The lack of organelle-targeted neurotransmitter probes limits understanding of their intracellular roles. Here we created an organelle-targeted neurotransmitter nanoprobe using specific molecule-trapped DNA nanostructures for multiple recognition effects. In particular, we designed phenylboronic acid derivatives for chemical reaction and hydroxymethyl groups for forming hydrogen bonding with norepinephrine (NE), which were confined into tetrahedral DNA nanostructures with the optimized spatial effects, achieving the specific and rapid NE identification. Moreover, cyanine 3 providing built-in correction was designed for accurate NE quantification and the HaloTag ligand was synthesized for HaloTag protein targeting onto the organelle membrane, which were bonded to tips of the DNA nanostructure. The developed nanotrap demonstrated high selectivity, fast response (~50 ms), good stability and biocompatibility for organelle NE imaging. Using this tool, we discovered that traumatic brain injury triggers NE bursts in the endoplasmic reticulum, inducing endoplasmic reticulum (ER) stress, altering ER-mitochondrial protein regulation, promoting mitophagy and mitochondrial dysfunction and ultimately causing neuronal death.

We performed a systems vaccinology analysis to investigate immune responses in humans to an H5N1 influenza vaccine, with and without the AS03 adjuvant, to identify factors influencing antibody response magnitude and durability. Our findings revealed a platelet and adhesion-related blood transcriptional signature on day 7 that predicted the longevity of the antibody response, suggesting a potential role for platelets in modulating antibody response durability. As platelets originate from megakaryocytes, we explored the effect of thrombopoietin (TPO)-mediated megakaryocyte activation on antibody response longevity. We found that TPO administration enhanced the durability of vaccine-induced antibody responses. TPO-activated megakaryocytes also promoted survival of human bone-marrow plasma cells through integrin β1/β2-mediated cell–cell interactions, along with survival factors APRIL and the MIF–CD74 axis. Using machine learning, we developed a classifier based on this platelet-associated signature, which predicted antibody response longevity across six vaccines from seven independent trials, highlighting a conserved mechanism for vaccine durability. Pulendran and colleagues define a molecular signature that can be used to predict the durability of antibody responses to vaccination and reveal important insights into the mechanisms by which vaccines induce durable immunity.

Many human diseases, including metabolic diseases, are intertwined with the immune system. The understanding of how the human immune system interacts with pharmaceutical drugs is still limited, and epidemiological studies only start to emerge. As the metabolomics technology matures, both drug metabolites and biological responses can be measured in the same global profiling data. Therefore, a new opportunity presents itself to study the interactions between pharmaceutical drugs and immune system in the high-resolution mass spectrometry data. We report here a double-blinded pilot study of seasonal influenza vaccination, where half of the participants received daily metformin administration. Global metabolomics was measured in the plasma samples at six timepoints. Metformin signatures were successfully identified in the metabolomics data. Statistically significant metabolite features were found both for the vaccination effect and for the drug-vaccine interactions. This study demonstrates the concept of using metabolomics to investigate drug interaction with the immune response in human samples directly at molecular levels.

Microbial interaction with the host through sensing receptors, including SIGNR1, sustains intestinal homeostasis against pathogenic inflammation. The newly discovered commensal Propionibacterium strain, P. UF1, regulates the intestinal immunity against pathogen challenge. However, the molecular events driving intestinal phagocytic cell response, including colonic dendritic cells (DCs), by this bacterium are still elusive. Here, we demonstrate that the glycosylation of bacterial large surface layer protein A (LspA) by protein O -mannosyltransferase 1 (Pmt1) regulates the interaction with SIGNR1, resulting in the control of DC transcriptomic and metabolomic machineries. Programmed DCs promote protective T cell response to intestinal Listeria infection and resist chemically induced colitis in mice. Thus, our findings may highlight a novel molecular mechanism by which commensal surface glycosylation interacting with SIGNR1 directs the intestinal homeostasis to potentially protect the host against proinflammatory signals inducing colonic tissue damage.

The immune response to live-attenuated Francisella tularensis vaccine and its host evasion mechanisms are incompletely understood. Using RNA-Seq and LC–MS on samples collected pre-vaccination and at days 1, 2, 7, and 14 post-vaccination, we identified differentially expressed genes in PBMCs, metabolites in serum, enriched pathways, and metabolites that correlated with T cell and B cell responses, or gene expression modules. While an early activation of interferon α/β signaling was observed, several innate immune signaling pathways including TLR, TNF, NF-κB, and NOD-like receptor signaling and key inflammatory cytokines such as Il-1α, Il-1β, and TNF typically activated following infection were suppressed. The NF-κB pathway was the most impacted and the likely route of attack. Plasma cells, immunoglobulin, and B cell signatures were evident by day 7. MHC I antigen presentation was more actively up-regulated first followed by MHC II which coincided with the emergence of humoral immune signatures. Metabolomics analysis showed that glycolysis and TCA cycle-related metabolites were perturbed including a decline in pyruvate. Correlation networks that provide hypotheses on the interplay between changes in innate immune, T cell, and B cell gene expression signatures and metabolites are provided. Results demonstrate the utility of transcriptomics and metabolomics for better understanding molecular mechanisms of vaccine response and potential host–pathogen interactions.

Structures of [Ce(GGG)]3+ and [Ce(GGG ? H)]2+ have been investigated by DFT calculations. The two lowest-energy structures of the triply charged metal complex have the peptide in either the iminol or conventional zwitterionic form, and these ions have almost identical energies. In the doubly charged complex, the iminol and charge-solvated structures are the best structures on the potential energy surface, but the latter is favored. In both iminol structures, the metal ion coordinates to the iminol oxygen rather than to the nitrogen, unlike in previously reported iminol-containing complexes. Triply charged [Ce(peptide)]3+ complexes are fragile and not easily isolated in a mass spectrometer, whereas the doubly charged [Ce(peptide ? H)]2+ complexes are more robust. Here, we studied the fragmentations of 37 [Ce(peptide ? H)]2+ and 30 [Ce(peptide)(peptide ? H)]2+ complexes and the results are systematically summarized. Losses of CO and/or H2O are the most commonly observed fragmentation channels for [Ce(peptide ? H)]2+ complexes and these dissociation pathways are modeled by DFT calculations. For [Ce(peptide)(peptide ? H)]2+ complexes the neutral peptide plays the role of a solvent molecule but, unlike in the dissociations of [Ce(CH3CN)(peptide ? H)]2+ complexes, the loss of the solvent molecule is not observed. Instead, fragmentation occurs by cleavage of the second amide bond of the solvating peptide molecule.

Purpose: To determine plasma metabolite and metabolic pathway differences between patients with type 2 diabetes with diabetic retinopathy (DR) and without retinopathy (diabetic controls), and between patients with proliferative DR (PDR) and nonproliferative DR (NPDR). Methods: Using high-resolution mass spectrometry with liquid chromatography, untargeted metabolomics was performed on plasma samples from 83 DR patients and 90 diabetic controls. Discriminatory metabolic features were identified through partial least squares discriminant analysis, and linear regression was used to adjust for age, sex, diabetes duration, and hemoglobin A1c. Pathway analysis was performed using Mummichog 2.0. Results: In the adjusted analysis, 126 metabolic features differed significantly between DR patients and diabetic controls. Pathway analysis revealed alterations in the metabolism of amino acids, leukotrienes, niacin, pyrimidine, and purine. Arginine, citrulline, glutamic γ-semialdehyde, and dehydroxycarnitine were key contributors to these pathway differences. A total of 151 features distinguished PDR patients from NPDR patients, and pathway analysis revealed alterations in the β-oxidation of saturated fatty acids, fatty acid metabolism, and vitamin D3 metabolism. Carnitine was a major contributor to the pathway differences. Conclusions: This study demonstrates that arginine and citrulline-related pathways are dysregulated in DR, and fatty acid metabolism is altered in PDR patients compared with NPDR patients.

[a3 + H]2+ ions generated from Ln3+/tripeptide complexes, where Ln = La or Ce, have similar structures to the linear [an]+ ions but with protonation at both the terminal NH2 and N═CH2 groups. Ion stability is favored by having the basic secondary amine of the proline residue at the N-terminus and by an amino acid residue accommodating one of the protons on the side chain. Dissociation of [a3 + H]2+ ions derived from peptides containing only aliphatic residues is by cleavage of the second amide bond to give [b2]+ or [a2]+ ions along with internal [a1]+ ions. For [a3 + H]2+ ions containing a tryptophan residue in the central location, in addition to cleavage of the amide bond, losses of neutrals NH3, HN═CHR, (NH3 + CO), and HNCO were observed. Dissociations of some unsolvated Ln3+/tripeptide complexes gave [b3 + H]2+ ions in low abundance; formation of these [b3 + H]2+ ions was favored by the presence of a proline residue at the N-terminus and by either a histidine or tryptophan residue in the central position. Dissociation of these [b3 + H]2+ ions was by the loss of (H2O + CO) and not only CO, indicating that these ions did not have the same type of oxazolone structure as found for [bn]+ ions. Density functional theory calculations suggest that the observed [b3 + H]2+ ions of ProGlyGly were formed from [Ce(ProGlyGly)]3+ complexes in which the peptide was bound to the metal ion as an enolate. Dissociation of the slightly lower-energy complex, where the peptide is bound in the keto form, would produce an oxazolone but the high barrier required to create this isomer of the [b3 + H]2+ ion would be sufficient to result in further dissociation. Two isomers of the [b3 + H]2+ ion of ProHisGly have been created, one from the [Ce(ProHisGly)]3+ complex that characteristically dissociates by the combined loss of (H2O + CO) and the other by the loss of glycine from [ProHisGlyGly + 2H]2+. The [b3 + H]2+ ion derived from [ProHisGlyGly + 2H]2+ dissociated by the loss of only CO.

Macrocyclization is commonly observed in large bn(+) (n≥ 4) ions and as a consequence can lead to incorrect protein identification due to sequence scrambling. In this work, the analogous [b5- H]˙(+) radical cations derived from aliphatic hexapeptides (GA5˙(+)) also showed evidence of macrocyclization under CID conditions. However, the major fragmentation for [b5- H]˙(+) ions is the loss of CO2 and not CO loss, which is commonly observed in closed-shell bn(+) ions. Isotopic labeling using CD3 and (18)O revealed that more than one common structure underwent dissociations. Theoretical studies found that the loss of CO2 is radical-driven and is facilitated by the radical being located at the Cα atom immediately adjacent to the oxazolone ring. Comparable energy barriers against macrocyclization, hydrogen-atom transfer, and fragmentations are found by DFT calculations and the results are consistent with the experimental observations that a variety of dissociation products are observed in the CID spectra.

The collision-induced dissociation (CID) of [b5 - H]˙(+) ions containing four alanine residues and one tryptophan give identical spectra regardless of the initial location of the tryptophan indicating that, as proposed for b5(+) ions, sequence scrambling occurs prior to dissociation. Cleavage occurs predominantly at the peptide bonds and at the N-Cα bond of the alanine residue that is attached to the N-terminus of the tryptophan residue. The product of the latter pathway, an ion at m/z 240, is the base peak in all the mass spectra. With the exception of one minor channel giving a b3(+) ion, the product ions retain both the tryptophan residue and the radical. Experiments with one trideuterated alanine established the sequences of loss of alanine residues. Formation of identical products implies a common intermediate, a [b5 - H]˙(+) ion that has a 'linear' structure in which the tryptophan residue is present as an α-radical located in the oxazolone ring, structure Ie. Density functional theory calculations show this structure to be at the global minimum, 14.6 kcal mol(-1) below the macrocyclic structure, ion II. Loss of CO from the [b5 - H]˙(+) ions is inhibited by the presence of the radical centre in the oxazolone ring and migration of the proton from the oxazolone ring onto the peptide backbone induces cleavage of an N-Cα or peptide bond. Three calculated structures for the ion at m/z 240 all have an oxazolone ring. Two of these structures may be formed from Ie, depending upon which proton migrates onto the peptide chain prior to the dissociation. The barrier to interconversion between these two structures requires a 1,3-hydrogen atom shift and is high (51.0 kcal mol(-1)), but both can convert into a third isomer that readily loses CO2 (barrier 38.7 kcal mol(-1)). The lowest barrier to the loss of CO, the usual fragmentation path observed for protonated oxazolones, is 47.0 kcal mol(-1).

The ubiquitous nature of microbes has made them the pioneers in radionuclides adsorption and transport. In this study, the radiation resistance and nuclide biosorption capacity of microbes isolated from the Lanyu low-level radioactive waste (LLRW) repository in Taiwan was assessed, the evaluation of the possibility of using the isolated strain as biosorbents for Co-60 and Co (II) from contaminated aquedus solution and the potential impact on radionuclides release.The microbial content of solidified waste and broken fragments of containers at the Lanyu LLRW repository reached 10(5) CFU/g. Two yeast strains, Candida guilliermondii (CT1) and Rhodotorula calyptogenae (RT1) were isolated. The radiation dose necessary to reduce the microbial count by one log cycle of CT1 and RT1 was 2.1 and 0.8 kGy, respectively. Both CT1 and RT1 can grow under a radiation field with dose rate of 6.8 Gy/h, about 100 times higher than that on the surface of the LLRW container in Lanyu repository.CT1 and RT1 had the maximum Co-60 biosorption efficiency of 99.7 +/- 0.1% and 98.3 +/- 0.2%, respectively in Co-60 aqueous solution (700 Bq/mL), and the Co-60 could stably retained for more than 30 days in CT 1. Nearly all of the Co was absorbed and reached equilibrium within 1 h by CT1 and RT1 in the 10 mu g/g Co (II) aqueous solution. Biosorption efficiency test showed almost all of the Co (II) was adsorbed by CT1 in 20 mu g/g Co (II) aqueous solution, the efficiency of biosorption by RT1 in 10 mu g/g of Co (II) was lower. The maximum Co (II) sorption capacity of CT1 and RT1 was 5324.0 +/- 349.0 mu g/g (dry wt) and 3737.6 +/- 86.5 mu g/g (dry wt), respectively, in the 20 mu g/g Co (II) aqueous solution.Experimental results show that microbial activity was high in the Lanyu LLRW repository in Taiwan. Two isolated yeast strains, CT1 and RT1 have high potential for use as biosorbents for Co-60 and Co (II) from contaminated aqueous solution, on the other hand, but may have the impact on radionuclides release from LLRW repository. (C) 2015 Elsevier Ltd. All rights reserved.

Peptide radical cations A(n)Y(•+) (where n = 3, 4, or 5) and A5W(•+) have been generated by collision-induced dissociation (CID) of [Cu(II)(tpy)(peptide)](•2+) complexes. Apart from the charge-driven fragmentation at the N-Cα bond of the hetero residue producing either [c + 2H](+) or [z - H](•+) ions and radical-driven fragmentation at the Cα-C bond to give a(+) ions, unusual product ions [x + H](•+) and [z + H](•+) are abundant in the CID spectra of the peptides with the hetero residue in the second or third position of the chain. The formation of these ions requires that both the charge and radical be located on the peptide backbone. Energy-resolved spectra established that the [z + H](•+) ion can be produced either directly from the peptide radical cation or via the fragment ion [x + H](•+). Additionally, backbone dissociation by loss of the C-terminal amino acid giving [b(n-1) - H](•+) increases in abundance with the length of the peptides. Mechanisms by which peptide radical cations dissociate have been modeled using density functional theory (B3LYP/6-31++G** level) on tetrapeptides AYAG(•+), AAYG(•+), and AWAG(•+).

Three-dimensional planning of a dental-implant site is critical when implants are to initially be used as orthodontic anchorage for tooth movement and to later serve as prosthetic abutments. The combined orthodontic and prosthetic implant restorative treatment of an adult patient with multiple missing teeth, an upper midline deviation, and malocclusion is described in this article. Plastic model bases combined with customized position plates allowed the precise transfer of the proposed implant position from the diagnostic arrangement to the original cast to avoid compromising subsequent orthodontic tooth movement.

A perception-aware motion-compensated frame interpolation algorithm has been proposed. There is no motion estimation process in this low cost frame rate up-conversion (FRUC) algorithm. With the proposed filter-based MV processing techniques and the motion compensation (MC) decision in a motion compensated interpolation (MCI) stage, the proposed algorithm reduces the memory and bandwidth requirement while maintains the visual quality of the interpolation results. The key factor is that most computations in the traditional interpolation are redundant and could be skipped without quality loss because properties of the human visual system.

This paper proposes a class of H∞ filter design for continue-time systems with time-varying delay. By using the Lyapunov functional function approach, some delay-dependent stability conditions can be obtained for the asymptotical stability of the H∞ filter systems, which are expressed as a set of Linear Matrix Inequalities (LMIs). Finally, a numerical example is given to demonstrate the effectiveness and the merit of the proposed method.

Objective: To explore correlations between mRNA (hA3G, hA3F, and hA3B) levels and CD4(+) T-cell counts and HIV-1 viral loads to evaluate their respective roles in disease progression.Methods: Real-time polymerase chain reaction was used to quantify the mRNA levels of hA3G, hA3B, and hA3F in peripheral blood mononuclear cells from slow progress patients (SP), asymptomatic HIV-infected patients (AS), AIDS patients, and HIV-negative controls.Results: The levels of hA3G and hA3B mRNA correlated positively with CD4(+) T-cell counts (r = 0.436, P = 0.002, r = 0.334, P = 0.025), and negatively with HIV-1 viral loads (r = -0.306, P = 0.038, r = -0.301 P = 0.044). The levels of hA3G and hA3B mRNA in HIV-infected subjects were lower than in HIV-negative controls (P < 0.05), and hA3G and hA3B mRNA levels were significantly higher in SP than in AIDS patients (P < 0.05). There was no correlation between the hA3F mRNA level and CD4(+) T-cell Counts or between the hA3F mRNA level and HIV-1 viral loads.Conclusions: Higher expression levels of hA3G and hA3B mRNA in the peripheral blood mononuclear cells of Chinese HIV-infected individuals were found to be associated with slower HIV disease progression, suggesting their potential roles in antiviral innate immunity.

This study investigates the feasibility of using gamma irradiation for photodegradation of a common residual fungicide, pentachloronitrobenzene (PCNB), in ginseng, and for microbial decontamination. American ginseng, Panax quinquefolius, was subjected to gamma irradiation. PCNB residues were analyzed by gas chromatography with electron capture detection and mass spectrometry. Eighty percent of PCNB (100 ppm) in a methanol aqueous solution was degraded by 5 kGy irradiation, and the primary degradation product was pentachloroaniline. Furthermore, contaminated PCNB (3.7 ppm) in ginseng were reduced to 0.2 ppm after 20 kGy irradiation. The IC50 for treatment of Sclerotium rolfsii with 20 kGy irradiated PCNB was about 2.7 times higher than that for treatment with unirradiated PCNB. The survival rate of mouse fibroblast L929 cells treated with 20 kGy irradiated PCNB was about 12.9% higher than that of L929 cells treated with unirradiated PCNB. Additionally, after 20 kGy irradiation, less than 5% reduction of contents of ginsenoside Rb1 and Re were observed, and amounts of ginsenosides Rc, Rd, and Rg1 were not reduced significantly. The minimal gamma dose for microbial decontamination was 10 kGy. Therefore, gamma irradiation can be used for both PCNB photodegradation and microbial decontamination of ginseng without obvious loses of ginsenoside contents.

The gel particle softness effect on cross-flow microfiltration performance is studied. Three kinds of soft gel particles with a similar size distribution but different softness and compression behaviors were prepared and used in these experiments. The particle softness is the most important factor affecting the pseudo-steady filtration flux compared to the other operating variables in the conditions of this study. A filter cake formed by softer particles is compressed into a more compact structure, consequently, resulting in higher filtration resistance and lower filtration flux. The particle deposition probability is analyzed using a force balance model. An increase in particle softness leads to lower particle deposition probability as well as lighter cake mass due to the drastic flux decay. The cake compression during filtration is described using the Voigt in series model. The cake porosity suddenly decreases and the specific cake filtration resistance drastically increases in the early filtration period. The skin cake layer formation in that period plays the most important role in determining the cross-flow microfiltration performance. The cake mass and average specific filtration resistance calculation results using the proposed methods agree fairly with the available experimental data.

Since the increasing populations of HIV-infected and immune-compromised patients due to immunosuppressive medical regimens,there was need to develop CD4+ T cell-independent therapeutic strategies to prevent opportunistic infection.There was a well-known inverse relationship between CD4+ lymphocyte count and the risk of pneumocystis infection and other bacterial infection,but the CD4+ T cells does not hold all of the answers to mechanisms of host defense against these infections.CD40 ligand(CD40L) was critical for host defense against pneumocystis as well as other opportunistic infections.To test whether CD40L could prime immune response in the deficiency of CD4+ T cells in mice,chicken ovalbumin(OVA) was used as the model antigen with the addition of CD40L in CD4-depleted mice.Anti-OVA IgG1 and IgG2a were measured by ELISA three weeks after the boost.The result showed significant increases in IgG2a and IgG1 in the CD40L co-transduced group compared to OVA alone.We also observed a significant increase in OVA-specific IFN-γ elaboration in splenocytes after stimulation with the SIINFEKL peptide in both CD4-replete mice and CD4-depleted mice that were co-transduced with CD40L.These data suggest that CD40L can prime both B-cell and CD8+ T-cell responses.

Objective To study the relationships between neutralizing antibody response against heterologous virus and disease progression in Chinese HIV-1 B'/C infected individuals. Methods Plasmas from HIV-1-infected individuals, grouped as HIV chronically infected or AIDS according to CD4+ count and clinical symptom, were tested for neutralizing activity against the three HIV-1 isolates with very low homology in vitro. Six two-fold dilutions of each plasma sample (from 1/10 to 1/320) were tested against each virus from the panel. Giving a 50% reduction in p24Ag compared with normal human plasma control wells was defined as positive. The breadth of the cross-neutralizing response was defined based on the number of viruses that were effectively neutralized by any given patient-derived plasma sample. The magnitude of the crossneutralizing response was defined based on the average neutralizing titer against all heterologous viruses. Resuits We found that there revealed a significant difference between HIV chronically infected and AIDS group in the breaths and magnitudes of neutralizing heterologous virus. There was higher prevalence for the frequency of neutralizing heterologous virus in HIV chronically infected than AIDS. The results showed that there was positive correlation between the breadths and magnitudes of neutralizing response against heterologous virus and the plasma HIV RNA level in HIV chronically infected group, while not in AIDS group. There was no association between the breadth of the neutralizing responses against heterologous virus and CD4 T cell counts. Conclusion The capacity of neutralizing antibodies against heterologous virus varied among different disease stage. There were higher titers of neutralizing antibodies in HIV chronically infected than AIDS group. The loss of neutralizing antibodies in plasma from AIDS group appears to be associated with a narrowing of the antibody response during disease progression. These suggest that the presence of neutralizing antibodies against hetreologous virus was associated with disease progression.