Heliobacteria Reveal Fermentation As a Key Pathway for Mercury Reduction in Anoxic Environments.

The accumulation of mercury (Hg) in rice, a dietary staple for over half of the world's population, is rapidly becoming a global food safety issue. Rice paddies support the anaerobic production of toxic methylmercury that accumulates in plant tissue, however the microbial controls of Hg cycling in anoxic environments remain poorly understood. In this study, we reveal a novel reductive Hg metabolism in a representative of the family Heliobacteria ( Heliobacterium modesticaldum Ice1) that we confirm in model chemotrophic anaerobes. Heliobacteria served as our initial model because they are a family of spore-forming fermentative photoheterotrophs commonly isolated from terrestrial environments. We observed that H. modesticaldum reduced up to 75% of HgII under phototrophic or fermentative conditions. Fermentative HgII reduction relied on the ability of cells to oxidize pyruvate whereas phototrophic HgII reduction could be supported even in the absence of a carbon source. Inhibiting pyruvate fermentation eliminated HgII reduction in all chemotrophic strains tested, whereas phototrophic cells remained unaffected. Here we propose a non mer-operon dependent mechanism for Hg0 production in anoxic environments devoid of light where external electron acceptors are limited. These mechanistic details provide the foundation for novel bioremediation strategies to limit the negative impacts of Hg pollution.

Up-regulation of glutamate receptors is associated with LTP defects in the early stages of diabetes mellitus.

AIMS/HYPOTHESIS: Recent studies involving electrophysiology and immunolabelling indicate that short-term insulin treatment of hippocampal neurons in culture induces changes in glutamate receptor function, suggesting that this receptor system can be altered on a relatively rapid time scale during diabetic conditions. To investigate this hypothesis, we examined whether brain glutamate receptors and long-term potentiation are altered in the early stages of diabetes mellitus in non-obese diabetic mice, a genetic model of Type I (insulin-dependent) diabetes mellitus. METHODS: In vitro receptor autoradiography and immunoblotting were used to study the impact of diabetes on brain glutamate receptors. From an electrophysiological point of view, field potential recordings were also examined in area CA1 of hippocampal slices to determine the influence of diabetes on long-term potentiation. RESULTS: Quantitative autoradiographic analysis revealed enhanced 3H-glutamate binding to several brain regions of diabetes mice, with maximal increases in the cerebral cortex and hippocampus. Saturation kinetics within the cerebral cortex disclosed that this change of 3H-glutamate was possibly due to an increase in the maximal number of N-methyl- D-aspartate binding sites, an interpretation that was corroborated by Western blot analysis of N-methyl- D-aspartate 2A subunits. Impairment in the expression of hippocampal long-term potentiation was also observed in diabetic mice, while the failure to elicit synaptic potentiation was prevented by insulin treatment. CONCLUSION/INTERPRETATION: Because glutamate receptors are thought to be involved in several degenerative processes, our results suggest that up-regulation of these receptors in the early stages of diabetes could represent an important mechanism underlying neurological complications within the brain of diabetic patients.

Effects of bilateral olfactory bulbectomy on N-methyl-D-aspartate receptor function: autoradiographic and behavioral studies in the rat.

Rat bilateral olfactory bulbectomy (OBX) serves as a useful model in the study of depression and the mechanisms of action of antidepressant treatments. Considering the evidence of NMDA receptors involvement in depression, the present study was undertaken in order to investigate the time-course effects of OBX on the NMDA receptor function. Following bilateral olfactory bulbectomy, rats display an increase in locomotor activity and changes in other types of behavior in a novel environment. Autoradiographic experiments using the noncompetitive NMDA antagonist [(125)I]-iodo-MK-801 as the labeling agent showed that this increase in behavioral activities corresponds to a decrease in [(125)I]-iodo-MK-801 binding in a number of brain regions. In most regions, this reduction reached significance by the third week following OBX. However, in some cortical areas-a nucleus of the thalamus (AV) and one of the amygdala (LA)-this reduction was already significant in the first or second week following OBX and lasted throughout the 4 weeks of the study. We also compared the behavioral modifications induced by a challenge injection of MK-801 (0.2 mg/kg i.p.) in OBX and sham-operated rats. This challenge is known to induce hyperlocomotion and a number of stereotypies in naive rats. These effects were drastically reduced in OBX as compared to sham-operated rats. These data are consistent with the above-mentioned decrease in cerebral binding of MK-801 to NMDA receptors.

The incidence of hematuria in middle distance track running.

The purpose of this study was to establish if middle distance track athletes experience hematuria during their competitive season interval workouts and, if so, what type of workout based on intensity and distance was most associated with hematuria. During a 4-week observational period, athletes (n = 10) underwent reagent strip urinalysis before and after their twice weekly interval sessions. Positive samples for hematuria were analyzed microscopically to accurately determine red blood cell (RBC) loss. Seventy-one individual interval workouts were observed, of which 32 cases of hematuria were reported. Nine cases of hematuria exhibited >100 RBC per High Power Field (Hpf). Furthermore, 90% of the athletes experienced post-workout hematuria at least once. The highest incidence of hematuria was observed after workouts run at 110% of VO(2peak) over short (600-1,500 m) to moderate (1,501-3,000 m) distances. All post-exercise cases of hematuria resolved within 2 hr of recovery.

Affinities of venlafaxine and various reuptake inhibitors for the serotonin and norepinephrine transporters.

In vitro radioligand binding studies were carried out in rat brain membranes to assess the affinity of various reuptake inhibitors for the serotonin (5-hydroxytryptamine, 5-HT) and the norepinephrine transporters using the selective ligands [3H]cyanoimipramine and [3H]nisoxetine, respectively. The selective 5-HT reuptake inhibitors paroxetine, indalpine and fluvoxamine displayed a high affinity for the 5-HT transporter, whereas the norepinephrine reuptake inhibitor desipramine had a high affinity for the norepinephrine transporter. Duloxetine, a dual 5-HT and norepinephrine reuptake inhibitor, displayed a high affinity for both the 5-HT and the norepinephrine transporters. Interestingly, venlafaxine, a dual 5-HT and norepinephrine reuptake inhibitor, displayed only a moderate affinity for the 5-HT transporter (Ki = 74 nM) and a very low affinity for the norepinephrine transporter (Ki = 1.26 microM). The relatively low affinities of venlafaxine contrast with its potent in vivo 5-HT and norepinephrine reuptake blocking properties. These results raise the possibility that the in vivo effects on the 5-HT and norepinephrine reuptake observed with venlafaxine may not be mediated solely by its binding to the [3H]cyanoimipramine and [3H]nisoxetine binding sites.

Antidepressants reverse the olfactory bulbectomy-induced decreases in splenic peripheral-type benzodiazepine receptors in rats.

The present study investigated the effects of 21-day administration of clorgyline (1 mg/kg/day), desipramine (10 mg/kg/day) or paroxetine (10 mg/kg/day) on peripheral-type benzodiazepine receptors in rat peripheral tissues following bilateral olfactory bulbectomy. Thymus and spleen weights decreased as a result of bulbectomy. Subsequent antidepressant drug administration had no further effects on the weights of thymus glands but increased those of spleens. In thymus glands, higher densities of peripheral-type benzodiazepine receptors were observed in medulla than in cortex; no significant variations were observed following bulbectomy or antidepressant drug administration. In spleen, higher densities were observed in white pulp than in red pulp. The bulbectomy-induced decreases in binding densities observed in both regions were reversed following administration of antidepressants. Adrenal peripheral-type benzodiazepine receptors were not altered by bulbectomy or subsequent treatment with clorgyline or desipramine while paroxetine upregulated these receptors. No changes in kidney peripheral-type benzodiazepine receptors were observed. The present study confirms that cell lines of the rat immune system possess high densities of peripheral-type benzodiazepine receptor binding sites and further support the contention that, following olfactory bulbectomy, rats may present an antidepressant-reversible immunitary dysfunction.

Antidepressant-induced modulation of GABAA receptors and beta-adrenoceptors but not GABAB receptors in the frontal cortex of olfactory bulbectomised rats.

The effects of prolonged administration of antidepressant drugs, belonging to three different classes, on high-affinity GABAA receptor, GABAB receptor and beta-adrenoceptor binding parameters were determined in the frontal cortex of olfactory bulbectomised rats. Clorgyline (1 mg/kg/day), paroxetine (10 mg/kg/day) or desipramine (10 mg/kg/day) were administered for 21 days via subcutaneous osmotic minipumps implanted in the scapular region 7 days after bulbectomy. Cortical GABAA receptor densities, defined with [3H]gamma-aminobutyric acid ([3H]GABA), were significantly increased following bulbectomy. This effect on Bmax values was reversed by all three antidepressant drugs. GABAB receptor densities decreased slightly after bulbectomy. Chronic antidepressant administration had no effect on GABAB receptor binding parameters. Olfactory bulbectomy did not induce any changes in cortical beta-adrenoceptor binding parameters determined with [3H]CGP-12177 ((-)-4-(3-t- butylamino-2-hydroxypropxy)- [5,7-3H]benzimidazol-2-one). However, prolonged administration of all three antidepressant drugs induced a downregulation of beta-adrenoceptors. The results of the present study confirm the involvement of cortical GABAA rather than GABAB receptors in the olfactory bulbectomy animal model of human depression. Moreover, the data further support the hypothesis that a decrease in function of the GABAA receptor complex could play a role in the therapeutic effects of antidepressant treatments.

Quantitative autoradiographic studies of the effects of bilateral olfactory bulbectomy in the rat brain: central- and peripheral-type benzodiazepine receptors.

We investigated the discrete regional effects of bilateral olfactory bulbectomy on central- and peripheral-type benzodiazepine receptors in rat brains at weekly intervals until one month after bulb ablation. Persistent increases in [3H]flunitrazepam binding to central benzodiazepine receptors were observed in the cingulum (27%) and in the frontal (15%) and parietal (14%) cortices. Progressive increases in central benzodiazepine receptors, reaching statistical significance four weeks after olfactory bulbectomy, were observed in the ventromedial thalamic nucleus (35%), the lateral hypothalamic region (22%), the basolateral amygdaloid nucleus (23%) and substantia nigra (25%). Persistent major increases (between four- and six-fold) in [3H]PK-11195 [eH]1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinoline carboxamide binding to peripheral-type benzodiazepine receptors were observed in all anterior olfactory nuclei. Similarly, throughout the time period studied, [3H]PK-11195 binding densities were increased two- to three-fold in the piriform cortex and lateral olfactory tract. These observations confirm the usefulness of [3H]PK-11195 binding as a marker of neuronal insult in the brain. Moreover, the persistent regional increases in [3H]flunitrazepam binding to central-type benzodiazepine receptors suggest that GABAergic transmission is altered following olfactory bulb ablation.

Differential effects of olfactory bulbectomy on GABAA and GABAB receptors in the rat brain.

GABAergic mechanisms have been implicated in the bilateral olfactory bulbectomy (OBX) animal model of depression, where GABAB receptor binding sites have been shown to decrease markedly at specific time points after OBX. However, as no detailed time course of events has been determined, the present study investigated the effects of OBX on high-affinity GABAA, GABAB, beta-adrenergic, and benzodiazepine receptor binding parameters in membrane preparations from rat brain regions at weekly intervals (1-4 weeks) after OBX. Persistent significant increases (40-60%) in Bmax values of high affinity GABAA receptors were observed in the frontal cortex throughout the period investigated following OBX. Bmax values in the hippocampus increased significantly after 1 week (53%) but were not statistically significant thereafter. No changes in GABAA binding parameters were observed in the hypothalamus or cerebellum. Conversely, GABAB receptor densities were significantly decreased in the frontal cortex after 1 (-38%) and 2 (-41%) weeks and moderately decreased 3 and 4 weeks (-27 and -23%, respectively) after OBX, while in the cerebellum they were significantly increased after 1 week (96%) and returned to sham-operated levels by 3 weeks. No changes in GABAB receptor binding parameters were observed in the hippocampus or hypothalamus. Binding parameters for benzodiazepine receptor binding sites or beta-adrenoceptors were not modified throughout the time course. GABAergic transmission, reflected by changes in GABAA and GABAB receptor density in the frontal cortex, may be altered in OBX rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption and assimilation of nitrate and ammonium ions by jack pine seedlings.

Jack pine (Pinus banksiana Lamb.) seedlings were grown in a shaded or unshaded light regime with either NO(3) (-)- or NH(4) (+)-N as the sole N source. After three months, seedlings grown with NH(4) (+)-N were larger than seedlings grown with NO(3) (-)-N. Irradiance had a greater effect on growth of ammonium-fed seedlings than on growth of nitrate-fed seedlings.At all times from 6 to 24 h following incorporation of (15)N, soluble, insoluble, and total (15)N contents of shoots and roots were higher in ammonium-fed seedlings than in nitrate-fed seedlings. The pattern of (15)N accumulation in shoots was similar to that in roots. After 6 and 24 h of (15)N incorporation, unshaded, ammonium-fed seedlings had 8.8 and 2.8 times greater total (15)N contents, respectively, than unshaded, nitrate-fed seedlings. In response to shading, ammonium-fed seedlings increased their total uptake of (15)N per unit root weight, whereas nitrate-fed seedlings did not. No nitrate or (15)NO(3) (-) was detected in any plant tissue. Nitrate-fed plants had higher NH(4) (+), Asp, and Gln concentrations in needles and higher gamma-aminobutyric acid and Arg concentrations in stems. Accumulation of (15)N in roots was not affected by the pH of the (15)N solution or by the N source fed to the seedlings before the period of (15)N incorporation. Thus NO(3) (-) transport into roots, rather than its reduction or transport within the plant, seems to be the factor limiting the growth of jack pine supplied with NO(3) (-)-N as the sole N source.

Type A behavior, aerobic power, and cardiovascular recovery from a psychosocial stressor.

Fifty-six male university students were tested for actual aerobic power and Type A behavior (Form T of the Jenkins Activity Survey) before performing the Stroop Color-Word Interference Test. Neither fitness nor Type A was related to magnitude of heart rate (HR) increase, and only Type A was related to speed of recovery following the test, with Type As being slower to return to baseline HR levels than Type Bs. These results are discussed in terms of the need to consider factors that mediate speed of recovery from stress.

Anaerobic testing using the Wingate and Evans-Quinney protocols with and without toe stirrups.

Recently, considerable attention has been focused on the measurement of maximal anaerobic power. The Evans-Quinney (1981) protocol which considers leg volume as well as body weight in establishing optimal load settings on the bicycle ergometer has been shown to result in significantly higher anaerobic power outputs than the body weight-relative Wingate protocol. The purpose of this investigation was to compare the anaerobic power outputs during bicycle ergometer work using the Wingate and Evans-Quinney protocols with and without toe stirrups. Fifty male physical education and varsity athletes (average age 22.6 years) volunteered to participate in this experiment. All subjects performed a total of four maximal 30-second anaerobic power tests utilizing the force settings established by the Wingate (load [L] = 0.075 kp/kg body weight) and Evans-Quinney (L = -0.4914-0.2151 (weight, kg) + 2.1124 (leg volume, litre) protocol. The subjects were randomly assigned to one of four counterbalanced orders of test administration. Analysis of data indicated significant differences (P less than or equal to .05) among the variables of: 5-second peak power (W), 30-second anaerobic capacity (W) and percent fatigue as a function of test protocol. Significantly higher values were obtained for all variables under the Evans-Quinney test procedure using toe stirrups while the Wingate test procedure without toe stirrups showed significantly lower values for all variables. It was concluded that the Evans-Quinney load setting protocol with toe stirrups resulted in significantly higher power measures than any of the other treatments tested.

Maleate-induced bicarbonaturia in the dog: a carbonic anhydrase-independene effect.

Studies were performed to characterize the renal effects of maleate in anesthetized dogs. Following the intravenous administration of maleate or maleic acid (50 mg/kg), mean fractional bicarbonate excretion (CHCO3/GFR) rose to as high as 26%. Na, K, and phosphate excretion also increased markedly, whereas C1 excretion remained low. An initial transient fall in urinary pH from 6.53 to 6.13 contrasted sharply with the rapid alkalinization of the urine induced by acetazolamide administration. During saline expansion CHCO3/GFR rose from 4 to 37% after maleate administration, whereas Cl excretion did not change significantly. During continuous carbonic anhydrase inhibition with acetazolamide, maleate administration resulted in a further rise in CHCO3/GFR from 22 to 35%. Whereas CPO4/GFR increased only from 1 to 3% during acetazolamide administration, this ratio reached 75% following the addition of maleate. Fumarate, the transisomer of maleate, and malonate, a well-known inhibitor of Krebs cycle, failed to affect bicarbonate excretion. This study demonstrates that maleate inhibits the fraction of bicarbonate reabsorption uncatalyzed by carbonic anhydrase. Impaired anionic reabsorption of bicarbonate or accelerated passive backflux of this ion into proximal tubular lumen are the two mechanisms that best explain the bicarbonaturia induced by maleate.