Search

The purpose of this study was to establish a novel nontoxic disulfide bond reducing method for lipophilic drug-loaded albumin nanoparticle preparation and make a systematic investigation on this method. Cysteine (Cys) was used to break the disulfide bond of albumin and introduce the self-assembly of drug and albumin. Paclitaxel (PTX) and bovine serum albumin (BSA) were selected to be the model lipophilic drug and albumin. The particle formation dynamics, influencing factors and formation mechanisms were investigated by determining the characteristics of particles including the particle size and yield. Nanoparticles with diameter of 50-400 nm and drug loading efficiency up to 18.3% were prepared successfully. pH 7-8 was suitable for nanoparticle preparation. Temperature, BSA concentration and Cys concentration had positive effects on the particle size and yield. When PTX added was less than the maximal amount of PTX that could bind to BSA, particles with a spherical structure could be formed; otherwise nanoparticles with a core-shell structure could be formed. This novel nontoxic disulfide bond reducing method provides a common and safe method for preparing various kinds of albumin-based nanocarriers for a wide range of applications, from drug (especially the lipophilic drug) delivery to diagnosis of disease.

The importance of pre-nucleation cluster stability as the key parameter controlling nucleation of atmospheric airborne ions is well-established. In this Article, large ternary ionic (HSO(4)(-))(H(2)SO(4))(m)(NH(3))(H(2)O)(n) clusters have been studied using Density Functional Theory (DFT) and composite ab initio methods. Twenty classes of clusters have been investigated, and thermochemical properties of common atmospheric (HSO(4)(-))(H(2)SO(4))(m)(NH(3))(0)(H(2)O)(k) and (HSO(4)(-))(H(2)SO(4))(m)(NH(3))(1)(H(2)O)(n) clusters (with m, k, and n up to 3) have been obtained. A large amount of new themochemical and structural data ready-to-use for constraining kinetic nucleation models has been reported. We have performed a comprehensive thermochemical analysis of the obtained data and have investigated the impacts of ammonia and negatively charged bisulfate ion on stability of binary clusters in some detail. The comparison of theoretical predictions and experiments shows that the PW91PW91/6-311++G(3df,3pd) results are in very good agreement with both experimental data and high level ab initio CCSD(T)/CBS values and suggest that the PW91PW91/6-311++G(3df,3pd) method is a viable alternative to higher level ab initio methods in studying large pre-nucleation clusters, for which the higher level computations are prohibitively expensive. The uncertainties in both theory and experiments have been investigated, and possible ways of their reduction have been proposed.

The compositional properties of water-extractable organic nitrogen (WEON) affect its behavior in lake ecosystems. This work is the first comprehensive study using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) for the characterization of the molecular composition of WEON in lake sediment. In sediments of Erhai Lake in China, this study found complex WEON species, with N-containing compounds in the northern, central, and southern regions contributing 34.47%, 42.44%, and 40.6%, respectively, of total compounds. Additionally, a van Krevelen diagram revealed that lignin components were dominant in sediment WEON structures (68% of the total), suggesting terrestrial sources. Furthermore, this study applied ESI-FT-ICR-MS to the examination of the environmental processes of lake sediment WEON on a molecular level. The results indicated that sediment depth impacted WEON composition and geochemical processes. Compared with other ecosystems, the double bond equivalent (DBE) value was apparently lower in Erhai sediment, indicating the presence of relatively fewer and smaller aromatic compounds. In addition, the presence of a large number of N-containing species and abundant oxidized nitrogen functional compounds that were likely to biodegrade may have further increased the potential releasing risk of WEON from Erhai sediment under certain environmental conditions.

Lysine adsorption at clay/aqueous interfaces plays an important role in the mobility, bioavailability, and degradation of amino acids in the environment. Knowledge of these interfacial interactions facilitates our full understanding of the fate and transport of amino acids. Here, X-ray diffraction (XRD) and attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR) measurements were used to explore the dynamic process of lysine adsorption on montmorillonite and the competition with Ca(2+) at the molecular level. Density functional theory (DFT) calculations were employed to determine the peak assignments of dissolved lysine in the solution phase. Three surface complexes, including dicationic, cationic, and zwitterionic structures, were observed to attach to the clay edge sites and penetrate the interlayer space. The increased surface coverage and Ca(2+) competition did not affect the interfacial lysine structures at a certain pH, whereas an elevated lysine concentration contributed to zwitterionic-type coordination at pH 10. Moreover, clay dissolution at pH 4 could be inhibited at a higher surface coverage with 5 and 10 mM lysine, whereas the inhibition effect was inconspicuous or undetected at pH 7 and 10. The presence of Ca(2+) not only could remove a part of the adsorbed lysine but also could facilitate the readsorption of dissolved Si(4+) and Al(3+) and surface protonation. Our results provide new insights into the process of lysine adsorption and its effects on montmorillonite surface sites.

Development of "smart" noninvasive bioimaging probes for trapping specific enzyme activities is highly desirable for cancer therapy in vivo. Given that β-galactosidase (β-gal) is an important biomarker for cell senescence and primary ovarian cancers, we design an enzyme-activatable ratiometric near-infrared (NIR) probe (DCM-βgal) for the real-time fluorescent quantification and trapping of β-gal activity in vivo and in situ. DCM-βgal manifests significantly ratiometric and turn-on NIR fluorescent signals simultaneously in response to β-gal concentration, which makes it favorable for monitoring dynamic β-gal activity in vivo with self-calibration in fluorescent mode. We exemplify DCM-βgal for the ratiometric tracking of endogenously overexpressed β-gal distribution in living 293T cells via the lacZ gene transfection method and OVCAR-3 cells, and further realize real-time in vivo bioimaging of β-gal activity in colorectal tumor-bearing nude mice. Advantages of our system include light-up ratiometric NIR fluorescence with large Stokes shift, high photostability, and pH independency under the physiological range, allowing for the in vivo real-time evaluation of β-gal activity at the tumor site with high-resolution three-dimensional bioimaging for the first time. Our work provides a potential tool for in vivo real-time tracking enzyme activity in preclinical applications.

Biomimetic multilayers based on layer-by-layer (LbL) assembly were prepared as functional films with compact structure by incorporating the mussel-inspired catechol cross-linking. Dopamine-modified poly(acrylic acid) (PAADopa) was synthesized as a polyanion to offer electrostatic interaction with the prelayer polyethylenimine (PEI) and consecutively cross-linked by zinc to generate compact multilayers with tunable physicochemical properties. In situ layer-by-layer growth and cross-linking were monitored by a quartz crystal microbalance with dissipation (QCM-D) to reveal the kinetics of the process and the influence of Dopa chemistry. Addition of Dopa enhanced the mass adsorption and led to the formation of a more compact structure. An increase of ionic strength induced an increase in mass adsorption in the Dopa-cross-linked multilayers. This is a universal approach for coating of various surfaces such as Au, SiO2, Ti, and Al2O3. Roughness observed by AFM in both wet and dry conditions was compared to confirm the compact morphology of Dopa-cross-linked multilayers. Because of the pH sensitivity of Dopa moiety, metal-chelated Dopa groups can be turned into softer structure at higher pH as revealed by reduction of Young's modulus determined by MFP-3D AFM. A deeper insight into the growth and mechanical properties of Dopa-cross-linked polyelectrolyte multilayers was addressed in the present study. This allows a better control of these systems for bioapplications.

Knowledge of the interfacial interactions between aspartate and minerals, especially its competition with phosphate, is critical to understanding the fate and transport of amino acids in the environment. Adsorption reactions play important roles in the mobility, bioavailability, and degradation of aspartate and phosphate. Attenuated total reflectance Fourier-transform infrared (ATR-FTIR) measurements and density functional theory (DFT) calculations were used to investigate the interfacial structures and their relative contributions in single-adsorbate and competition systems. Our results suggest three dominant mechanisms for aspartate: bidentate inner-sphere coordination involving both α- and γ-COO(-), outer-sphere complexation via electrostatic attraction and H-bonding between aspartate NH2 and goethite surface hydroxyls. The interfacial aspartate is mainly governed by pH and is less sensitive to changes of ionic strength and aspartate concentration. The phosphate competition significantly reduces the adsorption capacity of aspartate on goethite. Whereas phosphate adsorption is less affected by the presence of aspartate, including the relative contributions of diprotonated monodentate, monoprotonated bidentate, and nonprotonated bidentate structures. The adsorption process facilitates the removal of bioavailable aspartate and phosphate from the soil solution as well as from the sediment pore water and the overlying water.

This review discusses the possible relationship between protein charge anisotropy, protein binding affinity, polymer structure, and selective phase separation. We hope that a fundamental understanding of primarily electrostatically driven protein-polyelectrolyte (PE) interactions can enable the prediction of selective protein binding, and hence selective coacervation through non-specific electrostatics. Such research will partially challenge the assumption that specific binding has to be realized through specific binding sites with a variety of short-range interactions and some geometric match. More specifically, the recent studies on selective binding of proteins by polyelectrolytes were examined from different assemblies in addition to the electrostatic features of proteins and PEs. At the end, the optimization of phase separation based on binding affinity for selective coacervation and some considerations relevant to using PEs for protein purification were also overviewed.

The binding of bovine serum albumin (BSA) and β-lactoglobulin (BLG) to TTMA (a cationic gold nanoparticle coupled to 3,6,9,12-tetraoxatricosan-1-aminium, 23-mercapto-N,N,N-trimethyl) was studied by high-resolution turbidimetry (to observe a critical pH for binding), dynamic light scattering (to monitor particle growth), and isothermal titration calorimetry (to measure binding energetics), all as a function of pH and ionic strength. Distinctively higher affinities observed for BLG versus BSA, despite the lower pI of the latter, were explained in terms of their different charge anisotropies, namely, the negative charge patch of BLG. To confirm this effect, we studied two isoforms of BLG that differ in only two amino acids. Significantly stronger binding to BLGA could be attributed to the presence of the additional aspartates in the negative charge domain for the BLG dimer, best portrayed in DelPhi. This selectivity decreases at low ionic strength, at which both isoforms bind well below pI. Selectivity increases with ionic strength for BLG versus BSA, which binds above pI. This result points to the diminished role of long-range repulsions for binding above pI. Dynamic light scattering reveals a tendency for higher-order aggregation for TTMA-BSA at pH above the pI of BSA, due to its ability to bridge nanoparticles. In contrast, soluble BLG-TTMA complexes were stable over a range of pH because the charge anisotropy of this protein at makes it unable to bridge nanoparticles. Finally, isothermal titration calorimetry shows endoenthalpic binding for all proteins: the higher affinity of TTMA for BLGA versus BLGB comes from a difference in the dominant entropy term.

The impact of organic species which are present in the Earth’s atmosphere on the burst of new particles is critically important for the understanding of the molecular nature of atmospheric nucleation phenomena. Amines have recently been proposed as possible stabilizers of binary pre-nucleation clusters. In order to advance the understanding of atmospheric nucleation phenomena, a quantum-chemical study of hydrogen-bonded complexes of binary sulfuric acid-water clusters with methyl-, dimethyl- and trimethylamines representing common atmospheric organic species, vegetation products and laboratory impurities has been carried out. The thermochemical stability of the sulfuric acid-amines-water complexes was found to be higher than that of the sulfuric acid-ammonia-water complexes, in qualitative agreement with the previous studies. However, the enhancement in stability due to amines appears to not be large enough to overcome the difference in typical atmospheric concentrations of ammonia and amines. Further research is needed in order to address the existing uncertainties and to reach a final conclusion about the importance of amines for the atmospheric nucleation.

Nucleation in the Earth’s atmosphere plays a vital role in the aerosol radiative forcing associated directly with the production of ultrafine particles responsible for adverse public health impacts and global climate changes. In the present work, the interaction of the common organic oxalic acid with the atmospheric nucleation precursors and trace ionic species has been investigated using the Density Functional Theory (DFT). A comprehensive study of the hydrogen-bonded complexes of oxalic acid with neutral monomers and ionic clusters of the sulfuric acid, water and ammonia, the key atmospheric nucleation precursors, has been carried out. We found that the bonding of the oxalic acid with the above-mentioned neutrals is moderately weak, and thus, the oxalic acid cannot stabilize the binary sulfuric acid–water and ternary sulfuric acid–ammonia clusters. The interaction of the oxalic acid with positive ionic species is, in contrast, strong and significantly enhances stability of the clusters formed over the ions. The hydration of positively charged (C2H2O4)(H3O+), ( C 2 H 2 O 4 ) ( NH 4 + ) and (C2H2O4)(H3O+)(H2SO4) clusters is also strong and close to that of the sulfuric acid, the key atmospheric nucleation precursor. These considerations lead us to a logical conclusion that the oxalic acid can catalyse the production of positively charged pre-nucleation clusters and, thus, its role in nucleation of positive ions in the Earth’s troposphere should be studied in further details.

Atmospheric aerosols formed via nucleation in the Earth's atmosphere play an important role in the aerosol radiative forcing associated directly with global climate changes and public health. Although it is well-known that atmospheric aerosol particles contain organic species, the chemical nature of and physicochemical processes behind atmospheric nucleation involving organic species remain unclear. In the present work, the interaction of common organic acids with molecular weights of 122, 116, 134, 88, 136, and 150 (benzoic, maleic, malic, pyruvic, phenylacetic, and tartaric acids) with nucleation precursors and charged trace species has been investigated. We found a moderate strong effect of the organic species on the stability of neutral and charged ionic species. In most cases, the free energies of the mixed H(2)SO(4)-organic acid dimer formation are within 1-1.5 kcal mol(-1) of the (H(2)SO(4))(NH(3)) formation energy. The interaction of the organic acids with trace ionic species is quite strong, and the corresponding free energies far exceed those of the (H(3)O(+))(H(2)SO(4)) and (H(3)O(+))(H(2)SO(4))(2) formation. These considerations lead us to conclude that the aforementioned organic acids may possess a substantial capability of stabilizing both neutral and positively charged prenucleation clusters, and thus, they should be studied further with regard to their involvement in the gas-to-particle conversion in the Earth's atmosphere.

A fundamental question in multilingualism is whether the neural substrates are shared or segregated for the two or more languages spoken by polyglots. This study employs functional MRI to investigate the neural substrates underlying the perception of two sentence‐level prosodic phenomena that occur in both Mandarin Chinese (L1) and English (L2): sentence focus (sentence‐initial vs. ‐final position of contrastive stress) and sentence type (declarative vs. interrogative modality). Late‐onset, medium proficiency Chinese‐English bilinguals were asked to selectively attend to either sentence focus or sentence type in paired three‐word sentences in both L1 and L2 and make speeded‐response discrimination judgments. L1 and L2 elicited highly overlapping activations in frontal, temporal, and parietal lobes. Furthermore, region of interest analyses revealed that for both languages the sentence focus task elicited a leftward asymmetry in the supramarginal gyrus; both tasks elicited a rightward asymmetry in the mid‐portion of the middle frontal gyrus. A direct comparison between L1 and L2 did not show any difference in brain activation in the sentence type task. In the sentence focus task, however, greater activation for L2 than L1 occurred in the bilateral anterior insula and superior frontal sulcus. The sentence focus task also elicited a leftward asymmetry in the posterior middle temporal gyrus for L1 only. Differential activation patterns are attributed primarily to disparities between L1 and L2 in the phonetic manifestation of sentence focus. Such phonetic divergences lead to increased computational demands for processing L2. These findings support the view that L1 and L2 are mediated by a unitary neural system despite late age of acquisition, although additional neural resources may be required in task‐specific circumstances for unequal bilinguals. Hum. Brain Mapp, 2007. © 2006 Wiley‐Liss, Inc.

In this study, nanosized zero-valent iron-reduced graphene oxide (nZVI-rGO)-activated persulfate (PS) was used to investigate the generation of reactive oxygen species (ROS) for the degradation of trichloroethylene (TCE) in the aqueous solution. More than 98 % of TCE was degraded within 2 min under experimental conditions. The generation of ·OH increased when the pH was shifted toward the basic region while ·SO4 − radicals’ intensity increased in the acidic pH. Different scenarios have been observed in ·O2 − generation in the neutral and strong basic pH and decreased in acidic or slightly basic pH. In addition, the intensity of ·OH was increased with the addition of HCO3 − (10 mM) and NO3 − (100 mM) but decreased in the presence of Cl− (10 and 100 mM), HCO3 − (100 mM), and NO3 − (10 mM). The degradation of anisole, probe for both ·OH and ·SO4 −, was slightly enhanced by 10 mM NO3 − anions but decreased in 100 mM salt solution. ·O2 − intensity was increased while HCO3 − (10 and 100 mM) and NO3 − (100 mM) anions were used. nZVI-rGO-activated PS process could remove TCE in aqueous effectively, and the ROS generation and intensity were influenced by solution pH values and anions.

Transmission electron microscopy (TEM) was employed to obtain morphology, size, composition, and mixing state of background fine particles with diameter less than 1 μm in the Qinghai-Tibetan Plateau (QTP) during 15 September to 15 October 2013. Individual aerosol particles mainly contained secondary inorganic aerosols (SIA-sulfate and nitrate) and organics during clean periods (PM2.5: particles less than 2.5 μg m−3). The presence of KCl-NaCl associated with organics and an increase of soot particles suggest that an intense biomass burning event caused the highest PM2.5 concentrations (> 30 μg m−3) during the study. A large number fraction of the fly ash-containing particles (21.73 %) suggests that coal combustion emissions in the QTP significantly contributed to air pollutants at the median pollution level (PM2.5: 10–30 μg m−3). We concluded that emissions from biomass burning and from coal combustion both constantly contribute to anthropogenic particles in the QTP atmosphere. Based on size distributions of individual particles in different pollution levels, we found that gas condensation on existing particles is an important chemical process for the formation of SIA with organic coating. TEM observations show that refractory aerosols (e.g., soot, fly ash, and visible organic particles) likely adhere to the surface of SIA particles larger than 200 nm due to coagulation. Organic coating and soot on surface of the aged particles likely influence their hygroscopic and optical properties in the QTP, respectively. To our knowledge, this study reports the first microscopic analysis of fine particles in the background QTP air.

We report the simple one pot synthesis of iron-silver (FeAg) bimetallic nanoparticles with different compositions on graphene support. The nanoparticles are well dispersed on the graphene sheet as revealed by the TEM, XRD, and Raman spectra. The antibacterial activity of graphene-FeAg nanocomposite (NC) towards Bacillus subtilis, Escherichia coli, and Staphylococcus aureus was investigated by colony counting method. Graphene-FeAg NC demonstrates excellent antibacterial activity as compared to FeAg bimetallic without graphene. To understand the antibacterial mechanism of the NC, oxidative stress caused by reactive oxygen species (ROS) and the glutathione (GSH) oxidation were investigated in the system. It has been observed that ROS production and GSH oxidation are concentration dependent while the increase in silver content up to 50% generally enhances the ROS production while ROS decreases on further increase in silver content. Graphene loaded FeAg NC demonstrates higher GSH oxidation capacity than bare FeAg bimetallic nanocomposite. The mechanism study suggests that the antibacterial activity is probably due to membrane and oxidative stress produced by the nanocomposites. The possible antibacterial pathway mainly includes the non-ROS oxidative stress (GSH oxidation) while ROS play minor role.

Poly(2-(methacryloyloxy)ethyltrimethyl ammonium chloride) (PMATAC) modified magnetic nanoparticles (NPs) with a high zeta potential of ca. 50mV were synthesized by atom transfer radical polymerization (ATRP). The prepared NPs consist of a magnetic core around 13nm and a PMATAC shell around 20nm attached on the surface of magnetic nanoparticles. Thermodynamic binding parameters between β-lactoglobulin and these polycationic NPs were investigated at different ionic strengths by high-resolution turbidimetry, dynamic light scattering (DLS), and isothermal titration calorimetry (ITC). Both turbidity and ITC show that binding affinities for BLG display a non-monotonic ionic strength dependence trend and a maximum appears at ionic strength of 50mM. Such observation should arise from the coeffects of protein charge anisotropy visualized by DelPhi electrostatic modeling and the strong electrostatic repulsion among highly charged NPs at a variety of ionic strengths.

Amines, organic derivatives of NH3, are important common trace atmospheric species that can enhance new particle formation in the Earth’s atmosphere under favorable conditions. While methylamine (MA), dimethylamine (DMA) and trimethylamine (TMA) all efficiently enhance binary nucleation, MA may represent the lower limit of the enhancing effect of amines on atmospheric nucleation. In the present paper, we report new thermochemical data concerning MA-enhanced nucleation, which were obtained using the DFT PW91PW91/6-311++G (3df, 3pd) method, and investigate the enhancement in production of stable pre-nucleation clusters due to the MA. We found that the MA ternary nucleation begins to dominate over ternary nucleation of sulfuric acid, water and ammonia at [MA]/[NH3] &gt, ~10−3. This means that under real atmospheric conditions ([MA] ~ 1 ppt, [NH3] ~ 1 ppb) the lower limit of the enhancement due to methylamines is either close to or higher than the typical effect of NH3. A very strong impact of the MA is observed at low RH, however it decreases quickly as the RH grows. Low RH and low ambient temperatures were found to be particularly favorable for the enhancement in production of stable sulfuric acid-water clusters due to the MA.

Creativity, a multifaceted construct, can be studied in various ways, for example, investigating phases of the creative process, quality of the creative product, or the impact of expertise. Previous neuroimaging studies have assessed these individually. Believing that each of these interacting features must be examined simultaneously to develop a comprehensive understanding of creative behavior, we examined poetry composition, assessing process, product, and expertise in a single experiment. Distinct activation patterns were associated with generation and revision, two major phases of the creative process. Medial prefrontal cortex (MPFC) was active during both phases, yet responses in dorsolateral prefrontal and parietal executive systems (DLPFC/IPS) were phase‐dependent, indicating that while motivation remains unchanged, cognitive control is attenuated during generation and re‐engaged during revision. Experts showed significantly stronger deactivation of DLPFC/IPS during generation, suggesting that they may more effectively suspend cognitive control. Importantly however, similar overall patterns were observed in both groups, indicating the same cognitive resources are available to experts and novices alike. Quality of poetry, assessed by an independent panel, was associated with divergent connectivity patterns in experts and novices, centered upon MPFC (for technical facility) and DLPFC/IPS (for innovation), suggesting a mechanism by which experts produce higher quality poetry. Crucially, each of these three key features can be understood in the context of a single neurocognitive model characterized by dynamic interactions between medial prefrontal areas regulating motivation, dorsolateral prefrontal, and parietal areas regulating cognitive control and the association of these regions with language, sensorimotor, limbic, and subcortical areas distributed throughout the brain. Hum Brain Mapp 36:3351–3372, 2015. © 2015 The Authors. Human Brain Mapping Published by Wiley Periodicals, Inc..

Whether or not categorical perception results from the operation of a special, language-specific, speech mode remains controversial. In this cross-language (Mandarin Chinese, English) study of the categorical nature of tone perception, we compared native Mandarin and English speakers' perception of a physical continuum of fundamental frequency contours ranging from a level to rising tone in both Mandarin speech and a homologous (nonspeech) harmonic tone. This design permits us to evaluate the effect of language experience by comparing Chinese and English groups; to determine whether categorical perception is speech-specific or domain-general by comparing speech to nonspeech stimuli for both groups; and to examine whether categorical perception involves a separate categorical process, distinct from regions of sensory discontinuity, by comparing speech to nonspeech stimuli for English listeners. Results show evidence of strong categorical perception of speech stimuli for Chinese but not English listeners. Categorical perception of nonspeech stimuli was comparable to that for speech stimuli for Chinese but weaker for English listeners, and perception of nonspeech stimuli was more categorical for English listeners than was perception of speech stimuli. These findings lead us to adopt a memory-based, multistore model of perception in which categorization is domain-general but influenced by long-term categorical representations.

A series of star-shaped naphthalimide derivatives (TPA-NP) were synthesized with a triphenylamine (TPA) core and chromophores at the rim. Their steady-state absorption, fluorescence, and transient ...

Auditory nerve single-unit population studies have demonstrated that phase-locking plays a dominant role in the neural encoding of both the spectrum and voice pitch of speech sounds. Phase-locked neural activity underlying the scalp-recorded human frequency-following response (FFR) has also been shown to encode certain spectral features of steady-state and time-variant speech sounds as well as pitch of several complex sounds that produce time-invariant pitch percepts. By extension, it was hypothesized that the human FFR may preserve pitch-relevant information for speech sounds that elicit time-variant as well as steady-state pitch percepts. FFRs were elicited in response to the four lexical tones of Mandarin Chinese as well as to a complex auditory stimulus which was spectrally different but equivalent in fundamental frequency ( f 0 ) contour to one of the Chinese tones. Autocorrelation-based pitch extraction measures revealed that the FFR does indeed preserve pitch-relevant information for all stimuli. Phase-locked interpeak intervals closely followed f 0 . Spectrally different stimuli that were equivalent in F 0 similarly showed robust interpeak intervals that followed f 0 . These FFR findings support the viability of early, population-based ‘predominant interval’ representations of pitch in the auditory brainstem that are based on temporal patterns of phase-locked neural activity.

The Chinese language provides an optimal window for investigating both segmental and suprasegmental units. The aim of this cross‐linguistic fMRI study is to elucidate neural mechanisms involved in extraction of Chinese consonants, rhymes, and tones from syllable pairs that are distinguished by only one phonetic feature (minimal) vs. those that are distinguished by two or more phonetic features (non‐minimal). Triplets of Chinese monosyllables were constructed for three tasks comparing consonants, rhymes, and tones. Each triplet consisted of two target syllables with an intervening distracter. Ten Chinese and English subjects were asked to selectively attend to targeted sub‐syllabic components and make same‐different judgments. Direct between‐group comparisons in both minimal and non‐minimal pairs reveal increased activation for the Chinese group in predominantly left‐sided frontal, parietal, and temporal regions. Within‐group comparisons of non‐minimal and minimal pairs show that frontal and parietal activity varies for each sub‐syllabic component. In the frontal lobe, the Chinese group shows bilateral activation of the anterior middle frontal gyrus (MFG) for rhymes and tones only. Within‐group comparisons of consonants, rhymes, and tones show that rhymes induce greater activation in the left posterior MFG for the Chinese group when compared to consonants and tones in non‐minimal pairs. These findings collectively support the notion of a widely distributed cortical network underlying different aspects of phonological processing. This neural network is sensitive to the phonological structure of a listener's native language. Hum. Brain Mapping 20:185–200, 2003. © 2003 Wiley‐Liss, Inc.

Bacteria must contend with conditions of nutrient limitation in all natural environments. Complex programmes of gene expression, controlled in part by the alternative sigma factors sigmaS (sigma38, RpoS) and sigmaH (sigma32, RpoH), allow a number of bacterial species to survive conditions of partial or complete starvation. We show here that the alternative sigma factor sigmaE (sigma24, RpoE) also facilitates the survival of Salmonella typhimurium under conditions of nutrient deprivation. Expression of the sigmaE regulon is strongly induced upon entry of Salmonella into stationary phase. A Salmonella mutant lacking sigmaE has reduced survival during stationary phase as well as increased susceptibility to oxidative stress. A Salmonella strain lacking both sigmaE and sigmaS is non-viable after just 24 h in stationary phase, but survival of these mutants is completely preserved under anaerobic stationary-phase conditions, suggesting that oxidative injury is one of the major mechanisms of reduced microbial viability during periods of nutrient deprivation. Moreover, the attenuated virulence of sigmaE-deficient Salmonella for mice can be largely restored by genetic abrogation of the host phagocyte respiratory burst, suggesting that the sigmaE regulon plays an important antioxidant role during Salmonella infection of mammalian hosts.

A more efficient and convenient strategy was demonstrated to immobilize silver nanoparticles (NPs) with a crystalline structure into the spherical polyelectrolyte brushes (SPB) as an antibacterial material. The SPB used for surface coating (Ag immobilized PVK–PAA SPB) consists of a poly(N-vinylcarbazole) (PVK) core and poly(acrylic acid) (PAA) chain layers which are anchored onto the surface of PVK core at one end. Well-dispersed silver nanoparticles (diameter ∼ 3.5 nm) then formed and were electrostatically confined in the brush layer. Ag content is controlled by a repeated loading process. Thin film coatings were then constructed by layer-by-layer depositions of positive charged poly(diallyldimethylammonium chloride) (PDDA) and SPB. The multilayer composites display excellent stability as well as antibacterial performance but not for simple PVK–PAA coated surface. The results show that almost complete bacteria growth including both dispersed bacterial cells and biofilms was inhibited over a period of 24 h. This approach opens a novel strategy for stable and efficient immobilization of Ag NPs in fabrication of antibacterial materials.

A comprehensive set of methods based on spatial independent component analysis (sICA) is presented as a robust technique for artifact removal, applicable to a broad range of functional magnetic resonance imaging (fMRI) experiments that have been plagued by motion-related artifacts. Although the applications of sICA for fMRI denoising have been studied previously, three fundamental elements of this approach have not been established as follows: 1) a mechanistically-based ground truth for component classification; 2) a general framework for evaluating the performance and generalizability of automated classifiers; and 3) a reliable method for validating the effectiveness of denoising. Here we perform a thorough investigation of these issues and demonstrate the power of our technique by resolving the problem of severe imaging artifacts associated with continuous overt speech production. As a key methodological feature, a dual-mask sICA method is proposed to isolate a variety of imaging artifacts by directly revealing their extracerebral spatial origins. It also plays an important role for understanding the mechanistic properties of noise components in conjunction with temporal measures of physical or physiological motion. The potentials of a spatially-based machine learning classifier and the general criteria for feature selection have both been examined, in order to maximize the performance and generalizability of automated component classification. The effectiveness of denoising is quantitatively validated by comparing the activation maps of fMRI with those of positron emission tomography acquired under the same task conditions. The general applicability of this technique is further demonstrated by the successful reduction of distance-dependent effect of head motion on resting-state functional connectivity.

A type III protein secretion system encoded bySalmonellapathogenicity island 2 (SPI2) has been found to be required for virulence and survival within macrophages. Here, SPI2 was shown to allowSalmonella typhimuriumto avoid NADPH oxidase–dependent killing by macrophages. The ability of SPI2-mutant bacteria to survive in macrophages and to cause lethal infection in mice was restored by abrogation of the NADPH oxidase–dependent respiratory burst. Ultrastructural and immunofluorescence microscopy demonstrated efficient localization of the NADPH oxidase in the proximity of vacuoles containing SPI2-mutant but not wild-type bacteria, suggesting that SPI2 interferes with trafficking of oxidase-containing vesicles to the phagosome.

Salmonella enterica serotype Enteritidis ( S. enteritidis ) is a major food-borne pathogen, and its incidence among all Salmonella serotypes has increased dramatically in the last two decades. To study the virulence characteristics of clinical isolates of S. enteritidis , we determined the 50% lethal doses (LD 50 ) in mice of isolates of two major phage types (4 and 8). Isolates of both phage types showed a wide range of LD 50 after oral inoculation, varying from under 10 2 organisms to over 10 8 organisms. No significant difference in LD 50 was observed between the phage types. These observations indicated that clinical isolates of S. enteritidis are highly heterogeneous in their ability to cause death in mice. We compared the LD 50 s of these isolates to the results observed from in vitro pathogenicity assays. We also analyzed these isolates for recognized Salmonella virulence loci ( spv , sodCI , sopE , and sef ). The in vitro phenotypes of the isolates showed no obvious correlation with their LD 50 in any given assay, and the virulence genes tested were present in all isolates. However, the isolate with the lowest LD 50 (isolate 97A 2472) was resistant to acidified sodium nitrite (ASN). Moreover, the most acid-susceptible, macrophage-susceptible, and ASN-susceptible isolates were attenuated for virulence in mice. These results, based on extensive analysis of clinical isolates of S. enteritidis , demonstrate the complex nature of Salmonella pathogenesis in mice. Our results also indicate the limitation of in vitro assays in predicting in vivo virulence.

Periplasmic Cu, Zn-cofactored superoxide dismutase (SodC) protects Gram-negative bacteria from exogenous oxidative damage. The virulent Salmonella typhimurium strain ATCC 14028s has been found to contain two discrete periplasmic Cu, Zn-SOD enzymes that are only 57% identical at the amino acid level. SodCI is carried by a cryptic bacteriophage, and SodCII is closely related to the Cu, Zn-superoxide dismutase of Escherichia coli . All Salmonella serotypes appear to carry the sodCII locus, but the phage-associated sodCI gene is found only in certain strains belonging to the most highly pathogenic serotypes. Expression of either sodC locus appears to be enhanced during stationary phase, but only sodCII is regulated by the alternative sigma factor σ s (RpoS). Mutants lacking both sodC genes are less lethal for mice than mutants possessing either sodC locus alone, indicating that both Cu, Zn-SOD enzymes contribute to Salmonella pathogenicity. The evolutionary acquisition of an additional sodC gene has contributed to the enhanced virulence of selected Salmonella strains.

Two β-lactoglobulin (BLG) isoforms, BLGA and BLGB, were used as a test bed for the differentiation of proteins using electrostatics. In these studies, the BLGA and BLGB binding to a highly charged, cationic gold nanoparticle (GNP) modified surface was investigated by atomic force microscopy (AFM) and surface plasmon resonance (SPR) spectroscopy. The binding affinity, and more importantly, the selectivity of this surface towards these two almost identical protein isoforms were both significantly increased on the cationic GNP surface array relative to the values measured with the same free cationic GNP in solution. While protein recognition is traditionally achieved almost exclusively via orientation dependent short-range interactions such as hydrogen bonds and hydrophobic interactions, our results show the potential of protein recognition platforms based on enhanced electrostatic interactions.

Rapid eye movement (REM) sleep constitutes a distinct “third state” of consciousness, during which levels of brain activity are commensurate with wakefulness, but conscious awareness is radically transformed. To characterize the temporal and spatial features of this paradoxical state, we examined functional interactions between brain regions using fMRI resting-state connectivity methods. Supporting the view that the functional integrity of the default mode network (DMN) reflects “level of consciousness,” we observed functional uncoupling of the DMN during deep sleep and recoupling during REM sleep (similar to wakefulness). However, unlike either deep sleep or wakefulness, REM was characterized by a more widespread, temporally dynamic interaction between two major brain systems: unimodal sensorimotor areas and the higher-order association cortices (including the DMN), which normally regulate their activity. During REM, these two systems become anticorrelated and fluctuate rhythmically, in reciprocally alternating multisecond epochs with a frequency ranging from 0.1 to 0.01 Hz. This unique spatiotemporal pattern suggests a model for REM sleep that may be consistent with its role in dream formation and memory consolidation.

Levoglucosan is a typical molecular tracer of biomass-burning aerosols in the atmosphere. The mechanism for OH-initiated reaction with levoglucosan is studied at the level of MPWB1K/6-311+G(3df,2p)//MPWB1K/6-31+G(d,p). The possible subsequent reactions in the presence of O 2 , NO and H 2 O are also taken into consideration. The study shows that the H atom abstraction from the C 4 -position by the OH radical is an energetically favorable pathway, and that the OH-initiated products contribute to the formation of SOA and atmospheric acidity. The kinetic calculation is performed and the rate constants are calculated over the temperature range of 200–1500 K, using the Rice–Ramsperger–Kassel–Marcus (RRKM) theory. The rate constant of levoglucosan reacting with the OH radical at 298 K is 2.21 × 10 −13 cm 3 molecule −1 s −1 and the atmospheric lifetime is 26 days ([ OH] = 2.0 × 10 6 molecule cm −3 ). The equilibrium constants both in gas phase and aqueous are computed. The free energy Δ G indicates that, the subsequent reactions tend to take place more spontaneously once the reaction occurs. This work provides a comprehensive investigation about OH-initiated atmospheric reactions with levoglucosan, which is helpful for experiment and risk assessment.

The aggregation of β-lactoglobulin (BLG) near its isoelectric point was studied as a function of ionic strength and pH. We compared the behavior of native BLG with those of its two isoforms, BLG-A and BLG-B, and with that of a protein with a very similar pI, bovine serum albumin (BSA). Rates of aggregation were obtained through a highly precise and convenient pH/turbidimetric titration that measures transmittance to ±0.05 %T. A comparison of BLG and BSA suggests that the difference between pHmax (the pH of the maximum aggregation rate) and pI is systematically related to the nature of protein charge asymmetry, as further supported by the effect of localized charge density on the dramatically different aggregation rates of the two BLG isoforms. Kinetic measurements including very short time periods show well-differentiated first and second steps. BLG was analyzed by light scattering under conditions corresponding to maxima in the first and second steps. Dynamic light scattering (DLS) was used to monitor the kinetics, and static light scattering (SLS) was used to evaluate the aggregate structure fractal dimensions at different quench points. The rate of the first step is relatively symmetrical around pHmax and is attributed to the local charges within the negative domain of the free protein. In contrast, the remarkably linear pH dependence of the second step is related to the uniform reduction in global protein charge with increasing pH below pI, accompanied by an attractive force due to surface charge fluctuations.

Paradoxically, nitric oxide (NO) has been found to exhibit cytotoxic, antiproliferative, or cytoprotective activity under different conditions. We have utilized Salmonella mutants deficient in antioxidant defenses or peptide transport to gain insights into NO actions. Comparison of three NO donor compounds reveals distinct and independent cellular responses associated with specific redox forms of NO. The peroxynitrite (OONO-) generator 3-morpholinosydnonimine hydrochloride mediates oxygen-dependent Salmonella killing, whereas S-nitrosoglutathione (GSNO) causes oxygen-independent cytostasis, and the NO. donor diethylenetriamine-nitric oxide adduct has no antibacterial activity. GSNO has the greatest activity for stationary cells, a characteristic relevant to latent or intracellular pathogens. Moreover, the cytostatic activity of GSNO may best correlate with antiproliferative or antimicrobial effects of NO, which are unassociated with overt cell injury. dpp mutants defective in active dipeptide transport are resistant to GSNO, implicating heterolytic NO+ transfer rather than homolytic NO. release in the mechanism of cytostasis. This transport system may provide a specific pathway for GSNO-mediated signaling in biological systems. The redox state and associated carrier molecules are critical determinants of NO activity.

The neural correlates of creativity are poorly understood. Freestyle rap provides a unique opportunity to study spontaneous lyrical improvisation, a multidimensional form of creativity at the interface of music and language. Here we use functional magnetic resonance imaging to characterize this process. Task contrast analyses indicate that improvised performance is characterized by dissociated activity in medial and dorsolateral prefrontal cortices, providing a context in which stimulus-independent behaviors may unfold in the absence of conscious monitoring and volitional control. Connectivity analyses reveal widespread improvisation-related correlations between medial prefrontal, cingulate motor, perisylvian cortices and amygdala, suggesting the emergence of a network linking motivation, language, affect and movement. Lyrical improvisation appears to be characterized by altered relationships between regions coupling intention and action, in which conventional executive control may be bypassed and motor control directed by cingulate motor mechanisms. These functional reorganizations may facilitate the initial improvisatory phase of creative behavior.

The importance of pre-nucleation cluster stability as the key parameter controlling nucleation of atmospheric airborne ions is well-established. In this Article, large ternary ionic (HSO(4)(-))(H(2)SO(4))(m)(NH(3))(H(2)O)(n) clusters have been studied using Density Functional Theory (DFT) and composite ab initio methods. Twenty classes of clusters have been investigated, and thermochemical properties of common atmospheric (HSO(4)(-))(H(2)SO(4))(m)(NH(3))(0)(H(2)O)(k) and (HSO(4)(-))(H(2)SO(4))(m)(NH(3))(1)(H(2)O)(n) clusters (with m, k, and n up to 3) have been obtained. A large amount of new themochemical and structural data ready-to-use for constraining kinetic nucleation models has been reported. We have performed a comprehensive thermochemical analysis of the obtained data and have investigated the impacts of ammonia and negatively charged bisulfate ion on stability of binary clusters in some detail. The comparison of theoretical predictions and experiments shows that the PW91PW91/6-311++G(3df,3pd) results are in very good agreement with both experimental data and high level ab initio CCSD(T)/CBS values and suggest that the PW91PW91/6-311++G(3df,3pd) method is a viable alternative to higher level ab initio methods in studying large pre-nucleation clusters, for which the higher level computations are prohibitively expensive. The uncertainties in both theory and experiments have been investigated, and possible ways of their reduction have been proposed.

Cyanobacteria (Cyb), an environment waste, was first utilized as a carrier for controlled release of avermectin (Av) and improvement of pesticide photostability. Microalgae powder prepared from Cyb blooms was characterized by elemental analysis (EA), X-ray photoelectron spectroscopy (XPS), infrared spectroscopy (IR), optical microscopy, and scanning electron microscopy (SEM). The globular morphology and the size (~3.5μm) of Cyb do not change significantly, with increase of roughness, after Av adsorption and Carbopol coating. Av manifests considerable binding affinity to Cyb in isopropanol and adsorption decreases in more polar solvents at 25°C. An extended release of Av from Cyb-Av in ethanol/water (v/v, 4:1) mixture was obtained and the release was further controlled by the Carbopol resin coated on Cyb-Av. Thicker Carbopol coating results in larger lag time (t(0)), slower release rate, and less total release amount. The release process was found to be controlled by a diffusion mechanism, but was strongly affected by polymer matrix-Av interaction and polymer relaxation. Finally, adsorbed Av on Cyb exhibits much greater UV-stability than free Av.

The aggregation of insulin is complicated by the coexistence of various multimers, especially in the presence of Zn(2+). Most investigations of insulin multimerization tend to overlook aggregation kinetics, while studies of insulin aggregation generally pay little attention to multimerization. A clear understanding of the starting multimer state of insulin is necessary for the elucidation of its aggregation mechanism. In this work, the native-state aggregation of insulin as either the Zn-insulin hexamer or the Zn-free dimer was studied by turbidimetry and dynamic light scattering, at low ionic strength and pH near pI. The two states were achieved by varying the Zn(2+) content of insulin at low concentrations, in accordance with size-exclusion chromatography results and literature findings (Tantipolphan, R.; Romeijn, S.; Engelsman, J. d.; Torosantucci, R.; Rasmussen, T.; Jiskoot, W. J. Pharm. Biomed. 2010, 52, 195). The much greater aggregation rate and limiting turbidity (τ(∞)) for the Zn-insulin hexamer relative to the Zn-free dimer was explained by their different aggregation mechanisms. Sequential first-order kinetic regimes and the concentration dependence of τ(∞) for the Zn-insulin hexamer indicate a nucleation and growth mechanism, as proposed by Wang and Kurganov (Wang, K.; Kurganov, B. I. Biophys. Chem. 2003, 106, 97). The pure second-order process for the Zn-free dimer suggests isodesmic aggregation, consistent with the literature. The aggregation behavior at an intermediate Zn(2+) concentration appears to be the sum of the two processes.

The effect of polyelectrolyte binding affinity on selective coacervation of proteins with the cationic polyelectrolyte, poly(diallyldimethylammonium chloride) (PDADMAC), was investigated for bovine serum albumin/β-lactoglobulin (BSA/BLG) and for the isoforms BLG-A/BLG-B. High-sensitivity turbidimetric titrations were used to define conditions of complex formation and coacervation (pH(c) and pH(ϕ), respectively) as a function of ionic strength. The resultant phase boundaries, essential for the choice of conditions for selective coacervation for the chosen protein pairs, are nonmonotonic with respect to ionic strength, for both pH(c) and pH(ϕ). These results are explained in the context of short-range attraction/long-range repulsion governing initial protein binding "on the wrong side of pI" and also subsequent phase separation due to charge neutralization. The stronger binding of BLG despite its higher isoelectric point, inferred from lower pH(c), is shown to result from the negative "charge patch" on BLG, absent for BSA, as visualized via computer modeling (DelPhi). The higher affinity of BLG versus BSA was also confirmed by isothermal titration calorimetry (ITC). The relative values of pH(ϕ) for the two proteins show complex salt dependence so that the choice of ionic strength determines the order of coacervation, whereas the choice of pH controls the yield of the target protein. Coacervation at I = 100 mM, pH 7, of BLG from a 1:1 (w/w) mixture with BSA was shown by SEC to provide 90% purity of BLG with a 20-fold increase in concentration. Ultrafiltration was shown to remove effectively the polymer from the target protein. The relationship between protein charge anisotropy and binding affinity and between binding affinity and selective coacervation, inferred from the results for BLG/BSA, was tested using the isoforms of BLG. Substitution of glycine in BLG-B by aspartate in BLG-A lowers pH(c) by 0.2, as anticipated on the basis of DelPhi modeling. The stronger binding of BLG-A, confirmed by ITC, led to a difference in pH(ϕ) that was sufficient to provide enrichment by a factor of 2 for BLG-A in the coacervate formed from "native BLG".

Salmonella typhimurium zwf mutants lacking glucose 6-phosphate dehydrogenase (G6PD) activity have increased susceptibility to reactive oxygen and nitrogen intermediates as well as attenuated virulence in mice. Abrogation of the phagocyte respiratory burst oxidase during experimental infection with zwf mutant Salmonella causes a prompt restoration of virulence, while inhibition of inducible nitric oxide synthase results in delayed lethality. These observations suggest that G6PD-dependent bacterial antioxidant defenses play an important pathogenic role during early salmonellosis and additionally may help to antagonize NO-dependent antimicrobial mechanisms later in the course of infection.

This paper presents a new application of the dynamic iterated rippled noise (IRN) algorithm by generating dynamic pitch contours representative of those that occur in natural speech in the context of EEG and the frequency following response (FFR). Besides IRN steady state and linear rising stimuli, curvilinear rising stimuli were modeled after pitch contours of natural productions of Mandarin Tone 2. Electrophysiological data on pitch representation at the level of the brainstem, as reflected in FFR, were evaluated for all stimuli, static or dynamic. Autocorrelation peaks were observed corresponding to the fundamental period (tau) as well as spectral bands at the fundamental and its harmonics for both a low and a high iteration step. At the higher iteration step, both spectral and temporal FFR representations were more robust, indicating that both acoustic properties may be utilized for pitch extraction at the level of the brainstem. By applying curvilinear IRN stimuli to elicit FFRs, we can evaluate the effects of temporal degradation on 1) the neural representation of linguistically-relevant pitch features in a target population (e.g., cochlear implant) and 2) the efficacy of signal processing schemes in conventional hearing aids and cochlear implants to recover these features.

Crosslanguage comparisons of brainstem-evoked potentials have revealed experience-dependent plasticity in pitch representation for curvilinear f0 contours representative of Mandarin tones. To assess the tolerance limits of this experience-dependent selectivity, we evaluated cross-linguistically (Chinese, English) the pitch strength and tracking accuracy of linear rising and falling f0 ramps representative of Mandarin tones 2 and 4. No crosslanguage differences in pitch strength or accuracy were observed for either tone, indicating that stimuli with linear rising/falling ramps elicit homogeneous pitch representations at the level of the brainstem regardless of language experience. We conclude that pitch extraction at the brainstem level is critically dependent on specific dimensions of pitch contours that native speakers have been exposed to in natural speech contexts.

Implicit, abstract knowledge acquired through language experience can alter cortical processing of complex auditory signals. To isolate prelexical processing of linguistic tones (i.e., pitch variations that convey part of word meaning), a novel design was used in which hybrid stimuli were created by superimposing Thai tones onto Chinese syllables (tonal chimeras) and Chinese tones onto the same syllables (Chinese words). Native speakers of tone languages (Chinese, Thai) underwent fMRI scans as they judged tones from both stimulus sets. In a comparison of native vs. non‐native tones, overlapping activity was identified in the left planum temporale (PT). In this area a double dissociation between language experience and neural representation of pitch occurred such that stronger activity was elicited in response to native as compared to non‐native tones. This finding suggests that cortical processing of pitch information can be shaped by language experience and, moreover, that lateralized PT activation can be driven by top‐down cognitive processing. Hum Brain Mapp, 2005. © 2005 Wiley‐Liss, Inc.