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We recently reported a novel interleukin-10 (IL-10)-secreting CD21 B cell population in jejunal Peyer's patches (JPP) of sheep with a regulatory function (B) suppressing Toll-like receptor 9 (TLR9)-induced cytokine responses. However, little is known about the development of these cells. Therefore, we investigate their existence in JPP cells from fetal and newborn lambs. CD21 B cells were purified from JPP cells by magnetic cell sorting and subsequently stimulated with the TLR9 agonist, CpG ODN (CpG oligodeoxynucleotide). Lymphocyte proliferative responses, cytokine production (IL-10, IL-12 and interferon-γ [INF-γ]) and antibody secretion were assayed. We found that fetal and neonatal CD21 B cells spontaneously secreted high levels of IL-10 regardless of CpG stimulation but that these cells did not produce any IL-12 or INF-γ upon stimulation with CpG. The observed responses are consistent with those previously reported for B characterized in JPP of older lambs. Surprisingly, unlike in older lambs, fetal and neonatal JPP CD21 B cells proliferated in response to CpG stimulation. Our investigations of fetal and neonatal lambs provide evidence for the development of IL-10-secreting CD21 B cells in PPs prior to antigen exposure. [ABSTRACT FROM AUTHOR]

The use of highly reactive fuels in the lean premixed combustion systems employed in stationary gas turbines can lead to many practical problems, such as unwanted autoignition in regions not designed for combustion. In the present study, autoignition characteristics for hydrogen, diluted with up to 30 vol. % nitrogen, were investigated at conditions relevant to reheat combustor operation (p = 15 bar, T > IO00 K, hot flue gas, relevant residence times). The experiments were performed in a generic, optically accessible reheat combustor, by applying high-speed imaging and particle image velocimetry. Autoignition limits for different mixing section (temperature, velocity) and fuel jet (N2 dilution) parameters are described. The dominant factor influencing autoignition was the temperature, with an increase of around 2% leading to a reduction of the highest possible 142 concentration without "flame-stabilizing autoignition kernels" of approximately 16 vol. %. Furthermore, the onset and propagation of the ignition kernels were elucidated using the high-speed measurements. It was found that the ability of individual autoignition kernels to develop into stable fames depends on the initial position of the kernel and the corresponding axial velocity at that position. While unwanted autoignition occurred prior to reaching the desired operating point for most investigated conditions, for certain conditions the reheat combustor could be operated stably with up to 80 vol. % H2 in the fuel. [ABSTRACT FROM AUTHOR]

Peyer's patches (PPs) play an important role in the induction of immune responses in the intestine, but regulation of Toll-like receptor (TLR)-induced innate immune responses in PPs is not well understood. We investigated the responses of PPs and other immune cells to the TLR9 agonist, CpG oligodeoxynucleotide (ODN). Peripheral blood mononuclear cells and lymph node cells secreted significant amounts of interferon (IFN)-α, IFNγ, and interleukin (IL)-12 following stimulation with CpG ODN. In contrast, PP cells exhibited poor cytokine responses, despite abundant expression of TLR9 mRNA. PP cells spontaneously secreted high levels of IL-10, and the primary source of the IL-10 was resting CD5−CD11c−CD21+ B cells. Neutralization of the IL-10 or depletion of CD21+ B cells resulted in a significant increase in CpG-induced IFNα-response in PPs, suggesting that IL-10 from B cells regulate innate responses in PPs. These IL-10-secreting PP B cells may represent a novel subset of the recently proposed regulatory B cells (Bregs) in the intestine.Mucosal Immunology (2009) 2, 265–275; doi:10.1038/mi.2009.6; published online 4 March 2009 [ABSTRACT FROM AUTHOR]

Flame stability is a crucial issue in low NOx combustion systems operating at extremely lean conditions. Hydrogen enrichment seems to be a promising option to extend lean blowout limits (LBO) of natural gas combustion. This experimental study addresses flame stability enhancement and NOx reduction in turbulent, high-pressure, lean premixed methane/air flames in a generic combustor capable of a wide range of operating conditions. Lean blowout limits and NOx emissions are presented for pressures up to 14 bar, bulk velocities in the range of 32-80 m/s, two different preheating temperatures (673 K, 773 K), and a range of fuel mixtures from pure methane to 20% H2/80% CH4 by volume. The influence of turbulence on LBO limits is also discussed. In addition to the investigation of perfectly premixed H2-enriched flames, LBO and NOx are also discussed for hydrogen piloting. Experiments have revealed that a mixture of 20% hydrogen and 80% methane, by volume, can typically extend the lean blowout limit by ˜ 10% compared to pure methane. The flame temperature at LBO is ˜60 K lower resulting in the reduction of NOx concentration by ≈35% (0.5 → 0.3 ppm/15% 02). [ABSTRACT FROM AUTHOR]

Changes in B-cell phenotype during development of heal and jejunal Peyer's patches (PP) of sheep were investigated using flow cytometry and immunoperoxidase-stained cryosections. On Day 104 of gestation (term at 150 days) B-cell clusters were identified in the lamina propria of the ileum. These clusters were composed of cells that expressed surface lgM (sIgM), λ or κ light chain, and BAQ44A, a B-cell differentiation molecule. No cells in the clusters stained for terminal deoxynucleotide transferase. On Day 132 gestation, a change was evident in the phenotype of ileal PP B cells. Most B cells expressed a reduced level of slgM and 20% were BAQ44A . The B cells in the dome region were BAQ44A + but few BAQ44A+ cells were present in the follicles. At 6-8 weeks of age BAQ44A+ cells were restricted to the dome region of the ileal PP: flow cytometric analysis confirmed that 25% of B cells isolated from the dome/follicle complex were BAQ44A+. Thus, the primordial PP was populated with B cells that were phenotypically similar to circulating B cells (slgMhigh, BAQ44A+). After 132 days gestation, the predominant B-cell phenotype in the ileal PP changed to sIgMlow' and BAQ44A . This phenotypic change could be the result of either early immigrant B-cell differentiation or subsequent colonization by sIgMlow" BAQ44A B cells. The phenotypic changes of ileal PP lollicular B cells were not complete until after birth and different phenotypic changes were observed in follicles of the jejunal PP of young lambs. [ABSTRACT FROM AUTHOR]

We developed a method to isolate and culture cells from the lymphoid follicles of the ileal Peyer's (PP) patch of young sheep (6-12 weeks). These cells were 98% sIgM + B cells and 1% T cells. Cultured follicular cells were used to investigate B-cell proliferation and differentiation. Less than 50% of B cells were viable after 24 hr of culture and this decrease in B-cell viability also occurred following co-stimulation with pokeweed mitogen (PWM) and recombinant bovine interleukin-1 (rBoIL-1) or rBoIL-2. In contrast, co-stimulation with PWM and either rBoIL-1 or rBoIL-2 induced a marked proliferative response that was maximal on Day 4 of culture. Cytokine-induced proliferation of the B cells required PWM co-stimulation and proliferation induced by rBoIL-1 and rBoIL-2 was neither additive or synergistic. This suggests that PWM bound a molecule or molecules that signalled responsiveness to both rBoIL-1 and rBoIL-2. Culture of follicular cells with PWM and both rBoIL-1 and rBoIL-2 also resulted in B-cell differentiation. This differentiation was associated with decreased proliferation, an increased number of viable B cells, and increased expression of both surface IgM and non-Ig membrane molecules. Thus, co-stimulation of ileal PP follicular cells with PWM and r BoIL-1 and rBoIL-2 resulted in both B-cell proliferation and differentiation. [ABSTRACT FROM AUTHOR]

Non-major histocompatibility complex (MHC) restricted cytotoxicity is an important part of the immune reaction mounted in response to bovine herpes virus type 1 (BHV-1) infection. In this study, we evaluated the effect of BHV-1 infection on the ability of lung parenchyma leucocytes (LPL), cranial tracheobronchial lymph node cells (BLNC) and peripheral blood mononuclear leucocytes (PBML) to mediate this function. While LPL from non-infected calves mediated cytotoxicity against BHV-1-infected cells, a similar activity could not be detected in PBML or BLNC. in contrast, both LPL and PBML from naive calves could mediate cytotoxicity against K562 target cells but only after activation with interleukin-2 (IL-2). BLNC were unable to kill K562 cells. Infection of calves with BHV-1 enhanced the ability of LPL and PBML to kill BHV-1-infected cells. This enhancement was detected as early as Day 1 after infection in LPL whereas it could only be detected in PBML 8 days after infection. The results demonstrate that the leucocyte population present at the site of infection was able to mediate a potentially important antiviral function and that this function was enhanced rapidly in response to infection. Thus LPL-mediated cytotoxicity may be an important mechanism for the recovery from BHV-1 infection. [ABSTRACT FROM AUTHOR]

Availability of effective oral vaccine delivery vehicles should contribute to the success of oral immunization in domestic animals. To achieve this goal, we evaluated alginate microspheres for their capacity to induce mucosal immune responses following oral and enteric immunizations. Mice were immunized with either live porcine rotavirus (PRV) or its recombinant VP6 protein, encapsulated in alginate microspheres or unencapsulated. VP6-specific IgG (but no IgA) antibodies were detected in the sera of mice after a single intraperitoneal (i.p.) immunization with either VP6 in Incomplete Freund’s adjuvant (VP6-IFA), VP6 in alginate microspheres (VP6-MS) or with live PRV in incomplete Freund’s adjuvant (PRV-IFA). In contrast, VP6-specific IgA (but no IgG) was detected in culture supernatants of mesenteric lymph nodes from mice immunized i.p. with either VP6-IFA or with PRV-IFA. Oral immunization with VP6-MS induced the highest level of VP6-specific fecal IgA antibody, similar to responses induced by oral immunization with live PRV. Furthermore, the VP6-specific fecal IgA could be boosted by a secondary i.p. immunization with VP6. Further experiments were performed in a sheep intestinal ‘loop’ model to evaluate uptake of microspheres by Peyer’s patches. Microspheres containing colloidal carbon were specifically bound and transported by follicle-associated epithelium of Peyer’s patches. Additionally, mucosal immune responses were detected following enteric immunization with porcine serum albumin (PSA) encapsulated in alginate microspheres. Our results confirm that alginate microspheres are an effective oral delivery vehicle for protein antigens and intestinal IgA antibody responses are induced by antigens encapsulated in alginate microspheres without any additional mucosal adjuvant. These investigations confirm that alginate microspheres have the potential as an effective delivery vehicle for oral immunization of ruminants. [Copyright &y& Elsevier]

Abstract To better understand uterine inflammation in postpartum dairy cows we collected sequential cytobrush samples at 29–35 and at 49–55 d in milk (DIM). Based on the uterine cytology, cows were classified as Non-endometritic (n = 23; <18% neutrophils) or Endometritic (n = 12; ≥18% neutrophils) at 29–35 DIM and Non-endometritic (n = 17; <10% neutrophils) or Endometritic (n = 9; ≥10% neutrophils) at 49–55 DIM. Cows defined as Sham Controls (n = 6) were examined by vaginoscopy at 29–35 DIM and identified as Non-endometritic (<10% neutrophils) at 49–55 DIM. Cytokine gene expression in cytobrush samples was assessed using qRT-PCR. Sham Controls did not differ significantly (P > 0.17) from Non-endometritic cows at 49–55 DIM and these data were combined (n = 23). Uterine cytology-based classification using the aforementioned thresholds effectively separated cows into groups with Endometritic cows having significantly higher expression of pro-inflammatory (interleukin (IL)-1α, IL-1β, IL-6, IL-8, IL-17A CSF-1; P < 0.01) and regulatory (IL-1RA and IL-10; P < 0.03) cytokines, relative to Non-endometritic cows. Furthermore, Non-endometritic cows showed a significant decline (P < 0.03) in the expression of pro-inflammatory (IL-1α, IL-6, IL-8) and regulatory (IL-10) cytokine genes as the postpartum period progressed; whereas Endometritic cows exhibited a sustained elevation in transcript abundance throughout the sample period for both pro-inflammatory and regulatory cytokine genes. Expression of transforming growth factor (TGF) genes was more complex with TGF-β3 expression significantly (P < 0.01) lower at 29–35 DIM and TGF-β1 gene expression significantly (P < 0.03) increased at 49–55 DIM in Endometritic versus Non-endometritic cows. Expression of TGF-β2 gene was 2.7-fold higher (P < 0.01) at 29–35 DIM in cows that remained Endometritic when compared to cows recovering by 49–55 DIM. Some Non-endometritic cows (n = 4) at 29–35 DIM were reclassified as Endometritic at 49–55 DIM. The sampling procedures at 29–35 DIM did not alter either the cellular response (P > 0.43) or cytokine gene expression (P > 0.17) at 49–55 DIM. In conclusion, normal uterine involution is characterized by a progressive decline in pro-inflammatory and regulatory cytokine gene expression, while cows with endometritis show a dysregulated inflammatory process characterized by a sustained elevation in pro-inflammatory and regulatory cytokine gene expression. This analysis also shows that decreased TGF-β2 gene expression at 29–35 DIM may be an indicator of recovery from endometritis. Highlights • Neutrophil thresholds derived using uterine cytobrush cytology are reflective of the inflammatory state of the bovine uterus. • Cows with endometritis have a dysregulated expression of pro-inflammatory and regulatory cytokine genes. • Non-endometritic cows at 29–35 DIM can develop endometritis later in the postpartum period. • Spontaneous resolution of endometritis can occur during the first two months of postpartum period. • Spontaneous resolution of endometritis was associated with a lower expression of TGF-β2 gene expression at 29–35 DIM. [ABSTRACT FROM AUTHOR]

Summary: In the last decades, many regional and country‐wide control programmes for Johne's disease (JD) were developed due to associated economic losses, or because of a possible association with Crohn's disease. These control programmes were often not successful, partly because management protocols were not followed, including the introduction of infected replacement cattle, because tests to identify infected animals were unreliable, and uptake by farmers was not high enough because of a perceived low return on investment. In the absence of a cure or effective commercial vaccines, control of JD is currently primarily based on herd management strategies to avoid infection of cattle and restrict within‐farm and farm‐to‐farm transmission. Although JD control programmes have been implemented in most developed countries, lessons learned from JD prevention and control programmes are underreported. Also, JD control programmes are typically evaluated in a limited number of herds and the duration of the study is less than 5 year, making it difficult to adequately assess the efficacy of control programmes. In this manuscript, we identify the most important gaps in knowledge hampering JD prevention and control programmes, including vaccination and diagnostics. Secondly, we discuss directions that research should take to address those knowledge gaps. [ABSTRACT FROM AUTHOR]

It is generally accepted that combustion of hydrogen and natural gas mixtures will become more prevalent in the near future, to allow for a further penetration of renewables in the European power generation system. The current work aims at the demonstration of the advantages of steam dilution, when highly reactive combustible mixtures are used in a swirl-stabilized combustor. To this end, high-pressure experiments have been conducted with a generic swirl-stabilized combustor featuring axial air injection to increase flashback safety. The experiments have been conducted with two fuel mixtures, at various pressure levels up to 9 bar and at four levels of steam dilution up to 25% steam-to-air mass flow ratio. Natural gas has been used as a reference fuel, whereas a mixture of natural gas and hydrogen (10% hydrogen by mass) represented an upper limit of hydrogen concentration in a natural gas network with hydrogen enrichment. The results of the emissions measurements are presented along with a reactor network model. The latter is applied as a means to qualitatively understand the chemical processes responsible for the observed emissions and their trends with increasing pressure and steam injection. [ABSTRACT FROM AUTHOR]

Stresses imposed on livestock have significant impact on their health and productivity as well as public perceptions of animal welfare. Understanding stress responses in livestock may help refine management procedures and facilitate selection of stress-tolerant animals. In this study, behavioral (chute entry order, chute behavior, and exit velocity), physiological (serum cortisol), and biochemical (kinome) responses were evaluated in cattle (n = 20) subjected to three 5-min restraint periods with weekly intervals. Correlations among stress responses were assessed across all animals as well as for subgroups (n = 4) representing animals consistently displaying a high and low extreme of serum cortisol responses. Across all animals, entry order (P = 0.006) and exit velocity (P = 0.023) were positively correlated with serum cortisol; however, these correlations were not consistently reproducible for the high and low serum cortisol responders. Kinome profiling of peripheral blood mononuclear cells revealed distinct signaling events between the high and low cortisol responders. In particular, kinome profiling revealed significant differences in carbohydrate metabolism and apoptosis that were independently validated. Furthermore, changes in serum glucose levels provided a reliable, inexpensive indicator of serum cortisol levels and often had greater predictive value than cortisol for stress-related behavioral responses. Serum cortisol levels displayed a pattern consistent with sensitization, whereas no habituation or sensitization was observed for serum glucose levels or behavioral responses. Collectively, this investigation provides insight into correlations among physiological, behavioral, and biochemical responses of cattle subjected to a brief restraint that may provide biomarkers for selection of stress-tolerant animals. [ABSTRACT FROM AUTHOR]

Mixture fraction measurements in a jet-in-cross flow configuration at high pressures (15 bar) and temperatures (above 1000 K) were performed using planar laser induced fluorescence of nitric oxide (NO-PLIF) as trace species. The goal was the evaluation of this tracer LIF technique for the characterization of the mixing of fuel and hot exhaust gas in the mixing channel. The fuel (natural gas (NG) or H/N/NG mixture) along with the tracer were injected into the crossflow of the exhaust gas and PLIF measurements were performed in different planes. In order to relate the measured NO-LIF signal to fuel concentration and mixture fraction, effects of pressure, temperature and species concentration were taken into account. Numerical calculations and spectroscopic simulations that mimic the experimental conditions were performed to identify excitation schemes that give optimum correlations between the NO-LIF signal and the mixture fraction. The measured NO-PLIF images were transformed into mixture fraction plots using the computed correlations. The paper reports on the experimental challenges encountered during the measurements and the steps taken to overcome those difficulties. Examples of mixture fraction distributions are presented and discussed. The paper concludes with a detailed analysis on the accuracy of the measured mixture fraction values. [ABSTRACT FROM AUTHOR]

Today, environmental issues play an important role in the viability of modern low-emission power plants. As a consequence, gas turbine combustors need to be operated at lean premixed conditions. However, successful designs require a detailed knowledge of the combustion process under realistic operating conditions. The present study focuses on nonreacting and reacting jets at operating pressures from 1 to 14 bar. First, an isothermal confined jet has been studied experimentally and numerically using particle image velocimetry and large eddy simulation. The flow is highly turbulent and includes large-scale unsteady structures. The comparison of the numerical results and the experimental velocity data showed an excellent agreement that was generally below the numerical or experimental uncertainty. Second, the influences of combustion and operating pressure on the flowfield were investigated. The large eddy simulation results showed that the jet core was lengthened, due to the density jump across the flame. The effect of pressure on the flame was studied using planar laser-induced fluorescence and large eddy simulation at a constant Mach number. The flame brush and the velocity fields were found to be relatively insensitive to an increase of pressure from 1 to 14 bar (and, correspondingly, to an increase of Reynolds and Karlovitz numbers). The numerical results suggest that increasing pressure decreases the laminar flame speed and increases the flame-front wrinkling, causing the turbulent flame speed to be less sensitive to pressure. [ABSTRACT FROM AUTHOR]

Bacterial DNA contains a much higher frequency of CpG dinucleotides than are present in mammalian DNA. Furthermore, bacterial CpG dinucleotides are often not methylated. It is thought that these two features in combination with specific flanking bases constitute a CpG motif that is recognized as a “danger” signal by the innate immune system of mammals and therefore an immune response is induced when these motifs are encountered. These immunostimulatory activities of bacterial CpG DNA can also be achieved with synthetic CpG oligodeoxynucleotides (ODN). Recognition of CpG motifs by the innate immune system requires engagement of Toll-like receptor 9 (TLR-9), which induces cell signaling and subsequently triggers a pro-inflammatory cytokine response and a predominantly Th1-type immune response. CpG ODN-induced innate and adaptive immune responses can result in protection in various mouse models of disease. Based on these observations, clinical trials are currently underway in humans to evaluate CpG ODN therapies for cancer, allergy and infectious disease. However, potential applications for immunostimulatory CpG ODN in species of veterinary importance are just being explored. In this review, we will highlight what is presently known about the immunostimulatory effects of CpG ODN in domestic animals. [Copyright &y& Elsevier]

Oral immunization is the most effective way of inducing immune responses in the intestinal tract. Biodegradable microspheres have been used extensively for the delivery of antigens to the Peyer’s patches (PPs) within the gut-associated lymphoid tissue (GALT). We evaluated various formulations of alginate microspheres for their capacity to induce mucosal immune responses in vivo. Multiple intestinal “loops” each containing a single PP, were surgically prepared in lambs. We have previously showed that PP in individual intestinal loops function as independent sites for the induction of immune responses. This animal model provides a system for directly comparing different antigen formulations within the same animal. Individual intestinal loops were injected with a model antigen, porcine serum albumin (PSA) encapsulated in three different formulations of alginate micropsheres. Three weeks after immunization, PSA-specific immune responses were assayed with antibody secreting cell (ASC) ELISPOT, lymphocyte proliferative responses (LPRs), IFN-γ production and antibody secreted into intestinal loops. PSA encapsulated in alginate micropsheres or in saline induced humoral immune responses as indicated by the presence of numerous ASC. However, PSA-specific T-cell responses (LPR and IFN-γ production) were not induced. [Copyright &y& Elsevier]