Skewed Profile Research Articles

Inhibition of T Cell Alloreactivity by Bronchial Epithelium Is Impaired in Lung Transplant Recipients, Through Pathways Involving TGF-β, IL-10 and HLA-G.

Bronchiolitis obliterans syndrome (BOS) after lung transplantation (LTx) results from bronchial epithelial cell (BECs) damages, thought to be orchestrated by T cells primed by antigen-presenting cell presenting alloantigens. In this cell cross-talk, BECs are also suspected to play a pivotal immunosuppressive role in T cell alloreactivity. We studied the immunomodulating role of BECs in a human ex vivo model of allogeneic T cell response, both in healthy subjects and LTx recipients. BECs from 35 LTx recipients (n = 22 stable, n = 13 BOS) and healthy controls (n = 25) were cultured as primary cell cultures. Their inhibitory capacities through the involvement of tolerogenic molecules (HLA-G, TGF-β, and IL-10) were tested on a mixed lymphocyte reaction between antigen-presenting cells and recipient T cells. Control BECs inhibited T cell alloproliferation by a mean of 53 ± 7%. This inhibitory effect of BECs was significantly reduced in the stable LTx group (24 ± 8%, P = 0.009), but not in the BOS TxP group (53 ± 10%, P = 0.97). Neutralization of HLA-G, TGF-β, and IL-10 partially restored T cell alloproliferation, arguing for their involvement in the immunosuppressive effect of BECs. BECs culture supernatant from stable LTx patients with impaired BEC properties showed a skewed Th2-type secretion profile (high IL-4/IFN-γ ratio). The inhibitory properties of BECs are dysregulated in stable LTx recipients, which could suggest their instrumental role in the initiation of BOS process and potential targeted therapies.

Year-Long Vertical Velocity Statistics Derived from Doppler Lidar Data for the Continental Convective Boundary Layer

AbstractOne year of coherent Doppler lidar data collected at the U.S. Department of Energy’s Atmospheric Radiation Measurement site in Oklahoma was analyzed to provide profiles of vertical velocity variance, skewness, and kurtosis for cases of cloud-free convective boundary layers. The variance was normalized by the Deardorff convective velocity scale, which was successful when the boundary layer depth was stationary but failed in situations in which the layer was changing rapidly. In this study, the data are sorted according to time of day, season, wind direction, surface shear stress, degree of instability, and wind shear across the boundary layer top. The normalized variance was found to have its peak value near a normalized height of 0.25. The magnitude of the variance changes with season, shear stress, degree of instability, and wind shear across the boundary layer top. The skewness was largest in the top half of the boundary layer (with the exception of wintertime conditions). The skewness was also found to be a function of the season, shear stress, and wind shear across the boundary layer top. Like skewness, the vertical profile of kurtosis followed a consistent pattern, with peak values near the boundary layer top. The normalized altitude of the peak values of kurtosis was found to be higher when there was a large amount of wind shear at the boundary layer top.

Afferent motor feedback determines the perceived location of tactile stimuli in the external space presented to the moving arm.

People make systematic errors when localizing a brief tactile stimulus in the external space presented on the index finger while moving the arm. Although these errors likely arise in the spatiotemporal integration of the tactile input and information about arm position, the underlying arm position information used in this process is not known. In this study, we tested the contributions of afferent proprioceptive feedback and predictive arm position signals by comparing localization errors during passive vs. active arm movements. In the active trials, participants were instructed to localize a tactile stimulus in the external space that was presented to the index finger near the time of a self-generated arm movement. In the passive trials, each of the active trials was passively replayed in randomized order, using a robotic device. Our results provide evidence that the localization error patterns of the passive trials are similar to the active trials and, moreover, did not lag but rather led the active trials, which suggests that proprioceptive feedback makes an important contribution to tactile localization. To further test which kinematic property of this afferent feedback signal drives the underlying computations, we examined the localization errors with movements that had differently skewed velocity profiles but overall the same displacement. This revealed a difference in the localization patterns, which we explain by a probabilistic model in which temporal uncertainty about the stimulus is converted into a spatial likelihood, depending on the actual velocity of the arm rather than involving an efferent, preprogrammed movement.NEW & NOTEWORTHY We show that proprioceptive feedback of arm motion rather than efferent motor signals contributes to tactile localization during an arm movement. Data further show that localization errors depend on arm velocity, not displacement per se, suggesting that instantaneous velocity feedback plays a role in the underlying computations. Model simulation using Bayesian inference suggests that these errors depend not only on spatial but also on temporal uncertainties of sensory and motor signals.

The respiratory control of carbon dioxide in children and adolescents referred for treatment of psychogenic non-epileptic seizures

Psychogenic non-epileptic seizures (PNES) are a common problem in paediatric neurology and psychiatry that can best be understood as atypical responses to threat. Threats activate the body for action by mediating increases in arousal, respiration, and motor readiness. In previous studies, a range of cardiac, endocrine, brain-based, attention-bias, and behavioral measures have been used to demonstrate increases in arousal, vigilance, and motor readiness in patients with PNES. The current study uses respiratory measures to assess both the motor readiness of the respiratory system and the respiratory regulation of CO2. Baseline respiratory rates during clinical assessment and arterial CO2 levels during the hyperventilation component of routine video electroencephalogram were documented in 60 children and adolescents referred for treatment of PNES and in 50 controls. Patients showed elevated baseline respiratory rates [t(78) = 3.34, p = .001], with 36/52 (69%) of patients [vs. 11/28 (39%) controls] falling above the 75th percentile (χ2 = 6.7343; df = 1; p = .009). Twenty-eight (47%) of patients [vs. 4/50 (8%) controls] showed a skewed hyperventilation-challenge profile—baseline PCO2 <36 mmHg, a trough PCO2 ≤ 20 mmHg, or a final PCO2 <36 mmHg after 15 min of recovery—signaling difficulties with CO2 regulation (χ2 = 19.77; df = 1; p < .001). Children and adolescents with PNES present in a state of readiness-for-action characterized by high arousal coupled with activation of the respiratory motor system, increases in ventilation, and a hyperventilation-challenge profile shifted downward from homeostatic range. Breathing interventions that target arousal, decrease respiratory rate, and normalize ventilation and arterial CO2 may help patients shift brain–body state and avert PNES episodes.

Combination treatment with 6-mercaptopurine and allopurinol in HepG2 and HEK293 cells - Effects on gene expression levels and thiopurine metabolism.

Combination treatment with low-dose thiopurine and allopurinol (AP) has successfully been used in patients with inflammatory bowel disease with a so called skewed thiopurine metabolite profile. In red blood cells in vivo, it reduces the concentration of methylated metabolites and increases the concentration of the phosphorylated ones, which is associated with improved therapeutic efficacy. This study aimed to investigate the largely unknown mechanism of AP on thiopurine metabolism in cells with an active thiopurine metabolic pathway using HepG2 and HEK293 cells. Cells were treated with 6-mercaptopurine (6MP) and AP or its metabolite oxypurinol. The expression of genes known to be associated with thiopurine metabolism, and the concentration of thiopurine metabolites were analyzed. Gene expression levels were only affected by AP in the presence of 6MP. The addition of AP to 6MP affected the expression of in total 19 genes in the two cell lines. In both cell lines the expression of the transporter SLC29A2 was reduced by the combined treatment. Six regulated genes in HepG2 cells and 8 regulated genes in HEK293 cells were connected to networks with 18 and 35 genes, respectively, present at known susceptibility loci for inflammatory bowel disease, when analyzed using a protein-protein interaction database. The genes identified as regulated as well as the disease associated interacting genes represent new candidates for further investigation in the context of combination therapy with thiopurines and AP. However, no differences in absolute metabolite concentrations were observed between 6MP+AP or 6MP+oxypurinol vs. 6MP alone in either of the two cell lines. In conclusion; the effect of AP on gene expression levels requires the presence of 6MP, at least in vitro. Previously described AP-effects on metabolite concentrations observed in red blood cells in vivo could not be reproduced in our cell lines in vitro. AP’s effects in relation to thiopurine metabolism are complex. The network-identified susceptibility genes represented biological processes mainly associated with purine nucleotide biosynthetic processes, lymphocyte proliferation, NF-KB activation, JAK-STAT signaling, and apoptotic signaling at oxidative stress.

The WISSH quasars project

Models and observations suggest that both the power and effects of AGN feedback should be maximised in hyper-luminous (LBol > 1047 erg s-1) quasars, i.e. objects at the brightest end of the AGN luminosity function. In this paper, we present the first results of a multiwavelength observing programme, focusing on a sample of WISE/SDSS selected hyper-luminous (WISSH) broad-line quasars at z ≈ 1.5–5. The WISSH quasars project has been designed to reveal the most energetic AGN-driven outflows, estimate their occurrence at the peak of quasar activity, and extend the study of correlations between outflows and nuclear properties up to poorly investigated, extreme AGN luminosities, i.e. LBol ~ 1047 − 1048 erg s-1. We present near-infrared, long-slit LBT/LUCI1 spectroscopy of five WISSH quasars at z ≈ 2.3 − 3.5, showing prominent [OIII] emission lines with broad (FWHM ~1200–2200 km s-1) and skewed profiles. The luminosities of these broad [OIII] wings are the highest measured so far, with L[OIII]broad ≳ 5 × 1044 erg s-1, and reveal the presence of powerful ionised outflows with associated mass outflow rates Ṁ ≳ 1700M⊙ yr-1 and kinetic powers Ėkin ≳ 1045 erg s-1. Although these estimates are affected by large uncertainties because of the use of [OIII] as a tracer of ionised outflows and the very basic outflow model adopted here, these results suggest that in our hyper-luminous targets the AGN is highly efficient at pushing large amounts of ionised gas outwards. Furthermore, the mechanical outflow luminosities measured for WISSH quasars correspond to higher percentages (~1–3%) of LBol than those derived for AGN with lower LBol. Our targets host very massive (MBH ≳ 2 × 109M⊙) black holes that are still accreting at a high rate (i.e. a factor of ~0.4–3 of the Eddington limit). These findings clearly demonstrate that WISSH quasars offer the opportunity to probe the extreme end of both luminosity and supermassive black holes (SMBH) mass functions and revealing powerful ionised outflows that are able to affect the evolution of their host galaxies.

FTIR study of silicon carbide amorphization by heavy ion irradiations

We have measured at room temperature (RT) the Fourier-transform infra-red (FTIR) absorption spectra of ion-irradiated thin epitaxial films of cubic silicon carbide (3C–SiC) with 1.1 µm thickness on a 500 µm thick (1 0 0) silicon wafer substrate. Irradiations were carried out at RT with 2.3 MeV 28Si+ ions and 3.0 MeV 84Kr+ ions for various fluences in order to induce amorphization of the SiC film. Ion projected ranges were adjusted to be slightly larger than the film thickness so that the whole SiC layers were homogeneously damaged. FTIR spectra of virgin and irradiated samples were recorded for various incidence angles from normal incidence to Brewster’s angle. We show that the amorphization process in ion-irradiated 3C–SiC films can be monitored non-destructively by FTIR absorption spectroscopy without any major interference of the substrate. The compared evolutions of TO and LO peaks upon ion irradiation yield valuable information on the damage process.Complementary test experiments were also performed on virgin silicon nitride (Si3N4) self-standing films for similar conditions. Asymmetrical shapes were found for TO peaks of SiC, whereas Gaussian profiles are found for LO peaks. Skewed Gaussian profiles, with a standard deviation depending on wave number, were used to fit asymmetrical peaks for both materials. A new methodology for following the amorphization process is proposed on the basis of the evolution of fitted IR absorption peak parameters with ion fluence. Results are discussed with respect to Rutherford backscattering spectrometry channeling and Raman spectroscopy analysis.

Innate Immune Cytokines, Fibroblast Phenotypes, and Regulation of Extracellular Matrix in Lung

Chronic inflammation can be caused by adaptive immune responses in autoimmune and allergic conditions, driven by a T lymphocyte subset balance (TH1, TH2, Th17, Th22, and/or Treg) and skewed cellular profiles in an antigen-specific manner. However, several chronic inflammatory diseases have no clearly defined adaptive immune mechanisms that drive chronicity. These conditions include those that affect the lung such as nonatopic asthma or idiopathic pulmonary fibrosis comprising significant health problems. The remodeling of extracellular matrix (ECM) causes organ dysfunction, and it is largely generated by fibroblasts as the major cell controlling net ECM. As such, these are potential targets of treatment approaches in the context of ECM pathology. Fibroblast phenotypes contribute to ECM and inflammatory cell accumulation, and they are integrated into chronic disease mechanisms including cancer. Evidence suggests that innate cytokine responses may be critical in nonallergic/nonautoimmune disease, and they enable environmental agent exposure mechanisms that are independent of adaptive immunity. Innate immune cytokines derived from macrophage subsets (M1/M2) and innate lymphoid cell (ILC) subsets can directly regulate fibroblast function. We also suggest that STAT3-activating gp130 cytokines can sensitize fibroblasts to the innate cytokine milieu to drive phenotypes and exacerbate existing adaptive responses. Here, we review evidence exploring innate cytokine regulation of fibroblast behavior.

Perturbed CD8+ T cell TIGIT/CD226/PVR axis despite early initiation of antiretroviral treatment in HIV infected individuals

HIV-specific CD8+ T cells demonstrate an exhausted phenotype associated with increased expression of inhibitory receptors, decreased functional capacity, and a skewed transcriptional profile, which are only partially restored by antiretroviral treatment (ART). Expression levels of the inhibitory receptor, T cell immunoglobulin and ITIM domain (TIGIT), the co-stimulatory receptor CD226 and their ligand PVR are altered in viral infections and cancer. However, the extent to which the TIGIT/CD226/PVR-axis is affected by HIV-infection has not been characterized. Here, we report that TIGIT expression increased over time despite early initiation of ART. HIV-specific CD8+ T cells were almost exclusively TIGIT+, had an inverse expression of the transcription factors T-bet and Eomes and co-expressed PD-1, CD160 and 2B4. HIV-specific TIGIThi cells were negatively correlated with polyfunctionality and displayed a diminished expression of CD226. Furthermore, expression of PVR was increased on CD4+ T cells, especially T follicular helper (Tfh) cells, in HIV-infected lymph nodes. These results depict a skewing of the TIGIT/CD226 axis from CD226 co-stimulation towards TIGIT-mediated inhibition of CD8+ T cells, despite early ART. These findings highlight the importance of the TIGIT/CD226/PVR axis as an immune checkpoint barrier that could hinder future “cure” strategies requiring potent HIV-specific CD8+ T cells.

Histological remission of autoimmune hepatitis after the addition of allopurinol and azathioprine dose reduction.

The standard therapy for some autoimmune diseases consists of a combination of corticosteroids and thiopurines. In non-responders to thiopurine drugs, the measurement of the metabolites of azathioprine, 6-thioguanine, and 6-methylmercaptopurine, can be a useful tool. The measurement has been used during the treatment of inflammatory bowel diseases and, less commonly, in autoimmune hepatitis. Many patients preferentially metabolize thiopurines to 6-methylmercaptopurine (6-MMP), which is potentially hepatotoxic, instead of 6-thioguanine, the active immunosuppressive metabolite. The addition of allopurinol shifts the metabolism of thiopurine towards 6-thioguanine, improving the immunosuppressive effect. We present the case of a 51-year-old female with autoimmune hepatitis who had a biochemical response after azathioprine and prednisone treatment without histological remission, and who preferentially shunted to 6-MMP. After the addition of allopurinol, the patient’s 6-thioguanine levels increased, and she reached histological remission with a reduction of 67% of the original dose of azathioprine. The patient did not develop clinical manifestations as a consequence of her increased immunosuppressive state. We also review the relevant literature related to this issue. In conclusion, the addition of allopurinol to thiopurine seems to be an option for those patients who do not reach histological remission and who have a skewed thiopurine metabolite profile.

Measurements of Turbulent Jet Mixing in a Turbulent Co-Flow Including the Influence of Periodic Forcing and Heating

In this work, the turbulent mixing of a confined coaxial jet in air is investigated by means of simultaneous particle image velocimetry and planar laser induced fluorescence of the acetone seeded flow injection. The jet is injected into a turbulent duct flow at atmospheric pressure through a 90 ∘ pipe bend. Measurements are conducted in a small scale windtunnel at constant mass flow rates and three modes of operation: isothermal steady jet injection at a Dean number of 20000 (Red=32000), pulsed isothermal injection at a Womersley number of 65 and steady injection at elevated jet temperatures of ΔT=50 K and ΔT=100 K. The experiment is aimed at providing statistically converged quantities of velocity, mass fraction, turbulent fluctuations and turbulent mass flux at several downstream locations. Stochastic error convergence over the number of samples is assessed within the outer turbulent shear layer. From 3000 samples the statistical error of time-averaged velocity and mass fraction is below 1 % while the error of Reynolds shear stress and turbulent mass flux components is in the of range 5-6 %. Profiles of axial velocity and turbulence intensity immediately downstream of the bend exit are in good agreement with hot-wire measurements from literature. During pulsed jet injection strong asymmetric growing of shear layer vortices lead to a skewed mass fraction profile in comparison with steady injection. Phase averaging of single shot PLIF-PIV measurements allows to track the asymmetric shear layer vortex evolvement and flow breakdown during a pulsation cycle with a resolution of 10∘. Steady injection with increased jet temperature supports mixing downstream from 6 nozzle diameters onward.

B cells present skewed profile and lose the function of supporting T cell inflammation after Roux-en-Y gastric bypass

Bariatric surgeries, including Roux-en-Y gastric bypass (RYGB) are currently the best treatment for obesity and obesity-related comorbidities, such as type 2 diabetes. However, the underlying mechanism of bariatric surgeries is not entirely understood. Further investigations are needed to improve the success rate and achieve sustained health benefits. Given that B cell dysregulation is a critical component of etiology in inflammatory diseases, whereas obesity and type 2 diabetes represent two major inflammatory disorders, we investigated the effect of RYGB on B cell inflammation. We found that B cells after RYGB presented significantly elevated frequency of interleukin (IL)-10-producing cells and reduced frequency of IL-6-producing cells compared to those before RYGB. When grouping B cell subsets into regulatory (secreting IL-10 and transforming growth factor beta [TGF-β]) and effector (secreting IL-2, IL-4, IL-6, IL-12, interferon gamma [IFN-γ] and tumor necrosis factor alpha [TNF-α]) types, we found that after RYGB, the regulatory to effector B cell ratio was significantly increased. Function analyses showed that B cells before RYGB supported IL-17 secretion from T cells whereas these cells after RYGB lost such capacity. B cells after RYGB also gained the capacity to suppress T cell IFN-γ production through TGF-β-mediated effects, a feature not present in B cells before RYGB. Interestingly, the regulatory to effector B cell ratio was directly associated with the reductions in obesity markers following RYGB, such as BMI and fat mass percentage. Together, these results demonstrated a potential mechanism through which RYGB promoted amelioration of obesity and type 2 diabetes.

Custom-Designed Affinity Capture LC-MS F(ab')2 Assay for Biotransformation Assessment of Site-Specific Antibody Drug Conjugates.

Affinity capture liquid chromatography-mass spectrometry (LC-MS) intact antibody assay has been widely used for direct drug-to-antibody ratio (DAR) and catabolite characterization of antibody-drug conjugates (ADCs). However, the intact mass spectra of new ADCs, which incorporate new types of linkers and payloads other than maytansines and auristatins, are more complex than those examined previously. The current method has showed some limitations in elucidating certain structural modifications. Herein, we report an alternative analytical approach for ADCs, such as THIOMAB antibody-drug conjugates (TDCs), where the linker drugs are site-specifically conjugated in the Fab region. The newly developed affinity capture LC-MS F(ab')2 assay incorporates affinity capture of human IgGs via binding to the Fab region, followed by on-bead IdeS digestion to remove the Fc domain specifically and uniformly. The resulting F(ab')2 (∼100 kDa) fragments contain the key ADC biotransformation information, such as drug-to-antibody ratio and drug metabolism and are more readily analyzed by electrospray ionization LC-MS than the intact ADC (∼150 kDa). The reduced size of analytes results in improved mass spectral sensitivity and resolution. In addition, the reduced and optimized sample preparation time, for example, rapid removal of the Fc fragment by IdeS digestion, minimizes assay artifacts of drug metabolism and skewed DAR profiles that may result from the prolonged incubation times (e.g., overnight enzymatic treatment for Fc deglycosylation). The affinity capture LC-MS F(ab')2 assay provides more detailed and accurate information on ADC biotransformations in vivo, enabling analysis of low-dose, labile, and complex site-specific ADCs with linker-drug conjugated in the Fab region.

Potential of Translationally Controlled Tumor Protein-Derived Protein Transduction Domains as Antigen Carriers for Nasal Vaccine Delivery.

Nasal vaccination offers a promising alternative to intramuscular (i.m.) vaccination because it can induce both mucosal and systemic immunity. However, its major drawback is poor absorption of large antigens in the nasal epithelium. Protein transduction domains (PTDs), also called cell-penetrating peptides, have been proposed as vehicles for nasal delivery of therapeutic peptides and proteins. Here, we evaluated the potential of a mutant PTD derived from translationally controlled tumor protein (designated TCTP-PTD 13) as an antigen carrier for nasal vaccines. We first compared the l- and d-forms of TCTP-PTD 13 isomers (l- or d-TCTP-PTD 13) as antigen carriers. Studies in mice demonstrated that nasally administered mixtures of the model antigen ovalbumin (OVA) and d-TCTP-PTD 13 induced higher plasma IgG titers and secretory IgA levels in nasal washes than nasally administered OVA alone, OVA/l-TCTP-PTD 13, or i.m.-injected OVA. Plasma IgG subclass responses (IgG1 and IgG2a) of mice nasally administered OVA/d-TCTP-PTD 13 showed that the predominant IgG subclass was IgG1, indicating a Th2-biased immune response. We also used synthetic CpG oligonucleotides (CpG) as a Th1 immune response-inducing adjuvant. Nasally administered CpG plus OVA/d-TCTP-PTD 13 was superior in eliciting systemic and mucosal immune responses compared to those induced by nasally administered OVA/d-TCTP-PTD 13. Furthermore, the OVA/CpG/d-TCTP-PTD 13 combination skewed IgG1 and IgG2a profiles of humoral immune responses toward a Th1 profile. These findings suggest that TCTP-derived PTD is a suitable vehicle to efficiently carry antigens and to induce more powerful antigen-specific immune responses and a more balanced Th1/Th2 response when combined with a DNA adjuvant.

LZIFU: an emission-line fitting toolkit for integral field spectroscopy data

We present LZIFU (LaZy-IFU), an IDL toolkit for fitting multiple emission lines simultaneously in integral field spectroscopy (IFS) data. LZIFU is useful for the investigation of the dynamical, physical and chemical properties of gas in galaxies. LZIFU has already been applied to many world-class IFS instruments and large IFS surveys, including the Wide Field Spectrograph, the new Multi Unit Spectroscopic Explorer (MUSE), the Calar Alto Legacy Integral Field Area (CALIFA) survey, the Sydney-Australian-astronomical-observatory Multi-object Integral-field spectrograph (SAMI) Galaxy Survey. Here we describe in detail the structure of the toolkit, and how the line fluxes and flux uncertainties are determined, including the possibility of having multiple distinct kinematic components. We quantify the performance of LZIFU, demonstrating its accuracy and robustness. We also show examples of applying LZIFU to CALIFA and SAMI data to construct emission line and kinematic maps, and investigate complex, skewed line profiles presented in IFS data. The code is made available to the astronomy community through github. LZIFU will be further developed over time to other IFS instruments, and to provide even more accurate line and uncertainty estimates.

Unique Inflammatory Mediators and Specific IgE Levels Distinguish Local from Systemic Reactions after Anthrax Vaccine Adsorbed Vaccination.

Although the U.S. National Academy of Sciences concluded that anthrax vaccine adsorbed (AVA) has an adverse event (AE) profile similar to those of other adult vaccines, 30 to 70% of queried AVA vaccinees report AEs. AEs appear to be correlated with certain demographic factors, but the underlying immunologic pathways are poorly understood. We evaluated a cohort of 2,421 AVA vaccinees and found 153 (6.3%) reported an AE. Females were more likely to experience AEs (odds ratio [OR] = 6.0 [95% confidence interval {CI} = 4.2 to 8.7]; P < 0.0001). Individuals 18 to 29 years of age were less likely to report an AE than individuals aged 30 years or older (OR = 0.31 [95% CI = 0.22 to 0.43]; P < 0.0001). No significant effects were observed for African, European, Hispanic, American Indian, or Asian ancestry after correcting for age and sex. Additionally, 103 AEs were large local reactions (LLRs), whereas 53 AEs were systemic reactions (SRs). In a subset of our cohort vaccinated 2 to 12 months prior to plasma sample collection (n = 75), individuals with LLRs (n = 33) had higher protective-antigen (PA)-specific IgE levels than matched, unaffected vaccinated individuals (n = 50; P < 0.01). Anti-PA IgE was not associated with total plasma IgE, hepatitis B-specific IgE, or anti-PA IgG in individuals who reported an AE or in matched, unaffected AVA-vaccinated individuals. IP-10 was also elevated in sera of individuals who developed LLRs (P < 0.05). Individuals reporting SRs had higher levels of systemic inflammation as measured from C-reactive protein (P < 0.01). Thus, LLRs and SRs are mediated by distinct pathways. LLRs are associated with a vaccine-specific IgE response and IP-10, whereas SRs demonstrate increased systemic inflammation without a skewed cytokine profile.

Fine Filament Formation Behavior of Polymethylpentene and Polypropylene near Spinneret in Melt Blowing Process

Abstract Formation behavior of fine fibers of polymethylpentene (PMP) and polypropylene (PP) in the melt blowing process was investigated through the observation of the spin-line near the spinning nozzle using a high speed camera. Reduction of the polymer throughput rate and increase of the air flow rate were necessary to achieve fine diameter fibers, however these conditions generally cause the instability in the spinning process. Observation of the spin-line at the melt blowing die revealed the periodic accumulation of polymer flow near the spinning nozzle followed by the quick pulling down of the accumulated polymer by the air flow. This behavior caused the periodic fluctuation of fiber diameter as well as the intermittent breakage of the spin-line under extreme conditions. Because of high extrusion viscosity, PMP showed more stable spinning behavior than PP. Frequency of diameter fluctuation became higher with the increases of air flow rate and throughput rate, and the maximum frequency of about 60 Hz was observed for PP spinning with the throughput rate of 0.18 g/min. Diameter distribution of the fibers in the prepared web was also analyzed to compare with the spinning behavior. Fiber diameter distributions were narrow and symmetric under stable spinning conditions, whereas skewed diameter profiles with a maximum at low value and a long tail to the larger diameter region were observed under unstable conditions. Intermittent spin-line breakage caused flaws of “shot” and/or “fly”, and the skewed fiber diameter distribution with the presence of a small amount of fiber of extremely large diameter was confirmed.

Comparison of turbulent boundary layers over smooth and rough surfaces up to high Reynolds numbers

Turbulent boundary layer measurements above a smooth wall and sandpaper roughness are presented across a wide range of friction Reynolds numbers, ${\it\delta}_{99}^{+}$, and equivalent sand grain roughness Reynolds numbers, $k_{s}^{+}$ (smooth wall: $2020\leqslant {\it\delta}_{99}^{+}\leqslant 21\,430$, rough wall: $2890\leqslant {\it\delta}_{99}^{+}\leqslant 29\,900$; $22\leqslant k_{s}^{+}\leqslant 155$; and $28\leqslant {\it\delta}_{99}^{+}/k_{s}^{+}\leqslant 199$). For the rough-wall measurements, the mean wall shear stress is determined using a floating element drag balance. All smooth- and rough-wall data exhibit, over an inertial sublayer, regions of logarithmic dependence in the mean velocity and streamwise velocity variance. These logarithmic slopes are apparently the same between smooth and rough walls, indicating similar dynamics are present in this region. The streamwise mean velocity defect and skewness profiles each show convincing collapse in the outer region of the flow, suggesting that Townsend’s (The Structure of Turbulent Shear Flow, vol. 1, 1956, Cambridge University Press.) wall-similarity hypothesis is a good approximation for these statistics even at these finite friction Reynolds numbers. Outer-layer collapse is also observed in the rough-wall streamwise velocity variance, but only for flows with ${\it\delta}_{99}^{+}\gtrsim 14\,000$. At Reynolds numbers lower than this, profile invariance is only apparent when the flow is fully rough. In transitionally rough flows at low ${\it\delta}_{99}^{+}$, the outer region of the inner-normalised streamwise velocity variance indicates a dependence on $k_{s}^{+}$ for the present rough surface.

Atypical Activin A and IL-10 Production Impairs Human CD16+ Monocyte Differentiation into Anti-Inflammatory Macrophages.

Human CD14(++)CD16(-) and CD14(+/lo)CD16(+) monocyte subsets comprise 85 and 15% of blood monocytes, respectively, and are thought to represent distinct stages in the monocyte differentiation pathway. However, the differentiation fates of both monocyte subsets along the macrophage (Mϕ) lineage have not yet been elucidated. We have now evaluated the potential of CD14(++) CD16(-) and CD16(+) monocytes to differentiate and to be primed toward pro- or anti-inflammatory Mϕs upon culture with GM-CSF or M-CSF, respectively (subsequently referred to as GM14, M14, GM16, or M16). Whereas GM16 and GM14 were phenotypic and functionally analogous, M16 displayed a more proinflammatory profile than did M14. Transcriptomic analyses evidenced that genes associated with M-CSF-driven Mϕ differentiation (including FOLR2, IL10, IGF1, and SERPINB2) are underrepresented in M16 with respect to M14. The preferential proinflammatory skewing of M16 relative to M14 was found to be mediated by the secretion of activin A and the low levels of IL-10 produced by M16. In fact, activin A receptor blockade during the M-CSF-driven differentiation of CD16(+) monocytes, or addition of IL-10-containing M14-conditioned medium, significantly enhanced their expression of anti-inflammatory-associated molecules while impairing their acquisition of proinflammatory-related markers. Thus, we propose that M-CSF drives CD14(++)CD16- monocyte differentiation into bona fide anti-inflammatory Mϕs in a self-autonomous manner, whereas M-CSF-treated CD16(+) monocytes generate Mϕs with a skewed proinflammatory profile by virtue of their high activin A expression unless additional anti-inflammatory stimuli such as IL-10 are provided.

Intrinsic challenges in ancient microbiome reconstruction using 16S rRNA gene amplification.

To date, characterization of ancient oral (dental calculus) and gut (coprolite) microbiota has been primarily accomplished through a metataxonomic approach involving targeted amplification of one or more variable regions in the 16S rRNA gene. Specifically, the V3 region (E. coli 341–534) of this gene has been suggested as an excellent candidate for ancient DNA amplification and microbial community reconstruction. However, in practice this metataxonomic approach often produces highly skewed taxonomic frequency data. In this study, we use non-targeted (shotgun metagenomics) sequencing methods to better understand skewed microbial profiles observed in four ancient dental calculus specimens previously analyzed by amplicon sequencing. Through comparisons of microbial taxonomic counts from paired amplicon (V3 U341F/534R) and shotgun sequencing datasets, we demonstrate that extensive length polymorphisms in the V3 region are a consistent and major cause of differential amplification leading to taxonomic bias in ancient microbiome reconstructions based on amplicon sequencing. We conclude that systematic amplification bias confounds attempts to accurately reconstruct microbiome taxonomic profiles from 16S rRNA V3 amplicon data generated using universal primers. Because in silico analysis indicates that alternative 16S rRNA hypervariable regions will present similar challenges, we advocate for the use of a shotgun metagenomics approach in ancient microbiome reconstructions.