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Clinical trials of Treg therapy in transplantation are currently entering phases IIa and IIb, with the majority of these employing polyclonal Treg populations that harbor a broad specificity. Enhancing Treg specificity is possible with the use of chimeric antigen receptors (CARs), which can be customized to respond to a specific human leukocyte antigen (HLA). In this study, we build on our previous work in the development of HLA‐A2 CAR‐Tregs by further equipping cells with the constitutive expression of interleukin 10 (IL‐10) and an imaging reporter as additional payloads. Cells were engineered to express combinations of these domains and assessed for phenotype and function. Cells expressing the full construct maintained a stable phenotype after transduction, were specifically activated by HLA‐A2, and suppressed alloresponses potently. The addition of IL‐10 provided an additional advantage to suppressive capacity. This study therefore provides an important proof‐of‐principle for this cell engineering approach for next‐generation Treg therapy in transplantation., Chimeric antigen receptor (CAR) constructs can be modified to incorporate elements for enhanced activity. Here, human Tregs were modified to express an anti‐HLA‐A2 CAR as well as IL‐10 and an imaging reporter. These cells maintained a stable Treg phenotype after lentiviral transduction, were specifically activated by cells expressing HLA‐A2, and had enhanced in vitro suppressive potency compared with CAR Tregs without constitutive IL‐10 expression.

New vertebrate communities are emerging in Europe following the recovery of multiple native predators to highly anthropized landscapes where predator control is still prevalent. While the lack of reference points for these communities creates novel challenges for conservationists and wildlife managers, they also provide opportunities to further our understanding of species interactions. Despite a growing body of evidence, many aspects of interactions among predators remain poorly understood, impairing our ability to anticipate the effects of such changes in predator communities. Through a systematic literature review, we gathered all the available evidence concerning the existence, strength, and demographic impacts of lethal predator interactions among forest‐grouse predators in Europe. We found a highly interconnected predator community, with 44 pairwise lethal interactions among 12 taxa. Three of these resulted in some degree of population suppression of the victim, while another three did not. However, most interactions (38) have not been evaluated for population suppression. Additionally, we highlight how predators interact simultaneously with a large range of other predators and identified at least two further taxa possibly suppressed through the combined impacts of multiple predators. We propose that interactions causing demographic suppression are characterized by impacts on individuals with high survival elasticity and that they are motivated by food limitation and additionally, in mammals, by competition for carcasses. Predator interactions, and our still poor understanding of them, introduce large uncertainties to conservation actions based on the management of predator abundances, which should be carefully evaluated., The uncertainties surrounding emerging predator communities in Europe and the resulting networks of predator interactions create novel challenges for conservation and wildlife management but also offer new opportunities to improve our understanding of species ecology and interactions. Through a literature review of the interactions among the predators of forest grouse in Europe, we found a highly interconnected community of predators, with at least 44 interactions among 12 taxa, three of which result in some degree of population suppression. While the frequency of predator interactions is seemingly driven by food limitation and, in mammals, by competition for carcasses, we propose that whether they cause suppression or not is determined by whether impacts are on population classes whose survival has large effects on the population growth rate.

Background and aim Dietary strategies that contribute to reducing incidence of Helicobacter pylori infection without negative side effects are highly desirable owing to worldwide bacterial prevalence and carcinogenesis potential. The aim of this study was to determine dosage effect of daily cranberry consumption on H. pylori suppression over time in infected adults to assess the potential of this complementary management strategy in a region with high gastric cancer risk and high prevalence of H. pylori infection. Methods This double‐blind, randomized, placebo‐controlled trial on 522 H. pylori‐positive adults evaluated dose–response effects of proanthocyanidin‐standardized cranberry juice, cranberry powder, or their placebos on suppression of H. pylori at 2 and 8 weeks by 13C‐urea breath testing and eradication at 45 days post‐intervention. Results H. pylori‐negative rates in placebo, low‐proanthocyanidin, medium‐proanthocyanidin, and high‐proanthocyanidin cranberry juice groups at week 2 were 13.24%, 7.58%, 1.49%, and 13.85% and at week 8 were 7.35%, 7.58%, 4.48%, and 20.00%, respectively. Consumption of high‐proanthocyanidin juice twice daily (44 mg proanthocyanidin/240‐mL serving) for 8 weeks resulted in decreased H. pylori infection rate by 20% as compared with other dosages and placebo (P

Tumor microenvironment (TME) is formed as a result of interaction and cross-linking between the tumor cell and different types of surrounding cells. Recent studies have shown that the tumor reprograms the microenvironment so that TME promotes the development of primary tumors, their metastasis and becomes an important regulator of oncogenesis. Under the influence of the tumor, the immune profile in the TME undergoes significant changes, “editing". An immunosuppressive network is formed, which suppresses the activity of the main effector of cellular immunity — T lymphocytes. T cells in TMA are in a state of anergy and exhaustion. T cells in TME are characterized by increased expression of inhibitory receptors, decreased secretion of cytokines and cytolytic activity. Blocking inhibitory receptors with specific antibodies can lead to the restoration of the functions of exausted T cells. Therefore, the restoration of the functional activity of T lymphocytes is one of the important strategies in cancer immunotherapy. The formation of the immune profile is influenced by genetic aberrations accumulating in the tumor. They play an important role in creating a specific, characteristic only for this tumor immune environment in the TME. Genetic changes in tumor cells lead to phenotypic and functional rearrangements of lymphocytes, which allows the tumor to escape the reaction of immune cells. Since many tumors occur after prolonged inflammation or exhibit characteristics of chronic inflammation as they progress, inflammation is considered an important factor in the formation of immune profile in TME. Immune infiltrates from different human tumors associated with inflammation may contain valuable prognostic and pathophysiological information. Macrophages in the TME now began to be regarded as descriptive marker and as a therapeutic target. One of the main mechanisms by which tumor cells reprogram surrounding cells is the release of exosomes — small vesicles that carry and deliver proteins and nucleic acids to other cells. When exosomal cargo is absorbed, molecular, transcriptional and translational changes occur in the recipient non-tumor cells in the TME. Therefore, tumor exosomes are an effective means by which the functions of immune cells in TME are purposefully changed. Thus, along with individual molecular and genomic testing of the tumor, attention should be paid to a deeper analysis of the immune profile of TME. It is a large resource of biomarkers and targets for immunotherapy.

Objective To identify correlates of incident HIV infection in rapidly growing HIV molecular clusters. Design Phylogenetic analysis of HIV public health surveillance data. Methods High-priority HIV genetic transmission clusters with evidence of rapid growth in 2012 (i.e. clusters with a pairwise genetic distance ≤0.005 substitutions/site and at least three cases diagnosed in 2012) were identified using HIV-TRACE. Then, we investigated cluster growth, defined as HIV cases diagnosed in the following 5 years that were genetically linked to these clusters. For clusters that grew during the follow-up period, Bayesian molecular clock phylogenetic inference was performed to identify clusters with evidence of incident HIV infection (as opposed to diagnosis of previously infected cases) during this follow-up period. Results Of the 116 rapidly growing clusters identified, 73 (63%) had phylogenetic evidence for an incident HIV case during the 5-year follow-up period. Correlates of an incident HIV case arising in clusters included a greater number of diagnosed but virally unsuppressed cases in 2012, a greater number of inferred undiagnosed cases in the cluster in 2012, and a younger time of most recent common ancestor for the cluster. Conclusion These findings suggest that incident infections in rapidly growing clusters originate equally from diagnosed but unsuppressed cases and undiagnosed infections. These results highlight the importance of promoting retention in care and viral suppression as well as partner notification and other case-finding activities when investigating and intervening on high-priority molecular transmission clusters.

Aquaporin-5 (AQP5) plays a role in breast cancer cell migration. This study aimed to identify AQP5-targeting miRNAs and examine their effects on breast cancer cell migration through exosome-mediated delivery. Bioinformatic analyses identified miR-1226-3p, miR-19a-3p, and miR-19b-3p as putative regulators of AQP5 mRNA. Immunoblotting revealed a decrease of AQP5 protein abundance when each of these miRNAs was transfected into human breast cancer MDA-MB-231 cells. Quantitative real-time PCR demonstrated the reduction of AQP5 mRNA expression by the transfection of miR-1226-3p and a luciferase reporter assay revealed the reduction of AQP5 translation after the transfection of miR-19b-3p in MDA-MB-231 cells. Consistently, the transfection of each miRNA impeded cell migration. Pathway enrichment analyses showed that these three miRNAs regulate target genes, which were predominantly enriched in the gap junction pathway. For the efficient delivery of AQP5-targeting miRNAs to breast cancer cells, exosomes expressing both miRNAs and a peptide targeting interleukin-4 receptor, which is highly expressed in breast cancer cells, were bioengineered and their inhibitory effects on AQP5 protein expression and cell migration were demonstrated in MDA-MB-231 cells. Taken together, AQP5-regulating miRNAs are identified, which could be exploited for the inhibition of breast cancer cell migration via the exosome-mediated delivery.

This chapter deals with the odorants naturally contained in wine responsible for its specific aroma nuances and for a large fraction of its flavor. First, some basic concepts related to the global sensory perception and to the outstanding role played by olfaction will be summarized. Second, the limitations of the Odor Activity Value (OAV) concept to interpret the role played by aroma chemicals in the overall perception will also be addressed. Following, the two major concepts used to understand wine aroma, the wine buffer and the aroma vectors, are defined and their properties discussed. The 35 aroma vectors forming the core of the wine aroma perception are classified into 10 sensory categories and are further described, summarizing the present knowledge about the roles played in the sensory notes of different wines. Finally, the different perceptual integration processes which form the aroma vectors leading to the final sensory perception are discussed., Funding from the Spanish MINECO (projects AGL2017-59840 and RTC-2016-4935) and from European Union’s Horizon 2020 grant agreement No 643063. M.P.S.N. acknowledges the MINECO for her postdoctoral fellowship (Juan de la Cierva Incorporación 2015). LAAE acknowledges the continuous support of Diputación General de Aragón (T29) and European Social Fund. All the former LAAE members whose work has been essential to this chapter are gratefully acknowledged.

Summary: Huntington disease (HD) is a devastating neurodegenerative disorder characterized by aggregation of huntingtin (HTT) protein containing expanded polyglutamine (polyQ) tracts. DNAJB6, a member of the DNAJ chaperone family, was reported to efficiently inhibit polyQ aggregation in vitro, in cell models, and in vivo in flies, xenopus, and mice. For the delivery of exogenous DNAJB6 to the brain, the DNAJB6 needs to be protected against (enzymatic) degradation and show good penetration into brain tissue. Here, we tested the potential of small extracellular vesicles (sEVs) derived from neural stem cells (NSCs) for delivery of DNAJB6 as anti-amyloidogenic cargo. Administration of sEVs isolated from DNAJB6-overexpressing cells to cells expressing expanded polyQ tracts suppressed HTT aggregation. Furthermore, intrathecal injection of DNAJB6-enriched sEVs into R6/2 transgenic HD mice significantly reduced mutant HTT aggregation in the brain. Taken together, our data suggest that sEV-mediated molecular chaperone delivery may hold potential to delay disease onset in HD.

Background and purpose Research on demyelinating disorders aims to find novel molecules that are able to induce oligodendrocyte precursor cell differentiation to promote central nervous system remyelination and functional recovery. Delta(9)-Tetrahydrocannabinol (THC), the most prominent active constituent of the hemp plant Cannabis sativa, confers neuroprotection in animal models of demyelination. However, the possible effect of THC on myelin repair has never been studied. Experimental approach By using oligodendroglia-specific reporter mouse lines in combination with two models of toxin-induced demyelination, we analysed the effect of THC on the processes of oligodendrocyte regeneration and functional remyelination. Key results We show that THC administration enhanced oligodendrocyte regeneration, white matter remyelination and motor function recovery. THC also promoted axonal remyelination in organotypic cerebellar cultures. THC remyelinating action relied on the induction of oligodendrocyte precursor differentiation upon cell cycle exit and via CB1 cannabinoid receptor activation. Conclusions and implications Overall, our study identifies THC administration as a promising pharmacological strategy aimed to promote functional CNS remyelination in demyelinating disorders. Centro de Investigacion Biomedica en Red sobre Enfermedades Neurodegenerativas, Grant/Award Numbers: CB06/0005/0076, CB06/05/0005; Comunidad de Madrid, Grant/Award Numbers: 2016-T1/BMD-1060, 20205A/BMD-19728; Basque Government (Eusko Jaurlaritza), Grant/Award Numbers: IT1203-19, PIBA19-0059; Fondation pour l'Aide a la Recherche sur la Sclerose en Plaques, ARSEP Foundation; Fundacion Tatiana Perez de Guzman el Bueno; FEDER and Instituto de Salud Carlos III, Grant/Award Numbers: PI18-00941, PI18/00513; Ministerio de Economia y Competitividad, Grant/Award Numbers: PID2020-112640RB-I00, RTI2018095311-B-I00, SAF2016-75292-R, SAF2017-83516

The adoptive transfer of alloantigen-specific regulatory T cells (alloTregs) has been proposed as a therapeutic alternative in kidney transplant recipients to the use of lifelong immunosuppressive drugs that cause serious side effects. However, the clinical application of alloTregs has been limited due to their low frequency in peripheral blood and the scarce development of efficient protocols to ensure their purity, expansion, and stability. Here, we describe a new experimental protocol that allows the long-term expansion of highly purified allospecific natural Tregs (nTregs) from both healthy controls and chronic kidney disease (CKD) patients, which maintain their phenotype and suppressive function under inflammatory conditions. Firstly, we co-cultured CellTrace Violet (CTV)-labeled Tregs from CKD patients or healthy individuals with allogeneic monocyte-derived dendritic cells in the presence of interleukin 2 (IL-2) and retinoic acid. Then, proliferating CD4+CD25hiCTV− Tregs (allospecific) were sorted by fluorescence-activated cell sorting (FACS) and polyclonally expanded with anti-CD3/CD28-coated beads in the presence of transforming growth factor beta (TGF-β), IL-2, and rapamycin. After 4 weeks, alloTregs were expanded up to 2,300 times the initial numbers with a purity of >95% (CD4+CD25hiFOXP3+). The resulting allospecific Tregs showed high expressions of CTLA-4, LAG-3, and CD39, indicative of a highly suppressive phenotype. Accordingly, expanded alloTregs efficiently suppressed T-cell proliferation in an antigen-specific manner, even in the presence of inflammatory cytokines (IFN-γ, IL-4, IL-6, or TNF-α). Unexpectedly, the long-term expansion resulted in an increased methylation of the specific demethylated region of Foxp3. Interestingly, alloTregs from both normal individuals and CKD patients maintained their immunosuppressive phenotype and function after being expanded for two additional weeks under an inflammatory microenvironment. Finally, phenotypic and functional evaluation of cryopreserved alloTregs demonstrated the feasibility of long-term storage and supports the potential use of this cellular product for personalized Treg therapy in transplanted patients.

Regulatory T cells (Tregs) are indispensable for the control of immune homeostasis and have clinical potential as a cell therapy for treating autoimmunity. Tregs can lose expression of the lineage-defining Foxp3 transcription factor and acquire effector T cell (Teff) characteristics, a process referred to as Treg plasticity. The extent and reversibility of such plasticity during immune responses remain unknown. Here, using a murine genetic fate-mapping system, we show that Treg stability is maintained even during exposure to a complex microbial/antigenic environment. Furthermore, we demonstrate that the observed plasticity of Tregs after adoptive transfer into a lymphopenic environment is a property limited to only a subset of the Treg population, with the nonconverting majority of Tregs being resistant to plasticity upon secondary stability challenge. The unstable Treg fraction is a complex mixture of phenotypically distinct Tregs, enriched for naïve and neuropilin-1-negative Tregs, and includes peripherally induced Tregs and recent thymic emigrant Tregs These results suggest that a "purging" process can be used to purify stable Tregs that are capable of robust fate retention, with potential implications for improving cell transfer therapy. ispartof: SCIENCE IMMUNOLOGY vol:6 issue:61 ispartof: location:United States status: published

The presence of gastric cancer stem cells (GCSCs) marks the onset of gastric carcinoma. The sonic hedgehog (SHH) pathway plays a vital role in the maintenance of GCSC characteristics. Apatinib has been approved in China for advanced gastric cancer (GC) treatment. However, whether apatinib can target GCSCs and affect the SHH pathway remains unclear. The present study aimed to investigate the underlying mechanism of apatinib’s antitumor effects on GC. The expression levels of GCSC markers and number of CD133+ cells were significantly elevated in the sphere-forming cells. Apatinib effectively suppressed GCSC traits by inhibiting tumorsphere formation and cell proliferation, suppressing GCSC markers expression and CD133+ cell number, and inducing apoptosis. Apatinib downregulated the activation of the SHH pathway; while upregulation of the SHH pathway attenuated the inhibitory effects of apatinib on GCSCs. Moreover, apatinib treatment significantly delayed tumor growth and inhibited GCSC characteristics in the xenograft model. Our data suggested that apatinib exhibited inhibitory effects on GCSCs by suppressing SHH pathway both in vitro and in vivo, thus providing new insights into the therapeutic application of apatinib in GCSC suppression and advanced gastric cancer treatment.

This article is part of the Research Topic Ras and other GTPases in Cancer: From Basic to Applied Research https://doi.org/10.3389/978-2-88974-081-9 Recently, the highly mutated oncoprotein K-Ras4B (hereafter K-Ras) was shown to drive cancer cell stemness in conjunction with calmodulin (CaM). We previously showed that the covalent CaM inhibitor ophiobolin A (OphA) can potently inhibit K-Ras stemness activity. However, OphA, a fungus-derived natural product, exhibits an unspecific, broad toxicity across all phyla. Here we identified a less toxic, functional analog of OphA that can efficiently inactivate CaM by covalent inhibition. We analyzed a small series of benzazulenones, which bear some structural similarity to OphA and can be synthesized in only six steps. We identified the formyl aminobenzazulenone 1, here named Calmirasone1, as a novel and potent covalent CaM inhibitor. Calmirasone1 has a 4-fold increased affinity for CaM as compared to OphA and was active against K-Ras in cells within minutes, as compared to hours required by OphA. Calmirasone1 displayed a 2.5-4.5-fold higher selectivity for KRAS over BRAF mutant 3D spheroid growth than OphA, suggesting improved relative on-target activity. Importantly, Calmirasone1 has a 40-260-fold lower unspecific toxic effect on HRAS mutant cells, while it reaches almost 50% of the activity of novel K-RasG12C specific inhibitors in 3D spheroid assays. Our results suggest that Calmirasone1 can serve as a new tool compound to further investigate the cancer cell biology of the K-Ras and CaM associated stemness activities.

Although perceptual interactions are usually mentioned and blamed for the difficulties in understanding the relationship between odorant composition and aromatic sensory properties, they are poorly defined and categorised. Furthermore, old classifications refer mainly to effects on the odour intensity of the mixture of dissimilar non-blending odours and do not consider odour blending, which is one of the most relevant and influential perceptual interactions. Beginning with the results from classical studies about odour interaction, a new and simple systematic is proposed in which odour interactions are classified into four categories: competitive, cooperative, destructive and creative. The first categories are most frequent and display a mild level of interaction, being characterised mostly by analytical processing. The last two are less frequent and activate (or deactivate) configurational processes of object recognition with deep effects on the quality and intensity of the perception. These interactions can be systematically applied to interpret the formation of sensory descriptors from the odorant composition, suggesting that qualitatively the system works. However, there is a lack of quantitative data to work with odour intensities reliably, and a pressing need to systematise the effects of creative interactions., Spanish Ministry of Economy and Competitiveness (MINECO) (project AGL-2017-87373- C3-1-R). LAAE acknowledges the continuous support of Diputación General de Aragón (T29) and European Social Fund. MPSN acknowledges the Spanish National Research Agency, the Ministry of Science, Innovation, and Universities and the European Social Fund for her postdoctoral fellowship: Ramón y Cajal Program (RYC2019-027995-I/AEI/10.13039/501100011033).

Parthenium hysterophorus is an invasive weed species that competes aggressively with other plants and is also allelopathic. It poses a significant risk to human health, livestock, the environment, soil, and agriculture. However, given some clinical studies, its potential for antidiabetic, antioxidant, antitumor, herbicidal, pesticidal, and antimalarial therapies should be researched further in attempts to discover more relevant applications. It can be used as a nutrient-dense, readily available, and cheap fertilizer. Parthenium can also be used as an herbicide, an insecticide, and a phyto-remedial mediator to extract metals and dyes from agricultural waste. Here we provide basic information on the morphology, reproduction, environmental impacts, and management of this species. Effects of methanol, ethanol, hexane, acetone, and aqueous (water) Parthenium extracts are described. Because P. hysterophorus is said to be one of the world’s seven worst weeds, some control measures, including mechanical, chemical, cultural, and biological control, are discussed. The allelopathy of this weed is difficult to regulate, and there are both positive and negative interactions between Parthenium and other species due to allelochemical action. Several toxic phenolic compounds produced by P. hysterophorus are responsible for weed suppression, and we discuss details of their mode of action and potential applications.

This concise review summarizes results of the latest clinical trials that support the use of standardized cranberry juice intake as a complementary means of suppressing Helicobacter pylori infections. While not a replacement for antibiotic therapies, this tolerable and accessible dietary approach to help reduce inoculum levels in high‐risk populations could help curb infections and potentially reduce antibiotic use and subsequent stomach cancer prevalence.

QD LED has an ideal light source for growing crops and can also be used to control plant pathogenic microorganisms. The mycelial growth inhibition effect of QD LED light on Rhizoctonia solani, Phytophthora drechsleri, Sclerotinia sclerotiorum, Sclerotinia minor, Botrytis cinerea, Fusarium oxysporum, Pectobacterium carotovorum, and Xanthomonas campestris were investigated. According to the results, BLUE (450 nm) light, suppressed S. sclerotiorum by 16.7% at 50 cm height from the light source, and 94.1% mycelial growth at 30 cm height. Mycelial growth of Sclerotinia minor was inhibited by 80.4% at 50 cm height and 36.3% at 50 cm height in B. cinerea. S. minor, and B. cinerea was inhibited by 100% mycelial growth at a height of 30 cm from the light source. At 15 cm height, all three pathogens (B. cinerea, S. minor, and S. sclerotiorum) was inhibited by 100%. QD RED (M1) and QD RED (M2) light suppressed mycelial growth of S. minor and B. cinerea by 100% at 30 cm and 15 cm height from the light source. For S. sclerotiorum, QD RED (M1) and QD RED (M2) showed 75.2% and 100% inhibition, respectively. Further experiment was conducted to know the suppression effect of lights after inoculating the fungal pathogens on lettuce crop. According to the results, QD RED (M2) suppressed the S. sclerotiorum by 59.9%. In addition, Blue (450 nm), QD RED (M1), and QD RED (M2) light reduce the infestation by 59.9%. In case of B. cinerea, disease reduction was found 84% by BLUE (450 nm) light. Results : suggest that the growth inhibition of mycelium increases by Quantum dot LED light.

Microbiomes composition, diversity, and variability into a collection of suppressive composts were investigated for effective biological control of soil-borne phytopathogens. Pyrosequencing resulted be a reliable and faster method for characterizing fungal and bacterial microbiomes into composts derived from a varied feedstock of different composition, origin and provenience. Differences in taxonomic structure assessed by bioinformatics analyses were related to feedstock origin. Green composts derived from agro-waste and agroindustrial co/byproducts provided the most varied microbiomes either related to suppression of Rhizoctonia damping-off in bean and Verticillium wilt in eggplant, either to control of Phytium damping-off in cucumber and Phytophthora root rot in tomato. On the other hand, composted municipal solid wastes and co-composted cow manure with household waste prevalently given a most specific microbiota related to suppression of Fusarium wilt in melon.

BackgroundStreptococcal pyrogenic exotoxin (SPE)A expression is epidemiologically linked to streptococcal tonsillo-pharyngitis and outbreaks of scarlet fever, although the mechanisms by which superantigens confer advantageStreptococcus pyogenesare unclear.S. pyogenesis an exclusively human pathogen. As the leukocyte profile of tonsil differs from peripheral blood, the impact of SPEA production on human tonsil cell function was investigated.MethodsHuman tonsil cells from routine tonsillectomy, were co-incubated with purified streptococcal superantigens or isogenic streptococcal culture supernatants, differing only in superantigen content. Tonsil cell proliferation was quantified by tritium-incorporation, and cell surface characteristics assessed by flow-cytometry. Soluble mediators were measured using ELISA and quantitative (q)RT-PCR was performed for immunoglobulin gene expression.ResultsTonsil T cells proliferated in response to SPEA and demonstrated typical release of pro-inflammatory cytokines. When cultured in the absence of superantigen, tonsil preparations released large quantities of immunoglobulin over 7d. In contrast, marked B cell apoptosis and abrogation of total IgA, IgM, and IgG production occurred in the presence of SPEA and other superantigens. In SPEA-stimulated cultures, T follicular helper (TfH) cells showed a reduction in CXCR5 expression, but up-regulation of CD134 (OX40), CD278 (ICOS) and CD150 (SLAM) expression, indicative of a phenotypic change in the TfH population and associated with impaired chemotactic response to CXCL13.ConclusionsSPEA and other superantigens cause dysregulated tonsil immune function, driving T cells from TFH to a proliferating phenotype, with resultant loss of B cells and immunoglobulin production, providing superantigen-producing bacteria with a likely survival advantage.

Overexpression of the B7-H1 (PD-L1) molecule in the tumor microenvironment (TME) is a major immune evasion mechanism in some patients with cancer, and antibody blockade of the B7-H1/PD-1 interaction can normalize compromised immunity without excessive side-effects. Using a genome-scale T cell activity array, we identified Siglec-15 as a critical immune suppressor. While only expressed on some myeloid cells normally, Siglec-15 is broadly upregulated on human cancer cells and tumor-infiltrating myeloid cells, and its expression is mutually exclusive to B7-H1, partially due to its induction by macrophage colony-stimulating factor and downregulation by IFN-γ. We demonstrate that Siglec-15 suppresses antigen-specific T cell responses in vitro and in vivo. Genetic ablation or antibody blockade of Siglec-15 amplifies anti-tumor immunity in the TME and inhibits tumor growth in some mouse models. Taken together, our results support Siglec-15 as a potential target for normalization cancer immunotherapy.

Galangin (3,5,7‑trihydroxyflavone), a natural flavonoid present in plants, has been reported to possess anticancer properties in various types of cancers comprising glioma. The underlying mechanism, however, has not been fully elucidated. CD44, a hall marker in glioma, has been reported to be associated with epithelial-mesenchymal transition (EMT) and angiogenesis, which play important roles in glioma progression. In this study, we aimed to investigate whether galangin can inhibit EMT, angiogenesis and CD44 expression in glioma. We observed that galangin inhibited the proliferation, migration, invasion and angiogenesis of glioma cells in a dose-dependent manner, suppressed the expression of CD44 and inhibited angiogenesis of glioma cells through downregulating vascular endothelial growth factor (VEGF) in HUVECs. In addition, the overexpression of CD44 in U87 and U251 cells partly abolished the effects of galangin on glioma cells. Moreover, galangin suppressed tumor growth in an intracranial glioma mouse model. These results indicate that galangin is a potential novel drug for glioblastoma treatment due to its ability to suppress of CD44, EMT and angiogenesis.

Hsp70 chaperone systems are very versatile machines present in nearly all living organisms and in nearly all intracellular compartments. They function in many fundamental processes through their facilitation of protein (re)folding, trafficking, remodeling, disaggregation, and degradation. Hsp70 machines are regulated by co-chaperones. J-domain containing proteins (JDPs) are the largest family of Hsp70 co-chaperones and play a determining role functionally specifying and directing Hsp70 functions. Many features of JDPs are not understood; however, a number of JDP experts gathered at a recent CSSI-sponsored workshop in Gdansk (Poland) to discuss various aspects of J-domain protein function, evolution, and structure. In this report, we present the main findings and the consensus reached to help direct future developments in the field of Hsp70 research. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12192-018-0948-4) contains supplementary material, which is available to authorized users.

Background MicroRNAs (miRNAs), important regulators of gene expression, have been implicated in a variety of disorders. The expression pattern of miRNAs in paediatric atopic dermatitis (AD) has not been well studied. Objectives We sought to investigate miRNA expression profiles in different blood compartments of infants with AD. Methods Small RNA and analysis with the HTG EdgeSeq system were performed to identify differentially expressed miRNAs in peripheral blood mononuclear cells (PBMCs) and plasma of infants with AD vs. age-matched healthy controls, with reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) used for validation and measurement of miRNA targets. Logistic regression models with area under the receiving operating characteristic estimation was used to evaluate the diagnostic potential of chosen miRNAs for AD. Results RNA sequencing was performed to access miRNA expression profiles in paediatric AD. We identified 10 differentially expressed miRNAs in PBMCs and eight dysregulated miRNAs in plasma of infants with AD compared with controls. Upregulated miRNAs in PBMCs included miRNAs known to be involved in inflammation: miR-223-3p, miR-126-5p and miR-143-3p. Differential expression of only one miRNA, miR-451a, was observed in both PBMCs and plasma of children with AD. Dysregulation of three miRNAs (miR-451a, miR-143-3p and miR-223-3p) was validated in larger numbers of samples and miR-451a was identified as a predictive biomarker for the early diagnosis of the disease. Experimentally verified targets of miR-451a, interleukin 6 receptor (IL6R) and proteasome subunit beta type-8 (PSMB8), were increased in patients with AD, negatively correlated with miR-451a levels and upregulated following inhibition of miR-451a in PBMCs. Conclusions In infants with AD, a distinct peripheral blood miRNA signature is seen, highlighting the systemic effects of the disease. miR-451a is uniquely expressed in different blood compartments of patients with AD and may serve as a promising novel biomarker for the early diagnosis of AD.

Neutrophils that reside in the bone marrow are switly recruited from circulating blood to fight infections. For a long time, these first line defenders were considered as microbe killers. However their role is far more complex as cross talk with T cells or dendritic cells have been described for human or mouse neutrophils. In cattle, these new roles are not documented yet. We identified a new subset of regulatory neutrophils that is present in the mouse bone marrow or circulate in cattle blood under steady state conditions. These regulatory neutrophils that display MHC-II on the surface are morphologically indistinguishable from classical MHC-IInegneutrophils. However MHC-IIposand MHC-IInegneutrophils display distinct transcriptomic profiles. While MHC-IInegand MHC-IIposneutrophils display similar bacterial phagocytosis or killing activity, MHC-IIposonly are able to suppress T cell proliferation under contact-dependent mechanisms. Regulatory neutrophils are highly enriched in lymphoid organs as compared to their MHC-IInegcounterparts and in the mouse they express PDL-1, an immune checkpoint involved in T-cell blockade. Our results emphasize neutrophils as true partners of the adaptive immune response, including in domestic species. They open the way for discovery of new biomarkers and therapeutic interventions to better control cattle diseases.

Contains fulltext : 219551.pdf (Publisher’s version ) (Open Access) Despite an elevated recovery need, research indicates that athletes often exhibit relatively poor sleep. Timing and consolidation of sleep is driven by the circadian system, which requires periodic light-dark exposure for stable entrainment to the 24-hour day, but is often disturbed due to underexposure to light in the morning (e.g., low-level indoor lighting) and overexposure to light in the evening (e.g., environmental and screen-light). This study examined whether combining fixed sleep schedules with light regulation leads to more consolidated sleep. Morning light exposure was increased using light-emitting goggles, whereas evening light exposure was reduced using amber-lens glasses. Using a within-subject crossover design, twenty-six athletes (14 female, 12 male) were randomly assigned to start the intervention with the light-regulation-week or the no light-regulation-week. Sleep was monitored by means of sleep diaries and actigraphy. Due to low protocol adherence regarding the fixed sleep-wake schedules, two datasets were constructed; one including athletes who kept a strict sleep-wake schedule (N = 8), and one that also included athletes with a more lenient sleep-wake schedule (N = 25). In case of a lenient sleep-wake schedule, light regulation improved self-reported sleep onset latency (delta SOL = 8 minutes). This effect was stronger (delta SOL = 17 minutes) and complemented by enhanced subjective sleep quality in case of a strict sleep-wake schedule. None of the actigraphy-based estimates differed significantly between conditions. To conclude, light regulation may be considered a potentially effective strategy to improve subjective sleep, but less obtrusive methods should be explored to increase protocol compliance. 9 p.

Biological control of sugarcane smut (Ustilago scitaminea Syd.) using Trichoderma isolates is one of the untapped potentials in Ethiopian sugarcane plantations. To this end, this study was carried out with the objective to identify potential native Trichoderma isolates for the management of sugarcane smut. A total of 120 soil samples were collected from Kesem, Metehara, and Wonji sugarcane plantation fields. Consequently, 21 Trichoderma isolates were recovered from the samples, and in-vitro antagonist study of Trichoderma isolates against U. scitaminea was done using dual culture technique on fresh PDA. Among the native isolates, 12 of them revealed a pathogen suppression potential of >90% at 7 days after inoculation and most of them had higher pathogen suppression than the commercial isolate. However, five of the isolates were selected as potential candidates based on distinct suppression features, growth rate, and antagonistic performance. Identification of the five effective Trichoderma isolates (WI4-27, MKI2-k17, MI3-m33, WI12-25, and MKI3) was done based on macroscopic and microscopic characteristics. Thus, the isolates were identified as T. harzianum, T. asperellum, T. viride, T. viride, and T. harzianum, respectively. Accordingly, these isolates could be used as one component of integrated smut management practices once a study on compatibility with fungicides, in-vivo efficacy test against smut, and substrate selection for mass production of the isolates were commenced.

The pink mealybug Maconellicoccus hirsutus Green is a pest of economic importance in various crops around the world. The objective was to determine the biological effectiveness of organic and inorganic products for the management of M. hirsutus in the soursop crop. The experiment was carried out in the communities of “Lima de Abajo” and “El Capomo”, in the municipality of Compostela, Nayarit, Mexico, during the period from September to November 2018 and from July to August 2019. The treatments were Ara3, Inter-a-ch-c, Evolution, a complex of plant extracts (Capsicum chinense + Allium sativum + Allium cepa + Artemisia absinthium), Mos Blanc® (Imidacloprid), farmers’ control (Detergent Blanca Nieves® + Sodium Hypochlorite Cloralex®) and the absolute control (Water). The nymphs and adult females of M. hirsutus were counted after each application, in young shoots. The results showed that in “Lima de Abajo”, Ara 3 registered the highest percentage of suppression after 28 days. In “El Capomo”, Imidacloprid recorded the greatest suppression after 28 days of application. In both communities, the treatments Ara 3, Evolution and Imidacloprid registered the least number of living individuals. The results indicated that Ara 3 and Evolution were the most promising alternatives in the management against M. hirsutus in the crop.

Mirolysin is a secretory protease of Tannerella forsythia, a member of the dysbiotic oral microbiota responsible for periodontitis. In this study, we show that mirolysin latency is achieved by a “cysteine-switch” mechanism exerted by Cys23 in the N-terminal profragment. Mutation of Cys23 shortened the time needed for activation of the zymogen from several days to 5 min. The mutation also decreased the thermal stability and autoproteolysis resistance of promirolysin. Mature mirolysin is a thermophilic enzyme and shows optimal activity at 65 °C. Through NMR-based fragment screening, we identified a small molecule (compound (cpd) 9) that blocks promirolysin maturation and functions as a competitive inhibitor (Ki = 3.2 µM), binding to the S1′ subsite of the substrate-binding pocket. Cpd 9 shows superior specificity and does not interact with other T. forsythia proteases or Lys/Arg-specific proteases.

Natural killer (NK) cells are cytotoxic innate lymphocytes that protect against viral infection and tumor metastasis. Despite their inherent ability to kill a broad range of virally infected, stressed and transformed cells, low numbers of dysfunctional NK cells are often observed in many advanced solid human cancers. Here, we review the potential mechanisms that influence suboptimal mature NK cell recruitment and function in the tumor microenvironment (TME) of solid tumors. We further highlight current immunotherapy approaches aimed to circumvent NK cell dysfunction and discuss next‐generation strategies to enhance adoptive NK cell therapy through targeting intrinsic and extrinsic checkpoints the regulate NK cell functionality in the TME. Understanding the mechanisms that drive NK cell dysfunction in the TME will lead to novel immunotherapeutic approaches in the fight against cancer., In this review, we provide a summary of the current literature profiling intrinsic and extrinsic mechanisms of NK cells that promote dysfunctional responses in solid tumor microenvironments. We critically analyse the current literature and propose open questions as well as new directions for NK‐cell‐based immunotherapies in cancer.

Cytotoxic CD4+ T cells (CD4 CTL) are terminally differentiated T helper cells that contribute to autoimmune diseases, such as multiple sclerosis. We developed a novel triple co-culture transwell assay to study mutual interactions between CD4 CTL, conventional TH cells, and regulatory T cells (Tregs) simultaneously. We show that, while CD4 CTL are resistant to suppression by Tregs in vitro, the conditioned medium of CD4 CTL accentuates the suppressive phenotype of Tregs by upregulating IL-10, Granzyme B, CTLA-4, and PD-1. We demonstrate that CD4 CTL conditioned medium skews memory TH cells to a TH17 phenotype, suggesting that the CD4 CTL induce bystander polarization. In our triple co-culture assay, the CD4 CTL secretome promotes the proliferation of TH cells, even in the presence of Tregs. However, when cell−cell contact is established between CD4 CTL and TH cells, the proliferation of TH cells is no longer increased and Treg-mediated suppression is restored. Taken together, our results suggest that when TH cells acquire cytotoxic properties, these Treg-resistant CD4 CTL affect the proliferation and phenotype of conventional TH cells in their vicinity. By creating such a pro-inflammatory microenvironment, CD4 CTL may favor their own persistence and expansion, and that of other potentially pathogenic TH cells, thereby contributing to pathogenic responses in autoimmune disorders.

Indoleamine 2,3-dioxygenase 1 (IDO1) and tryptophan 2,3-dioxygenase (TDO) catalyze the rate-limiting step of tryptophan catabolism along the kynurenine pathway, which has important immuno suppressive properties, particularly in tumor cells and dendritic cells. The prominent expression of IDO1 in the placenta also suggested a role in preventing immune rejection of fetal tissues, and pharmacological inhibition of IDO1 induced abortion of allogeneic fetuses in mice. However, this was later challenged by the lack of rejection of allogeneic fetuses in IDO1-KO mice, suggesting that other mechanisms may compensate for IDO1 deficiency. Here we investigated whether TDO could contribute to feto-maternal tolerance and compensate for IDO1 deficiency in IDO1-KO mice. Expression of TDO mRNA was previously detected in placental tissues. We developed a new chimeric rabbit anti-TDO antibody to confirm TDO expression at the protein level and identify the positive cell type by immunohistochemistry in murine placenta. We observed massive TDO expression in decidual stromal cells, starting at day E3.5, peaking at day E6.5 then declining rapidly while remaining detectable until gestation end. IDO1 was also induced in decidual stromal cells, but only at a later stage of gestation when TDO expression declined. To determine whether TDO contributed to feto-maternal tolerance, we mated TDO-KO and double IDO1-TDO-KO females with allogeneic males. However, we did not observe reduced fertility. These results suggest that, despite its expression in decidual stromal cells, TDO is not a dominant mechanism of feto-maternal tolerance able to compensate for the absence of IDO1. Redundant additional mechanisms of immunosuppression likely take over in these KO mice. The massive expression of TDO during decidualization might suggest a role of TDO in angiogenesis or vessel tonicity, as previously described for IDO1. ispartof: FRONTIERS IN IMMUNOLOGY vol:11 ispartof: location:Switzerland status: published

Many studies have demonstrated that cancer stem cells (CSCs) or tumor-initiating cells (TICs) are responsible for tumor cell proliferation, chemotherapy resistance, metastasis, and relapse in various cancers. We, and others, have previously shown that the signal transducer and activator of transcription 3 (STAT3) signaling pathway is responsible for CSCs and TICs growth. Recent reports have indicated that the heat shock protein 90 (Hsp90) is also essential for the survival of CSCs and TICs. SNX-2112 is an Hsp90 inhibitor. However, it remains unclear whether proliferation of esophageal cancer stem-like cells (ECSLCs) is suppressed by SNX-2112 with knockdown of STAT3 (shSTAT3). Here, we explored the association between SNX-2112 with shSTAT3 and the suppression of ECSLCs growth. We found that the expression level of both STAT3 and p-STAT3 was higher in clinical esophageal cancer tissue than in the adjacent normal tissue, using western blot and qPCR analysis. Furthermore, differential expression analysis demonstrated that STAT3 was overexpressed in clinical specimens. We demonstrated that SNX-2112 inhibited cancer cell proliferation, decreased ABCB1 and ABCG2 gene expression levels and reduced the colony formation capacity of ECSLCs, which was enhanced by STAT3 silencing. Flow cytometry analysis revealed that the combination of SNX-2112 and shSTAT3 significantly induced apoptosis and cell cycle arrest at G2/M phase in ECSLCs. Levels of proliferation pathway proteins, including p38, c-Jun N-terminal kinase (JNK), and extracellular signal–regulated kinase (ERK) which were also client proteins of Hsp90, were also reduced. In addition, SNX-2112 with shSTAT3 inhibited the proliferation of ECSLCs in vivo. Finally, STAT3 overexpression eliminated the apoptotic and antiproliferative effects of SNX-2112 on ECSLCs. Hence, these results provide a rationale for the therapeutic potential of the combination of SNX-2112 with shSTAT3 in esophageal cancer, and may indicate new targets for clinical intervention in human cancer.

Global warming, pollution problems, and the demand for sustainable food production forced farmers and scientists to search for new solutions in biological plant protection. The usage of natural renewable sources around us seems to be an alternative. Compounds isolated from plants and distinguished with antifungal properties can be used to protect vegetables from the seed-borne pathogens. The study aimed to elucidate the ability of Juniperus communis L., Hyssopus officinalis L. and Thymus vulgaris L. essential oils to control the carrot seed-borne pathogen Alternaria spp. The agar-plate method was used for carrot seed infestation with micromycetes. Essential oils extracted from common juniper, hyssop and thyme were then separately mixed with potato dextrose agar media at different concentrations and the antifungal activity of each oil tested in vitro. The results revealed that the T. vulgaris essential oil (200–1000 μL L−1) significantly inhibited Alternaria spp. growth. The H. officinalis essential oil promoted seed-borne pathogens growth on the second and fifth days of the evaluation compared to the control, however, the concentration of 400 μL L−1 showed little suppression of micromycete development 7 days after inoculation. The in vitro experiments indicated that 600 μL L−1 of J. communis essential oil could control seed-borne pathogen viability. Overall, thyme essential oil expressed a high potential for application in biofungicide formulations.

We reviewed the research into the mechanisms of growth of triple negative breast cancer (TNBC) based on laboratory pre-clinical studies that have shaped understanding of the disease over the past decade. In response to these findings, we propose an approach to potentially prevent cancer metabolic adaptation and recurrence. This paper collates pre-clinical results, first to determine the tumor’s mechanisms of growth and then to source natural substances that could potentially suppress those mechanisms. The results from in vivo and in vitro studies of TNBC were combined first to select 10 primary mechanisms (Hypoxia-inducible factor 1α, Hedgehog, MAPK, MTAP, NF-κ B, Notch, P13K, STAT3, and Wnt signaling pathways plus p53 and POL2A gene expression) that promote TNBC growth, and second to propose a treatment array of 21 natural compounds that suppress laboratory models of TNBC via these mechanisms. We included BRCA mutations in the review process, but only pathways with the most preclinical studies utilizing natural products were included. Then we outlined potential biomarkers to assess the changes in the micro-environment and monitor biochemical pathway suppression. This suppression-centric aim targets these mechanisms of growth with the goal of potentially halting tumor growth and preventing cancer cell metabolic adaptation. We chose TNBC to demonstrate this 5-step strategy of supplementary therapy, which may be replicated for other tumor types.

Bacteriophages (phages) are critical players in the dynamics and function of microbial communities and drive processes as diverse as global biogeochemical cycles and human health. Phages tend to be predators finely tuned to attack specific hosts, even down to the strain level, which in turn defend themselves using an array of mechanisms. However, to date, efforts to rapidly and comprehensively identify bacterial host factors important in phage infection and resistance have yet to be fully realized. Here, we globally map the host genetic determinants involved in resistance to 14 phylogenetically diverse double-stranded DNA phages using two model Escherichia coli strains (K-12 and BL21) with known sequence divergence to demonstrate strain-specific differences. Using genome-wide loss-of-function and gain-of-function genetic technologies, we are able to confirm previously described phage receptors as well as uncover a number of previously unknown host factors that confer resistance to one or more of these phages. We uncover differences in resistance factors that strongly align with the susceptibility of K-12 and BL21 to specific phage. We also identify both phage-specific mechanisms, such as the unexpected role of cyclic-di-GMP in host sensitivity to phage N4, and more generic defenses, such as the overproduction of colanic acid capsular polysaccharide that defends against a wide array of phages. Our results indicate that host responses to phages can occur via diverse cellular mechanisms. Our systematic and high-throughput genetic workflow to characterize phage-host interaction determinants can be extended to diverse bacteria to generate datasets that allow predictive models of how phage-mediated selection will shape bacterial phenotype and evolution. The results of this study and future efforts to map the phage resistance landscape will lead to new insights into the coevolution of hosts and their phage, which can ultimately be used to design better phage therapeutic treatments and tools for precision microbiome engineering., To map the landscape of genetic determinants important in host-phage interactions, this study applies unbiased high-throughput loss-of-function and gain-of-function screening methods to two different strains of E. coli. These methods rapidly identify phage receptors and other host factors (both novel and previously described) involved in resistance across a wide panel of 13 dsDNA phages.

This project investigated how kairomone lures, camera traps, and counting software could together contribute to pest management. Images of cumulative daily catch of New Zealand Flower Thrips (NZFT) attracted to a ripe peach lactone (6-pentyl-2H-pyran-2-one, 6-PAP) were automatically loaded to the internet and compared with scanned bases checked weekly using in-house software and manual counting. Camera traps were able to provide thrips counts equivalent to delta traps, but daily and remotely. An 11-fold greater NZFT count occurred within 24 h in passive traps after polyethylene sachets loaded with 250 mg of 6-PAP were placed in trees. Intensive trapping, by placing 1, 2, 4, and 8 traps per tree (500 mg/trap), resulted in a maximum 32-fold increase in thrips per tree. While 6-PAP has proved to be a useful tool for monitoring NZFT numbers, our results suggest that it is not likely to be suitable for mass trapping. Future research should investigate NZFT behavior to better understand population movement on an area-wide basis. Camera traps can be a valuable tool for recording insect flight activity remotely, but the number of traps required for statistically reliable estimates may be prohibitive.

Background Vagus nerve (VN) stimulation is currently evaluated as a novel approach to treat immune‐mediated disorders. The optimal stimulation parameters, however, largely depend on the VN composition potentially impacting on its clinical translation. Hence, we evaluated whether morphological differences exist between the cervical and abdominal VNs across different species. Materials and methods The cervical and abdominal VNs of mouse, pig, and humans were stained for major basic protein and neurofilament F to identify the percentage and size of myelinated and non‐myelinated fibers. Results The percentage of myelinated fibers was comparable between species, but was higher in the cervical VN compared with the abdominal VN. The cervical VN contained 54 ± 4%, 47 ± 7%, and 54 ± 7% myelinated fibers in mouse, pig, and humans, respectively. The myelinated fibers consisted of small‐diameter (mouse: 71%, pig: 80%, and humans: 63%), medium‐diameter (mouse: 21%, pig: 18%, and humans: 33%), and large‐diameter fibers (mouse: 7%, pig: 2%, and humans: 4%). The abdominal VN predominantly contained unmyelinated fibers (mouse: 93%, pig: 90%, and humans: 94%). The myelinated fibers mainly consisted of small‐diameter fibers (mouse: 99%, pig: 85%, and humans: 74%) and fewer medium‐diameter (mouse: 1%, pig: 13%, and humans: 23%) and large‐diameter fibers (mouse: 0%, pig: 2%, and humans: 3%). Conclusion The VN composition was largely similar with respect to myelinated and unmyelinated fibers in the species studied. Human and porcine VNs had a comparable diameter and similar amounts of fibrous tissue and contained multiple fascicles, implying that the porcine VN may be suitable to optimize stimulation parameters for clinical trials., The cervical and abdominal vagus nerve have a similar fiber composition across the studied species. However, the cervical vagus nerve generally possess a higher level of myelination compared to its abdominal counterpart, suggesting that higher stimulation parameters are required to activate the abdominal vagus nerve.

Background: Plants synthesize compounds of specialized metabolism to defend themselves against biotic and abiotic stresses. These compounds could be used as models for pesticide development. Among the species that have not yet been studied for the potential to produce active compounds is the kidney leaf mud plantain (Heteranthera reniformis). Objective: The goal was to evaluate the effect of hexanic, ethyl acetate and methanolic extracts of H. reniformis leaves on cress (Lepidium sativum) seed germination to determine which one have the highest biological activity; and to evaluate different concentrations of extract with the highest biological activity on lettuce (Lactuca sativa), barnyard grass (Echinochloasp.) and giant arrowhead (Sagittaria montevidensis) germination. Methods: The seeds were placed on petri dishes containing two sheets of germitest paper, and after the application of the extract, they were sealed and placed in growth chambers with controlled temperature and photoperiod. The variables analyzed were 1st and 2nd germination count, germination speed index and length of the aerial part and roots. Results: The ethyl acetate extract provided greater phytotoxicity on cress than the other extracts. In general, the highest concentration of ethyl acetate extract was the most efficient in reducing variables for all species. Conclusions: The ethyl acetate extract of H. reniformis presents inhibitory activity on the seeds of cress, lettuce, giant arrowhead and barnyard grass, but this activity was dependent on the concentration of the extract and the species studied. H. reniformis synthesizes compounds with phytotoxic activity and purification of extracts is required to isolate, identify and characterize the action mechanism of the compounds with herbicide activity, so that in the future these can be used as models for the development of herbicides.

Heterodera avenae (cereal cyst nematode, CCN) infects wheat and other cereal crops and causes severe losses in their yield. Research has shown that CCN infestations can be mitigated by organic fertilization in wheat fields, but the mechanisms underlying this phenomenon are still largely unknown. In this study, the relationships among CCN, soil properties, and soil fungal communities with organic fertilizer (OF) or chemical fertilizer (CF) and without fertilizer (CK), were investigated for two years in a wheat field in Henan province, China. Our results showed that the concentrations of soil total N, total P, available P, available K, and organic matter were all promoted by the OF treatment at the jointing stage of wheat, coinciding with the peak in egg hatching and penetration of wheat root by CCNs. Soil total N correlated positively (R2 = 0.759, p < 0.05) with wheat yields but negatively (R2 = 0.663, p < 0.01) with Pf/Pi (index of cyst nematode reproduction), implying the regulated soil property contributes to suppressing CCN in the OF treatment. Furthermore, fungal community α-diversity (Shannon and Simpson) and β-diversity (PCoA) of rhizosphere soil was improved under the organic fertilizer treatment. The fungal genera negatively associated with the Pf/Pi of CCN were highly enriched, which included Mortierella and Chaetomium, two taxa already reported as being nematophagous fungi in many other studies. These two genera were heavily surrounded by much more related fungal genera in the constructs co-occurrence network. These results suggested that the OF treatment shifted soil fungal community functioning towards the suppression of CCN. Taken together, the suppressed cyst nematode reproduction with the assembly of fungal communities in the rhizosphere led to greater wheat yields under organic fertilization. These findings provide an in-depth understanding of the benefits provided by organic fertilization for developing sustainable agriculture.

Cluster of differentiation 226 (CD226) molecules play a crucial role in the activation of effector CD4+ T cells during the immune response process, but a cell-intrinsic function of CD226 in CD4+ T subsets is not clear. In this study, we showed that Cd226 -/- mice were resistant to myelin oligodendrocyte glycoprotein peptide 35-55 (MOG35-55)-induced experimental autoimmune encephalomyelitis (EAE) with highly expressed IL-10+CD4+ T cells and downregulated IL-17A+CD4+ T cells when compared with wild-type (WT) mice. Th17 cell infiltration into the central nervous system (CNS) was largely decreased in the absence of CD226 during EAE. CD226 deficiency facilitated the proliferation of regulatory T cells (Tregs), with increased numbers of Tregs observed in EAE mice, and supported the elevated induced regulatory T cell (iTregs) proliferation in vitro. The Akt and Erk signaling pathways were shown to be involved in Cd226 -/- Treg proliferation and function in vivo and in vitro. These findings collectively indicate that CD226 is a key molecule regulating the Treg-mediated suppression of autoimmune responses by inhibiting Treg proliferation. Thus, the results of this study identify additional mechanisms by which CD226 regulates Treg functions in EAE and supports the potential therapeutic effects of anti-CD226 molecules on autoimmune diseases.

Summary ESC- and iPSC-derived retinal transplantation is a promising therapeutic approach for disease with end-stage retinal degeneration, such as retinitis pigmentosa and age-related macular degeneration. We previously showed medium- to long-term survival, maturation, and light response of transplanted human ESC- and iPSC-retina in mouse, rat, and monkey models of end-stage retinal degeneration. Because the use of patient hiPSC-derived retina with a disease-causing gene mutation is not appropriate for therapeutic use, allogeneic transplantation using retinal tissue/cells differentiated from a stocked hESC and iPSC line would be most practical. Here, we characterize the immunological properties of hESC- and iPSC-retina and present their three major advantages: (1) hESC- and iPSC-retina expressed low levels of human leukocyte antigen (HLA) class I and little HLA class II in vitro, (2) hESC- and iPSC-retina greatly suppressed immune activation of lymphocytes in co-culture, and (3) hESC- and iPSC-retina suppressed activated immune cells partially via transforming growth factor β signaling. These results support the use of allogeneic hESC- and iPSC-retina in future clinical application., Graphical abstract, Highlights • hESC- and iPSC-retinas poorly express HLA molecules • hESC- and iPSC-retinas have immunosuppressive properties • hESC- and iPSC-retinas can suppress allogeneic lymphocytes • hESC- and iPSC-retinas suppress immune cells via TGF-β signaling, In this article, Yamasaki, Sugita, and colleagues show that the immunogenicity and immunosuppressive properties of hESC- and iPSC-retinas was differentiated in vitro outside the immune-privileged site. TGF-β secreted by these retinas played a critical role of immunosuppressive properties against activated lymphocytes. These features suggested a low risk of immune rejection after transplantation of stem cell-retina in clinical applications.

When plants detect herbivores they strengthen their defenses. As a consequence, some herbivores evolved the means to suppress these defenses. Research on induction and suppression of plant defenses usually makes use of particular life stages of herbivores. Yet many herbivorous arthropods go through development cycles in which their successive stages have different characteristics and lifestyles. Here we investigated the interaction between tomato defenses and different herbivore developmental stages using two herbivorous spider mites, i.e., Tetranychus urticae of which the adult females induce defenses and T. evansi of which the adult females suppress defenses in Solanum lycopersicum (tomato). First, we monitored egg-to-adult developmental time on tomato wild type (WT) and the mutant defenseless-1 (def-1, unable to produce jasmonate-(JA)-defenses). Then we assessed expression of salivary effector genes (effector 28, 84, SHOT2b, and SHOT3b) in the consecutive spider mite life stages as well as adult males and females. Finally, we assessed the extent to which tomato plants upregulate JA- and salicylate-(SA)-defenses in response to the consecutive mite developmental stages and to the two sexes. The consecutive juvenile mite stages did not induce JA defenses and, accordingly, egg-to-adult development on WT and def-1 did not differ for either mite species. Their eggs however appeared to suppress the SA-response. In contrast, all the consecutive feeding stages upregulated SA-defenses with the strongest induction by T. urticae larvae. Expression of effector genes was higher in the later developmental stages. Comparing expression in adult males and females revealed a striking pattern: while expression of effector 84 and SHOT3b was higher in T. urticae females than in males, this was the opposite for T. evansi. We also observed T. urticae females to upregulate tomato defenses, while T. evansi females did not. In addition, of both species also the males did not upregulate defenses. Hence, we argue that mite ontogenetic niche shifts and stage-specific composition of salivary secreted proteins probably together determine the course and efficiency of induced tomato defenses.

Gene-environment interaction (GxE) determines the vulnerability of an individual to a spectrum of stress-related neuropsychiatric disorders. Increased impulsivity, excessive aggression, and other behavioural characteristics are associated with variants within the tryptophan hydroxylase-2 (Tph2) gene, a key enzyme in brain serotonin synthesis. This phenotype is recapitulated in naïve mice with complete, but not with partial Tph2 inactivation. Tph2 haploinsufficiency in animals reflects allelic variation of Tph2 facilitating the elucidation of respective GxE mechanisms. Recently, we showed excessive aggression and altered serotonin brain metabolism in heterozygous Tph2-deficient male mice (Tph2+/−) after predator stress exposure. Here, we sought to extend these studies by investigating aggressive and anxiety-like behaviours, sociability, and the brain metabolism of dopamine and noradrenaline. Separately, Tph2+/− mice were examined for exploration activity in a novel environment and for the potentiation of helplessness in the modified swim test (ModFST). Predation stress procedure increased measures of aggression, dominancy, and suppressed sociability in Tph2+/− mice, which was the opposite of that observed in control mice. Anxiety-like behaviour was unaltered in the mutants and elevated in controls. Tph2+/− mice exposed to environmental novelty or to the ModFST exhibited increased novelty exploration and no increase in floating behaviour compared to controls, which is suggestive of resilience to stress and despair. High-performance liquid chromatography (HPLC) revealed significant genotype-dependent differences in the metabolism of dopamine, and norepinephrine within the brain tissue. In conclusion, environmentally challenged Tph2+/− mice exhibit behaviours that resemble the behaviour of non-stressed null mutants, which reveals how GxE interaction studies can unmask latent genetically determined predispositions. © 2020 The Authors. The authors' work reported here was supported by Deutsche Forschungsgemeinschaft (DFG:CRC TRR58A1/A5), DAAD (to ES), the European Union's Seventh Framework Programme (FP7/2007–2013) under Grant No.602805 (Aggressotype) and the Horizon 2020 Research and Innovation Programme under Grant No.728018 (Eat2beNICE) (to KPL and TS) and the President's program of PhD Exchange of RF-2017 (to TS and DA). We appreciate the valuable technical help of Natalia Bazhenova, Drs. Alexander Trofimov and Natalia Markova with this project.

Background Eukaryotic initiation factor 2B (eIF2B) initiates and regulates translation initiation in eukaryotes. eIF2B gene mutations cause leukoencephalopathy called vanishing white matter disease (VWM) in humans and slow growth (Slg−) and general control derepression (Gcd−) phenotypes in Saccharomyces cerevisiae. Results To suppress eIF2B mutations, S. cerevisiae genomic DNA library was constructed in high-copy vector (YEp24) and transformed into eIF2B mutant S. cerevisiae strains. The library was screened for wild-type genes rescuing S. cerevisiae (Slg−) and (Gcd−) phenotypes. A genomic clone, Suppressor-I (Sup-I), rescued S. cerevisiae Slg− and Gcd− phenotypes (gcd7-201 gcn2∆). The YEp24/Sup-I construct contained truncated TAN1, full length EMC4, full length YGL230C, and truncated SAP4 genes. Full length EMC4 (chaperone protein) gene was sub-cloned into pEG (KG) yeast expression vector and overexpressed in gcd7-201 gcn2∆ strain which suppressed the Slg− and Gcd− phenotype. A GST-Emc4 fusion protein of 47 kDa was detected by western blotting using α-GST antibodies. Suppression was specific to gcd7-201 gcn2∆ mutation in eIF2Bβ and Gcd1-502 gcn2∆ in eIF2Bγ subunit. Emc4p overexpression also protected the wild type and mutant (gcd7-201 gcn2∆, GCD7 gcn2∆, and GCD7 GCN2∆) strains from H2O2, ethanol, and caffeine stress. Conclusions Our results suggest that Emc4p is involved in eIF2B-mediated translational regulation under stress and could provide an amenable tool to understand the eIF2B-mediated defects.

Cortical inhibition plays an important role in information processing in the brain. However, the mechanisms by which inhibition and excitation are coordinated to generate functions in the six layers of the cortex remain unclear. Here, we measured laminar-specific responses to stimulus orientations in primary visual cortex (V1) of awake monkeys (male, Macaca mulatta). We distinguished inhibitory effects (suppression) from excitation, by taking advantage of the separability of excitation and inhibition in the orientation and time domains. We found two distinct types of suppression governing different layers. Fast suppression (FS) was strongest in input layers (4C and 6), and slow suppression (SS) was 3 times stronger in output layers (2/3 and 5). Interestingly, the two types of suppression were correlated with different functional properties measured with drifting gratings. FS was primarily correlated with orientation selectivity in input layers (r = −0.65, p < 10−9), whereas SS was primarily correlated with surround suppression in output layers (r = 0.61, p < 10−4). The earliest SS in layer 1 indicates the origin of cortical feedback for SS, in contrast to the feedforward/recurrent origin of FS. Our results reveal two V1 laminar subnetworks with different response suppression that may provide a general framework for laminar processing in other sensory cortices. SIGNIFICANCE STATEMENT This study sought to understand inhibitory effects (suppression) and their relationships with functional properties in the six different layers of the cortex. We found that the diversity of neural responses across layers in primary visual cortex (V1) could be fully explained by one excitatory and two suppressive components (fast and slow suppression). The distinct laminar distributions, origins, and functional roles of the two types of suppression provided a simplified representation of the differences between two V1 subnetworks (input network and output network). These results not only help to elucidate computational principles in macaque V1, but also provide a framework for general computation of cortical laminae in other sensory cortices.

The human ability to imagine motor actions without executing them (i.e., motor imagery) is crucial to a number of cognitive functions, including motor planning and learning, and has been shown to improve response times and accuracy of subsequent motor actions [1, 2]. Although these behavioral findings suggest the possibility that imagined movements directly influence primary motor cortex (M1), how this might occur remains unknown [3]. Here, we use a non-blood-oxygen-level-dependent (BOLD) method for collecting fMRI data, called vascular space occupancy (VASO) [4, 5], to measure neural activations across cortical laminae in M1 while participants either tapped their thumb and forefinger together or simply imagined doing so. We report that, whereas executed movements (i.e., finger tapping) evoked neural responses in both the superficial layers of M1 that receive cortical input and the deep layers of M1 that send output to the spinal cord to support movement, imagined movements evoked responses in superficial cortical layers only. Furthermore, we found that finger tapping preceded by both imagined and executed movements showed a reduced response in the super-ficial layers (repetition suppression) coupled with a heightened response in the deep layers (repetition enhancement). Taken together, our results provide evidence for a mechanism whereby imagined movements can directly affect motor performance and might explain how neural repetition effects lead to improvements in behavior (e.g., repetition priming).

&alpha, Gal syndrome (AGS) is a type of anaphylactic reaction to mammalian meat characterized by an immunoglobulin (Ig)E immune response to the oligosaccharide &alpha, Gal (Gal&alpha, 1-3Gal&beta, 1-4GlcNAc-R). Tick bites seems to be a prerequisite for the onset of the allergic disease in humans, but the implication of non-tick parasites in &alpha, Gal sensitization has also been deliberated. In the present study, we therefore evaluated the capacity of helminths (Toxocara canis, Ascaris suum, Schistosoma mansoni), protozoa (Toxoplasma gondii), and parasitic fungi (Aspergillus fumigatus) to induce an immune response to &alpha, Gal. For this, different developmental stages of the infectious agents were tested for the presence of &alpha, Gal. Next, the potential correlation between immune responses to &alpha, Gal and the parasite infections was investigated by testing sera collected from patients with AGS and those infected with the parasites. Our results showed that S. mansoni and A. fumigatus produce the terminal &alpha, Gal moieties, but they were not able to induce the production of specific antibodies. By contrast, T. canis, A. suum and T. gondii lack the &alpha, Gal epitope. Furthermore, the patients with T. canis infection had significantly decreased anti-&alpha, Gal IgE levels when compared to the healthy controls, suggesting the potential role of this nematode parasite in suppressing the allergic response to the glycan molecule. This rather intriguing observation is discussed in the context of the &lsquo, hygiene hypothesis&rsquo, Taken together, our study provides new insights into the relationships between immune responses to &alpha, Gal and parasitic infections. However, further investigations should be undertaken to identify T. canis components with potent immunomodulatory properties and to assess their potential to be used in immunotherapy and control of AGS.

Lateral posterior nucleus (LP) of thalamus, the rodent homologue of primate pulvinar, projects extensively to sensory cortices. However, its functional role in sensory cortical processing remains largely unclear. Here, bidirectional activity modulations of LP or its projection to the primary auditory cortex (A1) in awake mice reveal that LP improves auditory processing in A1 supragranular-layer neurons by sharpening their receptive fields and frequency tuning, as well as increasing the signal-to-noise ratio (SNR). This is achieved through a subtractive-suppression mechanism, mediated largely by LP-to-A1 axons preferentially innervating specific inhibitory neurons in layer 1 and superficial layers. LP is strongly activated by specific sensory signals relayed from the superior colliculus (SC), contributing to the maintenance and enhancement of A1 processing in the presence of auditory background noise and threatening visual looming stimuli respectively. Thus, a multisensory bottom-up SC-pulvinar-A1 pathway plays a role in contextual and cross-modality modulation of auditory cortical processing.

Insect antennae are important olfactory organs that house high concentrations of odorant-binding proteins (OBPs) in the sensillum lymph. Previous studies in other insects have shown that OBPs play important roles in transporting odorants and enhancing the sensitivity of the olfactory system. However, the functions of OBPs in the oriental fruit fly, Bactrocera dorsalis, especially those specifically expressed in antennae, have not been fully elucidated. In this study, cDNA libraries were constructed from both the male and female antennal transcriptome, and twenty OBPs were identified in total. The expression profiles of these OBPs were examined in the adult antenna, head, thorax, leg, and abdomen of both sexes. Seven of the identified OBP genes had significantly higher expression in both the male and female antennae than in other tissues, while the transcript levels of the remaining OBPs varied across different tissues. Regarding the function of antenna-specific OBPs, we targeted Bdorsobp2 as a representative for further RNA interference (RNAi) and identified via electrophysiology a decrease in detection of a potential species-specific a potent attractant, methyl eugenol. Moreover, subsequent behavioral assay data showed that the behavioral response of B. dorsalis toward this odorant decreased when Bdorobp2 was silenced with injection of double-stranded RNA (dsRNA). Combined, these results support our initial hypothesis that antennae-specific OBPs are of critical importance for insect odorant detection, sensitivity, and behavior.