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The ovalbumin gene family in Gallus gallus is composed of three homologous genes located within a 46 kb locus on chromosome 2: ovalbumin, ovalbumin-related protein Y (OVAY), and ovalbumin-related protein X (OVAX) genes. The expression of these genes in hen oviduct is under estrogen control, but their relative hormonal responsiveness and subsequent protein concentration in egg, is distinctive. Interestingly, all three proteins lack the classical signal peptide for secretion. Ovalbumin, OVAX, and OVAY belong to the serine protease inhibitor (serpin) family whose members share a common tertiary structure. Ovalbumin and OVAX are one of the few members of this family that do not express any protease inhibition activity whereas OVAY has been predicted to be inhibitory, by comparison with the consensus sequence for inhibitory serpins. In contrast to ovalbumin and OVAY, OVAX interacts with heparin, a negatively charged glycosaminoglycan, via a positively charged domain exposed at the surface of the molecule. Ovalbumin is the major egg white protein and might be a source of amino acids for the developing embryo. The physiological function of OVAY is not known, but recent data have revealed a possible role of this protein in early embryonic development. Considering the antibacterial activities of OVAX, this protein might play a role in egg defense. This review sheds light on the expression, biochemistry, and structural specificities of these three highly similar paralogs. It gives new clues in favor of diverging functions, which are likely to have arisen by duplication events from a common ancestral gene., (© 2015 by the Society for the Study of Reproduction, Inc.)

Patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) are threatened by potentially lethal viral manifestations like cytomegalovirus (CMV) reactivation. Because the success of today's virostatic treatment is limited by side effects and resistance development, adoptive transfer of virus-specific memory T cells derived from the stem cell donor has been proposed as an alternative therapeutic strategy. In this context, dose minimization of adoptively transferred T cells might be warranted for the avoidance of graft-versus-host disease (GVHD), in particular in prophylactic settings after T-cell-depleting allo-HSCT protocols. To establish a lower limit for successful adoptive T-cell therapy, we conducted low-dose CD8(+) T-cell transfers in the well-established murine Listeria monocytogenes (L.m.) infection model. Major histocompatibility complex-Streptamer-enriched antigen-specific CD62L(hi) but not CD62L(lo) CD8(+) memory T cells proliferated, differentiated, and protected against L.m. infections after prophylactic application. Even progenies derived from a single CD62L(hi) L.m.-specific CD8(+) T cell could be protective against bacterial challenge. In analogy, low-dose transfers of Streptamer-enriched human CMV-specific CD8(+) T cells into allo-HSCT recipients led to strong pathogen-specific T-cell expansion in a compassionate-use setting. In summary, low-dose adoptive T-cell transfer (ACT) could be a promising strategy, particularly for prophylactic treatment of infectious complications after allo-HSCT., (© 2014 by The American Society of Hematology.)

The use of monoclonal antibodies targeting the CD154 molecule remains one of the most effective means of promoting graft tolerance in animal models, but thromboembolic complications during early clinical trials have precluded their use in humans. Furthermore, the role of Fc-mediated deletion of CD154-expressing cells in the observed efficacy of these reagents remains controversial. Therefore, determining the requirements for anti-CD154-induced tolerance will instruct the development of safer but equally efficacious treatments. To investigate the mechanisms of action of anti-CD154 therapy, two alternative means of targeting the CD40-CD154 pathway were used: a nonagonistic anti-CD40 antibody and an Fc-silent anti-CD154 domain antibody. We compared these therapies to an Fc-intact anti-CD154 antibody in both a fully allogeneic model and a surrogate minor antigen model in which the fate of alloreactive cells could be tracked. Results indicated that anti-CD40 mAbs as well as Fc-silent anti-CD154 domain antibodies were equivalent to Fc-intact anti-CD154 mAbs in their ability to inhibit alloreactive T cell expansion, attenuate cytokine production of antigen-specific T cells and promote the conversion of Foxp3(+) iTreg. Importantly, iTreg conversion observed with Fc-silent anti-CD154 domain antibodies was preserved in the presence of CTLA4-Ig, suggesting that this therapy is a promising candidate for translation to clinical use., (© Copyright 2013 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Asthma is a chronic inflammatory disease of the small airways, with airway hyperresponsiveness (AHR) and inflammation as hallmarks. Recent studies suggest a role for arginase in asthma pathogenesis, possibly because arginine is the substrate for both arginase and NO synthase and because NO modulates bronchial tone and inflammation. Our objective was to investigate the importance of increased pulmonary arginase 1 expression on methacholine-induced AHR and lung inflammation in a mouse model of allergic asthma. Arginase 1 expression in the lung was ablated by crossing Arg1(fl/fl) with Tie2Cre(tg/-) mice. Mice were sensitized and then challenged with ovalbumin. Lung function was measured with the Flexivent. Adaptive changes in gene expression, chemokine and cytokine secretion, and lung histology were quantified with quantitative PCR, ELISA, and immunohistochemistry. Arg1 deficiency did not affect the allergic response in lungs and large-airway resistance, but it improved peripheral lung function (tissue elastance and resistance) and attenuated adaptive increases in mRNA expression of arginine-catabolizing enzymes Arg2 and Nos2, arginine transporters Slc7a1 and Slc7a7, chemokines Ccl2 and Ccl11, cytokines Tnfa and Ifng, mucus-associated epithelial markers Clca3 and Muc5ac, and lung content of IL-13 and CCL11. However, expression of Il4, Il5, Il10, and Il13 mRNA; lung content of IL-4, IL-5, IL-10, TNF-α, and IFN-γ protein; and lung pathology were not affected. Correlation analysis showed that Arg1 ablation disturbed the coordinated pulmonary response to ovalbumin challenges, suggesting arginine (metabolite) dependence of this response. Arg1 ablation in the lung improved peripheral lung function and affected arginine metabolism but had little effect on airway inflammation.

Staphylococcal enterotoxins A (SEA) and B (SEB) are classical models of superantigens (SAg), which induce potent T-cell-stimulating activity by forming complexes with MHC class II molecules on antigen-presenting cells. This large-scale activation of T-cells is accompanied by increased production of cytokines such as interferon-γ (IFN-γ). Additionally, as we previously reported, IFN-γ-producing CD8(+) T cells act as "helper cells," supporting the ability of dendritic cells to produce interleukin-12 (IL-12)p70. Here, we show that DC pulsed with SAg promote the enhancement of anti-tumor immunity. Murine bone marrow-derived dendritic cells (DC) were pulsed with OVA(257-264) (SIINFEKL), which is an H-2Kb target epitope of EG7 [ovalbumin (OVA)-expressing EL4] cell lines, in the presence of SEA and SEB and were subcutaneously injected into naïve C57BL/6 mice. SAg plus OVA(257-264)-pulsed DC vaccine strongly enhanced peptide-specific CD8(+) T cells exhibiting OVA(257-264)-specific cytotoxic activity and IFN-γ production, leading to the induction of protective immunity against EG7 tumors. Furthermore, cyclophosphamide (CY) added to SAg plus tumor-antigens (OVA(257-264), tumor lysate, or TRP-2) pulsed DC immunization markedly enhanced tumor-specific T-cell expansion and had a significant therapeutic effect against various tumors (EG7, 2LL, and B16). Superantigens are potential candidates for enhancing tumor immunity in DC vaccines.

Effective control of the intracellular protozoan parasite Toxoplasma gondii depends on the activation of antigen-specific CD8(+) T-cells that manage acute disease and prevent recrudescence during chronic infection. T-cell activation in turn, requires presentation of parasite antigens by MHC-I molecules on the surface of antigen presenting cells. CD8(+) T-cell epitopes have been defined for several T. gondii proteins, but it is unclear how these antigens enter into the presentation pathway. We have exploited the well-characterized model antigen ovalbumin (OVA) to investigate the ability of parasite proteins to enter the MHC-I presentation pathway, by engineering recombinant expression in various organelles. CD8(+) T-cell activation was assayed using 'B3Z' reporter cells in vitro, or adoptively-transferred OVA-specific 'OT-I' CD8(+) T-cells in vivo. As expected, OVA secreted into the parasitophorous vacuole strongly stimulated antigen-presenting cells. Lower levels of activation were observed using glycophosphatidyl inositol (GPI) anchored OVA associated with (or shed from) the parasite surface. Little CD8(+) T-cell activation was detected using parasites expressing intracellular OVA in the cytosol, mitochondrion, or inner membrane complex (IMC). These results indicate that effective presentation of parasite proteins to CD8(+) T-cells is a consequence of active protein secretion by T. gondii and escape from the parasitophorous vacuole, rather than degradation of phagocytosed parasites or parasite products.

In the present study, we examined the composition, amount, and uptake of yolk nutrients [fat, protein, water, and carbohydrates (COH)] during incubation of eggs from 30- and 50-wk-old broiler breeder hens. Eggs were sampled at embryonic d 0 (fresh eggs), 13, 15, 17, 19, and 21 (hatch). Egg, embryo, yolk content, and yolk sac membrane were weighed, and the yolk sac (YS; i.e., yolk content + yolk sac membrane) composition was analyzed. From 30 to 50 wk of age, the albumen weight increased by 13.3%, whereas the yolk increased by more than 40%. The proportion of fat in the fresh yolk of the 30-wk-old group was 23.8% compared with 27.4% in the 50-wk-old group, whereas the proportion of protein was 17.9% compared with 15.6%, respectively. During incubation, results indicated that water and protein infiltrated from other egg compartments to the YS. Accordingly, the calculated change in the content of water and protein between fresh yolk and sampled YS does not represent the true uptake of these components from the YS to the embryo, and only fat uptake from the YS can be accurately estimated. By embryonic d 15, fat uptake relative to embryo weight was lower in the 30-wk-old group than in the 50-wk-old group. However, by embryonic d 21, embryos of both groups reached similar relative fat uptake, suggesting that to hatch, embryos must attain a certain amount of fat as a source of energy for the hatching process. The amount of COH in the YS increased similarly during incubation in eggs from hens of both ages, reaching a peak at embryonic d 19, suggesting COH synthesis in the YS. At hatch, the amount of protein, water, and COH in the residual YS, relative to the weight of the yolk-free chick, was similar in eggs from young and old hens. However, chicks from the younger hens had less fat in the YS for their immediate posthatch nutrition compared with those from the older hens.

The mechanisms underlying the exaggerated distal airway inflammation and hyperresponsiveness that characterize acute exacerbations of asthma are largely unknown. Using BALB/c mouse experimental models, we demonstrated a potentially important role for alveolar macrophages (AM) in the development of an allergen-induced exacerbation of asthma. To induce features of airway inflammation and remodeling characteristic of mild chronic asthma, animals were systemically sensitized and exposed to low mass concentrations (≈3 mg/m(3)) of aerosolized ovalbumin for 30 minutes per day, 3 days per week, for 4 weeks. A subsequent single moderate-level challenge (≈30 mg/m(3)) was used to trigger an acute exacerbation. In chronically challenged animals, cytokine expression by AM was not increased, whereas after an acute exacerbation, AM exhibited significantly enhanced expression of proinflammatory cytokines, including interleukin (IL) 1β, IL-6, CXCL-1, and tumor necrosis factor α. In parallel, there was a marked increase in the expression of several cytokines by CD4(+) T-lymphocytes, notably the Th2 cytokines IL-4 and IL-13. Importantly, AM from an acute exacerbation stimulated the expression of Th2 cytokines when cocultured with CD4(+) cells from chronically challenged animals, and their ability to do so was significantly greater than AM from either chronically challenged or naïve controls. Stimulation was partly dependent on interactions involving CD80/86. We conclude that in an acute exacerbation of asthma, enhanced cytokine expression by AM may play a critical role in triggering increased expression of cytokines by pulmonary CD4(+) T-lymphocytes.

Allergic asthma, a major cause of morbidity and leading cause of hospitalizations, is an inflammatory disease orchestrated by T helper cells and characterized by the lung migration of eosinophils, which are important asthma effector cells. Lung migration of inflammatory cells requires, among other events, the chemokine receptor transduction of lung-produced inflammatory chemokines. Despite the widespread prevalence of this disease, the molecular mechanisms regulating chemokine production and receptor regulation in asthma are poorly understood. Previous work from our laboratory demonstrated that beta-arrestin-2 positively regulates the development of allergic airway disease in a mouse model, partly through positive regulation of T-lymphocyte chemotaxis to the lung. However, beta-arrestin-2 is expressed in many cell types, including other hematopoietic cells and lung structural cells, which are involved in the development and manifestation of allergic airway disease. To determine the cell types required for beta-arrestin-2-dependent allergic inflammation, we generated bone marrow chimera mice. Using the ovalbumin murine model of allergic airway disease, we show that eosinophilic and lymphocytic inflammation is restored in chimeric mice, with expression of beta-arrestin-2 exclusively on hematopoietic-derived cell types. In contrast, airway hyperresponsiveness is dependent on the expression of beta-arrestin-2 in structural cells. Our data demonstrate that the expression of beta-arrestin-2 in at least two divergent cell types contributes to the pathogenesis of allergic airway disease.

The effects of remodeling of airway smooth muscle (SM) by hyperplasia on airway SM contractility in vivo are poorly explored. The aim of this study was to investigate the relationship between allergen-induced airway SM hyperplasia and its contractile phenotype. Brown Norway rats were sensitized with ovalbumin (OVA) or saline on day 0 and then either OVA-challenged once on day 14 and killed 24 h later or OVA-challenged 3 times (on days 14, 19, and 24) and killed 2 or 7 days later. Changes in SM mass, expression of total myosin, SM myosin heavy chain fast isoform (SM-B) and myosin light chain kinase (MLCK), tracheal contractions ex vivo, and airway responsiveness to methacholine (MCh) in vivo were assessed. One day after a single OVA challenge, the number of SM cells positive for PCNA was greater than for control animals, whereas the SM mass, contractile phenotype, and tracheal contractility were unchanged. Two days after three challenges, SM mass and PCNA immunoreactive cells were increased (3- and 10-fold, respectively; P < 0.05), but airway responsiveness to MCh was unaffected. Lower expression in total myosin, SM-B, and MLCK was observed at the mRNA level (P < 0.05), and total myosin and MLCK expression were lower at the protein level (P < 0.05) after normalization for SM mass. Normalized tracheal SM force generation was also significantly lower 2 days after repeated challenges (P < 0.05). Seven days after repeated challenges, features of remodeling were restored toward control levels. Allergen-induced hyperplasia of SM cells was associated with a loss of contractile phenotype, which was offset by the increase in mass.

The immunoregulatory enzyme indoleamine 2,3-dioxygenase (IDO) is expressed by a subset of murine plasmacytoid DCs (pDCs) in tumor-draining lymph nodes (TDLNs), where it can potently activate Foxp3+ regulatory T cells (Tregs). We now show that IDO functions as a molecular switch in TDLNs, maintaining Tregs in their normal suppressive phenotype when IDO was active, but allowing inflammation-induced conversion of Tregs to a polyfunctional T-helper phenotype similar to proinflammatory T-helper-17 (TH17) cells when IDO was blocked. In vitro, conversion of Tregs to the TH17-like phenotype was driven by antigen-activated effector T cells and required interleukin-6 (IL-6) produced by activated pDCs. IDO regulated this conversion by dominantly suppressing production of IL-6 in pDCs, in a GCN2-kinase dependent fashion. In vivo, using a model of established B16 melanoma, the combination of an IDO-inhibitor drug plus antitumor vaccine caused up-regulation of IL-6 in pDCs and in situ conversion of a majority of Tregs to the TH17 phenotype, with marked enhancement of CD8+ T-cell activation and antitumor efficacy. Thus, Tregs in TDLNs can be actively reprogrammed in situ into T-helper cells, without the need for physical depletion, and IDO serves as a key regulator of this critical conversion.

Serine protease inhibitors (serpins) are a family of proteins that are important in the regulation of several biological processes. This mainly involves the inhibition of serine proteases, although some serpins inhibit a different class of proteases or even function without inhibitory activity. In contrast to other protease inhibitor families, serpins inhibit their target proteases by a specific mechanism, which depends on a change in conformation. This review primarily focuses on one subgroup of serpins--ovalbumin (ov)-serpins. Different than most members of the family, this group of serpins lacks secretion signal sequences and therefore, mainly functions intracellularly. In addition to expression in most normal tissues, ov-serpins can be found in multiple different cells of the immune system. Interestingly, expression of ov-serpins in these cells is tightly regulated, indicating a role for these serpins in the regulation of immune responses. The role of serpins in the immune response will be the topic of this review.

In a series of experiments, effects of storage of eggs in water on internal egg quality, embryonic development, and hatchling quality were investigated. In experiment 1, unfertilized eggs were stored for 4 to 14 d in water (W) or air (control; C). In experiment 2, fertilized eggs were stored for 3 to 14 d in water or air and thereafter incubated for 9 d. In experiment 3, eggs were stored for 16 d in water or air and incubated for 1 to 9 d thereafter. In experiment 4, eggs were stored for 14 d in water or air, incubated thereafter, and hatching time and hatchling quality were determined. In all experiments, egg weight loss in the C treatment increased with duration of storage, whereas W eggs gained weight during storage. Albumen and yolk pH after storage and during incubation were greater in the C eggs compared with the W eggs. In experiment 3, embryonic development at d 4 and 9 was advanced in the W eggs compared with the C eggs. In experiment 4, the number of viable embryonic cells after storage and after trypsinization was lower in the C treatment than in the W treatment (30,188 vs. 69,618; P < 0.001). Hatching time was postponed in the W treatment compared with the C treatment (501 vs. 495 h; P < 0.05). Hatchling length was greater in the C treatment (19.7 vs. 20.3 cm; P = 0.01), and residual yolk was less in the C treatment than in the W treatment (4.9 vs. 8.3 g; P < 0.001). We concluded that storage of eggs in water for a prolonged period positively affects internal egg characteristics and early embryonic development, but negatively affects hatchling quality. The reason for the loss of the head start with progressing incubation needs further investigation.

In avian species, maternal provisioning to the eggs is predicted to be more valuable for the offspring under adverse environmental conditions and intense sibling competition. However, studies manipulating both the amount of maternal pre-hatching resources and the harshness of post-hatching environment have seldom been performed to date. In this experimental study of Barn Swallow (Hirundo rustica) nestlings, we tested the consequences of a reduction in the albumen content of the eggs for fitness-related offspring traits, while performing an unbalanced partial cross-fostering soon after hatching, either increasing or decreasing brood size by one nestling. By molecular sexing of the chicks, we additionally tested for sex-specific sensitivity of individual nestlings to experimental treatments and to sex ratio variation in nestmates. We predicted that chicks hatching from albumen-deprived eggs should suffer more than control chicks from the harsher rearing conditions of enlarged broods. However, although albumen removal depressed chick body mass, chicks hatching from control eggs did not fare better than those hatching from eggs with reduced albumen content in enlarged vs. reduced broods. Albumen removal had sex-specific effects on immunity, with males, but not females, hatching from eggs with reduced albumen content showing a lower T-cell-mediated immune response than controls, suggesting that the two sexes were differentially susceptible to resource deprivation during early ontogeny. In addition, both immune response and chick body mass at age 7 days, when maximum growth rate is attained, declined with an increasing proportion of male nestmates. The effect of brood size manipulation on chick body mass at age 12 days, when peak body mass is attained, was also found to depend on brood sex composition, in that an increase in the proportion of male nestmates depressed offspring body mass in reduced broods, while the reverse was true in enlarged broods. On the whole, these findings suggest that sex differences may exist in environmental sensitivity and patterns of resource allocation among different body functions, and that brood size variation and sex composition may affect offspring fitness-related traits.

In this communication, the surface activity of the ovalbumin (OVA) at the air/water interface was studied to establish the nature of the interaction with the stearic acid (SA) monolayer, based on Langmuir-Blodgett (LB) technique. The interaction was monitored by studying the time (t) variation of surface pressure (pi) at constant area (A). The growth of pi with time indicates a positive association between the SA and the OVA molecules. The surface compressibility analysis has been performed to specify the phase transition of OVA-SA mixed monolayer. Incorporation/association of OVA within the SA monolayer led to noteworthy changes in surface compressibility and was surface pressure as well as protein concentration dependent. Both the hydrophobic and the Vander wall type interactions are found to be responsible for the association. The quenching of tyrosine band in tryptophan excitation spectrum is observed in steady-state fluorescence spectroscopy. This suggests that the tyrosine is the probable binding site with SA. Due to incorporation of SA, the energy transfer from tyrosine to tryptophan is hindered. At higher pressure, OVA tend to squeeze out from the SA monolayer. The high-resolution field emission scanning electron microscope (FE-SEM) image confirms this observation. Aggregated protein structure observed at high pressure indicates unfolding of protein.

This study was undertaken to determine the influence of storage time and temperature on the volume, weight, and pH of egg albumen, the physical strength of vitelline membrane, and the fate of Salmonella Enteritidis artificially inoculated into egg albumen. A fiber-optic probe was used for inoculation with Salmonella Enteritidis at 10(2), 10(4), or 10(6) cells per egg. Both fresh and inoculated eggs were stored at 4, 10, and 22 degrees C for 6 weeks. Five fresh uninoculated eggs from each storage group were collected each week, and the weight, volume, and pH of the egg albumen were measured. The forces, energies, and degrees of membrane deformation required to rupture the vitelline membranes also were determined from either albumen-free yolks or yolks surrounded by albumen. In separate experiments, five inoculated eggs were evaluated each week for populations of Salmonella Enteritidis. When the eggs were stored at 4 degrees C, the albumen retained significantly more volume and weight and had a relatively lower pH. The vitelline membranes from eggs stored at 4 and 10 degrees C required more force and energy for rupture. Salmonellae flourished at 22 degrees C, even in the albumen with the lowest initial population, 10(2) cells per egg. Storage at 4 and 10 degrees C inhibited the growth of salmonellae in the albumen of eggs with initial populations of 10(2), 10(4), or 10(6) cells per egg. In eggs with initial Salmonella populations of 10(6) cells per egg that were stored at 22 degrees C, the populations of reached as high as 10(10) cells per egg after 4 weeks of storage. Storage at 4 and perhaps 10 degrees C postponed the aging process of chicken eggs, preserved the antimicrobial agents of the albumen, and maintained the integrity of vitelline membrane. Low-temperature storage therefore had a significant impact on the safety and overall quality of the eggs.

A total of 2123 eggs obtained from Brown Leghorn hens (unselected since 1965, ISA Brown, commercial brown egg layer) and Babcock hens (commercial white egg layer) at 32, 50, and 68 wk of age were used to investigate relationships among measures of albumen quality and a functional property of albumen. The eggs were sampled fresh and after storage for 5 and 10 d. At sampling, eggs were weighed and broken, and albumen height, pH, and volume after whipping for 80 s were measured. Also, yolks were weighed, dried shells were weighed, and albumen weight was determined by difference. Egg weight and the weights of the 3 principal components of the egg all increased with increasing age of the hen, with yolk weights increasing proportionately more. With storage, egg and albumen weights decreased, whereas yolk weight increased. Eggs from Brown Leghorn hens were smallest but had proportionately the largest yolks. Albumen height decreased with time in storage, and albumen pH and whipping volume increased. Differences between lines suggested that selection has changed the proportion of the yolk, albumen, and shell and has increased albumen height. Albumen height and whipping volume were negatively correlated, and differences between lines suggest that selection could have decreased the foaming ability of albumen, a principal reason for including eggs in many processed food products.

The relationship of albumen height (AH) to the compositions of yolks and embryos in hatching eggs from a young (30 wk of age) broiler breeder flock was evaluated during incubation. On Day 2 of incubation, egg weight, yolk weight, and yolk moisture, lipid, and fatty acid contents were determined in eggs from broiler breeders previously identified as laying eggs of either low or high AH. In addition, egg weight, wet and dry embryo weight, and embryo moisture and protein contents were determined on Days 10, 12, and 16, and embryo lipid content was determined on Days 12 and 16. Yolk and embryo weights were expressed as percentages of sampled egg weight. Egg, yolk, and wet embryo weights, yolk moisture and lipid contents, and embryo moisture, protein, and lipid contents were not affected by AH; however, yolk myristic acid concentration was higher, and yolk linoleic acid concentration was lower, in low AH eggs on Day 2 of incubation. Furthermore, on Day 16, dry embryo weight was significantly higher in low AH eggs. Young breeder hens laying eggs of different AH may also produce egg yolks with different fatty acid compositions. Differences in yolk fatty acid profiles between AH groups during early incubation may impact subsequent embryo DM weight without associated effects on embryo moisture, protein, or lipid contents.

The objective of these two experiments was to determine the temporal changes in albumen during storage and early incubation as a means of understanding some of the effects of egg storage on early embryonic development. Eggs from 30- or 50-wk-old broiler breeder hens were incubated (37.5 C dry bulb, 30 C wet bulb) after storage for 0 (fresh) or 5 d (18 C, 75% RH) in Experiment 1. Albumen height, albumen pH, and egg weight loss were recorded at 2, 24, 48, and 66 h of incubation. The same measurements were taken on another group of eggs from 43-wk-old hens stored for 0 (fresh), 4, 8, or 12 d in Experiment 2. All hens were of the same strain. Egg weight loss during incubation was significantly greater in fresh eggs than in stored eggs in Experiment 1. Fresh eggs had significantly greater albumen height and significantly lower albumen pH than stored eggs in both experiments. These differences diminished with length of incubation. Because the blastoderm is located adjacent to the albumen, changes in the viscosity or pH of the albumen may play an integral role in determining the viability of the embryo during the very early stages of development. Incubation of fresh eggs without storage appears to expose the developing embryo to an inappropriate trans-vitelline membrane pH gradient and a thick albumen that may slow vital gas diffusion and limit nutrient availability. These conditions may cause an increased incidence of embryonic death.

1. To investigate the role of platelet activating factor (PAF) in the immediate asthmatic response, we examined the bronchial reactivity to histamine after administration of PAF to guinea-pigs or antigen challenge to passively sensitized guinea-pigs. 2. A bolus injection of PAF (20-40 ng kg-1), which did not cause a significant increase in intrathoracic pressure (ITP), augmented the bronchial response to histamine almost 8 fold. This airway hyperreactivity was observed even 1 min after PAF treatment. 3. A subthreshold dose of antigen (0.01 mg kg-1, i.v.) also provoked hyperreactivity to histamine, which became significant 6 and 11 min after the antigen treatment. 4. The specific PAF-antagonists, SM-10661 and CV-6209 (i.v.) dose-dependently inhibited both PAF- and antigen-induced airway hyperreactivities to histamine. 5. These results suggest that PAF plays an important role in antigen-induced acute airway responses by augmenting the activities of spasmogens.