eIF4EHP promotes Ldh mRNA translation in and fruit fly adaptation to hypoxia.

Hypoxia induces profound modifications in the gene expression program of eukaryotic cells due to lowered ATP supply resulting from the blockade of oxidative phosphorylation. One significant consequence of oxygen deprivation is the massive repression of protein synthesis, leaving a limited set of mRNAs to be translated. Drosophila melanogaster is strongly resistant to oxygen fluctuations; however, the mechanisms allowing specific mRNA to be translated into hypoxia are still unknown. Here, we show that Ldh mRNA encoding lactate dehydrogenase is highly translated into hypoxia by a mechanism involving a CA-rich motif present in its 3' untranslated region. Furthermore, we identified the cap-binding protein eIF4EHP as a main factor involved in 3'UTR-dependent translation under hypoxia. In accordance with this observation, we show that eIF4EHP is necessary for Drosophila development under low oxygen concentrations and contributes to Drosophila mobility after hypoxic challenge. Altogether, our data bring new insight into mechanisms contributing to LDH production and Drosophila adaptation to oxygen variations.

Stimulatory Toll-like receptor 2 suppresses restraint stress-induced immune suppression.

Stress can enhance or suppress immune functions depending on a variety of factors. Our previous studies observed that Toll-like receptor 2 (TLR2) participates in chronic restraint stress-induced immune dysfunction. However, the mechanism by which TLR2 prevents immune suppression remains elusive. Our investigation found that stimulation of TLR2 by peptidoglycan (PGN) significantly attenuates splenocyte apoptosis and markedly blocks alterations of anti-apoptotic and apoptotic proteins. Activation of TLR2 inhibits chronic stress-reduced phosphorylation of c-Jun N-terminal kinase (JNK) and diminishes chronic stress-induced up-regulation of corticosterone production. Additionally, our data show that chronic stress causes a dramatic decrease of cytokine IL-2 level but an increase of IL-4 and IL-17 in CD4(+) T cells. Interestingly, PGN could block these alterations of cytokine levels. Collectively, our studies demonstrate that stimulation of TLR2 attenuates chronic stress-induced immune suppression by modulating apoptosis-related proteins and immunoregulatory agents.

Pathogenicity of Bordetella avium under immunosuppression induced by Reticuloendotheliosis virus in specific-pathogen-free chickens.

Multiple infections of Bordetella avium (B. avium) with virus, especially immunosuppressive virus, have become more and more severe in chickens in China. The increasing morbidity and mortality of its complications have amplified concerns about the impact of B. avium on animal health. To evaluate the pathogenicity of B. avium under immunosuppression status, we developed four types of Reticuloendotheliosis virus (REV) infection models. After a comparison of body weight, relative immune organ index, Newcastle disease virus antibody titers and lymphocyte ratio, we chose the early age with low dose infection as our immunosuppressive model. To investigate the pathogenicity of B. avium under this model, a study was completed in which chickens were inoculated with REV-only, B. avium-only, both agents (REV -B. avium) or first inoculated with REV and 5 d later with B. avium (REV/B. avium). Results revealed that antibody titers to B. avium, concentrations of IFN-γ and SIgA were decreased in coinfected chickens when compared to the B. avium-only chickens, but the changing trend was similar between REV/B. avium and B. avium-only groups. Overall, REV did enhance the pathogenicity of B. avium. However, B. avium-only did not cause severe immune dysfunction unless chicks were coinfected with REV. REV preceding infection with B. avium showed mild impairment, which needs further exploration.

Exploring mRNA translation strategies for hypoxia adaptation across distantly related metazoans.

Adaptation to variations in oxygen concentration is a conserved mechanism in all metazoans. Extensive studies have been focused on the roles of hypoxia-inducible factors (HIFs) in response to hypoxia. However, HIF1α is not conserved in all animals, and HIF2α and HIF3α are only observed in vertebrates. In this review, we discuss mechanisms enabling three cross-species to adapt to hypoxic conditions. Using transcriptomic data of hypoxia-induced genes from distantly related metazoans, we analyzed the enrichment and conservation of mRNA determinants such as transcript, CDS, 5'UTR and 3'UTR size. We found that long genes are enriched in hypoxia-induced transcripts and might be translated in a 3'UTR-dependent manner. We hope our work can provide a new direction on investigating alternative mechanisms for mRNA translation under hypoxia.

Many studies have focused on a group of proteins called hypoxia-inducible factors (HIFs) that help animals adapt to different oxygen concentrations, but not all animals have the same HIF proteins. This means that some genes involved in oxygen adaptation work differently in different animals. In this review, we look at three different species and how they adapt to low oxygen levels, with a special focus on how genes are used to make proteins. We used information from these species to see if certain features of genes (like their size) are important for adaptation. We found that in response to low oxygen, longer genes become more active and may use a specific part of the gene to make proteins. However, some common features thought to be important for gene activity in low oxygen were not found to be significant. This research gives us new directions to explore how genes make proteins in low-oxygen conditions, which is important for understanding how animals survive in changing environments.

Verification of blood flow path reconstruction mechanism in distal sigmoid colon and rectal cancer after high IMA ligation through preoperative and postoperative comparison by manual subtraction CTA.

INTRODUCTION: We aimed to investigate manual subtraction computed tomography angiography (MS-CTA) to further confirm the distribution and classification of LCA (left colic artery) ascending/descending branches, then observe the postoperative blood flow path to illustrate how the above branches evolved to postoperative blood path. MATERIAL AND METHODS: 89 patients with distal sigmoid and rectal cancer were referred in our observation and underwent MS-CTA between June 2020 and March 2022. We classified the distribution of LCA and confirmed whether there exists AMCA (accessory middle colic artery). Then we planned blood flow path based on the classification of LCA branches before operation. High ligation was applied in regular radical surgery. During operation, we carefully protect the bifurcation of ascending and descending LCA. Then we compared the planned blood flow path with the actual postoperative blood flow path to verify the mechanism we proposed previously. RESULTS: Of 89 patients, 82 cases met our criteria, we summarized 6 distribution pattens of LCA ascending and descending branches. These preoperative pattens are consistent with the inspection during operation. The postoperative blood flow path of 6 pattens is evolved from the above adjacent anastomotic branches and is consistent with the planned blood flow path. We also found 2 cases with IMA stenosis and 1 case with SMA stenosis under pathological condition, and their compensatory blood flow path is in accordance with our theory. The rate of the anastomotic leakage in our study group is relatively low (7.3%). CONCLUSION: MS-CTA could confirm the distribution of LCA and AMCA, display accurate postoperative blood reconstruction path after IMA high ligation, and it further verified the mechanism we proposed previously, which is the proximal anastomotic branches forming new blood flow path from high-pressure area to the low-pressure area. This mechanism might be helpful for performing accurate laparoscopic sigmoid and rectal cancer surgery.

Monoclonal antibodies directed against the outer membrane protein of Bordetella avium.

Bordetella avium is the etiologic agent of coryza and rhinotracheitis in poultry. This respiratory disease is responsible for substantial economic losses in the poultry industry. Monoclonal antibodies (MAbs) were produced against the outer membrane proteins (OMPs) of B. avium isolated from diseased chickens. BALB/c mice were immunized with the extracted B. avium OMPs. Then the splenocytes from immunized mice and SP2/0 myeloma cells were fused using PEG 4000. Three stable hybridoma clones (designated as 3G10, 4A3, and 4E8) were produced via indirect ELISA and three rounds of subcloning. The MAbs were classified as IgG1, and can recognize the 58 kDa OMP band by Western blot assays. No MAb cross-reactivity with chicken Proteus mirabilis, Escherichia coli, and Salmonella was observed. A double antibody sandwich ELISA (DAS-ELISA) was developed using the rabbit polyclonal antibodies as the capture antibody and MAb 4A3 as the detection antibody. Under the DAS-ELISA, the minimum detectable concentration of B. avium was 1 × 10(4) CFU/mL, and no cross-reactivity occurred with chicken Proteus mirabilis, Escherichia coli, and Salmonella. Results showed that the DAS-ELISA has good sensitivity and specificity. Clinical application showed the DAS-ELISA was more sensitive than the plate agglutination test. This study may be used to develop a quick and specific diagnostic kit, analyze epitopes, and establish systems for typing B. avium.

Effects of Taishan Pinus massoniana pollen polysaccharide on the subunit vaccine of Proteus mirabilis in birds.

Three adjuvants, namely, Taishan Pinus massoniana pollen polysaccharide (TPPPS), white mineral oil (WO) and propolis (PP), were added to the outer membrane protein (OMP) of Proteus mirabilis (P. mirabilis) and their effects were compared. Three hundred 1-day-old chicks were randomly divided into five groups (I-V), with 60 chicks per group, and injected subcutaneously with WO-OMP vaccine (I), PP-OMP vaccine (II), TPPPS-OMP vaccine (III), OMP-only vaccine (IV) and physiological saline (V) at 3, 7 and 12 days old. On days 3, 7, 14, 21, 28, 35, 42 and 49 after the first vaccination, the antibody titers, interleukin-2 levels (IL-2) and T-lymphocyte proliferation rates in the peripheral blood as well as the secreting-type IgA levels (SIgA) in the duodenum were measured. On day 7 after the third vaccination, the chicks were challenged with P. mirabilis strain Q1 and the protective effects of each group were observed. The highest protective rate was observed in group III. Moreover, the antibody titers as well as IL-2, SIgA and T-lymphocyte proliferation rates in this group significantly increased and were significantly higher than those in the other groups at most time points. The results indicate that TPPPS could significantly enhance the effects of the subunit vaccine of P. mirabilis; induced stronger humoral, cellular and mucosal immunity as compared with WO and PP; and should be developed as a vaccine adjuvant.

Taishan Pinus massoniana pollen polysaccharides promote immune responses of recombinant Bordetella avium ompA in BALB/c mice.

To study the effects of Taishan Pinus massoniana pollen polysaccharides (TPPPS) on Bordetella avium outer membrane protein A (ompA) recombinant protein vaccine, ompA was expressed, confirmed by Western blotting and mixed with TPPPS. Female BALB/c mice were randomly divided into six groups (I-VI). Groups I, II, and III were treated with TPPPS-ompA at doses of 200, 400, and 800 mg/ml, respectively. Groups IV, V, and VI were treated with Freund's adjuvant-ompA, pure ompA, and physiological saline, respectively. On days 3, 7, 14, 21, 28, 35, 42, and 49 after the first vaccination, antibody titers, interleukin-2 (IL-2) levels, peripheral blood CD4+ and CD8+ levels, and T lymphocyte proliferation rates in peripheral blood, as well as secreting-type immunoglobulin A (SIgA) levels in the duodenum, were measured. The antibody titers against ompA, IL-2, T lymphocyte proliferation rate, CD4+, and CD8+ in Group II were significantly (P<0.05) higher than those in other groups. However, little difference in SIgA content was observed among Groups I, II, and IV. These results indicated that TPPPS strengthened humoral and cellular immune response against recombinant ompA vaccine and 400 mg/ml TPPPS showed significance (P<0.05) compared with Freund's adjuvant. Therefore, TPPPS can be developed into an adjuvant for recombinant protein vaccines or plant-derived medicine for animal husbandry.

Effects of Taishan Robinia pseudoacacia Polysaccharides on immune function in chickens.

To determine the immune function of Taishan Robinia pseudoacacia Polysaccharide (TRPPS) on chickens, 240 chickens were selected as experimental animals and treated with various doses of TRPPS by hypodermic injection before immunized NDV inactivated vaccine. The results indicated that any dose of TRPPS could significantly promote the development of the immune organs, increase the quantity of leukocyte and the ratio of lymphocyte, and improve the antibody titers against Newcastle disease. Meanwhile, it also increased the magnitude of SIgA in duodenum. However, the dose of 200 mg/ml showed to be the most effective. Therefore, in terms of improving immunologic function and production performance, TRPPS could be used as a vaccine immunopotentiator for immune responses.