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Several factors that affect production of cocoa (Theobroma cacao L.), one of which is the quality of seedlings. The quality of seedlings is affected by the type of clones and the composition of the planting media. This study aims to obtain the influence of clone types and types of manure, as well as the interaction between clone types and types of manure on the growth of cocoa seedlings. The study was conducted in the field of practices and Laboratory of the State Polytechnic of Lampung, in January to June 2017. The method used was a Randomized Block Design (RBD) factorial pattern consisting of two factors. The first factor is the type of clone consisting of three levels, namely K1 = TSH clone 858, K2 = ICS 60, and K3 = TSH 908. The second factor consists of four levels namely P0 = Control, P1 = Chicken Manure, P2 = Cow Manure , and P3 = Goat Manure. The results showed that the use of TSH 858, ICS 60, and TSH 908 clones had the same effect on all observed variables. However, the use of manure has a significant effect on each variable, but the amount of leaf chlorophyll is relatively the same between clone types and types of manure, there are interactions between clone types and types of manure.

Epichloë festucae forms a mutualistic symbiotic association with Lolium perenne. This biotrophic fungus systemically colonizes the intercellular spaces of aerial tissues to form an endophytic hyphal network and also grows as an epiphyte. However, little is known about the cell wall–remodeling mechanisms required to avoid host defense and maintain intercalary growth within the host. Here, we use a suite of molecular probes to show that the E. festucae cell wall is remodeled by conversion of chitin to chitosan during infection of L. perenne seedlings, as the hyphae switch from free-living to endophytic growth. When hyphae transition from endophytic to epiphytic growth, the cell wall is remodeled from predominantly chitosan to chitin. This conversion from chitin to chitosan is catalyzed by chitin deacetylase. The genome of E. festucae encodes three putative chitin deacetylases, two of which (cdaA and cdaB) are expressed in planta. Deletion of either of these genes results in disruption of fungal intercalary growth in the intercellular spaces of plants infected with these mutants. These results establish that these two genes are required for maintenance of the mutualistic symbiotic interaction between E. festucae and L. perenne. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

The authors wish to thank Chelsie Smith, Chelsie Money and staff from the Thad Cochran Marine Aquaculture Cen-ter for assistance with spawning and larval culture trials and husbandry of stocks, and C.S. Lee and Jason Hide forgreatly appreciated contributions to early spawning trials. The Mississippi Department of Marine Resources(Tidelands Trust Funds Program award #S-09-USM/GCRL-M201-13-01), NOAA and the Gulf States Marine Fisher-ies Commission (award #NA15NMF4720399, sub-award #ACQ-210-039-2020-USM2) and Perciformes Group LLCare acknowledged for funding support.

Objective Periodontitis is one the most common chronic inflammatory conditions, resulting in destruction of tooth-supporting tissues and leading to tooth loss. Porphyromonas gingivalis activates host macrophages to secrete pro-inflammatory cytokines and elicit tissue damage, in part by inducing NF-kappa-B transactivation. Since NFκB transactivation is negatively regulated by the Nicotinamide adenine dinucleotide (NAD)-dependent deacetylase enzyme Sirt1, we sought to assess if RAW264.7 macrophages exposed to P. gingivalis demonstrate impaired Sirt1 activity, to ultimately induce a pro-inflammatory response. Methods RAW264.7 macrophages were incubated with heat- killed P. gingivalis for 2, 4, 8, and 24 h. Stimulated RAW264.7 were assessed for TNFα expression via PCR, ELISA, and ChIP analysis. Following the activation of RAW264.7 macrophages, immunoblot analysis was executed to detect modifications in Sirt1 and the NFκB subunit RelA that is essential for NFκB transcriptional activity. Results TNFα expression was elevated 4 h after exposure to P. gingivalis. ChIP confirmed that RelA was enriched in the mouse TNFα promoter 4 h following stimulation, which correlated with the increased TNFα mRNA levels. Preceding TNFα expression, we detected Phosphoserine 536 and acetylated lysine 310 of RelA after 2 hours exposure with P. gingivalis. Moreover, reduced Sirt1 activity was associated with its cleavage in RAW264.7 protein extracts, after 2 hours of P. gingivalis exposure. Blocking TLR2/4 signaling prevented Sirt1 cleavage, loss of deacetylase activity, and TNFα secretion, while co-administering CA074Me (a cathepsin B inhibitor) with P. gingivalis reduced RelA promoter enrichment, resulting in impaired TNFα expression. Conclusions Together, the results suggest that P. gingivalis induces TNFα expression, at least in part, by enhancing cleavage of Sirt1 via a TLR-dependent signaling circuit.

1. Aphids are capable of adapting to parasitoid pressure in the absence of sexual reproduction using epigenetically controlled polyphenism. Asexual lineages of aphids could thus become resistant to parasitoids over time and with repeated exposure, hampering biocontrol efforts that rely on parasitoid wasps. 2. Prior to this study, wing polyphenism and reproductive polyphenism had been reported as parasitoid‐adaptive responses in asexual lineages of at least two species of aphids, but it remained unclear whether parasitoid exposure could induce other defensive polyphenisms. Using a rigorous, replicated experimental evolution design, we aimed to induce adaptive polyphenisms in the green peach aphid, Myzus persicae Sulzer, through serial exposures of single asexual lines to the parasitoid Aphidius colemani Viereck over four consecutive generations. 3. We measured changes in parasitoid susceptibility, reproductive schedule, body size, and production of pink or alate offspring compared to aphid control lines not exposed to parasitoids. Despite this consistently strong selective parasitoid pressure, there was no evidence that asexual lineages of M. persicae adapt to parasitoid pressure using adaptive polyphenism. 4. Our data indicate that pseudo‐crowding stress – where aphids disturbed by parasitoids or predators wander within a colony, increasing aphid‐to‐aphid contact and creating the illusion of a more crowded environment – may be necessary for M. persicae to produce defensive polyphenisms in response to parasitoid attack.

Unrefined non-centrifugal sugars in Southeast Asia emanate from various plant sources including sugarcane, palm, and coconut. To assess food and flavor characteristic differences of these unrefined sugars, seven cane, three palm, and two coconut sugars were evaluated in term of their physicochemical, antioxidant, and volatile flavor properties. Results indicated that unrefined sugars from cane had significantly higher ICUMSA color value than palm and coconut sugars, yet they possessed comparable total soluble solid levels. Cane sugars also presented superior phenolic and flavonoid contents and antioxidant activities, particularly in Thai and Japanese products. Each sugar had distinct volatile flavor composition, comprising acids, Maillard reaction products (furanone, pyrazine, furan, and pyrrole), aldehydes, alcohols, and sulfurs. All traditionally made unrefined sugars had significantly greater Maillard reaction products and less acetic acid than in industrially manufactured cane sugars. Then, propanoic acid and [R-(R*,R*)]-2,3-butanediol were solely major volatiles in palm sugars from Thailand. Additionally, mass spectrometry-based electronic-nose (e-nose) profiling detection provided specific ion masses from various volatile components as potential chemical markers in monitoring aroma characteristic of unrefined sugars and differentiated these sugars into nine statistical clusters based on their materials and origins, wherein palm sugars from Thailand and Indonesia, as well as cane sugars from Thailand and Japan, had comparable volatile profiles.

Somatic mutations in driver genes may ultimately lead to the development of cancer. Understanding how somatic mutations accumulate in cancer genomes and the underlying factors that generate somatic mutations is therefore crucial for developing novel therapeutic strategies. To understand the interplay between spatial genome organization and specific mutational processes, we studied 3,000 tumor-normal-pair whole-genome datasets from 42 different human cancer types. Our analyses reveal that the change in somatic mutational load in cancer genomes is co-localized with topologically-associating-domain boundaries. Domain boundaries constitute a better proxy to track mutational load change than replication timing measurements. We show that different mutational processes lead to distinct somatic mutation distributions where certain processes generate mutations in active domains, and others generate mutations in inactive domains. Overall, the interplay between three-dimensional genome organization and active mutational processes has a substantial influence on the large-scale mutation-rate variations observed in human cancers.

Satureja punctata Benth. Briq. (Lamiaceae) is a small shrub known by its vernacular name “Lomishet”. It is used in Ethiopian traditional medicine for the treatment of hypertension, diabetes, liver disease, headache and febrile illness. Mean Arterial Blood Pressure (MABP) lowering effect and vasorelaxant activity of the aqueous extract of aerial parts of the plant were studied. Intravenous administration of the aqueous extract at different doses brought a fall in MABP in normotensive guinea pigs and also in renovascular hypertensive guinea pigs. The extract also caused a dose-dependent relaxation of precontracted aorta; with a maximum relaxation of 98.19% achieved at 40 mg/ml. Preparative reversed-phase HPLC analyses of the extract resulted in the isolation of two phenolic compounds, namely, rosmarinic acid and linarin. The structures of these compounds were elucidated using UV, IR, MS, and NMR (1D and 2D) techniques. Furthermore, rosmarinic acid showed significant (p < 0.01) reduction of MABP in normotensive guinea pigs at doses of 0.75, 1.50 and 3.00 mg/kg. The findings of this study suggest that the aerial parts of S. punctata have genuine antihypertensive activity.

The field of fish gonadotropin-releasing hormones (GnRHs) is also celebrating its 50th anniversary this year. This review provides a chronological history of fish GnRH biology over the past five decades. It demonstrates how discoveries in fish regarding GnRH and GnRH receptor multiplicity, dynamic interactions between GnRH neurons, and additional neuroendocrine factors acting alongside GnRH, amongst others, have driven a paradigm shift in our understanding of GnRH systems and functions in vertebrates, including mammals. The role of technological innovations in enabling scientific discoveries is portrayed, as well as how fundamental research in fish GnRH led to translational outcomes in aquaculture. The interchange between fish and mammalian GnRH research is discussed, as is the value and utility of using fish models for advancing GnRH biology. Current challenges and future perspectives are presented, with the hope of expanding the dialogue and collaborations within the neuroendocrinology scientific community at large, capitalizing on diversifying model animals and the use of comparative strategies.

SummaryThe conserved hemagglutinin stalk domain is an attractive target for broadly effective antibody-based therapeutics and next generation universal influenza vaccines. Protection provided by hemagglutinin stalk binding antibodies is principally mediated through activation of immune effector cells. Titers of stalk-binding antibodies are highly variable on an individual level, and tend to increase with age as a result of increasing exposures to influenza virus. In our study, we show that stalk-binding antibodies cooperate with neuraminidase inhibitors to protect against influenza virus infection in an Fc-dependent manner. These data suggest that the effectiveness of neuraminidase inhibitors is likely influenced by an individual’s titers of stalk-binding antibodies, and that neuraminidase inhibitors may enhance the effectiveness of future stalk-binding monoclonal antibody-based treatments.

What are the key topological features of connectivity critically relevant for generating the dynamics underlying efficient cortical function? A candidate feature that has recently emerged is that the connectivity of the mammalian cortex follows an exponential distance rule, which includes a small proportion of long-range high-weight anatomical exceptions to this rule. Whole-brain modeling of large-scale human neuroimaging data in 1,003 participants offers the unique opportunity to create two models, with and without long-range exceptions, and explicitly study their functional consequences. We found that rare long-range exceptions are crucial for significantly improving information processing. Furthermore, modeling in a simplified ring architecture shows that this improvement is greatly enhanced by the turbulent regime found in empirical neuroimaging data. Overall, the results provide strong empirical evidence for the immense functional benefits of long-range exceptions combined with turbulence for information processing. G.D. is supported by a Spanish national research project (reference PID2019-105772GB-I00 MCIU AEI), funded by the Spanish Ministry of Science, Innovation and Universities (MCIU), State Research Agency (AEI); HBP SGA3 Human Brain Project Specific grant agreement 3 (grant agreement 945539), funded by the EU H2020 FET Flagship program; SGR Research Support Group (reference 2017 SGR 1545), funded by the Catalan Agency for Management of University and Research Grants (AGAUR); Neurotwin Digital twins for model-driven non-invasive electrical brain stimulation (grant agreement 101017716), funded by the EU H2020 FET Proactive program; euSNN European School of Network Neuroscience (grant agreement 860563), funded by the EU H2020 MSCA-ITN Innovative Training Networks; The Emerging Human Brain Cluster (CECH) (001-P-001682) within the framework of the European Research Development Fund Operational Program of Catalonia 2014–2020; Brain-Connects: Brain Connectivity during Stroke Recovery and Rehabilitation (201725.33), funded by the Fundacio La Marato TV3; and Corticity, FLAG-ERA JTC 2017 (reference PCI2018-092891), funded by the MCIU, AEI. M.L.K. is supported by the Center for Music in the Brain, funded by the Danish National Research Foundation (DNRF117), and the Centre for Eudaimonia and Human Flourishing, funded by the Pettit Foundation and Carlsberg Foundation.

Little is known about the genetic architecture of antifungal immunity in natural populations. Using two population genetic approaches, quantitative trait locus (QTL) mapping and evolve and resequence (E&R), we explored D. melanogaster immune defense against infection with the fungus Beauveria bassiana. The immune defense was highly variable both in the recombinant inbred lines from the Drosophila Synthetic Population Resource used for our QTL mapping and in the synthetic outbred populations used in our E&R study. Survivorship of infection improved dramatically over just 10 generations in the E&R study, and continued to increase for an additional nine generations, revealing a trade-off with uninfected longevity. Populations selected for increased defense against B. bassiana evolved cross resistance to a second, distinct B. bassiana strain but not to bacterial pathogens. The QTL mapping study revealed that sexual dimorphism in defense depends on host genotype, and the E&R study indicated that sexual dimorphism also depends on the specific pathogen to which the host is exposed. Both the QTL mapping and E&R experiments generated lists of potentially causal candidate genes, although these lists were nonoverlapping.

Digital, Background Guidelines recommend nonstatin lipid-lowering agents in patients at very high risk for major adverse cardiovascular events (MACE) if low-density lipoprotein cholesterol (LDL-C) remains ≥70 mg/dL on maximum tolerated statin treatment. It is uncertain if this approach benefits patients with LDL-C near 70 mg/dL. Lipoprotein(a) levels may influence residual risk. Objectives In a post hoc analysis of the ODYSSEY Outcomes (Evaluation of Cardiovascular Outcomes After an Acute Coronary Syndrome During Treatment With Alirocumab) trial, the authors evaluated the benefit of adding the proprotein subtilisin/kexin type 9 inhibitor alirocumab to optimized statin treatment in patients with LDL-C levels near 70 mg/dL. Effects were evaluated according to concurrent lipoprotein(a) levels. Methods ODYSSEY Outcomes compared alirocumab with placebo in 18,924 patients with recent acute coronary syndromes receiving optimized statin treatment. In 4,351 patients (23.0%), screening or randomization LDL-C was 13.7 mg/dL or ≤13.7 mg/dL; corresponding adjusted treatment hazard ratios were 0.82 (95% CI: 0.72-0.92) and 0.89 (95% CI: 0.75-1.06), with Pinteraction = 0.43. Conclusions In patients with recent acute coronary syndromes and LDL-C near 70 mg/dL on optimized statin therapy, proprotein subtilisin/kexin type 9 inhibition provides incremental clinical benefit only when lipoprotein(a) concentration is at least mildly elevated. (ODYSSEY Outcomes: Evaluation of Cardiovascular Outcomes After an Acute Coronary Syndrome During Treatment With Alirocumab; NCT01663402), Ciencias Médicas y de la Salud

Herein we report the high mortality of larval Hurter's spadefoot toad (Scaphiopus hurterii) tadpoles when fed dry powdered feeds. The evidence implies that dry powders remain suspended in the water column where they enter and clog the filtering apparatus of the tadpoles. When the same foods were wetted to a paste and dropped into the aquaria as a moist pellet, the ration fell directly to the substrate and did not become suspended in the water column. Mortalities of tadpoles fed dry powdered rations were severe, whereas mortalities when fed wetted feed were inconsequential. We recommend that that rations made of ground or powdered components not be fed unless first wetted. Further, we speculate that sediment and siltation of natural waters may contribute to amphibian losses via the same mechanism.

Paramount to guide the diagnostic strategy, the prevalence of pulmonary embolism in patients with syncope and no other symptoms is uncertain. We aimed to assess the pulmonary embolism prevalence in patients that present to the emergency department (ED) with syncope and no chest pain nor dyspnea.Multicenter prospective cohort study in seven EDs in France. Adult patients who presented to the ED with syncope (transient loss of consciousness) were included. Patients with chest pain or dyspnea were excluded. Included patients underwent formal work-up for pulmonary embolism, including D-dimer testing and further imaging if positive. Cases of pulmonary embolism were adjudicated by two independent expert radiologists with the review of imaging studies. Secondary objectives included the pulmonary embolism prevalence in the subgroup of patients with cancer, in patients with negative Pulmonary Embolism Rule Out criteria (PERC) score, and according to the clinical probability assessed either by Wells or revised Geneva score.We analyzed 411 patients, in whom 128 (31%) underwent a computed tomographic pulmonary angiogram and 9 (2%) a ventilation-perfusion scan. A pulmonary embolism was confirmed in nine patients (prevalence of 2.2%, 95% confidence interval, 1.1-4.3%), including one subsegmental pulmonary embolism. This prevalence was of 18% (3-52%) in patients with cancer and 0.7% (0.1-4%) in PERC negative patients.In our sample of patients with isolated syncope, the prevalence of pulmonary embolism was 2.2%. This value is not sufficiently low to negate the requirement for a formal work up in the ED, even in the absence of chest pain or dyspnea. However, the large confidence interval precludes any strong conclusion.

Over the past decade, a new strategy was developed to bypass the difficulties to genetically engineer some microbial species by transferring (or "cloning") their genome into another organism that is amenable to efficient genetic modifications and therefore acts as a living workbench. As such, the yeast Saccharomyces cerevisiae has been used to clone and engineer genomes from viruses, bacteria, and algae. The cloning step requires the insertion of yeast genetic elements in the genome of interest, in order to drive its replication and maintenance as an artificial chromosome in the host cell. Current methods used to introduce these genetic elements are still unsatisfactory, due either to their random nature (transposon) or the requirement for unique restriction sites at specific positions (TAR cloning). Here we describe the CReasPy-cloning, a new method that combines both the ability of Cas9 to cleave DNA at a user-specified locus and the yeast's highly efficient homologous recombination to simultaneously clone and engineer a bacterial chromosome in yeast. Using the 0.816 Mbp genome of Mycoplasma pneumoniae as a proof of concept, we demonstrate that our method can be used to introduce the yeast genetic element at any location in the bacterial chromosome while simultaneously deleting various genes or group of genes. We also show that CReasPy-cloning can be used to edit up to three independent genomic loci at the same time with an efficiency high enough to warrant the screening of a small (

Many studies on Gnrh1, and the teleost Gnrh3, have elucidated the roles of these peptides in reproductive regulation. However, the role of the midbrain population of Gnrh, Gnrh2, has long been a mystery, despite its ubiquitous conservation in all jawed vertebrates except rodents. Previous behavioral studies in sparrows, musk shrews, mice, zebrafish, and goldfish show that Gnrh2 administrations both increase spawning behaviors and decrease feeding behaviors, suggesting a role of this peptide in metabolism regulation along with the canonical role in regulating reproduction. In order to more deeply explore the roles of Gnrh2, we used a cyprinid teleost, zebrafish, which has 2 forms of Gnrh, Gnrh2 and Gnrh3, to generate a knockout zebrafish line which contains a frameshift mutation and subsequent disruption of the coding for the functional Gnrh2 peptide. We examined differences in reproduction, feeding, growth, and mobility in this line, and discovered major differences in feeding and growth parameters, suggesting that Gnrh2 is a potent anorexigen in zebrafish. Additionally, there were no differences in mobility except for increased distances swam during feeding periods. There were no major differences in reproductive success, however, female gnrh2−/− zebrafish exhibited smaller oocytes and increased embryo mortality, indicating slightly decreased oocyte quality. Additionally, there were changes in the expression levels of many feeding, growth, and reproductive neuropeptides in gnrh2−/− zebrafish. Taken together, these findings suggest a role for Gnrh2 in controlling satiation in zebrafish along with a minor role in maintaining optimal oocyte quality in females.

Studies on the regulation of fungal secondary metabolism highlight the importance of histone H3K4 methylation regulators Set1, CclA (Ash2) and KdmB (KDM5), but it remains unclear whether these proteins act by direct modulation of H3K4me3 at the target genes. In filamentous fungi, secondary metabolite genes are frequently located near telomeres, a site where H3K4 methylation is thought to have a repressive role. Here we analyzed the role of CclA, KdmB and H3K4me3 in regulating the subtelomeric EAS and LTM cluster genes in Epichloë festucae. Depletion of H3K4me3 correlated with transcriptional activation of these genes in ΔcclA, similarly enrichment of H3K4me3 correlated with transcriptional repression of the genes in ΔkdmB which was accompanied by significant reduction in the levels of the agriculturally undesirable lolitrems. These transcriptional changes could only be explained by the alterations in H3K4me3 and not in the subtelomerically-important marks H3K9me3/K27me3. However, H3K4me3 changes in both mutants were not confined to these regions but occurred genome-wide, and at other subtelomeric loci there were inconsistent correlations between H3K4me3 enrichment and gene repression. Our study suggests that CclA and KdmB are crucial regulators of secondary metabolite genes, but these proteins likely act via means independent to, or in conjunction with the H3K4me3 mark.

BACKGROUND: Drug resistance to malaria is still a problem in various regions, and there have even been developments in resistance to the ACTs (artemisinin-based combination therapies) as standard antimalarial drugs included to artemisinin’s partner drugs. Ethyl acetate fraction of G. mangostana L rind, containing xanthones as an antioxidant, has antimalarial activity in vitro which has a synergistic effect with artemisinin. That’s why the activities of this fraction are needed to be studied in vivo. AIM: To explore the antimalarial and antioxidant activity of ethyl acetate fraction of Garcinia mangostana L rind in mice. METHODS: This was a complete randomised design true experimental study. Six groups of mice: a healthy mice group and 5 groups of Plasmodium berghei inoculated mice treated with various doses of the sample for 3 days compared to artemisinin. Parasitemia and total antioxidant status were examined and analysed using ANOVA, and probit analysis were done.RESULTS: The parasitemia level in all of the treatment groups were lower than the positive control group without treatment (p < 0.01) and the parasitemia level was the lowest in artemisinin group which was not significantly different from the 100 mg/kg body weight dose group (p > 0.05). The parasitemia level in 20 and 4 mg/kg body weight dose groups were higher than the artemisinin group (p < 0.01). Parasite growth inhibition rate from the highest to the lowest consecutively was: artemisinin, 100 mg/kg body weight, 20 mg/kg body weight, 4 mg/kg body weight, and positive control group (p < 0.05) and ED50 was 3.396 mg/kg body weight. Total antioxidant status was the highest in 20 mg/ kg body weight dose and higher than the negative control group (p < 0.05) while the lowest total antioxidant status was in the positive control group. CONCLUSION: Ethyl acetate fraction of G. mangostana L rind potentially showed antimalarial and antioxidant activity in vivo. Further study is needed to explore the detail of its mechanism of action and its quantitative phytochemical analysis to find the leading compound in it.

This study was conducted to investigate changes in P-fractions, bio-available P (CAL-P), citric acid extractable P, acid phosphatase activity and microbial biomass C and N during incubation...

Human oral mucosa stem cells (hOMSCs) arise from the neural crest, they can self-renew, proliferate, and differentiate to several cell lines and could represent a good source for application in tissue engineering. Because of their anatomical location, hOMSCs are easy to isolate, have multilineage differentiation capacity and express embryonic stem cells markers such as—Sox2, Oct3/4 and Nanog. We have used SHEM (supplemented hormonal epithelial medium) media and cultured hOMSCs over human amniotic membrane and determined the cell’s capacity to differentiate to an epithelial-like phenotype and to express corneal specific epithelial markers—CK3, CK12, CK19, Pan-cadherin and E-cadherin. Our results showed that hOMSCs possess the capacity to attach to the amniotic membrane and express CK3, CK19, Pan-Cadherin and E-Cadherin without induction with SHEM media and expressed CK12 or changed the expression pattern of E-Cadherin to a punctual-like feature when treated with SHEM media. The results observed in this study show that hOMSCs possess the potential to differentiate toward epithelial cells. In conclusion, our results revealed that hOMSCs readily express markers for corneal determination and could provide the ophthalmology field with a therapeutic alternative for tissue engineering to achieve corneal replacement when compared with other techniques. Nevertheless, further studies are needed to develop a predictable therapeutic alternative for cornea replacement.

The Alpine goat Capra aegagrus hircus is parasitized by the barber pole worm (Haemonchus contortus). This relationship results in changes that affect the gene expression of the host, the pest, and the microbiome of both. Hematological parameters indicating genes that are expressed and/or the % Composition of abundant and diverse microbial flora are reflective of infestation. We explored the similarity/dissimilarity between and among blood samples of non-infected, infected, infected zoledronic acid-treated, and infected antibody (anti-γσ T cells) treated wethers under controlled conditions. We identified responses to barber pole worms using blood-based analysis of transcripts and the microbiome. Seven (7) days post-inoculation (dpi) we identified 7,627 genes associated with different treatment types. Across all treatments we identified fewer raw read counts and a reduced diversity in microbial flora on 7 dpi than in 21 dpi wethers. We also identified that there were differences in % Composition of microbial flora known to be associated with inflammation. This study identifies treatment specific genes, and an increase in microflora abundance and diversity as wethers age post infestation. Further, Firmicutes/Bacteriodetes (F/B) ratio reflect metabolic health, based on depression or elevation above thresholds defined by the baseline of non-infected hosts depending on the type of intrusion exhibited by the pest.

Many of the molecular mechanisms for antifungal immunity inDrosophila melanogasterhave been defined, but relatively little is known about the genetic basis for variation in antifungal immunity in natural populations. Using two population genetic approaches, Quantitative Trait Locus (QTL) Mapping and Evolve and Resequence (E&R), we explored the genetics underlyingD. melanogasterimmune defense against infection with the fungusBeauveria bassiana. Immune defense was highly variable both in the recombinant inbred lines from the Drosophila Synthetic Population Resource used for our QTL Mapping and in the synthetic outbred populations used in our E&R study. Survivorship of infection improved dramatically over just 10 generations in the E&R study, and continued to increase for an additional 9 generations, revealing a trade-off with uninfected longevity. Populations selected for increased defense againstB. bassianaevolved cross resistance to a second, distinctB. bassianastrain but not to bacterial pathogens. The QTL mapping study revealed that sexual dimorphism in defense depends on host genotype, and the E&R study indicated that dimorphism also depends on the specific pathogen to which the host is exposed. Both the QTL Mapping and E&R experiments generated lists of potentially causal candidate genes, although these lists were non-overlapping.

Restricted food intake, either from lack of food sources or endogenous fasting, during reproductive periods is a widespread phenomenon across the animal kingdom. Considering previous studies show the canonical upstream regulator of reproduction in vertebrates, the hypothalamic Gonadotropin-releasing hormone (Gnrh), is inhibited in some fasting animals, we sought to understand the neuroendocrine control of reproduction in fasted states. Here, we explore the roles of the midbrain neuropeptide, Gnrh2, in inducing reproduction via its pituitary prevalence, gonadotropin synthesis, gametogenesis, and reproductive outputs in the zebrafish model undergoing different feeding regimes. We discovered a fasting-induced four-fold increase in length and abundance of Gnrh2 neuronal projections to the pituitary and in close proximity to gonadotropes, whereas the hypothalamic Gnrh3 neurons are reduced by six-fold in length. Subsequently, we analyzed the functional roles of Gnrh2 by comparing reproductive parameters of a Gnrh2-depleted model, gnrh2−/−, to wild-type zebrafish undergoing different feeding conditions. We found that Gnrh2 depletion in fasted states compromises spawning success, with associated decreases in gonadotropin production, oogenesis, fecundity, and male courting behavior. Gnrh2 neurons do not compensate in other circumstances by which Gnrh3 is depleted, such as in gnrh3−/− zebrafish, implying that Gnrh2 acts to induce reproduction specifically in fasted zebrafish.

RSUD dr. Soehadi Prijonegoro Sragen produces liquid waste containing microorganisms, toxic and radioactive chemicals. The results of the laboratory examination showed that the liquid waste level always exceeded the quality standard, namely the parameter of phosphate. The phosphate level in September 2013 was 4.80 mg / L, while in December 2013 the concentration was 2.60 mg / L, so additional treatment is needed, one of which is phytoremediation using white daffodils (Crinum asiaticum Linn.). This research method is an experimental research design with pre-post Test with Control Group Design where the wastewater will pass through a wet tank containing white lily plants with a residence time of 24 hours/day and a treatment rate of 1100 liters/day. Th research aimed to determine the effect of phytoremediation of white daffodils on the reduction of phosphate levels and the average percentage of its reduction. The results of this study in the ninth and tenth sample collection met the quality standards of 1.99 mg / L and 1.89 mg / L. Phytoremediation of white daffodils (Crinum asiaticum linn.) may reduce phosphate levels by 45.59%.

Background There is widespread interkingdom signalling between insects and microbes. For example, microbes found in floral nectar may modify its nutritional composition and produce odorants that alter the floral odor bouquet which may attract insect pollinators. Mosquitoes consume nectar and can pollinate flowers. We identified microbes isolated from nectar of common tansy, Tanacetum vulgare, elucidated the microbial odorants, and tested their ability to attract the common house mosquito, Culex pipiens. Results We collected 19 microbial isolates from T. vulgare nectar, representing at least 12 different taxa which we identified with 16S or 26S rDNA sequencing as well as by biochemical and physiological tests. Three microorganisms (Lachancea thermotolerans, Micrococcus lactis, Micrococcus luteus) were grown on culture medium and tested in bioassays. Only the yeast L. thermotolerans grown on nectar, malt extract agar, or in synthetic nectar broth significantly attracted Cx. pipiens females. The odorant profile produced by L. thermotolerans varied with the nutritional composition of the culture medium. All three microbes grown separately, but presented concurrently, attracted fewer Cx. pipiens females than L. thermotolerans by itself. Conclusions Floral nectar of T. vulgare contains various microbes whose odorants contribute to the odor profile of inflorescences. In addition, L. thermotolerans produced odorants that attract Cx. pipiens females. As the odor profile of L. thermotolerans varied with the composition of the culture medium, we hypothesize that microbe odorants inform nectar-foraging mosquitoes about the availability of certain macro-nutrients which, in turn, affect foraging decisions by mosquitoes.

Background and Aim: In the male reproductive system, the aging process can lead to infertility. Recently, placenta and its derivatives have been researched as regenerative agents. This study aimed to describe the basic components of dried bovine placenta powder and its potential effects as a regenerative agent in a rat model of male reproductive aging with D-galactose induction. Materials and Methods: We divided 15 male Wistar rats, 2 months of age, into three groups: A, the health control group; B, the D-galactose induction group, and C, the D-galactose induction and 10% dried bovine placenta supplementation group. We measured epididymal sperm concentration and testicular weight and volume and analyzed these using one-way analysis of variance. Results: Dried bovine placenta was rich in nutrients, with 61.98% protein, 21.25±2.07 carbohydrates, 8.58% water, 4.93% ash, and 3.27% fat. The mean epididymal spermatozoa concentration of the rats in Groups A, B, and C was 3026×106/mL, 1492.8×106/mL, and 2732.5×106/mL, respectively. The average total testicle weights were 2.44 g, 2.72 g, and 2.57 g, respectively. The average total testicle volumes were 2.29 cm3, 2.49 cm3, and 2.33cm3, respectively. Conclusion: Dried bovine placenta powder is rich in nutrients, especially protein. Supplementation with dried bovine placenta can improve epididymal spermatozoa concentration that is important in fertility.

Throughout the world, compost application is viewed as a long-term strategy to improve the availability of phosphorus (P) for agricultural crops. Limited information is available on the legacy effects of P in compost-amended soils with respect to the subsequent effects of mineral P fertilizer on crop growth. We therefore conducted two cycles of maize growth experiments in pots (replicates; n = 4) under greenhouse conditions, with each cycle lasting 84 days. We used two soils that had been fertilized with compost over the past 10 years. The first cycle of maize growth was carried out under full P fertilization conditions, i.e. 150 mg P kg−1 soil (300 kg P ha−1), as calcium dihydrogen phosphate. The second cycle of maize was grown in the same soil and pots after harvesting the first cycle. Plant and soil measurements included soil Hedley P fractions and plant P uptake. Readily available plant P NaHCO3-Pi ranged from 93 to 221 mg P kg−1, increasing by 31% due to compost and 76% due to Ca-P fertilization compared with no compost or Ca–P fertilization, respectively. However, the application of compost and fertilizer only increased the production of dry matter by 7–9%, and plant P uptake by 11% and 17% compared to previous compost and Ca–P applications, respectively. Compost protected against the depletion of the NaHCO3–Po pool and was therefore helpful to initiating a P legacy stock, enhancing soil resilience and limiting the depletion of P by future crop rotations with high P requirements. However, compost mixed with mineral P provided the best availability of P for both immediate crop requirements and the long-term preservation of soil fertility.

International audience; Mycoplasmas are small, genome-reduced bacteria. They are obligate parasites that can be found in a wide range of host species, including the majority of livestock animals and humans. Colonization of the host can result in a wide spectrum of outcomes. In many cases, these successful parasites are considered commensal, as they are found in the microbiota of asymptomatic carriers. Conversely, mycoplasmas can also be pathogenic, as they are associated with a range of both acute and chronic inflammatory diseases which are problematic in veterinary and human medicine. The chronicity of mycoplasma infections and the ability of these bacteria to infect even recently vaccinated individuals clearly indicate that they are able to successfully evade their host's humoral immune response. Over the years, multiple strategies of immune evasion have been identified in mycoplasmas, with a number of them aimed at generating important antigenic diversity. More recently, mycoplasma-specific anti-immunoglobulin strategies have also been characterized. Through the expression of the immunoglobulin-binding proteins protein M or mycoplasma immunoglobulin binding (MIB), mycoplasmas have the ability to target the host's antibodies and to prevent them from interacting with their cognate antigens. In this review, we discuss how these discoveries shed new light on the relationship between mycoplasmas and their host's immune system. We also propose that these strategies should be taken into consideration for future studies, as they are key to our understanding of mycoplasma diseases' chronic and inflammatory nature and are probably a contributing factor to reduce vaccine efficacy.

Curcuma amada Roxb. (Zingiberaceae), commonly known as mango ginger because its rhizome and foliar parts have a similar aroma to mango. The rhizome has been widely used in food industries and alternative medicines to treat a variety of internal diseases such as cough, bronchitis, indigestion, colic, loss of appetite, hiccups, and constipation. The composition of the volatile constituents in a fresh rhizome of C. amada is not reported in detail. The present study aimed to screen and characterize the composition of volatile organic compound (VOC) in a fresh rhizome of three C. amada (ZO45, ZO89, and ZO114) and one C. longa (ZO138) accessions originated from Myanmar. The analysis was carried out by means of headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-time-of-flight-mass spectrometry (GC-TOF-MS). As a result, 122 VOCs were tentatively identified from the extracted 373 mass spectra. The following compounds were the ten most highly abundant and broadly present ones: ar-turmerone, α-zingiberene, α-santalene, (E)-γ-atlantone, cuparene, β-bisabolene, teresantalol, β-sesquiphellandrene, trans-α-bergamotene, γ-curcumene. The intensity of ar-turmerone, the sesquiterpene which is mainly characterized in C. longa essential oil (up to 15.5–27.5%), was significantly higher in C. amada accession ZO89 (15.707 ± 5.78a) compared to C. longa accession ZO138 (0.300 ± 0.08b). Cis-α-bergamotene was not detected in two C. amada accessions ZO45 and ZO89. The study revealed between-species variation regarding identified VOCs in the fresh rhizome of C. amada and C. longa.

Although several oropharyngeal cancer (OPC) susceptibility loci have been identified, most previous studies lacked detailed information on human papillomavirus (HPV) status. We conduct a genome-wide analysis by HPV16 serology status in 4,002 oral cancer cases (OPC and oral cavity cancer (OCC)) and 5,256 controls. We detect four susceptibility loci pointing to a distinct genetic predisposition by HPV status. Our most notable finding in the HLA region, that is now confirmed to be specific of HPV(+)OPC risk, reveal two independent loci with strong protective effects, one refining the previously reported HLA class II haplotype association. Antibody levels against HPV16 viral proteins strongly implicate the protective HLA variants as major determinants of humoral response against L1 capsid protein or E6 oncoprotein suggesting a natural immune response against HPV(+)OPC promoted by HLA variants. This indicates that therapeutic vaccines that target E6 and attenuate viral response after established HPV infections might protect against HPV(+)OPC., Genetic susceptibility loci for oropharyngeal cancer have been reported but these studies have not always examined human papillomavirus (HPV) status. Here, the authors perform genome-wide analysis taking into account HPV16 serology status and report two independent loci in the HLA region, suggesting the protective role of HLA variants against HPV infection.

Recent studies have identified key genes that control the symbiotic interaction between Epichloe festucae and Lolium perenne. Here we report on the identification of specific E. festucae genes that control host infection. Deletion of setB, which encodes a homolog of the H3K36 histone methyltransferase Set2/KMT3, reduced histone H3K36 trimethylation and led to severe defects in colony growth and hyphal development. The E. festucae ΔclrD mutant, which lacks the gene encoding the homolog of the H3K9 methyltransferase KMT1, displays similar developmental defects. Both mutants are completely defective in their ability to infect L. perenne. Alleles that complement the culture and plant phenotypes of both mutants also complement the histone methylation defects. Co-inoculation of either ΔsetB or ΔclrD with the wild-type strain enables these mutants to colonize the host. However, successful colonization by the mutants resulted in death or stunting of the host plant. Transcriptome analysis at the early infection stage identified four fungal candidate genes, three of which encode small-secreted proteins, that are differentially regulated in these mutants compared to wild-type. Deletion of crbA, which encodes a putative carbohydrate binding protein, resulted in significantly reduced host infection rates by E. festucae. This article is protected by copyright. All rights reserved.

MIB and MIP bind to antibodies and disrupt the antigen binding site to promote the dissociation of antibody-antigen interactions., Mycoplasma immunoglobulin binding (MIB) and mycoplasma immunoglobulin protease (MIP) are surface proteins found in the majority of mycoplasma species, acting sequentially to capture antibodies and cleave off their VH domains. Cryo–electron microscopy structures show how MIB and MIP bind to a Fab fragment in a “hug of death” mechanism. As a result, the orientation of the VL and VH domains is twisted out of alignment, disrupting the antigen binding site. We also show that MIB-MIP has the ability to promote the dissociation of the antibody-antigen complex. This system is functional in cells and protects mycoplasmas from antibody-mediated agglutination. These results highlight the key role of the MIB-MIP system in immunity evasion by mycoplasmas through an unprecedented mechanism, and open exciting perspectives to use these proteins as potential tools in the antibody field.

Recent studies have identified key genes in Epichloë festucae that control the symbiotic interaction of this filamentous fungus with its grass host. Here we report on the identification of specific fungal genes that determine its ability to infect and colonize the host. Deletion of setB, which encodes a homolog of the H3K36 histone methyltransferase Set2/KMT3, specifically reduced histone H3K36 trimethylation and led to severe defects in colony growth and hyphal development. The E. festucae ΔclrD mutant, which lacks the gene encoding the homolog of the H3K9 methyltransferase KMT1, displays similar developmental defects. Both mutants are completely defective in their ability to infect the host grass, and mutational studies of key residues in the catalytic SET domains from these proteins show that these phenotypes are dependent on the methyltransferase activities of SetB and ClrD. A comparison of the differences in the host transcriptome between seedlings inoculated with wild-type versus mutants suggests that the inability of these mutants to infect the host was not due to an aberrant host defense response. Co-inoculation of either ΔsetB or ΔclrD with the wild-type strain enables these mutants to colonize the host. However, successful colonization by the mutants resulted in death or stunting of the host plant. Transcriptome analysis at the early infection stage identified four fungal candidate genes, three of which encode small-secreted proteins, that are differentially regulated in these mutants compared to wild-type. Deletion of crbA, which encodes a putative carbohydrate binding protein, resulted in significantly reduced host infection rates by E. festucae.Author SummaryThe filamentous fungus Epichloë festucae is an endophyte that forms highly regulated symbiotic interactions with the perennial ryegrass. Proper maintenance of such interactions is known to involve several signalling pathways, but much less is understood about the infection capability of this fungus in the host. In this study, we uncovered two epigenetic marks and their respective histone methyltransferases that are required for E. festucae to infect perennial ryegrass. Null mutants of the histone H3 lysine 9 and lysine 36 methyltransferases are completely defective in colonizing the host intercellular space, and these defects are dependent on the methyltransferase activities of these enzymes. Importantly, we observed no evidence for increased host defense response to these mutants that can account for their non-infection. Rather, these infection defects can be rescued by the wild-type strain in co-inoculation experiments, suggesting that failure of the mutants to infect is due to altered expression of genes encoding infection factors that are under the control of the above epigenetic marks that can be supplied by the wild-type strain. Among genes differentially expressed in the mutants at the early infection stage is a putative small-secreted protein with a carbohydrate binding function, which deletion in E. festucae severely reduced infection efficiency.

Alport syndrome (AS) is a genetic disorder involving mutations in the genes encoding collagen IV α3, α4 or α5 chains, resulting in the impairment of glomerular basement membrane. Podocytes are responsible for production and correct assembly of collagen IV isoforms; however, data on the phenotypic characteristics of human AS podocytes and their functional alterations are currently limited. The evident loss of viable podocytes into the urine of patients with active glomerular disease enables their isolation in a non-invasive way. Here we isolated, immortalized, and subcloned podocytes from the urine of three different AS patients for molecular and functional characterization. AS podocytes expressed a typical podocyte signature and showed a collagen IV profile reflecting each patient's mutation. Furthermore, RNA-sequencing analysis revealed 348 genes differentially expressed in AS podocytes compared with control podocytes. Gene Ontology analysis underlined the enrichment in genes involved in cell motility, adhesion, survival, and angiogenesis. In parallel, AS podocytes displayed reduced motility. Finally, a functional permeability assay, using a podocyte-glomerular endothelial cell co-culture system, was established and AS podocyte co-cultures showed a significantly higher permeability of albumin compared to control podocyte co-cultures, in both static and dynamic conditions under continuous perfusion. In conclusion, our data provide a molecular characterization of immortalized AS podocytes, highlighting alterations in several biological processes related to extracellular matrix remodelling. Moreover, we have established an in vitro model to reproduce the altered podocyte permeability observed in patients with AS. © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.. ispartof: JOURNAL OF PATHOLOGY vol:252 issue:1 pages:88-100 ispartof: location:England status: published

Background & Aims Understanding the mechanisms by which tumors adapt to therapy is critical for developing effective combination therapeutic approaches to improve clinical outcomes for patients with cancer. Methods To identify promising and clinically actionable targets for managing colorectal cancer (CRC), we conducted a patient-centered functional genomics platform that includes approximately 200 genes and paired this with a high-throughput drug screen that includes 262 compounds in four patient-derived xenografts (PDXs) from patients with CRC. Results Both screening methods identified exportin 1 (XPO1) inhibitors as drivers of DNA damage-induced lethality in CRC. Molecular characterization of the cellular response to XPO1 inhibition uncovered an adaptive mechanism that limited the duration of response in TP53-mutated, but not in TP53-wild-type CRC models. Comprehensive proteomic and transcriptomic characterization revealed that the ATM/ATR-CHK1/2 axes were selectively engaged in TP53-mutant CRC cells upon XPO1 inhibitor treatment and that this response was required for adapting to therapy and escaping cell death. Administration of KPT-8602, an XPO1 inhibitor, followed by AZD-6738, an ATR inhibitor, resulted in dramatic antitumor effects and prolonged survival in TP53-mutant models of CRC. Conclusions Our findings anticipate tremendous therapeutic benefit and support the further evaluation of XPO1 inhibitors, especially in combination with DNA damage checkpoint inhibitors, to elicit an enduring clinical response in patients with CRC harboring TP53 mutations.

The emissions of volatile organic compounds (VOCs) strongly depend on the plant species and are differently represented in specific taxa. VOCs have a degree of chemical diversity and also can serve as chemotaxonomic markers. Zingiber barbatum Wall. is a wild medicinal ginger plant endemic to Myanmar whose VOC composition has never been screened before. In this study, we screened the rhizome of Z. barbatum to identify the VOC composition by the application of gas chromatography combined with time-of-flight-mass spectrometry (GC-TOF-MS). The resulting VOC profile of Z. barbatum showed that it consists mainly of monoterpenes (21%) and sesquiterpenes (30%). Intraspecific similarities and dissimilarities were found to exist between Z. barbatum genotypes in terms of VOC composition. Four accessions (ZO191, ZO223, ZO217, and the control accession ZO105) collected from the Shan State and Mandalay region of Myanmar were found to share a similar VOC profile, while two accessions (ZO64 and ZO160) collected from the Bago region were found to vary in their VOC profiles compared with the control accession. The two identified compounds, i.e., &alpha, bergamotene and &beta, (E)-guaiene may serve as discriminative chemical markers for the characterization of Z. barbatum species collected in these three geographical regions of Myanmar. This study represents a first attempt to identify and describe the VOCs in the medicinal species Z. barbatum that have not been reported to date.

Highlights • Fish reproductive biology made large advances in five decades in both basic research and applications. • Understanding the brain-pituitary-gonadal axis enabled predictable spawning in farmed fish. • Dissecting the mechanisms of sex control advanced production of monosex and sterile fish. • Molecular genetics and model fish revolutionized functional analysis and bridged to biomedicine., Driven by the broad diversity of species and physiologies and by reproduction-related bottlenecks in aquaculture, the field of fish reproductive biology has rapidly grown over the last five decades. This review provides my perspective on the field during this period, integrating fundamental and applied developments and milestones. Our basic understanding of the brain-pituitary-gonadal axis led to overcoming the failure of farmed fish to ovulate and spawn in captivity, allowing us to close the fish life cycle and establish a predictable, year-round production of eggs. Dissecting the molecular and hormonal mechanisms associated with sex determination and differentiation drove technologies for producing better performing mono-sex and reproductively-sterile fish. The growing contingent of passionate fish biologists, together with the availability of innovative platforms such as transgenesis and gene editing, as well as new models such as the zebrafish and medaka, have generated many discoveries, also leading to new insights of reproductive biology in higher vertebrates including humans. Consequently, fish have now been widely accepted as vertebrate reproductive models. Perhaps the best testament of the progress in our discipline is demonstrated at the International Symposia on Reproductive Physiology of Fish (ISRPF), at which our scientific family has convened every four years since the grandfather of the field, the late Ronald Billard, organized the inaugural 1977 meeting in Paimpont, France. As the one person who has been fortunate enough to attend all of these meetings since their inception, I have witnessed first-hand the astounding evolution of our field as we capitalized on the molecular and biotechnological revolutions in the life sciences, which enabled us to provide a higher resolution of fish reproductive and endocrine processes, answer more questions, and dive into deeper comprehension. Undoubtedly, the next (five) decades will be similarly exciting as we continue to integrate physiology with genomics, basic and translational research, and the small fish models with the aquacultured species.

This paper reports an exploratory study based on quantitative genomic analysis in dairy traits of American Alpine goats. The dairy traits are quality-determining components in goat milk, cheese, ice cream, etc. Alpine goat phenotypes for quality components have been routinely recorded for many years and deposited in the Council on Dairy Cattle Breeding (CDCB) repository. The data collected were used to conduct an exploratory genome-wide association study (GWAS) from 72 female Alpine goats originating from locations throughout the U.S. Genotypes were identified with the Illumina Goat 50K single-nucleotide polymorphisms (SNP) BeadChip. The analysis used a polygenic model where the dropping criterion was a call rate≥0.95. The initial dataset was composed of ~60,000 rows of SNPs and 21 columns of phenotypic traits and composed of 53,384 scaffolds containing other informative data points used for genomic predictive power. Phenotypic association with the 50K BeadChip revealed 26,074 reads of candidate genes. These candidate genes segregated as separate novel SNPs and were identified as statistically significant regions for genome and chromosome level trait associations. Candidate genes associated differently for each of the following phenotypic traits: test day milk yield (13,469 candidate genes), test day protein yield (25,690 candidate genes), test day fat yield (25,690 candidate genes), percentage protein (25,690 candidate genes), percentage fat (25,690 candidate genes), and percentage lactose content (25,690 candidate genes). The outcome of this study supports elucidation of novel genes that are important for livestock species in association to key phenotypic traits. Validation towards the development of marker-based selection that provides precision breeding methods will thereby increase the breeding value.

Background Bark beetles are major pests of conifer forests, and their behavior is primarily mediated via olfaction. Targeting the odorant receptors (ORs) may thus provide avenues towards improved pest control. Such an approach requires information on the function of ORs and their interactions with ligands, which is also essential for understanding the functional evolution of these receptors. Hence, we aimed to identify a high-quality complement of ORs from the destructive spruce bark beetle Ips typographus (Coleoptera, Curculionidae, Scolytinae) and analyze their antennal expression and phylogenetic relationships with ORs from other beetles. Using 68 biologically relevant test compounds, we next aimed to functionally characterize ecologically important ORs, using two systems for heterologous expression. Our final aim was to gain insight into the ligand-OR interaction of the functionally characterized ORs, using a combination of computational and experimental methods. Results We annotated 73 ORs from an antennal transcriptome of I. typographus and report the functional characterization of two ORs (ItypOR46 and ItypOR49), which are responsive to single enantiomers of the common bark beetle pheromone compounds ipsenol and ipsdienol, respectively. Their responses and antennal expression correlate with the specificities, localizations, and/or abundances of olfactory sensory neurons detecting these enantiomers. We use homology modeling and molecular docking to predict their binding sites. Our models reveal a likely binding cleft lined with residues that previously have been shown to affect the responses of insect ORs. Within this cleft, the active ligands are predicted to specifically interact with residues Tyr84 and Thr205 in ItypOR46. The suggested importance of these residues in the activation by ipsenol is experimentally supported through site-directed mutagenesis and functional testing, and hydrogen bonding appears key in pheromone binding. Conclusions The emerging insight into ligand binding in the two characterized ItypORs has a general importance for our understanding of the molecular and functional evolution of the insect OR gene family. Due to the ecological importance of the characterized receptors and widespread use of ipsenol and ipsdienol in bark beetle chemical communication, these ORs should be evaluated for their potential use in pest control and biosensors to detect bark beetle infestations.

ObjectiveThis study was conducted to isolate and characterize lytic bacteriophages for pathogenicEscherichia colifrom chicken and beef offal, and analyze their capability as biocontrol for several foodborne pathogens. Methods done in this research are bacteriophage isolation, purification, titer determination, application, determination of host range and minimum multiplicity of infection (miMOI), and bacteriophage morphology.ResultsSix bacteriophages successfully isolated from chicken and beef offal using EPEC and EHEC as host strain. Bacteriophage titers observed between 109and 1010PFU mL−1. CS EPEC and BL EHEC bacteriophage showed high efficiency in reduction of EPEC or EHEC contamination in meat about 99.20% and 99.04%. The lowest miMOI was 0.01 showed by CS EPEC bacteriophage. CI EPEC and BL EPEC bacteriophage suspected asMyoviridaefamily based on its micrograph from Transmission Electron Microscopy (TEM). Refers to their activity, bacteriophages isolated in this study have a great potential to be used as biocontrol against several foodborne pathogens.

Yeast cells have long been used as hosts to propagate exogenous DNA. Recent progress in genome editing opens new avenues in synthetic biology. These developments allow the efficient engineering of microbial genomes in Saccharomyces cerevisiae that can then be rescued to yield modified bacteria/viruses. Recent examples show that the ability to quickly synthesize, assemble and/or modify viral and bacterial genomes may be a critical factor to respond to emerging pathogens. However, this process has some limitations. DNA molecules much larger than two megabase pairs are complex to clone, bacterial genomes have proven difficult to rescue, and the dual-use potential of these technologies must be carefully considered. Regardless, the use of yeast as a factory has enormous appeal for biological applications., Graphical abstract Image 1, Highlights • Saccharomyces cerevisiae is a robust and efficient host for cloning microbial genomes. • Recent developments allow the efficient engineering of microbial genomes in yeast that can then be rescued using transplantation/transfection to yield modified bacteria/viruses. • Overcoming bottlenecks is key to expand use of yeast as a factory and to offer new possibilities in the synthetic biology field. • Rapid synthesis, assembly or modification of viral and bacterial genomes may be a critical factor to respond to emerging pathogens.

The CAPUA study was supported by FIS-FEDER/Spain grant numbers FIS-01/310, FIS-PI03-0365, and FIS-07-BI060604, FICYT/Asturias grant numbers FICYT PB02-67 and FICYT IB09-133, and the University Institute of Oncology (IUOPA), of the University of Oviedo and the Ciber de Epidemiologia y Salud Pública. CIBERESP, SPAIN (...), Ji, X., Mukherjee, S., Landi, M.T., Bosse, Y., Joubert, P., Zhu, D., Gorlov, I., Xiao, X., Han, Y., Gorlova, O., Hung, R.J., Brhane, Y., Carreras-Torres, R., Christiani, D.C., Caporaso, N., Johansson, M., Liu, G., Bojesen, S.E., Le Marchand, L., Albanes, D., Bickeböller, H., Aldrich, M.C., Bush, W.S., Tardon, A., Rennert, G., Chen, C., Byun, J., Dragnev, K.H., Field, J.K., Kiemeney, L.F., Lazarus, P., Zienolddiny, S., Lam, S., Schabath, M.B., Andrew, A.S., Bertazzi, P.A., Pesatori, A.C., Diao, N., Su, L., Song, L., Zhang, R., Leighl, N., Johansen, J.S., Mellemgaard, A., Saliba, W., Haiman, C., Wilkens, L., Fernandez-Somoano, A., Fernandez-Tardon, G., Heijden, E.H.F.M., Kim, J.H., Davies, M.P.A., Marcus, M.W., Brunnström, H., Manjer, J., Melander, O., Muller, D.C., Overvad, K., Trichopoulou, A., Tumino, R., Goodman, G.E., Cox, A., Taylor, F., Woll, P., Wichmann, E., Muley, T., Risch, A., Rosenberger, A., Grankvist, K., Johansson, M., Shepherd, F., Tsao, M.-S., Arnold, S.M., Haura, E.B., Bolca, C., Holcatova, I., Janout, V., Kontic, M., Lissowska, J., Mukeria, A., Ognjanovic, S., Orlowski, T.M., Scelo, G., Swiatkowska, B., Zaridze, D., Bakke, P., Skaug, V., Butler, L.M., Offit, K., Srinivasan, P., Bandlamudi, C., Hellmann, M.D., Solit, D.B., Robson, M.E., Rudin, C.M., Stadler, Z.K., Taylor, B.S., Berger, M.F., Houlston, R., McLaughlin, J., Stevens, V., Nickle, D.C., Obeidat, M., Timens, W., Artigas, M.S., Shete, S., Brenner, H., Chanock, S., Brennan, P., McKay, J.D., Amos, C.I.

Germline telomere maintenance defects are associated with an increased incidence of inflammatory diseases in humans, yet whether and how telomere dysfunction causes inflammation are not known. Here, we show that telomere dysfunction drives pATM/c-ABL-mediated activation of the YAP1 transcription factor, up-regulating the major pro-inflammatory factor, pro-IL-18. The colonic microbiome stimulates cytosolic receptors activating caspase-1 which cleaves pro-IL-18 into mature IL-18, leading to recruitment of interferon (IFN)-γ-secreting T cells and intestinal inflammation. Correspondingly, patients with germline telomere maintenance defects exhibit DNA damage (γH2AX) signaling together with elevated YAP1 and IL-18 expression. In mice with telomere dysfunction, telomerase reactivation in the intestinal epithelium or pharmacological inhibition of ATM, YAP1, or caspase-1 as well as antibiotic treatment, dramatically reduces IL-18 and intestinal inflammation. Thus, telomere dysfunction-induced activation of the ATM-YAP1-pro-IL-18 pathway in epithelium is a key instigator of tissue inflammation., How telomere dysfunction is directly linked to inflammation in humans is currently unclear. Here the authors reveal that telomere dysfunction drives activation of the YAP1 transcription factor, up-regulating the pro inflammatory factor, pro-IL-18 thus revealing a link between telomere dysfunction and initiation of intestinal inflammation.

Fish have been of paramount importance to our understanding of vertebrate comparative neuroendocrinology and the mechanisms underlying the physiology and evolution of gonadotropin-releasing hormones (GnRH) and their genes. This review integrates past and recent knowledge on the Gnrh system in the fish model. Multiple Gnrh isoforms (two or three forms) are present in all teleosts, as well as multiple Gnrh receptors (up to five types), which differ in neuroanatomical localization, pattern of projections, ontogeny and functions. The role of the different Gnrh forms in reproduction seems to also differ in teleost models possessing two versus three Gnrh forms, Gnrh3 being the main hypophysiotropic hormone in the former and Gnrh1 in the latter. Functions of the non-hypothalamic Gnrh isoforms are still unclear, although under suboptimal physiological conditions (e.g. fasting), Gnrh2 may increase in the pituitary to ensure the integrity of reproduction under these conditions. Recent developments in transgenesis and mutagenesis in fish models have permitted the generation of fish lines expressing fluorophores in Gnrh neurons and to elucidate the dynamics of the elaborate innervations of the different neuronal populations, thus enabling a more accurate delineation of their reproductive roles and regulations. Moreover, in combination with neuronal electrophysiology, these lines have clarified the Gnrh mode of actions in modulating Lh and Fsh activities. While loss of function and genome editing studies had the premise to elucidate the exact roles of the multiple Gnrhs in reproduction and other processes, they have instead evoked an ongoing debate about these roles and opened new avenues of research that will no doubt lead to new discoveries regarding the not-yet-fully-understood Gnrh system.

We tested the recent hypothesis that the "fly factor" phenomenon (food currently or previously fed on by flies attracts more flies than the same type of food kept inaccessible to flies) is mediated by bacterial symbionts deposited with feces or regurgitated by feeding flies. We allowed laboratory-reared black blow flies, Phormia regina (Meigen), to feed and defecate on bacterial Luria-Bertani medium solidified with agar, and isolated seven morphologically distinct bacterial colonies. We identified these using matrix-assisted laser desorption/ionization mass spectrometry and sequencing of the 16S rRNA gene. In two-choice laboratory experiments, traps baited with cultures of Proteus mirabilis Hauser, Morganella morganii subsp. sibonii Jensen, or Serratia marcescens Bizio, captured significantly more flies than corresponding control jars baited with tryptic soy agar only. A mixture of seven bacterial strains as a trap bait was more attractive to flies than a single bacterial isolate (M. m. sibonii). In a field experiment, traps baited with agar cultures of P. mirabilis and M. m. sibonii in combination captured significantly more flies than traps baited with either bacterial isolate alone or the agar control. As evident by gas chromatography-mass spectrometry, the odor profiles of bacterial isolates differ, which may explain the additive effect of bacteria to the attractiveness of bacterial trap baits. As "generalist bacteria," P. mirabilis and M. m. sibonii growing on animal protein (beef liver) or plant protein (tofu) are similarly effective in attracting flies. Bacteria-derived airborne semiochemicals appear to mediate foraging by flies and to inform their feeding and oviposition decisions.

A potential therapeutic strategy for diabetes is the transplantation of induced-insulin secreting cells. Based on the common embryonic origin of liver and pancreas, we studied the potential of adult human liver stem-like cells (HLSC) to generate in vitro insulin-producing 3D spheroid structures (HLSC-ILS). HLSC-ILS were generated by a one-step protocol based on charge dependent aggregation of HLSC induced by protamine. 3D aggregation promoted the spontaneous differentiation into cells expressing insulin and several key markers of pancreatic β cells. HLSC-ILS showed endocrine granules similar to those seen in human β cells. In static and dynamic in vitro conditions, such structures produced C-peptide after stimulation with high glucose. HLSC-ILS significantly reduced hyperglycemia and restored a normo-glycemic profile when implanted in streptozotocin-diabetic SCID mice. Diabetic mice expressed human C-peptide and very low or undetectable levels of murine C-peptide. Hyperglycemia and a diabetic profile were restored after HLSC-ISL explant. The gene expression profile of in vitro generated HLSC-ILS showed a differentiation from HLSC profile and an endocrine commitment with the enhanced expression of several markers of β cell differentiation. The comparative analysis of gene expression profiles after 2 and 4 weeks of in vivo implantation showed a further β-cell differentiation, with a genetic profile still immature but closer to that of human islets. In conclusion, protamine-induced spheroid aggregation of HLSC triggers a spontaneous differentiation to an endocrine phenotype. Although the in vitro differentiated HLSC-ILS were immature, they responded to high glucose with insulin secretion and in vivo reversed hyperglycemia in diabetic SCID mice. Electronic supplementary material The online version of this article (10.1007/s12015-018-9845-6) contains supplementary material, which is available to authorized users.

In many animal species, females and males differ in physiology, lifespan, and immune function. The magnitude and direction of the sexual dimorphism in immune function varies greatly and the genetic and mechanistic bases for this dimorphism are often unknown. Here we show that Drosophila melanogaster females are more likely than males to die from infection with several strains of the fungal entomopathogen Beauveria bassiana. The sexual dimorphism is not exclusively due to barrier defenses and persists when flies are inoculated by injection as well as by surface exposure. Loss of function mutations of Toll pathway genes remove the dimorphism in survivorship. Surprisingly, loss of function mutation of relish, a gene in the Imd pathway, also removes the dimorphism, but the dimorphism persists in flies carrying other Imd pathway mutations. The robust sexual dimorphism in D. melanogaster survival to B. bassiana presents opportunities to further dissect its mechanistic details, with applications for biological control of insect vectors of human disease and insect crop pests.