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[Display omitted] • Agronomic quality of digestate was improved by adding Agricultural Jiaosu (AJ). • Adding AJ reducing free ammonia of digestate helps to reduce the phytotoxicity. • The abundance of some agricultural beneficial genera was increased by adding AJ. • We revealed links between chemical, plant, and microbial indicators in digestate. Anaerobic digestate disposal is challenging, hampering the development of large-scale biogas projects. Currently, digestate is primarily stabilized naturally and subsequently applied to farmland. However, during this process, the number of beneficial microorganisms decreases, free ammonia nitrogen increases, and pH rises, decreasing the benefits of digestate to soil and plants. In this study, digestate was treated by adding agricultural Jiaosu (AJ). The agronomic quality of the digestate was explored by conducting a pot experiment. Subsequently, the mechanisms of the changes in digestate properties were investigated based on chemical and microbial parameters. The results showed that the digestate with a 10% AJ supplement significantly improved the plant characteristics such as plant height, fresh weight, and leaf area. Regarding chemical properties, the addition of 10% AJ decreased the total ammonia nitrogen and free ammonia nitrogen in the digestate to below the toxicity threshold. With respect to microorganisms, the addition of AJ improved the microbial diversity of the digestate and increased the relative abundances of Advenella , Trichococcus, and Comamonas spp., which are beneficial to both soil and plants. Moreover, AJ decreased the abundance of potentially threatening microorganisms such as Thiopseudomonas and Papillibacter spp. Collaborative analysis showed that the improvement in digestate agronomic quality was closely related to its effects on microorganisms. Based on these results, the present study suggests an approach to the post-treatment of anaerobic digestate that can improve its fertilizer value while reducing its phytotoxicity. Furthermore, this study provides a model for a more comprehensive evaluation of the agronomic quality of digestate. [ABSTRACT FROM AUTHOR]

Abstract: The Brugia malayi endosymbiont Wolbachia has recently been shown to be essential for its host’s survival and development. However, relatively little is known about Wolbachia proteins that interact with the filarial host and which might be important in maintaining the obligate symbiotic relationship. The Wolbachia surface proteins (WSPs) are members of the outer membrane protein family and we hypothesise that they might be involved in the Wolbachia–Brugia symbiotic relationship. Notably, immunolocalisation studies of two WSP members, WSP-0432 and WSP-0284 in B. malayi female adult worms showed that the corresponding proteins are not only present on the surface of Wolbachia but also in the host tissues, with WSP-0284 more abundant in the cuticle, hypodermis and the nuclei within the embryos. These results confirmed that WSPs might be secreted by Wolbachia into the worm’s tissue. Our present studies focus on the potential involvement of WSP-0284 in the symbiotic relationship of Wolbachia with its filarial host. We show that WSP-0284 binds specifically to B. malayi crude protein extracts. Furthermore, a fragment of the hypothetical B. malayi protein (Bm1_46455) was found to bind WSP-0284 by panning of a B. malayi cDNA library. The interaction of WSP-0284 and this protein was further confirmed by ELISA and pull-down assays. Localisation by immunoelectron microscopy within Wolbachia cells as well as in the worm’s tissues, cuticle and nuclei within embryos established that both proteins are present in similar locations within the parasite and the bacteria. Identifying such specific interactions between B. malayi and Wolbachia proteins should lead to a better understanding of the molecular basis of the filarial nematode and Wolbachia symbiosis. [Copyright &y& Elsevier]

Abstract: Transient transfection of isolated Brugia malayi embryos by biolistics has proven to be useful in defining promoter structure and function in this parasite. However, isolated transfected embryos are developmentally incompetent. A method of producing developmentally competent transfected parasites is therefore needed. We report that L3 parasites can be chemically transfected in situ in the peritoneal cavity of a gerbil with a construct consisting of a secreted luciferase reporter gene containing a promoter, the 3′ untranslated region and first intron derived from the B. malayi 70kDa heat shock protein gene. The in situ chemically transfected parasites are developmentally competent, producing adult parasites with an efficiency similar to that obtained from implanted untreated L3s. Cultured adult parasites and progeny microfilariae (mf) derived from L3s transfected with this construct secreted luciferase into the culture medium. When the transfected mf were fed to mosquitoes and the resulting L3s collected, the L3s also secreted luciferase into the culture medium. Progeny mf from transgenic adult parasites contained transgenic DNA, and the transgenic mRNA produced in these parasites was found to be correctly cis- and trans-spliced. In situ chemical transformation thus results in developmentally competent transfected B. malayi in which the transgenic sequences remain transcriptionally active in all life cycle stages and are present in the subsequent generation. [Copyright &y& Elsevier]

Abstract: LFA-1 signaling is required for the generation of central memory CD8+ T cells in priming phase. However, its role for effector and memory CD8+ T cell survival in transition and maintenance phases is elusive. We transferred effector and memory CD8+ T cells into C57BL/6 and CD54−/− mice, which were generated by cultivation of ovalbumin (OVA)-pulsed dendritic cells (DCOVA) with naïve CD8+ T cells derived from transgenic OT I mice and purified from effector CD8+ T cell-transferred C57BL/6 mice, respectively. We then assessed kinetics of T cell survival using PE-H2-Kb/OVAI tetramer and FITC-CD8 staining by flow cytometry. We found that survival of transferred effector and memory CD8+ T cells in CD54−/− mice significantly decreased (p <0.05) compared to that in C57BL/6 mice due to an increased T cell apoptosis, which is mediated via downregulation of proapoptotic Bid, anti-apoptotic Bcl-2, Bcl-XL and pro-Caspase-8, and up-regulation of apoptotic Bax and cleaved Caspase-3 and -7 by RNA array and Western blotting analyses. Decreased expression of CD27 and IL-15R on transferred CD8+ T cells with less survival was found to be associated with increased T cell apoptosis, which was confirmed by silencing CD27 with siRNA transfection or using CD8+ (IL-15R−/−)T cells for adoptive transfer into C57BL/6 mice. These data indicate that LFA-1 signal defect-induced CD8+ T cell apoptosis is associated with reduced CD27 costimulation and IL-15R survival signal. Therefore, our study provides important evidence on and elucidates the molecular mechanism associated with the role LFA-1 signaling plays in effector and memory CD8+ T cell survival. [ABSTRACT FROM AUTHOR]

• We evaluated the immunogenicity of a rDBPII vaccine with different variables. • Significant variations in immune responses were observed with different variables. • No specific pattern in immune responses with respect to sex or adjuvant observed. • Sex and immune genes are critical variables to consider in vaccine study design. Relapsing malaria caused by Plasmodium vivax is a neglected tropical disease and an important cause of malaria worldwide. Vaccines to prevent clinical disease and mosquito transmission of vivax malaria are needed to overcome the distinct challenges of this important public health problem. In this vaccine immunogenicity study in mice, we examined key variables of responses to a P. vivax Duffy binding protein vaccine, a leading candidate to prevent the disease-causing blood-stages. Significant sex-dependent differences were observed in B cell (CD80+) and T cell (CD8+) central memory subsets, resulting in significant differences in functional immunogenicity and durability of anti-DBP protective efficacy. These significant sex-dependent differences in inbred mice were in the CD73+CD80+ memory B cell, H2KhiCD38hi/lo, and effector memory subsets. This study highlights sex and immune genes as critical variables that can impact host responses to P. vivax antigens and must be taken into consideration when designing clinical vaccine studies. [ABSTRACT FROM AUTHOR]

A cascadic sensing system was developed for detection of DNA target at ultralow concentration by a combination of magnetic nanoparticles (MNPs) and exonuclease III (Exo III)-induced cascade two-stage isothermal amplification in the study. An ingeniously designed capture hairpin probe (CHP) that integrates target-binding and signal transduction sequences within one multifunctional design was assembled on MNPs. Upon sensing of the analyte nucleic acid, the hairpin probe on MNPs could be opened and stepwise removed by Exo III accompanied by the releasing of target DNA for the successive hybridization and cleavage process and the generation of bare signal transduction sequences of CHP as a new trigger for next circular reaction. The new DNA triggers initiate hybridizing with hairpin DNA probe that contains a partially "caged" G-quadruplex sequence (GHP), forming a duplex structure and liberating the active G-quadruplex structure. Then, Exo III digests the resulting duplex domain, leading to the recycling of new DNA trigger and simultaneously generating numerous ZnPPIX/G-quadruplex supramolecular complexes with the help of the zinc (II)-protoporphyrin IX (ZnPPIX), as an optical label for amplified fluorescence sensing event. Finally, numerous liberated cascade ZnPPIX/G-quadruplex supramolecular complexes give a remarkable fluorescence response. Because of two-stage autocatalytic recycling amplification and the specifically catalyzed formation of ZnPPIX/G-quadruplex supramolecular complexes, this newly designed protocol provides a high sensitivity with a detection limit of 0.75 fM, can discriminate mismatched DNA from perfectly matched target DNA, and gives low matrix effect due to using MNPs as the separation and amplification elements in the real samples. Therefore, it holds great potential for early diagnosis in gene-related diseases. [ABSTRACT FROM AUTHOR]