Human auditory frequency-following responses to a missing fundamental

Both a complex tone perceived as a 365-hertz "missing fundamental" and a 365-hertz pure tone evoked 365-hertz far-field frequency-following responses. Narrow-band masking noise centered at 365 hertz attenuated the responses to the pure tone but not to the complex tone. Results support the concept that perception of the missing fundamental is based on periodic neural activity.     

Disulfide bond engineered into T4 lysozyme: stabilization of the protein toward thermal inactivation

By recombinant DNA techniques, a disulfide bond was introduced at a specific site in T4 lysozyme, a disulfide-free enzyme. This derivative retained full enzymatic activity and was more stable toward thermal inactivation than the wild-type protein. The derivative, T4 lysozyme (Ile3----Cys), was prepared by substituting a Cys codon for an Ile codon at position 3 in the cloned lysozyme gene by means of oligonucleotide-dependent, site-directed mutagenesis. The new gene was expressed in Escherichia coli under control of the (trp-lac) hybrid tac promoter, and the protein was purified. Mild oxidation generated a disulfide bond between the new Cys3 and Cys97, one of the two unpaired cysteines of the native molecule. Oxidized T4 lysozyme (Ile3----Cys) exhibited specific activity identical to that of the wild-type enzyme when measured at 20 degrees C in a cell-clearing assay. The cross-linked protein was more stable than the wild type during incubation at elevated temperatures as determined by recovered enzymatic activity at 20 degrees C.     

蛋壳中提取溶菌酶的研究

[目的]为了进行蛋壳中提取溶菌酶的试验。[方法]从药物原料的角度和废物利用的角度,在前人的研究基础上,以减轻环境污染和废物回收利用为原则提取的方法进行加工提取,同时利用晴纶毛线成功合成了聚丙烯酸,在250 g蛋壳中提出溶菌酶0.21 g,用比浊法测定其活性。[结果]该提取方法从250 g新鲜鸡蛋壳中可以提取出0.200 3 g溶菌酶,通过对其活力测定可知其为目标产物,酶活力单位数为10 U/mg,回收率测定达到99.5%。按此计算1 kg新鲜鸡蛋壳可以得到溶菌酶0.801 2 g。[结论]该研究为从蛋壳中提取溶菌酶提供了一种有效的方法。     

A processive single-headed motor: kinesin superfamily protein KIF1A

A single kinesin molecule can move "processively" along a microtubule for more than 1 micrometer before detaching from it. The prevailing explanation for this processive movement is the "walking model," which envisions that each of two motor domains (heads) of the kinesin molecule binds coordinately to the microtubule. This implies that each kinesin molecule must have two heads to "walk" and that a single-headed kinesin could not move processively. Here, a motor-domain construct of KIF1A, a single-headed kinesin superfamily protein, was shown to move processively along the microtubule for more than 1 micrometer. The movement along the microtubules was stochastic and fitted a biased Brownian-movement model.     

胞壁质酶的性质研究

[目的]以提取的胞壁质酶为材料,研究胞壁质酶的性质。[方法]通过测定胞壁质酶在595nm处光吸收值的增量,测定该酶活力和蛋白质浓度。通过测定米氏常数Km,研究酶促反应的速度及影响速度的因素。在不同pH值缓冲液中测定胞壁质酶活力,同时测定该酶的最适pH值。酶促反应的速度在酶的最适温度时达到最大。利用SDS-PAGE电泳法,测定胞壁质酶的分子量及纯度。[结果]胞壁质酶在219.00g/L光吸收下降最快,活力最高。Km为0.57。脲对胞壁质酶为抑制作用,其抑制类型为竞争性抑制。胞壁质酶在pH值为8.0时酶活力最高,属于弱碱性,在40℃时酶活力最高,其Kd为13。[结论]该研究可为今后采用生物工程技术对胞壁质酶进行克隆、提取以及制取提供参考。     

Fluorescence-force spectroscopy maps two-dimensional reaction landscape of the holliday junction

Despite the recent advances in single-molecule manipulation techniques, purely mechanical approaches cannot detect subtle conformational changes in the biologically important regime of weak forces. We developed a hybrid scheme combining force and fluorescence that allowed us to examine the effect of subpiconewton forces on the nanometer scale motion of the Holliday junction (HJ) at 100-hertz bandwidth. The HJ is an exquisitely sensitive force sensor whose force response is amplified with an increase in its arm lengths, demonstrating a lever-arm effect at the nanometer-length scale. Mechanical interrogation of the HJ in three different directions helped elucidate the structures of the transient species populated during its conformational changes. This method of mapping two-dimensional reaction landscapes at low forces is readily applicable to other nucleic acid systems and their interactions with proteins and enzymes.     

1株降酚菌的原生质体制备和再生研究

为了优化组合降酚菌原生质体制备与再生的条件。以从合肥钢厂焦化废水处理池的活性污泥中驯化筛选得到1株高效降酚假单胞菌为材料,通过正交实验,探讨蔗糖浓度、溶菌酶浓度E、DTA浓度和酶解时间对该菌原生质体制备与再生的影响。原生质体制备与再生的最优条件:蔗糖浓度20%,溶菌酶浓度0.5 mg/ml,EDTA浓度0.2%,酶解时间40 min。在此条件下,它的菌株形成率为75.3%,再生率为7.8%。影响降酚假单胞菌原生质体形成率的因素顺序为蔗糖浓度>溶菌酶浓度>作用时间>EDTA浓度。     

蛋壳残留溶菌酶的磁性亲和分离

采用包埋法制备了具有磁响应性能的几丁质凝胶亲和吸附介质,并对其颗粒表观结构、吸附特性等物化特性进行了分析．试验表明：介质中的Fe3O4颗粒对杂蛋白存在非特异性吸附,而含醋酸铵的磷酸盐缓冲液的充分洗涤可有效消除非特异性吸附．利用此介质所建立的方法对蛋壳清洗液中的溶菌酶进行了分离．结果表明：通过外加磁场可快速地实现亲和吸附介质与粘稠清洗液及其中蛋壳碎片和泥沙等固形物杂质的分离,大大简化了操作步骤,酶活力回收可达26％,纯化酶对蛋白的比活性为50894U／mg,经SDS-聚丙烯酰胺凝胶电泳显示单一条酶蛋白带．     

Intrinsic oscillations of neocortex generated by layer 5 pyramidal neurons

Rhythmic activity in the neocortex varies with different behavioral and pathological states and in some cases may encode sensory information. However, the neural mechanisms of these oscillations are largely unknown. Many pyramidal neurons in layer 5 of the neocortex showed prolonged, 5- to 12-hertz rhythmic firing patterns at threshold. Rhythmic firing was due to intrinsic membrane properties, sodium conductances were essential for rhythmicity, and calcium-dependent conductances strongly modified rhythmicity. Isolated slices of neocortex generated epochs of 4- to 10-hertz synchronized activity when N-methyl-D-aspartate receptor-mediated channels were facilitated. Layer 5 was both necessary and sufficient to produce these synchronized oscillations. Thus, synaptic networks of intrinsically rhythmic neurons in layer 5 may generate or promote certain synchronized oscillations of the neocortex.     

Control of enzyme activity by an engineered disulfide bond

A novel approach to the control of enzyme catalysis is presented in which a disulfide bond engineered into the active-site cleft of bacteriophage T4 lysozyme is capable of switching the activity on and off. Two cysteines (Thr21----Cys and Thr142----Cys) were introduced by oligonucleotide-directed mutagenesis into the active-site cleft. These cysteines spontaneously formed a disulfide bond under oxidative conditions in vitro, and the catalytic activity of the oxidized (cross-linked) T4 lysozyme was completely lost. On exposure to reducing agent, however, the disulfide bond was rapidly broken, and the reduced (non-cross-linked) lysozyme was restored to full activity. Thus an enzyme has been engineered such that redox potential can be used to control catalytic activity.     

A radio pulsar spinning at 716 Hz

We have discovered a 716-hertz eclipsing binary radio pulsar in the globular cluster Terzan 5 using the Green Bank Telescope. It is the fastest spinning neutron star found to date, breaking the 24-year record held by the 642-hertz pulsar B1937+21. The difficulty in detecting this pulsar, because of its very low flux density and high eclipse fraction (approximately 40% of the orbit), suggests that even faster spinning neutron stars exist. If the pulsar has a mass less than twice the mass of the Sun, then its radius must be constrained by the spin rate to be <16 kilometers. The short period of this pulsar also constrains models that suggest that gravitational radiation, through an r-mode (Rossby wave) instability, limits the maximum spin frequency of neutron stars.     

溶菌酶产生菌的筛选及产酶条件研究

为了从微生物中获取溶菌酶,本研究采用双层平板法从土壤中筛选获得一株产溶菌酶活性较高的菌株S2-6b.利用摇瓶发酵优化产酶条件,液体发酵培养基成分为可溶性淀粉15%,牛肉膏25%,玉米浆05%,NaCl 05%,起始pH值为70,培养温度为28 ℃,接种量为30%,摇瓶的装液量为40 mL/300 mL,转速为200 r...     

河北省财政支农生产性支出与非生产性支出的对比分析

对河北省近20 a来财政支农资金中生产性支出与非生产性支出进行了对比分析,并建立回归模型分析了生产性支出与非生产性支出对农业增长及农民增收的作用.认为非生产性支出增长过快在一定程度上影响了支农资金的使用效益,应在保持财政支农资金增长的前提下,不断提高生产性支出的比重.     

溶菌酶活力测定及其体外抑菌试验

将溶菌酶与中草药提取液配制成复合制剂,测定复合制剂中溶菌酶的活力。结果与单一溶菌酶活力比较差异不显著(P>0.05)。用不同来源的溶菌酶对金黄色葡萄球菌和大肠杆菌进行体外抑菌试验,并与常用抗生素进行比较。结果溶菌酶对金黄色葡萄球菌抑菌效果明显,与青霉素钠及硫酸丁胺卡那霉素比较无明显差异。溶菌酶对大肠杆菌的抑菌效果因来源不同而有所差异。     

Protein conformation in solution: cross-linking of lysozyme

The cross-linking of lysozyme by reaction with phenol-2,4-disulfonyl chloride has been effected. The cross-linked protein retained enzymatic activity, has approximately the same molecular weight as native lysozyme, and has essentially the same conformation as native lysozyme as judged by optical rotatory dispersion analysis. The positions of sulfonylation were assigned by a standard degradation sequence; the presence of sulfonamide bonds was confirmed by infrared spectroscopy. Cross-links may thus be introduced without incurring major structural changes in the protein, and certain intramolecular distances that are allowed in the active enzyme may be deduced.     

Gamma oscillations and object processing in the infant brain

An enduring controversy in neuroscience concerns how the brain "binds" together separately coded stimulus features to form unitary representations of objects. Recent evidence has indicated a close link between this binding process and 40-hertz (gamma-band) oscillations generated by localized neural circuits. In a separate line of research, the ability of young infants to perceive objects as unitary and bounded has become a central focus for debates about the mechanisms of perceptual development. Here we demonstrate that binding-related 40-hertz oscillations are evident in the infant brain around 8 months of age, which is the same age at which behavioral and event-related potential evidence indicates the onset of perceptual binding of spatially separated static visual features.     

脲对麦芽酸性磷酸酶活力与构象的影响

应用荧光光谱、紫外差光谱等检测手段研究麦芽酸性磷酸酶经脲变性后的分子构象与活力的变化情况,结果表明,低浓度的脲(＜2 mol/L)微扰了酶的活性部位而使其局部构象发生变化,使酶的荧光强度略有下降,荧光发射峰位红移,酶活力下降.继续增大脲浓度,酶的整体构象被破坏,酶荧光强度增加,发射峰位红移,酶活力仍然呈下降趋势.荧光强度的增加和最大发射峰位的红移是酶变性失活的一个指标,酶活力的变化快于酶分子整体构象的变化,说明麦芽酸性磷酸酶的活性部位位于柔性较大的区域.     

Active human-yeast chimeric phosphoglycerate kinases engineered by domain interchange

Phosphoglycerate kinase (PGK) is a monomeric protein composed of two domains of approximately equal size, connected by a hinge. Substrate-induced conformational change results in the closure of the active site cleft, which is situated between these two domains. In a study of the relations between structure and function of this enzyme, two interspecies hybrids were constructed, each composed of one domain from the human enzyme and one domain from the yeast enzyme. Despite a 35% difference in the amino acid composition between human and yeast PGK, catalytic properties of the hybrid enzymes are very similar to those of the parental proteins. This result demonstrates that the evolutionary substitutions within these two distantly related molecules do not significantly affect formation of the active site cleft, mechanism of domain closure, or enzyme activity itself.