Conformations and folding of lysozyme ions in vacuo

Proton transfer reactivity of isolated charge states of the protein hen egg-white lysozyme shows that multiple distinct conformations of this protein are stable in the gas phase. The reactivities of the 9+ and 10+ charge state ions, formed by electrospray ionization of "native" (disulfide-intact) and "denatured" (disulfide-reduced) solutions, are consistent with values calculated for ions in their crystal structure and fully denatured conformations, respectively. Charge states below 8+ of both forms, formed by proton stripping, have similar or indistinguishable reactivities, indicating that the disulfide-reduced ions fold in the gas phase to a more compact conformation.     

Identification of two distinct properties of class II major histocompatibility complex-associated peptides

We have examined the interactions of various peptides with the mouse class II major histocompatibility complex molecule I-Ak. The peptides were derived from the model protein hen egg white lysozyme (HEL). The immunodominant peptide of HEL is a 10-mer, residues 52-61. Our previous work established that this sequence contains the key residues for binding and presentation to T cells. Now we show that the binding of this 10-mer sequence resulted in complexes of I-Ak and peptide that, in SDS/PAGE (without boiling the protein), rapidly dissociated from the component alpha and beta chains. The binding interactions were studied in vitro, by incubating purified I-Ak and radiolabeled peptide, or ex vivo, by using antigen-presenting cells incubated with peptides. Peptides with additional residues at either the amino or carboxyl terminus behaved dramatically differently. Complexes of I-Ak with the longer peptides were stable to SDS/PAGE. Very few amino acid additions result in the change from unstable to stable complexes. The important issue here is that when cultured with HEL, antigen-presenting cells selected the HEL peptides containing the 52-61 sequences that favored stability [Nelson, C. A., Roof, R. W., McCourt, D. W. & Unanue, E. R. (1992) Proc. Natl., Acad. Sci. USA 89, 7380-7383]. Also, from other studies, such sequences correlate with a high immunogenicity of the peptide. We conclude that there are structural features of peptides that change the stability of the class II molecule and that are independent of the "core" peptide seen by the T cells.     

Inhibition of HeLa cell messenger RNA translation by 7-methylguanosine 5'-monophosphate

Translation of HeLa cell RNA containing poly(A) in a wheat germ cell-free system is markedly but incompletely inhibited by 7-methylguanosine 5'-monophosphate (m7G5'p). We have analyzed the translation products synthesized in the presence of different concentrations of m7G5'p and find that translation of all mRNAs is equally inhibited. To demonstrate the specificity of the inhibitor for RNAs with 5'-terminal m7G5' ppp... we show that specific translation products of satellite tobacco necrosis virus RNA, which does not have this 5' terminus, are synthesized in the presence of m7G5' p. Protein synthesis programmed by endogenous mRNA in a HeLa cell-free system is inhibited after a 10-min lag by m7G5' p. Other guanosine nucleotides without the 7-methyl group or with the phosphate in a different position are not inhibitor. We show that translation of all mRNAs is inhibited to a similar extent by m7G5'p in the HeLa cell-free system, by synthesizing 35S-labeled proteins in the presence of different inhibitory concentrations of this nucleotide and analyzing the translation products by electrophoresis and autoradiography. Translation of encephalomyocarditis virus RNA added to the HeLa cell-free system is not inhibited by m7"g5p; this viral RNA does not have this nucleotide at the 5' terminus. This indicates that m7G5'p specifically inhibits translation of mRNAs with the 5' terminus m7G5'ppp... and suggests that initiation of translation of picornavirus RNA may proceed via a mechanism different from that of cellular mRNAs.     

Occurrence of gastric ulcer after Nissen fundoplication

Of 160 patients who underwent Nissen fundoplication for treatment of symptomatic peptic reflux esophagitis, five patients (3.1%) developed gastric ulcers. Four of these five patients experienced the "gas-bloat" syndrome. All ulcers were located on the lesser curvature of the stomach. Analyses of our experience with use of various types of hiatal hernia repair suggests that creation of the valvuloplastic mechanism unique to the Nissen procedure may be of etiologic significance in the development of gastric ulcers following this procedure.     

DFNN及其在非线性动态系统中的应用

为了更好地辨识和控制非线性动态系统,在FNN基础上对其进行优化和改进,形成了动态模糊神经网络(DFNN)。给出了基于BP梯度算法的参数迭代学习算法,并应用于某非线性动态系统仿真试验中。仿真试验表明,该网络比单纯的FNN具有更强的辨识和控制能力,应用于非线性动态系统的控制中可以有效解决系统的非线性和不确定性,提高系统的跟踪性能,并且控制系统具有很强的鲁棒性。     

Analysis and neural network modeling of the nonlinear correlates of habituation in the vestibulo-ocular reflex

Through the process of habituation, continued exposure to low-frequency (0.01 Hz) rotation in the dark produced suppression of the low-frequency response of the vestibulo-ocular reflex (VOR) in goldfish. The response did not decay gradually, as might be expected from an error-driven learning process, but displayed several nonlinear and nonstationary features. They included asymmetrical response suppression, magnitude-dependent suppression for lower- but not higher-magnitude head rotations, and abrupt-onset suppressions suggestive of a switching mechanism. Microinjection of lidocaine into the vestibulocerebellum of habituated goldfish resulted in a temporary dishabituation. This suggests that the vestibulocerebellum mediates habituation, presumably through Purkinje cell inhibition of vestibular nuclei neurons. The habituated VOR data were simulated with a feed-forward, nonlinear neural network model of the VOR in which only Purkinje cell inhibition of vestibular nuclei neurons was varied. The model suggests that Purkinje cell inhibition may switch in to introduce nonstationarities, and cause asymmetry and magnitude-dependency in the VOR to emerge from the essential nonlinearity of vestibular nuclei neurons.     

In vitro displacement by rat serum of adsorbed radiolabeled poloxamer and poloxamine copolymers from model and biodegradable nanospheres

Poloxamer 407 and poloxamine 908 have been used by many research groups to modify the surface of both model latex and biodegradable nanospheres, thereby producing nanospheres that have shown reduced protein adsorption in vitro and extended circulation times in vivo. A potential limitation of such systems is the desorption of the copolymer coating layer. We describe a two-stage process to radiolabel poloxamer 407 and poloxamine 908 that has facilitated an investigation into this potential desorption, in vitro. The first stage of the labeling procedure involved the substitution of the terminal hydroxyl groups in each poly(ethylene oxide) (PEO) chain of poloxamer 407 and poloxamine 908 with an amino group. The aminated copolymers were then radiolabeled with 125Iodine Bolton-Hunter reagent. The efficiency of labeling was calculated to be approximately 20% for the tetramine poloxamine 908 and approximately 33% for the diamine poloxamer 407. Remaining free amino groups were then either acetylated, using acetic anhydride, or left in the free amino form. Covalent linkage of the radiolabel to the copolymer was confirmed by nuclear magnetic resonance (NMR) and infrared (IR) spectroscopy. The stability of the link between radiolabel and copolymer to hydrolysis was also confirmed; <4% loss of radiolabel occurred from poloxamine 908 after incubation in phosphate-buffered saline (PBS) at 37 degrees C for 8 days. The radiolabeled copolymers (with the free amino groups acetylated) were then used in experiments that have given the first direct evidence that adsorbed copolymers can be displaced by serum proteins in significant amounts from the surface of model and biodegradable nanospheres. The displacement was highly dependent on copolymer-nanosphere compatibility, with up to 78% of 125I tetramine poloxamine 908 being displaced from poly(lactide-co-glycolide) (PLGA) nanospheres in 24 h, compared with 20% displacement of 125I tetramine poloxamine 908 in 24 h from polystyrene nanospheres. These results have direct implication for the future design of drug delivery systems based on coated nanospheres.     

Improved localization in a generalized Hough scheme for the detection of straight edges

Hough transforms are widely used for the location of straight edges in digital images, yet most common line parametrization schemes give no information on longitudinal localization. The generalized Hough transform goes some way to overcoming this problem. This paper studies how to improve the situation further. A trade-off between sensitivity and localization is found; in practical situations this results in significantly greater accuracy, but the important gain is a reduction in the number of ambiguities introduced by interactions between the transforms of unrelated straight edges.     

高精度机械伺服系统的一种新型自适应滑模控制方法

针对高精度机械伺服系统,提出一种高性能的新型自适应滑模控制方法,使闭环系统渐近跟踪给定的参考模型.该方法对转动惯量的大范围变化及非线性摩擦等外干扰均具有很强的鲁棒性.该方法的主要思想是用滑模方法抑制系统中的外部力矩扰动,对系统参数进行自适应估计,用估计值来补偿转动惯量的变化.对于控制算法的全局稳定性,采用李雅普诺夫直接法给出了严格的证明.该算法简单,其实现不需要误差的高阶微分信号,适于实时控制.本文方法以某高精度飞行仿真转台为例,对提出方法进行了实验研究,结果表明了该方法具有良好的跟踪性能,暂态响应和鲁棒性.     

雷达寻的制导半实物仿真误差研究

在介绍雷达寻的制导控制基本原理、半实物基本方法及仿真原理的基础上,详细介绍了雷达寻的制导控制半实物仿真系统的组成、各关键设备的工作原理并进行了主要误差因素分析,同时对关键仿真设备的误差进行研究,并建立数学模型,进一步推导出此类雷达仿真系统具有一般意义的仿真误差模型.最后通过型号半实物仿真试验对仿真系统的仿真精度进行验证.仿真试验表明,该半实物仿真系统的仿真精度满足控制系统半实物仿真试验的要求.