Peyer’s Patches and Mesenteric Lymph Nodes Cooperatively Promote Enteropathy in a Mouse Model of Food Allergy

Background and Objective

To improve the efficacy and safety of tolerance induction for food allergies, identifying the tissues responsible for inducing intestinal inflammation and subsequent oral tolerance is important. We used OVA23-3 mice, which express an ovalbumin-specific T-cell receptor, to elucidate the roles of local and systemic immune tissues in intestinal inflammation.

Methods and Results

OVA23-3 mice developed marked enteropathy after consuming a diet containing egg white (EW diet) for 10 days but overcame the enteropathy (despite continued moderate inflammation) after receiving EW diet for a total of 28 days. Injecting mice with anti-IL-4 antibody or cyclosporine A confirmed the involvement of Th2 cells in the development of the enteropathy. To assess the individual contributions of Peyer’s patches (PPs), mesenteric lymph nodes (MLNs), and the spleen to the generation of effector CD4⁺ T-cells, we analyzed the IL-4 production, proliferation in response to ovalbumin, and CD4⁺ T-cell numbers of these tissues. EW feeding for 10 days induced significant IL-4 production in PPs, the infiltration of numerous CD4⁺ T-cells into MLNs, and a decrease in CD4⁺ T-cell numbers in spleen. On day 28, CD4⁺ T-cells from all tissues had attenuated responses to ovalbumin, suggesting tolerance acquisition, although MLN CD4⁺ T-cells still maintained IL-4 production with proliferation. In addition, removal of MLNs but not the spleen decreased the severity of enteropathy and PP-disrupted mice showed delayed onset of EW-induced inflammatory responses. Disruption of peripheral lymphoid tissues or of both PPs and MLNs almost completely prevented the enteropathy.

Conclusions

PPs and MLNs coordinately promote enteropathy by generating effector T-cells during the initial and exacerbated phases, respectively; the spleen is dispensable for enteropathy and shows tolerogenic responses throughout EW-feeding. The regulation of PPs may suppress the initiation of intestinal inflammation, subsequently restricting MLNs and inhibiting the progression of food-allergic enteropathy.     

Effective distance of volatile cues for plant–plant communication in beech

In response to volatiles emitted from a plant infested by herbivorous arthropods, neighboring undamaged conspecific plants become better defended against herbivores; this is referred to as plant‒plant communication. Although plant‒plant communication occurs in a wide range of plant species, most studies have focused on herbaceous plants. Here, we investigated plant‒plant communication in beech trees in two experimental plantations in 2018 and one plantation in 2019. Approximately 20% of the leaves of a beech tree were clipped in half in the spring seasons of 2018 and 2019 (clipped tree). The damage levels to leaves in the surrounding undamaged beech trees were evaluated 90 days after the clipping (assay trees). In both years, the damage levels decreased with a reduction in the distance from the clipped tree. In 2019, we also recorded the damage levels of trees that were not exposed to volatiles (nonexposed trees) as control trees and found that those that were located <5 m away from clipped trees had significantly less leaf damage than nonexposed trees. By using a gas chromatograph–mass spectrometer, ten and eight volatile compounds were detected in the headspaces of clipped and unclipped leaves, respectively. Among them, the amount of (Z)‐3‐hexenyl acetate in clipped leaves was significantly higher than that in nonclipped leaves. Our result suggests that green leaf volatiles such as (Z)‐3‐hexenol and (Z)‐3‐hexenyl acetate and other volatile organic compounds emitted from clipped trees induced defenses in the neighboring trees within the 5 m radius. The effective distances of plant‒plant communication in trees were discussed from the viewpoint of the arthropod community structure in forest ecosystems.     

Synthesis and structure–activity relationships of 8-substituted-2-aryl-5-alkylaminoquinolines: Potent,orally active corticotropin-releasing factor-1 receptor antagonists

We previously reported a series of 8-methyl-2-aryl-5-alkylaminoquinolines as a novel class of corticotropin-releasing factor-1 (CRF₁) receptor antagonists. A critical issue encountered for this series of compounds was low aqueous solubility at physiological pH (pH 7.4). To address this issue, derivatization at key sites (R², R³, R⁵, R^5′, and R⁸) was performed and the relationships between structure and solubility were examined. As a result, it was revealed that introduction of a methoxy substituent at the C₈ position had a positive impact on the solubility of the derivatives. Consequently, through in vivo and in vitro biological studies, compound 21d was identified as a potent, orally active CRF₁ receptor antagonist with improved physicochemical properties.     

Involvement of penicillin-binding proteins in the metabolism of a bacterial peptidoglycan containing a non-canonical d-amino acid

Amino Acids - Bacteria produce various d-amino acids, including non-canonical d-amino acids, to adapt to environmental changes and overcome a variety of threats. These d-amino acids are largely...     

Spatial pattern changes of grazing cattle population I

A cattle population presents three spatial patterns according to their behaviour. These patterns are as follows: the random spatial pattern which occurs when the cattle move during grazing (feeding), the aggregated spatial pattern which occurs when they rest, and the regular or quasi-regular spatial pattern which occurs when they separate into two or more subgroups. In this study, these three states of behaviour were represented by a mathematical model, and a method to test the pattern statistically was derived.     

Tyrosinase‐catalyzed metabolism of rhododendrol (RD) in B16 melanoma cells: production of RD‐pheomelanin and covalent binding with thiol proteins

RS‐4‐(4‐Hydroxyphenyl)‐2‐butanol (rhododendrol, RD) was reported to induce leukoderma of the skin. To explore the mechanism underlying that effect, we previously showed that oxidation of RD with mushroom tyrosinase produces RD‐quinone, which is converted to secondary quinone products, and we suggested that those quinones are cytotoxic because they bind to cellular proteins and produce reactive oxygen species. We then confirmed that human tyrosinase can oxidize both enantiomers of RD. In this study, we examined the metabolism of RD in B16F1 melanoma cells in vitro. Using 4‐amino‐3‐hydroxy‐n‐butylbenzene as a specific indicator, we detected moderate levels of RD‐pheomelanin in B16F1 cells exposed to 0.3 to 0.5 mM RD for 72 h. We also confirmed the covalent binding of RD‐quinone to non‐protein thiols and proteins through cysteinyl residues. The covalent binding of RD‐quinone to proteins was 20‐ to 30‐fold greater than dopaquinone. These results suggest that the tyrosinase‐induced metabolism of RD causes melanocyte toxicity.     

Characterization of hemoglobin binding to Actinobacillus actinomycetemcomitans

In this study, binding of hemoglobin to Actinobacillus actinomycetemcomitans was characterized. The ability of A. actinomycetemcomitans to utilize hemoglobin as an iron source was examined by growth studies. Although the bacterial growth was limited almost completely by adding 400 microM 2, 2'-dipyridyl to culture medium, addition of hemoglobin recovered the growth in a dose-dependent manner, revealing that hemoglobin can be utilized effectively as an iron source by A. actinomycetemcomitans. Binding of A. actinomycetemcomitans to hemoglobin was examined by dot-blot assay. Optimal hemoglobin-binding activity occurred at pH 6 and activity under acidic conditions was found to be higher than that under alkaline conditions. Hemoglobin-binding activity was higher under anaerobic conditions than under aerobic conditions, and iron restriction in culture medium decreased the activity by 55%. Heat and trypsin treatments of the bacterial components reduced the activity by 28% and 60%, respectively. Globin inhibited the activity by 49%, while transferrin, lactoferrin and tested amino acids and sugars had little or no inhibitory effects. These results indicate that proteinaceous components of the bacterial cells may be involved in hemoglobin binding and that globin moiety of the hemoglobin molecule may be essential for the binding. In order to identify hemoglobin-binding proteins, the bacterial cell components extracted with n-octyl-beta-D-thioglucoside were subjected to SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was incubated with hemoglobin and bound hemoglobin was detected with anti-hemoglobin antibodies. About 40- and 65-kDa proteins from A. actinomycetemcomitans reacted with hemoglobin. The 65-kDa protein was detected despite the iron concentration in culture medium, whereas expression of the 40-kDa protein was enhanced only when the organism was grown in iron-restricted culture medium. From these results, it is suggested that 40- and 65-kDa proteins of A. actinomycetemcomitans may be involved in hemoglobin binding.     

Climate change and the effect on the biomass growth in steppe communities in Kherlon River Basin, Northern China

The bacterial community composition and diversity in rock varnish of Turpan Basin were investigated by restriction fragment length polymorphism (RFLP) and clone library of the 16S rRNA gene. 114 positive clones were screened, which could be grouped into 28 phylotypes and then further divided into 23 different operational taxonomic units (OTUs). These were affiliated into 5 phyla (Acidobacteria, Proteobacteria, Chloroflexi, Firmicutes and Cyanobacteria). Clones from actinobacteria were the dominant, accounting for 67.5% of total clones in the library, followed by Proteobacteria (15.8%), Chloroflexi (13.2%), Firmicutes (2.6%) and Cyanobacteria (0.9%). Rubrobacter (accounts for 35%) in the phylum Actinobacteria was the dominant genus and contained many species which might be resistant to gamma radiation. A 70% of the library clone sequences showed less 97% similarity to 16S rRNA gene sequences of standard strains obtained by pure culture. Shannon–Wiener index value of this study is 2.52 and is lower than deep-sea sediments, soils, lakes and other environments. Results of this study showed that bacterial diversity in rock varnishes of Turpan Basin was low, but maybe exist a large number of new unknown taxons, especially species that could well adapted to drought and resist radiation.