Cellular properties and modulation of the stomatogastric ganglion neurons of a stomatopod,Squilla oratoria

Cellular properties and modulation of the identified neurons of the posterior cardiac plate-pyloric system in the stomatogastric ganglion of a stomatopod, Squilla oratoria, were studied electrophysiologically. Each class of neurons involved in the cyclic bursting activity was able to trigger an endogenous, slow depolarizing potential (termed a driver potential) which sustained bursting. Endogenous oscillatory properties were demonstrated by the phase reset behavior in response to brief stimuli during ongoing rhythm. The driver potential was produced by membrane voltage-dependent activation and terminated by an active repolarization. Striking enhancement of bursting properties of all the cell types was induced by synaptic activation via extrinsic nerves, seen as increases in amplitude or duration of driver potentials, spiking rate during a burst, and bursting rate. The motor pattern produced under the influence of extrinsic modulatory inputs continued for a long time, relative to that in the absence of activation of modulatory inputs. Voltage-dependent conductance mechanisms underlying postinhibitory rebound and driver potential responses were modified by inputs. It is concluded that endogenous cellular properties, as well as synaptic circuitry and extrinsic inputs, contribute to generation of the rhythmic motor pattern, and that a motor system and its component neurons have been highly conserved during evolution between stomatopods and decapods.Abbreviations AB anterior burster neuron - CoG commissural ganglion - CPG central pattern generator - lvn lateral ventricular nerve - OG oesophageal ganglion - pcp posterior cardiac plate - PCP pcp constrictor neuron - PD pyloric dilator neuron - PY pyloric constrictor neuron - son superior oesophageal nerve - STG stomatogastric ganglion - stn stomatogastric nerve     

Eu3+-activated red phosphor Ca3YAl3B4O15 with low thermal quenching behaviour

This paper reports a sequence of a Ca₃YAl₃B₄O₁₅:xEu³⁺ red phosphor prepared using a high-temperature solid-state reaction. At the excitation of 396 nm, the samples emitted intense red emission centred at ~623 nm, which could be attributed to the ⁵D₀→⁷F₂ transition of the Eu³⁺ ion. The results showed that the optimum Eu³⁺ doping concentration of Ca₃YAl₃B₄O₁₅:Eu³⁺ phosphor was x = 80 mol%, and the concentration quenching mechanism of Ca₃YAl₃B₄O₁₅:Eu³⁺ red phosphor belonged to the exchange coupling between Eu³⁺ ions. The Commission Internationale de l'éclairage (CIE) coordinates and colour purity of Ca₃Y_0.2Al₃B₄O₁₅:0.8Eu³⁺ were calculated as (0.6375, 0.3476) and 95.5%, respectively. Moreover, the red emission of the obtained phosphor Ca₃YAl₃B₄O₁₅:0.8Eu³⁺ exhibited a low thermal quenching behaviour with an intensity retention rate of 92.85% at 150°C. The above results manifest that the Eu³⁺-activated Ca₃YAl₃B₄O₁₅ phosphor is predicted to be a promising red luminescent component for white light-emitting diodes.     

Synthetic chimeras of mouse growth factor-associated glandular kallikreins. I. Kinetic properties.

A series of six chimeric proteins, composed of fragments corresponding to either one or the other of the growth factor-associated mouse glandular kallikreins-epidermal growth factor binding protein (EGF-BP) and the gamma-subunit of nerve growth factor (gamma-NGF)--were expressed in Escherichia coli and isolated, and their kinetic properties were characterized. The assembly of these synthetic proteases involved the substitution of regions of the proteins containing four specific surface loops that have been postulated to influence both kinetic specificity and the formation of growth factor complexes. The substrates utilized in the kinetic characterization of these chimeric kallikreins were tripeptide nitroanilides representing carboxyl termini of both the EGF and beta-NGF mature hormones, putative processing sites for these kallikreins in the precursors. Characterization of these hybrid enzymes demonstrates that Km and kcat kinetic constants may be independently affected by the regions utilized in construction of these chimeric kallikreins. Specifically, loop 1, located in the amino terminal region (Bode, W., et al., J. Mol. Biol. 164, 237-282, 1983), in gamma-NGF enhanced the kcat for substrates containing threonine in the P2 position, as is the case during the processing of the carboxy terminus of the beta-NGF precursor. Also, the central regions of the kallikreins containing loop 2 and the kallikrein loop dictated the generally inverted Km and kcat kinetic constants observed between EGF-BP and gamma-NGF. Finally, in gamma-NGF the autolysis loop, found in the carboxyl terminal region, functions to lower the Km kinetic constant for a variety of substrates. The results allow previously characterized kinetic differences between EGF-BP and gamma-NGF to be interpreted in terms of specific regions of the proteins and identify a subset of amino acid positions responsible for these functional characteristics.     

Representing an ensemble of NMR-derived protein structures by a single structure.

The usefulness of representing an ensemble of NMR-derived protein structures by a single structure has been investigated. Two stereochemical properties have been used to assess how a single structure relates to the ensemble from which it was derived, namely the distribution of phi psi torsion angles and the distribution of chi 1 torsion angles. The results show that the minimized average structure derived from the ensemble (a total of 11 ensembles from the Brookhaven Protein Data Bank were analyzed) does not always correspond well with this ensemble, particularly for those ensembles generated with a smaller number of experimentally derived restraints per residue. An alternative method that selects the member of the ensemble which is closest to the "average" of the ensemble has been investigated (a total of 23 ensembles from the Brookhaven Protein Data Bank were analyzed). Although this method selected a structure that on the whole corresponded more closely to the ensemble than did the minimized average structure, this is still not a totally reliable means of selecting a single structure to represent the ensemble. This suggests that it is advisable to study the ensemble as a whole. A study has also been made of the practice of selecting the "best" rather than the most representative member of the ensemble. This too suggests that the ensemble should be studied as a whole. A study of the conformational space occupied by the ensemble also suggests the need to consider the ensemble as a whole, particularly for those ensembles generated with a smaller number of experimentally derived restraints per residue.     

来源于嗜热氢化杆菌的新型对苯二甲酸双(羟乙)酯水解酶的表达纯化与酶学性质

石化来源的聚对苯二甲酸乙二酯(polyethylene terephthalate,PET)被广泛用于矿泉水瓶、食品包装和纺织品等领域,因其在自然界中不易分解,大量使用后的PET废弃物造成了严重的环境污染与资源浪费。使用生物酶法对PET废弃物进行解聚,并对解聚产物进行升级循环利用是进行塑料污染治理的重要方向之一,其中关键的是PET水解酶的解聚效率。对苯二甲酸双(羟乙基)酯(bis(hydroxyethyl)terephthalate,BHET)是PET生物酶解的中间产物,其累积是限制PET水解酶催化效率的一个重要因素,BHET水解酶和PET水解酶的联用能提升PET的整体水解效率。来源于嗜热氢化杆菌(Hydrogenobacter thermophilus)的双烯内酯酶(HtBHETase)对BHET有显著水解效果,将该酶在大肠杆菌(Escherichia coli)中进行重组表达并纯化后,对其酶学性质进行了研究。结果显示,HtBHETase对短碳链的酯类如对硝基苯酚乙酸酯催化活性较高,HtBHETase以BHET为底物时的最适反应pH值和最适反应温度分别为5.0和55℃;该酶有较好的热稳定性,经80℃的条件处理1 h仍能保持80%以上活性,显示出了良好的热稳定性,HtBHETase有在PET塑料生物解聚中使用的潜力,本研究为推动生物酶法降解PET提供了新的参考。     

Arbuscular Mycorrhizal Fungal Colonization at Different Succession Stages in Songnen Saline-Alkali Grassland

Arbuscular mycorrhizal (AM) fungi can form symbiosis with 90% of the vascular plants and play important roles in ecosystem. To realize the AM fungal colonization at different succession stages in saline-alkali land and screen AM fungi species with great functions, roots and soil samples were collected from the three succession stages of Songnen saline-alkali grassland. The soil properties and AM fungal colonization were measured, and the fungus distributed extensively in three stages was annotated by sequencing for AML1/AML2 target, subsequently, maize was selected as the host to verify its colonization. The results showed that the soil properties improved with the succession of saline-alkali grassland. The plants’ communities of the three stages could be colonized by AM fungi, and the colonization rate of Leymus chinensis (the third stage) ranged from 66.67% to 100%, Puccinellia tenuiflora (the second stage) ranged from 50% to 80%, while the Suaeda glauca (the first stage) was only 35%–60%. Glomeraceae sp1 was identified as the dominant AM fungi species which occurred frequently in the succession of saline-alkali land with the isolation frequency, relative abundance, and importance value of 100%, 18.1%, and 59.1%, respectively. The colonization rate of Glomeraceae sp1 in maize ranged from 80% to 87% and similar mycorrhizal characteristics were detected in the roots of P. tenuiflora, S. glauca, and L. chinensis, indicating that Glomeraceae sp1 colonized the samples in the field. The correlation matrix indicated that colonization rate, colonization intensity, and vesicle abundance were closely related to soil conditions most, and they were related significantly to all the soil properties except cellulase activity. Besides, redundancy analysis (RDA) showed that soil properties drove the changes of AM fungal colonization and sporulation. These results will provide theoretical support for realizing the relationship between AM fungal colonization and soil conditions, and also for the exploration of AM fungi species with great functions.

     

Protonation,alkylation, addition and redox reactions of 16, 17 and 18 valence electron [Mo(L)(NO)('S4')] complexes [L = SPh,PMe3, NO; 'S4'2– = 1,2-Bis-(2-mercaptophenylthio)ethane(2-)]

In the quest for complexes modelling functional characteristics of metal sulfur oxidoreductases, a series of molybdenum nitrosyl complexes with sulfur-dominated coordination sphere was synthesized. Treatment of the 16, 17 and 18 valence electron (VE) complexes [Mo(L)(NO)('S₄')] (1–3) [L?=?SPh (1), PMe₃ (2), NO (3), 'S₄'^2–?=?1,2-bis-(2-mercaptophenylthio) ethane(2-)] with the Brönsted acid HBF₄ resulted in formation of different types of products. 1 and 3 were reversibly protonated at one thiolate atom of the 'S₄'^2– ligand;2, however, yielded the phosphonium salt [HPMe₃]BF₄ and the dinuclear [Mo(NO)('S₄')]₂. Alkylation of 1, 2 and 3 by Me₃OBF₄ or Et₃OBF₄ uniformly resulted in high yields of [Mo(L)(NO)(R-'S₄')]BF₄ complexes [L?=?SPh: R?=?Me (5), Et (6); L?=?PMe₃: R?=?Me (7); L?=?NO: R?=?Me (8), Et (9)] in which one thiolate atom of the 'S₄'^2– ligand had become alkylated; the NMR spectra of 5, 6, 8 and 9 indicated that only one out of four theoretically possible diastereoisomers had formed. 5 and 6 were characterized also by single-crystal X-ray structure analyses. A comparison of ν(NO) bands and redox potentials (cyclic voltammetry) of parent complexes and alkylated derivatives showed that alkylation leads to a decrease in electron density at the molybdenum center and to a positive shift in redox potentials. The 16 VE complex 1 could be reduced, also chemically, to give the corresponding 17 VE anion [1]^–, and inserted elemental sulfur into the Mo-SPh bond, forming the 18 VE phenylperthio complex [Mo(η²–SSPh)(NO)('S₄')] (11) which, upon reaction with PPh₃, gave SPPh₃ and regenerated the parent complex 1. These results are discussed with regard to the sequence of proton and electron transfer steps occurring in substrate conversions catalyzed by metal sulfur oxidoreductases.     

Dynamic properties of the advertisement calls of gray tree frogs: patterns of variability and female choice

We assessed the potential for several acoustic properties ofthe advertisement calls of male gray tree frogs to affect relativemating success by relating patterns of variation in these propertiesto minimum differences required to elicit female choice. Dynamicproperties (pulse number, PN; call rate, CR; and duty cycle,DC, the ratio of call duration to call period) varied much morewithin bouts of calling than a static property (dominant frequency,DF) but nevertheless exhibited significant between male variationin three of four breeding seasons. Many multiply recorded malesconsistently produced calls with values substantially aboveor below mean values of males recorded on the same nights. Nightlyranges of variation in PN and CR were often greater than theminimum differences required to elicit female preferences inthe laboratory. In most experiments, females chose high-PN orfast-CR calls over low-PN or slow-CR alternatives, respectively,even if the preferred stimuli were farther away or 6-10 dB lowerin sound pressure level (SPL), provided that differences inPN or CR were 100%. Consistent with these results, females didnot always choose the closer of two calling males in the field.Nightly ranges of variation in DF rarely equaled the minimumdifference required to elicit SPL independent preferences. Femalespreferred a stimulus of high-PN and slow-CR over an alternativeof low-PN or fast-CR with the same acoustic on-time; in twoexperiments, females chose calls of high-PN over low-PN alternativeseven though the playback of the high-PN call was interruptedand the low-PN call was broadcast continuously. Thus, femalepreferences were not merely based on the total time of acousticstimulation. Responses of females tested twice in the same experimentsuggest that phenotypic variation in preference was limitedin our study populations.     

Characterization of chicken gizzard calcyclin and examination of its interaction with caldesmon

Using a procedure developed to purify calcyclin from mouse Ehrlich ascites tumor cells calcyclin was purified from smooth muscle of chicken gizzard. Chicken gizzard calcyclin bound to phenyl-Sepharose in a calcium dependent manner as did mouse EAT cells and rabbit lung calcyclin but appeared to be more acidic than its mammalian counterparts as revealed by ion exchange chromatography on Mono Q. Chicken gizzard calcyclin bound ⁴⁵Ca²⁺ on nitrocellulose filters and exhibited a shift in electrophoretic mobility on urea-PAGE depending on Ca²⁺ concentration. Crosslinking experiments with BS³ showed that chicken gizzard calcyclin was able to form noncovalent dimers. As indicated by a decrease in maximum tryptophan fluorescence emission of caldesmon (about 14% at 1:1 molar ratio) and displacement of calmodulin from its complex with caldesmon, chicken gizzard calcyclin binds caldesmon. This binding was, however, much weaker than that of calmodulin and could not influence the interaction of caldesmon with actin. In consequence, calcyclin was unable to reverse the inhibitory effect of caldesmon on actin-activated Mg²⁺-ATPase activity of myosin in the presence of Ca²⁺.     

Radiative properties of hardwood leaves to ultraviolet irradiation

Spectral reflectance and transmittance of leaves to ultraviolet irradiation were determined under laboratory conditions for seven species of hardwood trees, namely red oak (Quercus rubra, L), black oak (Q. velutina, Lamarch), white oak (Q. alba, L.), sugar maple (Acer saccharum), Norway maple (A. plantanoides), hickory (Carya tomemtosa), sweetgum (Liquidambar styraciflua), and black oak litter. The experimental system consisted of a solar simulator, an integrating sphere, and a spectroradiometer. Experiments were repeated three to five times for both adaxial and abaxial surfaces of fresh leaves chosen at randomly. The spectral distributions and simple averages of the radiative properties in the wavelength ranges of ultraviolet-B (UV-B, 280–320 nm) and ultraviolet-A (UV-A, 320–400 nm) were determined. The spectral distributions of reflectance were similar between adaxial and abaxial surfaces, although the magnitude varied among tree species. Leaf reflectance was very low for the ultraviolet spectrum in general and varied among species and between adaxial and abaxial surfaces. It was generally higher over the UV-A waveband compared to UV-B, and higher on the abaxial than adaxial surface. The broadband reflectance in the UV-A range (over all species) was 5.0 and 3.9% for abaxial and adaxial surface, respectively, compared to 3.5 and 2.8% in UV-B. The transmittance through leaves was extremely small in the UV-B (<0.1%) and nearly zero in the UV-A spectral range. Consequently, the absorptance of ultraviolet radiation by leaves, as determined from the measured reflectance and transmittance, was quite high, being more than 90% for all the combinations of species and wavebands examined. The reported results are useful for studies requiring spectral radiative properties of the examined leaves with respect to ultraviolet irradiation.