[Regulation of androgen on singing behavior and song control system of songbirds].

Androgen plays an important role in singing of songbirds. Recent studies have shown that androgen levels in vivo not only affect the external morphology of songbirds, but also affect their singing behavior. Androgens (including derivatives) affect singing behavior and singing system in many ways. Based mainly on the results from our research group in the zebra finch, this review summarizes the effects of androgen on singing behavior, excitability and synaptic transmission of projection neurons of singing system, and the interaction of androgen with other neurotransmitter receptors in the brain of songbirds.

[Central cholinergic system innervates song control system of songbirds and regulates song behavior].

Songbird has become an ideal model for studying motor learning due to its unique learned song behavior. It has been proved that song behavior is directly regulated by song control system in the forebrain of songbirds. There are lines of evidence to show that cholinergic transmitters and their receptors are distributed in song control system, and vocal control nuclei in song control system are innervated by cholinergic nerves from the central cholinergic system in basal forebrain, which can affect activities of vocal control nuclei through cholinergic transmitters, and then affect song behavior. Studies in mammals have confirmed that the central cholinergic system is involved in the regulation of motor behavior and neural process of motor learning. Elucidation of regulation of songbirds' song behavior by central cholinergic system would shed light on the neural mechanism of song motor control and song learning and memory in songbirds, and provide theoretical insights for researches on other animals' sensorimotor processes and human language learning. This review summarized recent progresses, including the research work of our laboratory, in the studies on the selectivity of cholinergic transmitters to their receptors and their effects on neuronal activities in vocal control nuclei of songbirds and provided valuable clues for revealing the regulation mechanism of central cholinergic system on songbirds' song behavior.

[Effects of androgens on the long-term depression of HVC-RA pathway in adult male zebra finches].

Androgens can affect the singing behavior via regulating the song control system. In the present study, the effect of androgen on the synaptic plasticity of high vocal center (HVC)-robust nucleus of the arcopallium (RA) pathway was investigated through electrophysiological recording in vivo. We divided the adult male zebra finches into control, castration and castration plus testosterone implantation groups. The changes of long-term depression (LTD) and the paired-pulse facilitation in HVC-RA pathway induced by high-frequency (400 Hz, 2 s) stimulation of HVC were recorded, respectively. The results showed that high-frequency stimulation could effectively induce LTD in control group, but only evoke short-term depression in the castration group. In castration plus testosterone implantation group, LTD was restored. The paired-pulse facilitation was not obvious in the castration group, whereas it was significantly improved in the control and castration plus testosterone implantation groups. These results suggest that androgens may maintain the stability of song by influencing the level of LTD in HVC-RA pathway in adult male zebra finches, and androgens can affect the short-term synaptic plasticity of HVC-RA pathway.

Carbachol-Induced Reduction in the Activity of Adult Male Zebra Finch RA Projection Neurons.

Cholinergic mechanism is involved in motor behavior. In songbirds, the robust nucleus of the arcopallium (RA) is a song premotor nucleus in the pallium and receives cholinergic inputs from the basal forebrain. The activity of projection neurons in RA determines song motor behavior. Although many evidences suggest that cholinergic system is implicated in song production, the cholinergic modulation of RA is not clear until now. In the present study, the electrophysiological effects of carbachol, a nonselective cholinergic receptor agonist, were investigated on the RA projection neurons of adult male zebra finches through whole-cell patch-clamp techniques in vitro. Our results show that carbachol produced a significant decrease in the spontaneous and evoked action potential (AP) firing frequency of RA projection neurons, accompanying a hyperpolarization of the membrane potential, an increase in the evoked AP latency, afterhyperpolarization (AHP) peak amplitude, and AHP time to peak, and a decrease in the membrane input resistance, membrane time constant, and membrane capacitance. These results indicate that carbachol reduces the activity of RA projection neurons by hyperpolarizing the resting membrane potential and increasing the AHP and the membrane conductance, suggesting that the cholinergic modulation of RA may play an important role in song production.

[Study on the extraction of mycelium polysaccharides from selenium-rich Morchella esculenta with neutral proteinase method].

OBJECTIVE: To study the extraction conditions of mycelium polysaccharides from selenium-rich Morchella esculenta with neutral proteinase for increasing of yield. METHODS: On the basis of single factor tests, orthogonal experiment design was applied to analyze the influence of factors including enzyme dosage, enzymolysis time, enzymolysis temperature and material/liquid ratio on the extraction rate of the polysaccharides. RESULTS: The optimal extraction conditions of mycelium polysaccharides from selenium-rich Morchella esculenta was as follows: enzyme dosage was 1.5%, enzymolysis time was 2 h, enzymolysis temperature was 40 degrees C, and material/liquid ratio was 1:15. The extraction rate of polysaccharide was 11.26% under the extraction conditions. CONCLUSION: The process is simple, stable and practicable, and can be used for the extraction of mycelium polysaccharides from selenium-rich Morchella esculenta.

Hydrogen sulfide alleviates aluminum toxicity in germinating wheat seedlings.

Protective role of hydrogen sulfide (H(2)S) on seed germination and seedling growth was studied in wheat (Triticum) seeds subjected to aluminum (Al(3+)) stress. We show that germination and seedling growth of wheat is inhibited by high concentrations of AlCl(3). At 30 mmol/L AlCl(3) germination is reduced by about 50% and seedling growth is more dramatically inhibited by this treatment. Pre-incubation of wheat seeds in the H(2)S donor NaHS alleviates AlCl(3)-induced stress in a dose-dependant manner at an optimal concentration of 0.3 mmol/L. We verified that the role of NaHS in alleviating Al(3+) stress could be attributed to H(2)S/HS(-) by showing that the level of endogenous H(2)S increased following NaHS treatment. Furthermore, other sodium salts containing sulfur were ineffective in alleviating Al(3+) stress. NaHS pretreatment significantly increased the activities of amylases and esterases and sustained much lower levels of MDA and H(2)O(2) in germinating seeds under Al(3+) stress. Moreover, NaHS pretreatment increased the activities of guaiacol peroxidase, ascorbate peroxidase, superoxide dismutase and catalase and decreased that of lipoxygenase. NaHS pretreatment also decreased the uptake of Al(3+) in AlCl(3)-treated seed. Taken together these results suggest that H(2)S could increase antioxidant capability in wheat seeds leading to the alleviation of Al(3+) stress.

The Influence of the Degradation of Tetracycline by Free Radicals from Riboflavin-5'-Phosphate Photolysis on Microbial Viability.

Tetracycline (TC) is a broad-spectrum antibiotic compound. Wastewater with TC may have an adverse effect on ecosystems. Riboflavin-5'-phosphate (FMN or flavin mononucleotide) is a non-toxic product of the phosphorylation of vitamin B2 and is required for the proper functioning of the humans. FMN is sensitized to ultraviolet (UV) and blue light radiation, as evidenced by the generation of reactive oxygen species (ROS). This study inspects feasible applications of blue light on FMN so as to develop a valid way of degrading TC by FMN photolysis. We used the increased rate of bacterial survival as a practical indicator of antibiotic degradation. TC in the presence of FMN solution decomposed completely after 20 W/m2 of blue light irradiation (TCF treatment), and the degradation of TC (D-TCF) occurred after the photolytic process. After TCF treatment, colony-forming units (CFUs) of Escherichia coli (E. coli) were determined for the D-TCF solution. The CFU of E. coli preservation was 93.2% of the D-TCF solution (50 µg/mL of TC in the presence of 114 µg/mL of FMN solution treated with 20 W/m2 of blue light irradiation at 25 °C for 1 h) cultivation. The mass spectrum of D-TCF showed diagnostic ion signals at m/z 431.0 and 414.0 Da. The molecular formula of D-TCF was C21H22N2O8, and the exact mass was 430.44 g/mol. TC degradation by FMN photolysis can significantly decrease the antimicrobial ability of TC. The results expressed here regarding the influence of FMN photolysis on TC degradation offer an environmentally sound wastewater treatment method.

Hydrogen sulfide promotes wheat seed germination and alleviates oxidative damage against copper stress.

With the enhancement of copper (Cu) stress, the germination percentage of wheat seeds decreased gradually. Pretreatment with sodium hydrosulfide (NaHS), hydrogen sulfide (H(2)S) donor alleviated the inhibitory effect of Cu stress in a dose-dependent manner; whereas little visible symptom was observed in germinating seeds and radicle tips cultured in NaHS solutions. It was verified that H(2)S or HS(-) rather than other sulfur-containing components derived from NaHS attribute to the potential role in promoting seed germination against Cu stress. Further studies showed that NaHS could promote amylase and esterase activities, reduce Cu-induced disturbance of plasma membrane integrity in the radicle tips, and sustain lower levels of malondialdehyde and H(2)O(2) in germinating seeds. Furthermore, NaHS pretreatment increased activities of superoxide dismutase and catalase and decreased that of lipoxygenase, but showed no significant effect on ascorbate peroxidase. Alternatively, NaHS prevented uptake of Cu and promoted the accumulation of free amino acids in seeds exposed to Cu. In addition, a rapid accumulation of endogenous H(2)S in seeds was observed at the early stage of germination, and higher level of H(2)S in NaHS-pretreated seeds. These data indicated that H(2)S was involved in the mechanism of germinating seeds' responses to Cu stress.

Sulfur Dioxide Enhances Endogenous Hydrogen Sulfide Accumulation and Alleviates Oxidative Stress Induced by Aluminum Stress in Germinating Wheat Seeds.

Aluminum ions are especially toxic to plants in acidic soils. Here we present evidences that SO2 protects germinating wheat grains against aluminum stress. SO2 donor (NaHSO3/Na2SO3) pretreatment at 1.2 mM reduced the accumulation of superoxide anion, hydrogen peroxide, and malondialdehyde, enhanced the activities of guaiacol peroxidase, catalase, and ascorbate peroxidase, and decreased the activity of lipoxygenase in germinating wheat grains exposed to Al stress. We also observed higher accumulation of hydrogen sulfide (H2S) in SO2-pretreated grain, suggesting the tight relation between sulfite and sulfide. Wheat grains geminated in water for 36 h were pretreated with or without 1 mM SO2 donor for 12 h prior to exposure to Al stress for 48 h and the ameliorating effects of SO2 on wheat radicles were studied. SO2 donor pretreatment reduced the content of reactive oxygen species, protected membrane integrity, and reduced Al accumulation in wheat radicles. Gene expression analysis showed that SO2 donor pretreatment decreased the expression of Al-responsive genes TaWali1, TaWali2, TaWali3, TaWali5, TaWali6, and TaALMT1 in radicles exposed to Al stress. These results suggested that SO2 could increase endogenous H2S accumulation and the antioxidant capability and decrease endogenous Al content in wheat grains to alleviate Al stress.

[Effects of syringeal denervation on the vocalization in red-billed Leiothrix (Leiothrix lutea)].

In this study, the unilateral and bilateral syrinx nerve (NXIIts) were resected in the red-billed Leiothrix (Leiothrix lutea) to assess the roles of NXIIts in vocalization. Wavesurfer and Sound Analysis Pro were used to analyze pre- and postsurgical acoustic changes. After resecting of unilateral NXIIts, red-billed Leiothrix produced the common calls with lengthened syllable interval, shortened duration and a declined FM index. The effects of left NXIIts resction on temporal and spectral characteristics of syllables were more significant than that of right NXIIts. These results indicated that the neural control of NXIIts was unilateral and left-side dominated. Moreover, the left NXIIts could produce high frequency components and harmonic waves. After resecting of bilateral NXIIts, the calls were characterized by monotone, reduced loudness and increased syllable pulse numbers.

SK channels modulate the excitability and firing precision of projection neurons in the robust nucleus of the arcopallium in adult male zebra finches.

OBJECTIVE: Motor control is encoded by neuronal activity. Small conductance Ca(2+)-activated K(+) channels (SK channels) maintain the regularity and precision of firing by contributing to the afterhyperpolarization (AHP) of the action potential in mammals. However, it is not clear how SK channels regulate the output of the vocal motor system in songbirds. The premotor robust nucleus of the arcopallium (RA) in the zebra finch is responsible for the output of song information. The temporal pattern of spike bursts in RA projection neurons is associated with the timing of the acoustic features of birdsong. METHODS: The firing properties of RA projection neurons were analyzed using patch clamp whole-cell and cell-attached recording techniques. RESULTS: SK channel blockade by apamin decreased the AHP amplitude and increased the evoked firing rate in RA projection neurons. It also caused reductions in the regularity and precision of firing. RA projection neurons displayed regular spontaneous action potentials, while apamin caused irregular spontaneous firing but had no effect on the firing rate. In the absence of synaptic inputs, RA projection neurons still had spontaneous firing, and apamin had an evident effect on the firing rate, but caused no significant change in the firing regularity, compared with apamin application in the presence of synaptic inputs. CONCLUSION: SK channels contribute to the maintenance of firing regularity in RA projection neurons which requires synaptic activity, and consequently ensures the precision of song encoding.

[Effects of copper stress on Medicago sativa seedlings leaf antioxidative system].

This paper studied the effects of different concentration (0, 10, 30, 50, and 100 micromol x L(-1)) CuSO4 on the leaf physiological and biochemical characteristics of Medicago sativa seedlings cultured with 1/4-strength Hoagland nutrient solution. In treatments 30, 50, and 100 micromol x L(-1) of CuSO4, the leaf H2O2, OH., and MDA contents and Fe-SOD and EST activities increased, and GSH and AsA contents increased significantly. With increasing concentration Cu, the POD, GR, and APX activities increased gradually, and the CAT and G6PDH activities decreased after an initial increase. In treatments >10 micromol x L(-1) of Cu, the capacity of leaf antioxidative system in reactive oxygen species scavenging increased to prevent the injury from copper-induced oxidative stress.

Purification, properties, and application of a novel acid urease from Enterobacter sp.

It has been demonstrated that acid urease is capable of decomposing urea in fermented beverage and foods. As urea is a precursor of ethylcarbamate, a potential carcinogenic compound, measures must be taken to control the level of urea. We herein describe the purification and characterization of a novel acid urease from Enterobacter sp. R-SYB082 and its application to the removal of urea in Chinese rice wine. The enzyme was purified to electrophoretic homogeneity using ethanol precipitation, Superdex 200 and Mono Q with a fold purification of 21.1 and a recovery of 49%. The molecular weight of the enzyme was 430,000 Da by gel filtration and 72,000 Da by sodium dodecyl sulfate polyacrylamide gel electrophoresis, suggesting that it was a hexamer. The activity of this purified enzyme was optimal at pH 4.5 and 35 degrees C. The temperature stability was under 55 degrees C, and the pH stability was 4.0~5.0. The enzyme exhibited an apparent K (m) of 19.5 micromol/l and a V (max) of 109 micromol urea/mg.min at 35 degrees C and pH 4.5. When incubating two different kinds of Chinese rice wine with the enzyme (0.08 U/ml) at 35 degrees C for 7 days, over 85% of urea was decomposed, and at 20 degrees C, above 78% was removed. The result showed that the enzyme is applicable to elimination of urea in Chinese rice wine.

[Effects of cadmium stress on the antioxidative system in Ganoderma lucidum mycelia].

The study on the effects of different concentration (0, 10, 50, 100, 200 and 400 micromol x L(-1)) cadmium (Cd) on the antioxidative system in Ganoderma lucidum mycelia indicated that with increasing concentration of Cd, the fresh mass and the proline, total polysaccharides, and reduced polysaccharides contents of G. lucidum mycelia decreased, but non-protein thiol (NPT) content increased. At 400 micromol x L(-1) of Cd, the NPT content increased dramatically, being 5.7 times higher than control. Within the range of test Cd concentrations, the activities of CAT, GR and POD increased first and decreased then, with the peak at 100 micromol x L(-1) of Cd, while the activities of LOX and SOD increased with increasing Cd concentration, with the maximum at 400 micromol x L(-1) of Cd. Polyacrylamide gel electrophoresis analysis revealed that 100-400 micromol x L(-1) of Cd induced two additional isozymes bands of Mn-SOD, 10-200 micromol x L(-1) of Cd increased the intensity of constitutive isozymes of CAT, POD, SOD and LOX, while 400 micromol x L(-1) of Cd decreased the intensity of isozymes of POD significantly.

[Anti-oxidative responses of Agrocybe aegerita mycelia to cadmium stress].

This paper studied the effects of different concentration Cd on the anti-oxidative enzyme activities and glutathione content in Agrocybe aegerita cultivated in liquid medium. The results indicated that at low concentrations of Cd, the test enzyme activities increased with increasing Cd concentration, being the maximum at 0.1 mmol x L(-1) for CAT, at 0.2 mmol x L(-1) for POD, GR and LOX, and at 0.4 mmol x L(-1) for SOD. At 1.6 mmol x L(-1) of Cd, the activities of POD, CAT and SOD were inhibited markedly. 0.4-1.6 mmol x L(-1) of Cd resulted in an increase of glutathione content, but glutathione disulfide content was less affected. The ascorbate acid content and APX activity were too low to be detectable. The PAGE analysis revealed that 0.1-0.8 mmol x L(-1) of Cd induced the additional isozyme bands of POD, EST and LOX, and increased the intensity of the constitutive isozymes of CAT and SOD. 1.6 mmol x L(-1) of Cd decreased the intensity of the isozymes of POD, CAT and SOD significantly.

Salicylic acid-induced aluminum tolerance by modulation of citrate efflux from roots of Cassia tora L.

Aluminum-induced exudation of organic acids from roots has been proposed as a mechanism for Al tolerance in plants. To better understand the regulatory process leading to efflux of organic acids, the possible involvement of salicylic acid (SA) in regulating Al-induced citrate release in Cassia tora L. was identified. The response of citrate efflux to exogenous SA was concentration-dependent. Application of SA at 5 microM in solution containing 20 microM Al increased citrate efflux to levels 1.76-fold higher than in controls (20 microM Al alone). However, inhibition of citrate release was observed when SA concentrations increased to more than 20 microM. Increased citrate efflux due to the SA treatment was associated with decreased inhibition of root growth and Al content in root tips, suggesting that exogenous SA could confer Al tolerance by increasing citrate efflux. We also examined citrate synthase activities (EC 4.1.3.7) and citrate concentrations in root tips exposed to Al and/or SA. However, both citrate synthase activities and citrate accumulation remained unaffected. These results indicate that SA-promotion of Al-induced citrate efflux is not correlated with increase in citrate production. Total endogenous SA concentrations were measured in root tips and the SA concentrations were significantly enhanced by Al at levels of 10-50 microM.