for by ethanol. Dose response curves indicate the maximum ethanol induced increase in hypoxic tolerance was approximately 335% of control at a dose of 2.4 g/kg. Buffers, etc. in the diluents evidently add to the protective effect of ethanol. Our data clearly indicate ethanol is the important component of some treatments which protect mice from hypoxia. The pharmacological activity of ethanol, even when used in a diluent, should not be ignored in evaluating therapeutic intervention for protection from hypoxia.

Animals↗

Photosynthetic responses of the tropical spiny shrub Lycium nodosum (Solanaceae) to drought, soil salinity and saline spray.

Water relations and photosynthetic characteristics of plants of Lycium nodosum grown under increasing water deficit (WD), saline spray (SS) or saline irrigation (SI) were studied. Plants of this perennial, deciduous shrub growing in the coastal thorn scrubs of Venezuela show succulent leaves which persist for approx. 1 month after the beginning of the dry season; leaf succulence is higher in populations closer to the sea. These observations suggested that L. nodosum is tolerant both to WD and salinity. In the glasshouse, WD caused a marked decrease in the xylem water potential (psi), leaf osmotic potential (psi(s)) and relative water content (RWC) after 21 d; additionally, photosynthetic rate (A), carboxylation efficiency (CE) and stomatal conductance (gs) decreased by more than 90 %. In contrast, in plants treated for 21 d with a foliar spray with 35 per thousand NaCl or irrigation with a 10 % NaCl solution, psi and RWC remained nearly constant, while psi(s) decreased by 30 %, and A, CE and gs decreased by more than 80 %. An osmotic adjustment of 0.60 (SS) and 0.94 MPa (SI) was measured. Relative stomatal and mesophyll limitations to A increased with both WD and SS, but were not determined for SI-treated plants. No evidence of chronic photoinhibition due to any treatment was observed, since maximum quantum yield of PSII, Fv/Fm, did not change with either drought in the field or water or salinity stress in the glasshouse. Nevertheless, WD and SI treatments caused a decrease in the photochemical (qP) and an increase in the non-photochemical (qN) quenching coefficients relative to controls; qN was unaffected by the SS treatment. The occurrence of co-limitation of A by stomatal and non-stomatal factors in plants of L. nodosum may be associated with the extended leaf duration under water or saline stress. Additionally, osmotic adjustment may partly explain the relative maintenance of A and gs in the SS and SI treatments and the tolerance to salinity of plants of this species in coastal habitats.

Disasters↗

Tolerance of mannitol-accumulating transgenic wheat to water stress and salinity.

Previous work with model transgenic plants has demonstrated that cellular accumulation of mannitol can alleviate abiotic stress. Here, we show that ectopic expression of the mtlD gene for the biosynthesis of mannitol in wheat improves tolerance to water stress and salinity. Wheat (Triticum aestivum L. cv Bobwhite) was transformed with the mtlD gene of Escherichia coli. Tolerance to water stress and salinity was evaluated using calli and T(2) plants transformed with (+mtlD) or without (-mtlD) mtlD. Calli were exposed to -1.0 MPa of polyethylene glycol 8,000 or 100 mM NaCl. T(2) plants were stressed by withholding water or by adding 150 mM NaCl to the nutrient medium. Fresh weight of -mtlD calli was reduced by 40% in the presence of polyethylene glycol and 37% under NaCl stress. Growth of +mtlD calli was not affected by stress. In -mtlD plants, fresh weight, dry weight, plant height, and flag leaf length were reduced by 70%, 56%, 40%, and 45% compared with 40%, 8%, 18%, and 29%, respectively, in +mtlD plants. Salt stress reduced shoot fresh weight, dry weight, plant height, and flag leaf length by 77%, 73%, 25%, and 36% in -mtlD plants, respectively, compared with 50%, 30%, 12%, and 20% in +mtlD plants. However, the amount of mannitol accumulated in the callus and mature fifth leaf (1.7-3.7 micromol g(-1) fresh weight in the callus and 0.6-2.0 micromol g(-1) fresh weight in the leaf) was too small to protect against stress through osmotic adjustment. We conclude that the improved growth performance of mannitol-accumulating calli and mature leaves was due to other stress-protective functions of mannitol, although this study cannot rule out possible osmotic effects in growing regions of the plant.

In Vitro Techniques↗

Photosynthetic capacity is related to the cellular and subcellular partitioning of Na+, K+ and Cl- in salt-affected barley and durum wheat.

The capacity of plants to tolerate high levels of salinity depends on the ability to exclude salt from the shoot, or to tolerate high concentrations of salt in the leaf (tissue tolerance). It is widely held that a major component of tissue tolerance is the capacity to compartmentalize salt into safe storage places such as vacuoles. This mechanism would avoid toxic effects of salt on photosynthesis and other key metabolic processes. To test this, the relationship between photosynthetic capacity and the cellular and subcellular distribution of Na+, K+ and Cl- was studied in salt-sensitive durum wheat (cv. Wollaroi) and salt-tolerant barley (cv. Franklin) seedlings grown in a range of salinity treatments. Photosynthetic capacity parameters (Vcmax, Jmax) of salt-stressed Wollaroi decreased at a lower leaf Na+ concentration than in Franklin. Vacuolar concentrations of Na+, K+ and Cl- in mesophyll and epidermal cells were measured using cryo-scanning electron microscopy (SEM) X-ray microanalysis. In both species, the vacuolar Na+ concentration was similar in mesophyll and epidermal cells, whereas K+ was at higher concentrations in the mesophyll, and Cl- higher in the epidermis. The calculated cytoplasmic Na+ concentration increased to higher concentrations with increasing bulk leaf Na+ concentration in Wollaroi compared to Franklin. Vacuolar K+ concentration was lower in the epidermal cells of Franklin than Wollaroi, resulting in higher cytoplasmic K+ concentrations and a higher K+ : Na+ ratio. This study indicated that the maintenance of photosynthetic capacity (and the resulting greater salt tolerance) at higher leaf Na+ levels of barley compared to durum wheat was associated with the maintenance of higher K+, lower Na+ and the resulting higher K+ : Na+ in the cytoplasm of mesophyll cells of barley.

Chlorides↗

[Sequence analysis of bacterial transposon in NHX gene of Populus euphratica].

The United Nations Environment Program estimates that approximately 20% of agricultural land and 50% of cropland in the world is salt-stressed. The gene NHX (Na+/H+ exchanger) encodes functional protein that catalyzes the countertransport of Na+ and H+ across membranes and may play an important role in plant salt tolerance. To clone the NHX from the wild plant Populus euphratica collected in Tarim basin and Xinjiang Wujiaqu district into a T-vector, designed primer was used to amplify 1kb NHX cDNA fragment with RT-PCR. Total RNA was extracted from Populus euphratica tissue (plant tissue was collected from Tarim basin and Xinjiang Wujiaqu district and stored in liquid nitrogen) according to the Plant RNA Mini Kits of Omega. First cDNAs were synthesized from 1 microg total RNA of Populus euphratica seedling. A pair of primers were used to perform RT-PCR. The amplified DNA fragment was purified and cloned into pMD18-T vector. However, 1kb and 2.3kb fragment were obtained from Tarim basin and Xinjiang Wujiaqu district and named as PtNHX and PwNHX, respectively. Sequence analysis reveals that the cloned PtNHX fragment of Populus euphratica contains partial NHX coding region with 98%, 86%, 84% and 80% identity comparing with Atriplex gemelini, Suaeda maritima, Arabidopsis thaliana and Oryza sativa, respectively. This analysis suggests that NHX gene would be highly conserved in terms of evolution in plant; and it also suggests that the NHX gene of Populus euphratica also would have the similarity with that of Arabidopsis. It may be of great importance in improvement of the plant salt tolerance and breed of crop. At the same time, sequence analysis shows that PwNHX gene includes a coding region about 1350bp with 99% identity comparing with transposon Tn10 IS10-left transposase of Shigella flexneri. On the one hand, the NHX gene may lose its function because it was inserted a fragment in coding region. On the other hand, its product may play a important role in salt tolerance. Populus grow in saline soil. It speculates that it may have other salt tolerance mechanism in Populus. The transposon can be used as transposon tagging to clone other genes and it will help us to understand farther the salt tolerance mechanism.

Amino Acid Sequence↗

Systemic immunization of mature mice by the oral route.

1. Mice of several strains which are susceptible to the induction of oral tolerance by a single gavage with 20 mg of ovalbumin (Ova) when young adults (7-8 weeks old) become less susceptible or refractory to tolerance induction when mature (20-40 weeks old). The antibody-forming capacity of these mature animals remains invariant compared to young adults (8-10 weeks old). 2. Mature mice of several strains display significant serum antibody responses to 3 gavages (days 0, 7 and 28) with Ova; as assessed by ELISA titers, these responses are similar in magnitude to those elicited by standard ip immunization with small doses of Ova. 3. In mature H-III mice, gavages on days 0, 7 and 28 induced significant antibody formation. On the contrary, the ingestion of the same amounts of Ova on days 0, 7 and 28 induced oral tolerance. Concomitant gavage with saline on the days of Ova ingestion failed to inhibit tolerance induction. 4. In H-III mice displaying circulating antibodies induced by repeated gavage with Ova, additional ip injections of Ova failed to increase the antibody titers.

Administration, Oral↗