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The waste produced by the shrimp industry can be a great source for the recovery of important bioactive compounds with potential applications in food industry. The present study aimed to characterize extracted proteases of Aspergillus flavus, Aspergillus niger and Penicillium sp. to be utilized as potential sources to produce shrimp waste protein hydrolysates with degraded antigenic proteins. The extracellular fungal acid protease enzymes were extracted under solid substrate fermentation and partially characterized the enzyme activities, concerning the reaction temperature and pH of the reaction medium. Then, the capabilities of extracted fungal protease enzymes to reduce the potential allergens of shrimp waste protein hydrolysates were examined by SDS-PAGE. After incubation of all three fungal species at 37°C and pH 3.0 for 5 days, the highest protease yield, 527.40 μg/mL was achieved from A. flavus. The optimum temperature for A. flavus for the highest protease activity was 45°C, while for other two microorganisms the optimum temperature was 50°C. The optimum pH values for all three proteases were pH 3. According to the SDS-PAGE protein hydrolyzing patterns, the crude acid protease extracted from A. flavus was the best protease to degrade the potential antigenic proteins over the other two acid proteases extracted from A.niger and Penicillium sp., while all three enzymes had shown the potential to reduce the allergenic capacity of shrimp waste proteins through enzymatic hydrolysis.

Background/Purpose: The global coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) resulted in significant mortality, social disorder and economic hardships. We set out to understand why certain individuals could resolve the disease successfully without serious complications by immune profiling of PBMCs from convalescent COVID-19 patients (n = 14) with mild disease trajectory. Methods: We designed 7 peptides targeting the receptor binding region (RBD) of the spike protein of SARS-CoV-2, that encompass broad HLA class I / II alleles, with the aid of the Immune Epitope Database and Analysis Resource (IEDB) website. The RBD site was chosen as it was shown to be protective when neutralising antibodies against this region could negate binding of viral spike protein to host cell ACE-2 receptor. Convalescent COVID-19 patients enrolled in this study were screened and selected for positive antibody titre against the RBD region. PBMCs from convalescent COVID-19 patients were stimulated with/without the peptides for 72hrs and immune profiled (n = 70 markers across two panels) with the high dimensional single cell mass cytometry platform (CyToF). Results: Convalescent COVID-19 patients elicited a robust recall memory T follicular helper response (CD3 + CD4 + CD45RO + CXCR5 + Tigit + ;∗∗∗∗ p < 0.0001) demonstrating peptide efficacy. Unsupervised clustering (FlowSOM) of the CD4 T cell immune landscape reaffirmed increase in memory T follicular subsets and additionally CD4 + CD45RO + CXCR5 -subsets. Further gating of antigen specific memory cells (CD45RO + CD69 + ) revealed an increase in Tbet + CXCR3 + T effectors in both CD4 and CD8 compartments. Strikingly we detected a parallel increase in CD4 + Treg (CD25 + FoxP3 + ) CXCR3 + Tbet + CD45RO + CD152 + Tigit + expression. Conclusions: COVID-19 patients that successfully resolve the viral infection not only mount a robust T effector and follicular response but also in tandem a similar T regulatory profile.

Bats are reservoir hosts for various zoonotic viruses with pandemdic potential in humans and livestock.

The rate and mechanism of the elimination of N2O from trans-R3Sn-O-N═N-O-SnR3 (R = Ph (1Ph) and R = Cy (1Cy)) to form R3Sn-O-SnR3 (R = Ph (2Ph) and R = Cy (2Cy)) have been studied using both NMR and IR techniques to monitor the reactions in the temperature range of 39-79 °C in C6D6. Activation parameters for this reaction are ΔH⧧ = 15.8 ± 2.0 kcal·mol-1 and ΔS⧧ = -28.5 ± 5 cal·mol-1·K-1 for 1Ph and ΔH⧧ = 22.7 ± 2.5 kcal·mol-1 and ΔS⧧ = -12.4 ± 6 cal·mol-1·K-1 for 1Cy. Addition of O2, CO2, N2O, or PPh3 to sealed tube NMR experiments did not alter in a detectable way the rate or product distribution of the reactions. Computational DFT studies of elimination of hyponitrite from trans-Me3Sn-O-N═N-O-SnMe3 (1Me) yield a mechanism involving initial migration of the R3Sn group from O to N passing through a marginally stable intermediate product and subsequent N2O elimination. Reactions of 1Ph with protic acids HX are rapid and lead to formation of R3SnX and trans-H2N2O2. Reaction of 1Ph with the metal radical •Cr(CO)3C5Me5 at low concentrations results in rapid evolution of N2O. At higher •Cr(CO)3C5Me5 concentrations, evolution of CO2 rather than N2O is observed. Addition of 1 atm or less CO2 to benzene or toluene solutions of 2Ph and 2Cy resulted in very rapid reaction to form the corresponding carbonates R3Sn-O-C(═O)-O-SnR3 (R = Ph (3Ph) and R = Cy (3Cy)) at room temperature. Evacuation results in fast loss of bound CO2 and regeneration of 2Ph and 2Cy. Variable temperature data for formation of 3Cy yield ΔHo = -8.7 ± 0.6 kcal·mol-1, ΔSo = -17.1 ± 2.0 cal·mol-1·K-1, and ΔGo298K = -3.6 ± 1.2 kcal·mol-1. DFT studies were performed and provide additional insight into the energetics and mechanisms for the reactions.

Chronic obstructive pulmonary disease (COPD) is characterised by airflow limitation and infective exacerbations, however, in-vitro model systems for the study of host-pathogen interaction at the individual level are lacking. Here, we describe the establishment of nasopharyngeal and bronchial organoids from healthy individuals and COPD that recapitulate disease at the individual level. In contrast to healthy organoids, goblet cell hyperplasia and reduced ciliary beat frequency were observed in COPD organoids, hallmark features of the disease. Single-cell transcriptomics uncovered evidence for altered cellular differentiation trajectories in COPD organoids. SARS-CoV-2 infection of COPD organoids revealed more productive replication in bronchi, the key site of infection in severe COVID-19. Viral and bacterial exposure of organoids induced greater pro-inflammatory responses in COPD organoids. In summary, we present an organoid model that recapitulates the in vivo physiological lung microenvironment at the individual level and is amenable to the study of host-pathogen interaction and emerging infectious disease. Ministry of Health (MOH) National Medical Research Council (NMRC) National Research Foundation (NRF) Published version This research was supported by the National Research Foundation Singapore under its COVID-19 Research Fund administered by the Singapore Ministry of Health’s National Medical Research Council (MOH000409) (to S.H.C) and National Medical Research Council COVID19RF2-0006 (to L-F.W and D.E.A.) and by the Singapore Ministry of Health’s National Medical Research Council under its Clinician Scientist Award (CSA) (MOH-000710) (S.H.C).

Understanding antiviral immune responses in bats, which are reservoirs for many emerging viruses, could aid the response to future epidemics. Here, we discuss five key areas in which greater consensus among the bat research community is necessary to drive breakthroughs in the field., The COVID-19 pandemic has stressed the importance of understanding species such as bats that can serve as reservoirs for emerging viral threats. Here, Wang and colleagues call for greater consensus among the bat immunology community in five key areas.

The reaction of Ru3(CO)12 with Pt(IPr)(SnBut3)(H) at 68 °C yielded the trigonal bipyramidal cluster Ru3Pt2(IPr)2(CO)12, 1, in 32% yield. The reaction of Os3(CO)12 with Pt(IPr)(SnBut3)(H) at 68 °C furnished the triosmium–monoplatinum cluster complex Os3Pt(IPr)(CO)12, 2, in 51% yield and the triosmium–diplatinum cluster complex Os3Pt2(IPr)2(CO)12, 3, in 19% yield. The reaction of Fe3(CO)12 with Pt(IPr)(SnBut3)(H) at room temperature resulted in cluster fragmentation to afford the diiron–monoplatinum cluster complex Fe2Pt(IPr)(CO)9, 4, in 44% yield. All four compounds were structurally characterized by single-crystal X-ray diffraction analyses.

The rate and mechanism of the elimination of N

Understanding and improving obstetrical quality and safety is an important goal of professional societies, and many interventions such as checklists, safety bundles, educational interventions, or other culture changes have been implemented to improve the quality of care provided to obstetrical patients. Although many factors contribute to delivery decisions, a reduced workload has addressed how provider issues such as fatigue or behaviors surrounding impending shift changes may influence the delivery mode and outcomes.The objective was to assess whether intrapartum obstetrical interventions and adverse outcomes differ based on the temporal proximity of the delivery to the attending's shift change.This was a secondary analysis from a multicenter obstetrical cohort in which all patients with cephalic, singleton gestations who attempted vaginal birth were eligible for inclusion. The primary exposure used to quantify the relationship between the proximity of the provider to their shift change and a delivery intervention was the ratio of time from the most recent attending shift change to vaginal delivery or decision for cesarean delivery to the total length of the shift. Ratios were used to represent the proportion of time completed in the shift by normalizing for varying shift lengths. A sensitivity analysis restricted to patients who were delivered by physicians working 12-hour shifts was performed. Outcomes chosen included cesarean delivery, episiotomy, third- or fourth-degree perineal laceration, 5-minute Apgar score of4, and neonatal intensive care unit admission. Chi-squared tests were used to evaluate outcomes based on the proportion of the attending's shift completed. Adjusted and unadjusted logistic models fitting a cubic spline (when indicated) were used to determine whether the frequency of outcomes throughout the shift occurred in a statistically significant, nonlinear pattern RESULTS: Of the 82,851 patients eligible for inclusion, 47,262 (57%) had ratio data available and constituted the analyzable sample. Deliveries were evenly distributed throughout shifts, with 50.6% taking place in the first half of shifts. There were no statistically significant differences in the frequency of cesarean delivery, episiotomy, third- or fourth-degree perineal lacerations, or 5-minute Apgar scores of4 based on the proportion of the shift completed. The findings were unchanged when evaluated with a cubic spline in unadjusted and adjusted logistic models. Sensitivity analyses performed on the 22.2% of patients who were delivered by a physician completing a 12-hour shift showed similar findings. There was a small increase in the frequency of neonatal intensive care unit admissions with a greater proportion of the shift completed (adjusted P=.009), but the findings did not persist in the sensitivity analysis.Clinically significant differences in obstetrical interventions and outcomes do not seem to exist based on the temporal proximity to the attending physician's shift change. Future work should attempt to directly study unit culture and provider fatigue to further investigate opportunities to improve obstetrical quality of care, and additional studies are needed to corroborate these findings in community settings.

The Raman Lidar for Meteorological Observations (RALMO) is operated at the MeteoSwiss station of Payerne (Switzerland) and provides, amongst other products, continuous measurements of temperature since 2010. The temperature profiles are retrieved from the pure rotational Raman (PRR) signals detected around the 355 nm Cabannes line. The transmitter and receiver systems of RALMO are described in detail, and the reception and acquisition units of the PRR channels are thoroughly characterized. The Fast- Com P7888 card used to acquire the PRR signal, the calculation of the dead time and the desaturation procedure are also presented. The temperature profiles retrieved from RALMO PRR data during the period going from July 2017 to the end of December 2018 have been validated against two reference operational radiosounding systems (ORSs) co-located with RALMO, i.e. the Meteolabor SRS-C50 and the Vaisala RS41. The ORSs have also served to perform the calibration of the RALMO temperature during the validation period. The maximum bias (1Tmax), mean bias ( ) and mean standard deviation ( ) of RALMO temperature Tral with respect to the reference ORS, Tors, are used to characterize the accuracy and precision of Tral along the troposphere. The daytime statistics provide information essentially about the lower troposphere due to lower signal-to-noise ratio. The 1Tmax, and of the differences 1T D Tral-Tors are, respectively, 0.28, 0:02 0:1 and 0:62 0:03 K. The nighttime statistics provide information for the entire troposphere and yield 1Tmax D 0:29 K, D 0:05 0:34K and D 0:66 0:06 K. The small 1Tmax, and values obtained for both daytime and nighttime comparisons indicate the high stability of RALMO that has been calibrated only seven times over 18 months. The retrieval method can correct for the largest sources of correlated and uncorrelated errors, e.g. signal noise, dead time of the acquisition system and solar background. Especially the solar radiation (scattered into the field of view from the zenith angle 8) affects the quality of PRR signals and represents a source of systematic error for the retrieved temperature. An imperfect subtraction of the background from the daytime PRR profiles induces a bias of up to 2K at all heights. An empirical correction f .8/ ranging from 0.99 to 1 has therefore been applied to the mean background of the PRR signals to remove the bias. The correction function f .8/ has been validated against the numerical weather prediction model COSMO (Consortium for Smallscale Modelling), suggesting that f .8/ does not introduce any additional source of systematic or random error to Tral. A seasonality study has been performed to help with understanding if the overall daytime and nighttime zero bias hides seasonal non-zero biases that cancel out when combined in the full dataset., Swiss National Science Foundation (SNSF) European Commission PZ00P2_168114

Bats are reservoir hosts of many zoonotic viruses with pandemic potential. We utilized single-cell transcriptome sequencing (scRNA-seq) to analyze the immune response in bat lungs upon in vivo infection with a double-stranded RNA virus, Pteropine orthoreovirus PRV3M. Bat neutrophils were distinguished by high basal IDO1 expression. NK cells and T cells were the most abundant immune cells in lung tissue. Three distinct CD8

The complex Pt(IPr)(SnBut3)(H), 1 [IPr=N-heterocylic carbene ligand N,N’-bis-(2,6-(diisopropyl)phenyl)imidazol-2-ylidene] reacts with trimethylsilylacetylene to yield the C-H activation product Pt(IPr)(SnBut3)(κ1-C≡CSiMe3)(H)2, 2. Complex 2 isomerizes to form the unique complex Pt(IPr)[η2-E-(SnBut3)(H)C=C(SiMe3)(H)], 3. This demonstrates that in addition to functioning as a bulky ligand the SnBut3 group can also be actively involved in catalysis. Compound 1 catalyzes the reaction of trimethylsilylacetylene and tri-t-butylstannane at room temperature to afford the selective hydrostannylated product E-(SnBut3)(H)C=C(SiMe3)(H), 4. Structures of all compounds have been determined by single crystal x-ray diffraction analyses.

Transition metal complexes containing the sterically encumbered SnBut3 group are the focus of this brief review. The ability of tin compounds to modify both heterogeneous and homogeneous catalysts has been known for some time. Our recent efforts center on utilizing the reagent But3SnH to investigate the influence of this ligand, which combines a steric profile similar to that of PBut3, with functional reactivity at the Sn-H bond. These properties of the SnBut3 ligand allow preparation of new complexes and comparison of their structures and reactivities with less encumbered trialkyl stannanes. This has allowed fine-tuning of the strained molecular geometry at the coordinatively unsaturated site and study of how that influences small molecule activation. Reversible H2 binding and activation, H2–D2 scrambling to form HD, C-H activation of bound ligands and solvents, catalytic hydrogenation, and catalytic hydrostannylation have all been observed for select complexes. In addition, a range of metal cluster complexes incorporating stannane moieties have been prepared and structurally characterized. The But3SnH ligand provides a new dimension for preparation of new molecular architectures with potential applications in homogeneous and heterogeneous catalysis.

The anti-proliferative agent hexamethylene bisacetamide (HMBA) belongs to a class of hybrid bipolar compounds developed more than 30 y ago for their ability to induce terminal differentiation of transformed cells. Recently, HMBA has also been shown to trigger HIV transcription from latently infected cells, via a CDK9/HMBA inducible protein-1 dependent process. However, the effect of HMBA on the immune response has not been explored. We observed that pretreatment of human peripheral blood mononuclear cells with HMBA led to a markedly increased production of IL-12 and IFN-γ, but not of TNF-α, IL-6, and IL-8 upon subsequent infection with Burkholderia pseudomallei and Salmonella enterica. HMBA treatment was also associated with better intracellular bacterial control. HMBA significantly improved IL-12p70 production from CD14+ monocytes during infection partly via the induction of type I IFN in these cells, which primed an increased transcription of the p35 subunit of IL-12p70 during infection. HMBA also increased early type I IFN transcription in human monocytic and epithelial cell lines, but this was surprisingly independent of its previously reported effects on positive transcription elongation factor b and HMBA inducible protein-1. Instead, the effect of HMBA was downstream of a calcium influx, and required the pattern recognition receptor and adaptor STING but not cGAS. Our work therefore links the STING-IRF3 axis to enhanced IL-12 production and intracellular bacterial control in primary monocytes. This raises the possibility that HMBA or related small molecules may be explored as therapeutic adjuvants to improve disease outcomes during intracellular bacterial infections.

Bats are asymptomatic reservoir hosts for several highly pathogenic viruses. Understanding this enigmatic relationship between bats and emerging zoonotic viruses requires tools and approaches which enable the comparative study of bat immune cell populations and their functions. We show that bat genomes have a conservation of immune marker genes which delineate phagocyte populations in humans, while lacking key mouse surface markers such as Ly6C and Ly6G. Cross-reactive antibodies against CD44, CD11b, CD14, MHC II, and CD206 were multiplexed to characterize circulating monocytes, granulocytes, bone-marrow derived macrophages (BMDMs) and lung alveolar macrophages (AMs) in the cave nectar bat Eonycteris spelaea. Transcriptional profiling of bat monocytes and BMDMs identified additional markers – including MARCO, CD68, CD163, CD172α, and CD88 – which can be used to further characterize bat myeloid populations. Bat cells often resembled their human counterparts when comparing immune parameters that are divergent between humans and mice, such as the expression patterns of certain immune cell markers. A genome-wide comparison of immune-related genes also revealed a much closer phylogenetic relationship between bats and humans compared to rodents. Taken together, this study provides a set of tools and a comparative framework which will be important for unravelling viral disease tolerance mechanisms in bats.

The complex [Au(IPr)(2-oxazolidinone)] (1 = IKa) was prepared from reaction of [Au(IPr)Cl] (2), K2CO3, and propargyl amine (PPA). Kinetic studies have been performed for acid cleavage of 1 to yield the oxazolidinone product and an [Au(IPr)(X)] adduct. The fastest rates of cleavage were found to occur for the hydrogen chloride salt of PPA (PPA-HCl) and for the CO2 adduct of PPA, PPA-CO2 = the carbamic acid (CA). This transformation was studied as a function of [CA], pressure of CO2 as well as temperature. Detailed computational studies support the formation of a key intermediate and are also in agreement with a rapid carbonylation/decarbonylation reaction. The computed reactions mechanisms for addition of PPA-HCl and CA are also presented as well as the crystal structure of 1.

It is reported that individuals have less willingness to start their own business. Lack of entrepreneurial intentions impact adversely for the economic development of the country as entrepreneurship is a major source of employment generation and economic development. Thus, scholars emphasize on investigating the factors stimulating the interest of undergraduates to become an entrepreneur. Among the factors, social capital of individuals plays a vital role. On the above backdrop, present study was undertaken to understand the impact of social capital affecting the entrepreneurial intentions of the undergraduates involved in Business Administration and Entrepreneurship courses in Sri Lankan Universities. Findings of the study suggests that the number of social ties, trustworthy relationships and the social norms shared among the undergraduates within their community affect their willingness to start a new business. Thus, the study provides significant insights for the university administrators highlighting importance of facilitating a platform for the undergraduates to network with the fellow undergraduates. R H G W P K Henegedara | D G M P M Gamage "An Investigation of the Impact of Social Capital for the Entrepreneurial Intentions" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-5 , August 2019, URL: https://www.ijtsrd.com/papers/ijtsrd25227.pdf

Universities play a major role in producing entrepreneurs. Until recently, fostering innovations and new product development through entrepreneurship has not been regarded as a primary task of universities. Although the graduates are given the education on entrepreneurship, it is reported that they have less willingness to start their own business. Lack of entrepreneurial intentions among the undergraduates impact adversely for the economic development of the country as entrepreneurship is a major source of employment generation and economic development. Thus, scholars emphasize more on investigating the factors stimulating the interest of undergraduates to become an entrepreneur. On the above backdrop, present study was undertaken to understand the factors affecting the entrepreneurial intentions of the undergraduates involved in Business Administration and Entrepreneurship courses in Sri Lankan Universities. D G M P M Gamage | R H G W P K Henegedara "The Entrepreneurial Intentions among the Undergraduates Involved in Business Administration and Entrepreneurship Courses in Sri Lanka" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-4 , June 2019, URL: https://www.ijtsrd.com/papers/ijtsrd25175.pdf

The reaction of Pt(COD)2 with one equivalents of IBut and PBut3 at room temperature yields the unsaturated platinum mixed N-heterocyclic carbene and phosphine complex Pt(IBut)(PBut3), 1. Complex 1 reacts rapidly with O2 and CO to form the dioxygen adduct Pt(IBut)(PBut3)(η2-O2), 2 and Pt3 trimer cluster complex [Pt(IBut)(CO)]3, 3, respectively. Complex 2 further reacts with CO2 at room temperature to give the peroxycarbonate complex Pt(IBut)(PBut3)[η2-OOC(O)O], 4, and the first example characterized by single crystal X-ray diffraction. The reaction of complex 2 with acetone also resulted in the formation of the peroxoacetonate complex Pt(IBut)(PBut3)[η2-OOC(CH3)2O], 5. All the structures for the complexes 1–5 were characterized by X-ray crystallographic studies.

Bacteria use various endogenous antioxidants for protection against oxidative stress associated with environmental survival or host infection. Although glutathione (GSH) is the most abundant and widely used antioxidant in Proteobacteria, ergothioneine (EGT) is another microbial antioxidant, mainly produced by fungi and Actinobacteria. The Burkholderia genus is found in diverse environmental niches. We observed that gene homologs required for the synthesis of EGT are widely distributed throughout the genus. By generating gene-deletion mutants and monitoring production with isotope-labeled substrates, we show that pathogenic Burkholderia pseudomallei and environmental B. thailandensis are able to synthesize EGT de novo. Unlike most other bacterial EGT synthesis pathways described, Burkholderia spp. use cysteine rather than γ-glutamyl cysteine as the thiol donor. Analysis of recombinant EgtB indicated that it is a proficient sulfoxide synthase, despite divergence in the active site architecture from that of mycobacteria. The absence of GSH, but not EGT, increased bacterial susceptibility to oxidative stresses in vitro. However, deletion of EGT synthesis conferred a reduced fitness to B. pseudomallei, with a delay in organ colonization and time to death during mouse infection. Therefore, despite the lack of an apparent antioxidant role in vitro, EGT is important for optimal bacterial pathogenesis in the mammalian host.-Gamage, A. M., Liao, C., Cheah, I. K., Chen, Y., Lim, D. R. X., Ku, J. W. K., Chee, R. S. L., Gengenbacher, M., Seebeck, F. P., Halliwell, B., Gan, Y.-H. The proteobacterial species Burkholderia pseudomallei produces ergothioneine, which enhances virulence in mammalian infection.

BACKGROUND: While there are well-accepted standards for the diagnosis of arrested active-phase labor, the definition of a "failed" induction of labor remains less certain. One approach to diagnosing a failed induction is based on the duration of the latent phase. However, a standard for the minimum duration that the latent phase of a labor induction should continue, absent acute maternal or fetal indications for cesarean delivery, remains lacking. OBJECTIVE: The objective of this study was to determine the frequency of adverse maternal and perinatal outcomes as a function of the duration of the latent phase among nulliparous women undergoing labor induction. METHODS: This study is based on data from an obstetric cohort of women delivering at 25 U.S. hospitals from 2008-2011. Nulliparous women who had a term singleton gestation in the cephalic presentation were eligible for this analysis if they underwent a labor induction. Consistent with prior studies, the latent phase was determined to begin once cervical ripening had ended, oxytocin was initiated and rupture of membranes (ROM) had occurred, and was determined to end once 5 cm dilation was achieved. The frequencies of cesarean delivery, as well as of adverse maternal (e.g., cesarean delivery, postpartum hemorrhage, chorioamnionitis) and perinatal outcomes (e.g., a composite frequency of either seizures, sepsis, bone or nerve injury, encephalopathy, or death), were compared as a function of the duration of the latent phase (analyzed with time both as a continuous measure and categorized in 3-hour increments). RESULTS: A total of 10,677 women were available for analysis. In the vast majority (96.4%) of women, the active phase had been reached by 15 hours. The longer the duration of a woman's latent phase, the greater her chance of ultimately undergoing a cesarean delivery (P

Type 2 diabetic patients have increased susceptibility to melioidosis, an infectious disease caused by Burkholderia pseudomallei. We had previously shown that peripheral blood mononuclear cells (PBMCs) from diabetic patients with poor glycemic control had a defective IL-12 and IFNγ response to B. pseudomallei infection, resulting in poor intracellular bacterial control. The impaired IL-12 response was due to glutathione (GSH) deficiency characterized by a low reduced to oxidized glutathione ratio (GSH ratio) and could be restored by the addition of reduced GSH to the infected cells. Our goal is to determine whether N-acetyl cysteine (NAC, a GSH pro-drug) supplementation in diabetic patients could improve their immune control of B. pseudomallei. Type 2 diabetic patients with poor glycemic control were given oral supplementation of NAC for six weeks at 1200 mg daily. Their PBMCs and subsets of immune cells showed a significant increase in free GSH concentration. However, the GSH ratio, IL-12 and IFNγ production, and intracellular bacterial killing upon ex-vivo infection did not improve. Thus, oral NAC supplementation in diabetic patients is sufficient to increase intracellular GSH content in blood cells. However, modulating the free GSH content is not sufficient to improve infection outcome as it is the GSH ratio that regulates the IL-12 response in monocytes.

Mass spectrometry imaging (MSI) performed under ambient conditions is a convenient and information-rich method that allows for the comprehensive mapping of chemical species throughout biological tissues with typical spatial resolution in the 40-200 μm range. Ambient MSI methods such as desorption electrospray ionization (DESI) eliminate necessary sample preparation but suffer from lower spatial resolution than laser-based and vacuum techniques. In order to take advantage of the benefits of ambient imaging and to compensate for the somewhat limited spatial resolution, a secondary orthogonal separation nested in the imaging scheme was implemented for more selective discernment of tissue features in the spectral domain. Differential mobility spectrometry (DMS), an ion mobility-based separation that selectively transmits ions based on their high-to-low electric field mobility differences, can significantly reduce background chemical interferences, allowing for increased peak capacity. In this work, DESI DM-MSI experiments on biological tissue samples such as sea algae and mouse brain tissue sections were conducted using fixed DMS compensation voltages that selectively transferred one or a class of targeted compounds. By reducing chemical noise, the signal-to-noise ratio was improved 10-fold and the image contrast was doubled, effectively increasing image quality.

Although fibrous dysplasia (FD) is a benign fibro-osseous lesion, locally aggressive behaviour has rarely been described but is poorly characterised. In this study, we document clinical, radiological and pathological (including molecular genetics) findings in three cases of locally aggressive FD, two of which involved the ribs. Lesions in these cases, one of which was a recurrent lesion, were followed up for 2-7 years. All of the lesions showed typical histological features of FD but were characterised by extension through the bone cortex into the extra-osseous soft tissue. The lesions did not exhibit overexpression/amplification of CDK4 and MDM2; in two of the cases, a GNAS mutation was identified. Our findings confirm that FD can rarely exhibit locally aggressive behaviour with extension beyond the bone compartment into the surrounding soft tissue; these lesions can be distinguished from low-grade intramedullary osteosarcoma by lack of amplification/overexpression of CDK4 and MDM2 and the presence of a GNAS mutation.

We present omniSpect, an open source web- and MATLAB-based software tool for both desorption electrospray ionization (DESI) and matrix-assisted laser desorption ionization (MALDI) mass spectrometry imaging (MSI) that performs computationally intensive functions on a remote server. These functions include converting data from a variety of file formats into a common format easily manipulated in MATLAB, transforming time-series mass spectra into mass spectrometry images based on a probe spatial raster path, and multivariate analysis. OmniSpect provides an extensible suite of tools to meet the computational requirements needed for visualizing open and proprietary format MSI data.

Evaporation of water from extensively hydrated protons and peptides formed by electrospray ionization (ESI) has been examined for the first time by cryogenic ion mobility-mass spectrometry (IM-MS). The extent of hydration was controlled using a heated capillary inlet operated between 340 and 391 K. Cold cluster ions formed in the source region were transported into a low temperature (∼80 K) IM drift tube using an electrostatic ion guide where they were separated on the basis of size-to-charge via low-energy collisions with helium gas. The eluting IM profile was subsequently pulsed into an orthogonal time-of-flight (TOF) mass spectrometer for mass-to-charge (m/z) identification of the cluster ion species. Key parameters that influence the cluster distributions were critically examined including the inlet temperature, drift tube temperature, and IM field strength. In agreement with previous studies, our findings indicate that water evaporation is largely dependent upon the particular charge-carrying species within the cluster. IM-MS results for protonated water clusters suggest that the special stability of H(+)(H(2)O)(n) (n = 21) is attributed to the presence of a compact isomer (assigned to a clathrate cage) that falls below the trendline produced by adjacent clusters in the n = 15 to 35 size range. Peptide studies are also presented in which specific and nonspecific solvation is observed for gramicidin S [GS + 2H](2+)(H(2)O)(n) (n = 0 to ∼26) and bradykinin [BK + 2H](2+)(H(2)O)(n) (n = 0 to ∼73), respectively.