Nucleotide and actin binding properties of the isolated motor domain from Dictyostelium discoideum myosin

Nucleotide and actin binding properties of the truncated myosin head (S1dC) from Dictyostelium myosin II were studied in solution using rabbit skeletal myosin subfragment 1 as a reference material. S1dC and subfragment 1 had similar affinities for ADP analogues, epsilon ADP and TNP-ADP. The complexes of epsilon ADP and BeFx or AIF4- were less stable with S1dC than with subfragment 1. Stern-Volmer constants for acrylamide quenching of S1dC complexes with epsilon ADP, epsilon ADP.AIF4- and epsilon ADP.BeFx were 2.6, 2.9 and 2.2 M-1, respectively. The corresponding values for subfragment 1 were 2.6, 1.5 and 1.1 M-1. The environment of the nucleotide binding site was probed by using a hydrophobic fluorescent probe, PPBA. PPBA was a competitive inhibitor of S1dC Ca(2+)-ATPase (Ki = 1.6 microM). The binding of nucleotides to subfragment 1 enhanced PPBA fluorescence and caused blue shifts in the wavelength of its maximum emission in the order: ATP approximately ADP.AIF4- approximately ADP.BeFx > ATP gamma S > ADP > PPi. In the case of S1dC, the effects of different nucleotides were smaller and indistinguishable from each other. S1dC bound actin tighter than S1 (Kd = 7 nM and 60 nM, respectively). The actin activated MgATPase activity of S1dC varied between preparations, and the Vmax and K(m) values ranged between 3 and 7 s-1 and 60 and 190 microM, respectively. S1dC showed lower structural stability than S1 as revealed by their thermal inactivations at 35 degrees C. These results show that the nucleotide and actin binding of S1dC and subfragment 1 are similar but there are some differences in nucleotide and phosphate analogue-induced changes and the communication between the nucleotide and actin binding sites in these proteins.     

机器人工作空间快速可靠数值算法

本文根据机器人的结构特点,选定机器人工作空间的中心点,利用机器人手臂端部在各个方向上到中心点的极限距离,设计出一种快速可靠的机器人工作空间数值算法.该算法比其他数值算法计算量少一个量级,且可靠性好.     

Nitric oxide synthase in neuroepithelial brain tumors

Immunohistochemical investigation of NO-synthase in brain astrocytic tumors revealed intense reaction in many tumor cells as well as direct correlation in the intensity of reaction and the degree of tumor anaplasia. Grade I astrocytomas did not show immunoreactivity in contrast to high anaplastic tumors where many cells had positive reaction with a different degree of intensity. Positive immunoreaction was shown in many giant cells. Small cell glioblastomas and oligodendrogliomas were immunonegative. There was a direct correlation between NO-synthase expression and glial fibrillar acidic protein.     

Cross-reactive cellular immune response to circumsporozoite proteins of Plasmodium vivax and P. falciparum in malaria-exposed individuals

The present review offers a new look at capillary isoelectric focusing (cIEF) by centering on the most troublesome aspects of the technique, namely: 1) how to modulate the slope of the pH gradient, for increasing resolution (equivalent to pH gradient engineering, as easily available in immobilized pH gradients); and 2) how to keep proteins in solution at (and in the proximity of) the pl value. A simple solution is offered in the first case: addition, to the standard 2-pH-units interval, of separators or spacers, i.e., of amphoteric molecules (either single or in combination) able to locally flatten the pH and increment resolution. Examples of the separation of fetal and glycated hemoglobins are provided. In the second case, a unique solubilization power (while maintaining full protein integrity and enzyme activity) is obtained if class I solubilizers are used. They consist of mixtures of sugars (e.g., sucrose and sorbitol) at ca. 1 M concentration, with zwitterions (up to 1 M) such as the class of nondetergent sulfobetaines, but also taurine and some of the Good's buffers (e.g., CAPS). In these solvents, the protein exists in a state of superhydration and its solubility is greatly augmented. The review ends with an excursus on the use of isoelectric buffers in zone electrophoretic separations. Such isoelectric buffers offer unique advantages: They permit very-high-voltage gradients (up to 1000 V/cm) and thus minimize analysis times (down to a few min in 30-35 cm long capillaries). This results in a marked increase in resolution, due to minimal diffusion-driven peak spreading. Such buffers are finding unique applications for generating peptide maps of tryptic digests of proteins and also in the analysis of oligonucleotides.     

Intelnet Services—A global data distribution and collection scheme

Responding to the recent availability of very small earth-stations (commonly called micro terminals) for data/voice communications, INTELSAT has introduced a new service called INTELNET. INTELNET services are designed to operate with two different modulation and coding techniques, i.e. spread spectrum and binary phase-shift keying (BPSK) with rate 1/2 forward error correction (FEC). The INTELNET I service is a data distribution service, whereas the INTELNET II service is a data collection service. These services are facilitated by the use of micro terminals at remote locations (i.e. outposts) which will be linked through the INTELSAT space segment to relatively large INTELSAT earth-stations (e.g. Standard A, B, C and E-3). INTELNET services are leased transponder services offered on a range of INTELSAT satellites in C- or Ku-bands on a pre-emptible or non-pre-emptible basis. These services are offered on a full-time or occasional-use basis, by assigning predetermined satellite resources of bandwidth and power with a smallest allotment of 1 MHz. In this paper, the system design and network architecture for the INTELNET I and II services are addressed. Representative transmission parameters have also been developed for the INTELNET I and II services. System cost considerations as a result of the use of micro terminals are analysed.     

Recommendations for reducing mortality among workers of mining enterprises

The authors elaborated measures aimed to lower mortality and higher life expectancy among workers of able-bodied age. The recommendations involve changes in technology of drift mining, improved electric safety of technologic equipment, annual analysis of mortality parameters among miners, accomplishment of prophylactic recommendations on treatment of functional cardiovascular disorders, provision of workers with pectin as a therapeutic and prophylactic nutrient.     

Chronic cold stress alters the basal and evoked electrophysiological activity of rat locus coeruleus neurons

In vivo extracellular single-unit recording techniques revealed that chronic cold stress significantly alters both the basal and the evoked electrophysiological activity of noradrenergic neurons in the locus coeruleus of the anaesthetized rat. Following 17-21 days of chronic cold exposure (5 degrees C), the single-unit activity of histologically-identified locus coeruleus neurons in chloral hydrate-anaesthetized rats was recorded and analysed in terms of their basal firing rate and pattern of spike activity, as well as their response to footshock stimulation. There was no significant difference in the incidence of spontaneously active cells/electrode track between cold-stressed rats and control rats. However, the basal spike activity of locus coeruleus cells recorded from cold-stressed rats differed significantly from that of control rats along two dimensions: i) they displayed significantly higher basal firing rates (mean = 1.88 Hz vs 1.20 Hz, respectively); and ii) they frequently exhibited spontaneous burst-firing activity that was not observed in control rats (observed in 15/17 cold-stressed rats vs 1/26 control rats). The evoked spike activity of locus coeruleus cells in cold-stressed rats also differed significantly from that of control rats along two dimensions: i) they were more likely to respond to footshock stimulation (mean = 90.3% vs 74.4%, respectively); and ii) these responses were more likely to consist of multispike bursts of action potentials (mean = 8 bursts/50 stimulations vs 1 burst/50 stimulations, respectively). These results indicate that alterations in the electrophysiological activity of noradrenergic locus coeruleus neurons may contribute to the phenomenon of stress-induced sensitization of norepinephrine release that is thought to underlie some of the neuropathological changes that accompany long-term stress.