Two-component systems are signal transduction systems which enable bacteria to regulate cellular functions in response to changing environmental conditions. The unicellular Synechocystis sp. PCC 6803 has become a model organism for a range of biochemical and molecular biology studies aiming at investigating environmental stress response. The publication of the complete genome sequence of the cyanobacterium Synechocystis sp. PCC 6803 provided a tremendous stimulus for research in this field, and at least 80 open reading frames were identified as members of the two-component signal transduction systems in this single species of cyanobacteria. To date, functional roles have been determined for only a limited number of such proteins. This review summarizes our current knowledge about the two-component signal transduction systems in Synechocystis sp. PCC 6803 and describes recent achievements in elucidating the functional roles of these systems.

This study investigated the inhibitory effect of a high efficiency diuretic, furosemide, on carbonic anhydrase (CA). First, comparing the inhibitory effect of acetazolamide, a low efficiency diuretic, on CA, shows that furosemide or acetazolamide can quickly make CA inactive when its concentration is close to the enzyme concentration, different from the usual inhibitory kinetics in which the concentration of the inhibitor is far higher than the enzyme concentration. Secondly, the reaction of the enzyme indicates that the inhibitory effect of furosemide or acetazolamide on carbonic anhydrase is quickly reversible. Finally, the degree of the inhibitory effect of furosemide and of acetazolamide on CA are compared. The results show that furosemide inhibits CA less than acetazolamide.

The stability of GCN4 leucine zipper and its four mutants in guanidine hydrochloride was detected to verify the contributions of different a position amino acid residues in polypeptide sequences to the forming and stability of parallel coiled coils. The changes of the circular dichroism spectra show that the displacement of the a position polar asparagine and the increase of asparagine in the GCN4 leucine zipper can reduce the α-helix content of the coiled coil structure. The mutants are less stable than the natural peptide in guanidine hydrochloride. The results show that the interaction between the polar asparagine contributes to the conformational stability of the coiled coil. Both the conformation and the number of polar residues in the coiled coil also affect the α-helix content and its resistance to the denaturant. The conclusions provide evidence describing the folding process of proteins including coiled coils in vivo.

The changes of the activity and conformation of Aspergillus niger phytase in urea were detected by far-ultraviolet circular dichroism (CD) spectra, fluorescence spectra, and enzyme activity assays. The results show that no enzyme activity can be detected after phytase is incubated for 10 h in 3.0 mol/L urea, even though at this urea concentration, less than 20% of the tertiary and secondary structures in the native enzyme changed. The inactivation reaction kinetics is found to be a monophasic first-order reaction, but the unfolding is a biphasic process consisting of two first-order reactions. The inactivation rates of the free enzyme and the substrate-enzyme complex are much faster than the conformational changes during urea denaturation. All of the results indicate that, as a glycoprotein, phytase's activity is strongly dependent on its conformational integrity. The phytase active sites seem to be located in a limited region in the molecule and display more conformational fragility and flexibility to denaturants than enzyme molecular structure as a whole.

The mechanism of apoptosis induced by nicotinamide was investigated by treating mouse myeloma cells (Sp2/0) with various concentrations of nicotinamide. The typical hallmarks of apoptosis, including chromatin condensation and DNA fragmentation, were detected when cells were treated with nicotinamide at concentrations of 30, 40, 50, and 60 mmol/L. The apoptosis percentage increased with increasing nicotinamide concentration. Interestingly, the strong antioxidant melatonin did not restrain the apoptosis induced by nicotinamide in mouse myeloma cells but greatly increased the induction of nicotinamide on apoptosis. When cells were preincubated with 0.1, 1, and 10 mmol/L melatonin before nicotinamide induction, the percentage of apoptosis induced by 50 mmol/L nicotinamide markedly increased with increasing melatonin concentration. These results suggest that apoptosis induced by nicotinamide has no relationship with oxidative stress and melatonin could enhance nicotinamide-induced apoptosis in mouse myeloma cells by stimulating cell division in a certain manner. Nicotinamide may provide a new method to treat some kinds of tumors with no damage to normal tissues.

Neural stem cells (NSCs) are currently considered as powerful candidate seeding cells for regeneration of both spinal cords and peripheral nerves. In this study, NSCs derived from fetal rat cortices were co-cultured with chitosan to evaluate the cell affinity of this material. The results showed that NSCs grew and proliferated well on chitosan films and most of them differentiated into neuron-like cells after 4 days of culture. Then, molded and braided chitosan conduits were fabricated and characterized for their cytotoxicity, swelling, and mechanical properties. Both types of conduits had no cytotoxic effects on fibroblasts (L929 cells) or neuroblastoma (Neuro-2a) cells. The molded conduits are much softer and more flexible while the braided conduits possess much better mechanical properties, which suggests different potential applications.

Chitosan has been shown to be a promising material for various applications in tissue engineering. Recently, adipose tissue derived stromal cells (ADSCs) have been investigated as an alternative source of seed cells for tissue engineering. The compatibility of chitosan and chitosan-gelatin complexes with ADSCs is not known. In the present study, ADSCs were isolated and characterized by phenotype using fluorescence-activated cell sorting (FACS). The morphology, viability, and the ability of the ADSCs to differentiate on chitosan and chitosan-gelatin composite films with 60 wt.% gelatin were evaluated. Results show that the ADSCs are positive for CD29, CD44, and CD105, but negative for CD31, CD34, and CD45. ADSCs adhere and grow better on the composite films than on the chitosan films. The ability of ADSCs to differentiate into osteogenic and adipogenic lineage cells is not affected by their being cultured on chitosan-gelatin composite films. Therefore, chitosan-gelatin composite films are compatible with ADSCs and do not impair the ability of ADSCs to differentiate into osteogenic and adipogenic lineage cells.

Porous chitosan microspheres with diameters ranging from 180 μm to 280 μm were successfully prepared, using an anti-phase suspension method combined with temperature controlled freeze-extraction. The mean pore diameter could be regulated from 5 μm to 60 μm by varying the freezing temperature through the cooling rate. Results with in vitro chondrocyte cultures showed that cells could attach, proliferate, and spread on these porous microspheres as well as inside the microcarriers. The materials and cell co-cultures were characterized using both optical and scanning electron microscopy. These results show that the porous chitosan microspheres are promising candidates for tissue culture for use as an injectable tissue engineering scaffold.

Spontaneous secretions occur in both neurons and non-neuronal cells, and calcium is important for these secretion processes. However, the detailed roles of calcium on the secretions have not yet been identified. In the present study, cultured Xenopus myocytes loaded with exogenous acetylcholine (ACh) into the cytoplasm in the absence of extracellular Ca2+ undergo spontaneous quantal ACh secretion as detected by the appearance of pulsatile miniature endplate currents. Analysis of the frequencies, amplitudes, and time courses of these currents suggests that similar cellular mechanisms are involved in the secretions of ACh in normal medium and Ca2+-free solution. Various doses of ryanodine were used to regulate the intracellular Ca2+ to different levels. The spontaneous ACh secretion from myocytes in Ca2+-free medium was decreased by reducing intracellular Ca2+ levels and enhanced by increasing cytosolic Ca2+ levels. These observations demonstrate that the spontaneous secretion from isolated myocytes and the effect of ryanodine on ACh-loaded cells are both independent of extracellular Ca2+ while Ca2+ in the sarcoplasmic reticulum plays a crucial role in the secretions.

Calcium ions are important in many vital neuron processes, including spontaneous neurotransmitter release. Extracellular calcium has long been known to be related to spontaneous neurotransmitter release, but the detailed mechanism for the effect of intracellular Ca2+ on synaptic release has not yet been understood. In this research, 1,2-bis-(o-aminophenoxy)-ethane-N, N, N’, N’-tetraacetic acid tetraacetoxy-methyl ester (BAPTA-AM) was used to combine with cytosolic free Ca2+ in a calcium free medium of cultured Xenopus neuromuscular junctions (NMJ). The spontaneous synaptic current (SSC) frequency was obviously reduced. Then, drugs were applied to interrupt and activate the Ca2+ release channels in the endoplasmic reticulum (ER) membrane, but the SSC frequency was not affected. The results show that spontaneous neurotransmitter release depends on intracellular rather than ER calcium in cultured Xenopus NMJ without extracellular calcium.

Deposition of amyloid in the brain is a critical step in the pathogenesis of Alzheimer's disease. The endocytosis of β-amyloid peptide (Aβ) is an important factor among the many factors that contribute to the genesis of amyloid deposits. Since cholesterol participates in many important physiological processes, the present work investigated the relationship between the cellular cholesterol content and the endocytosis of the exogenic Aβ, and found that reduction of the cholesterol content by methyl-β-cyclodextrin could reduce the endocytosis of Aβ. The study indicates that the endocytosis of Aβ is partly mediated by cholesterol.

For a long time, lipid rafts have been thought to participate in regulating neurotransmitter release. However, the existence of lipid rafts on synaptic vesicles (SVs) and the mechanism by which exocytosis-relative proteins distribute on this structure have not been fully investigated. There is also much controversial data concerning rafts on SVs and synaptic vesicle proteins which makes the results difficult to interpret. This study systematically analyzed the existence and properties of lipid rafts on purified SVs by sucrose density gradient centrifugation, cholesterol depletion, and temperature variation. The data reveals that typical lipid rafts on SVs are both cholesterol dependent and temperature sensitive. Previous confusing results may have been caused by improper treatment or side effects of particular reagent. We also screened the lateral distribution of major exocytosis-related SV proteins and found that only the synaptobrevin (syb) and synaptotagmin (syt) produce detectable association with lipid rafts in 1% Triton X-100.

Baicalin facilitates the effectiveness of both penicillin and Notopterygium ethanol extracts (NEE) on Staphylococcus aureus. Two Staphylococcus aureus strains, ATCC29213 and 03-21 (a clinical isolate), were tested. When combined with 128 μg ? mL?1 of baicalin, the minimum inhibitory concentration (MIC) of penicillin was reduced from 0.25 μg ? mL?1 to 0.06 μg ? mL?1, while that of NEE was reduced from 32 μg ? mL?1 to 8 μg ? mL?1 against ATCC29213. Against the 03-21 strain, the MIC of penicillin was reduced from 4 μg ? mL?1 to 1 μg ? mL?1, while that of NEE was reduced from 32 μg ? mL?1 to 4 μg ? mL?1. Viable number counting results show that combination of baicalin and penicillin acts on the killing mechanism, while the baicalin and NEE acts on the inhibition. These results show that baicalin acts as a synergistic ally with both penicillin and NEE but has different modes of action.

The spatial distribution patterns of species are always scale-dependent and spatially self-similar in ecological systems. In this work, vegetation distribution data collected from the vegetation map of the Xigaz?region was analyzed using a box-counting method. The power law of the box-counting dimension (DB) across a range of scales (5-160 km) confirms the fractal patterns for most vegetation formations, while the fluctuations of the scale-specific DB among the different abundance groups indicate limitations of fractal coherence. The fractal method is shown to be a useful tool for measuring the distribution patterns of vegetation formations across scales, which provides important information for both species and habitat conservation, especially in landscape management.

The ionizing groups at the active site of alkaline phosphatase (ALP, EC 3.1.3.1) from Ostrea cucullate were studied using kinetic methods. The ionization constant, pKe, of the ionizing groups at the active site of the enzyme was found to be 10.11 at 37.0°C and the organic solvent, dioxin, had no effect on the pKe. The standard dissociation enthalpy (ΔHo) was determined to be 10.57 kcal/mol (1 cal=4.18 J). The results show that the dissociation group of the enzyme active center is the ?-NH3+ of lysine. Chemical modification of the enzyme by acetic anhydride and succinic anhydride demonstrates that the amino group is one of the enzyme's functional groups.

In order to remove background noise and improve the quality of speech for digital hearing aids, a single-channel speech enhancement algorithm is proposed. The algorithm is implemented and assessed on a digital hearing aid platform based on the TI DSP TMS320VC5502 chip. Assuming that background noise is stationary or varies slowly, an energy-based voice activity detection algorithm is adopted by adaptively tracking the minima and maxima of the power envelope in noisy speech. The target speech is then enhanced by using a Wiener filter, on the basis of a short-term power spectral estimation. To deal with the distracting musical noise of the processed speech, phase randomization, along with adjacent spectral averaging, is adopted. Objective measures and an informal hearing test both show an improved performance as well as obvious attenuation of residual noise. The low power consumption and high efficiency render the whole algorithm very applicable for use in digital hearing aids.

To produce high-quality sweet potato plantlets rapidly at low production costs, single nodal leafy cuttings of sweet potato (Ipomoea batatas (L.) Lam.) plantlets were cultured in vitro for 14 days on sugar- and phytohormone-free Murashige and Skoog (MS) liquid medium, at a photosynthetic photon flux (PPF) of 200 μmol ? m?2 ? s?1 and a CO2 concentration (v/v) of 1.8×10?3. A factorial experiment was conducted with two levels of the number of air exchanges of the vessel (NAE), 8.7-12.2 h?1 and ≥12.2 h?1, and two types of supporting material, vermiculite and Florialite (a porous material). The control treatment consisted of a photomixotrophic culture using a sugar- and NAA-containing agar MS medium with an NAE of 2.4 h?1. PPF and CO2 concentrations were the same as the photoautotrophic treatments. In comparison with the control treatment, the photoautotrophic treatments with NAE of 8.7–12.2 h?1 (MF treatment) and ≥12.2 h?1 (HF treatment) were, respectively, 2.2 and 2.8 times in dry weight, and 3.7 and 4.2 times in net photosynthetic rate, when Florialite was used. The survival percentages of the plantlets in the field were, respectively, 86% and 97% in the MF and HF treatments, and 35% and 46% higher than that in the control treatment. The plantlets cultured with Florialite showed greater growth, compared with those cultured with vermiculite. Photoautotrophic micropropagation system with high NAE and the use of porous supporting material can produce high-quality plantlets and make it possible to reduce production costs.

The spike protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) mediates cell fusion by binding to target cell surface receptors. This paper reports a simple method for dissecting the viral protein and for searching for foldable fragments in a random but systematic manner. The method involves digestion by DNase I to generate a pool of short DNA segments, followed by an additional step of reassembly of these segments to produce a library of DNA fragments with random ends but controllable lengths. To rapidly screen for discrete folded polypeptide fragments, the reassembled gene fragments were further cloned into a vector as N-terminal fusions to a folding reporter gene which was a variant of green fluorescent protein. Two foldable fragments were identified for the SARS-CoV spike protein, which coincide with various anti-SARS peptides derived from the hepated repeat (HR) region 2 of the spike protein. The method should be applicable to other viral proteins to isolate antigen or vaccine candidates, thus providing an alternative to the full-length proteins (subunits) or linear short peptides.