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Plants synthesize a plethora of chemical defence compounds, which vary between evolutionary lineages. We hypothesize that plants evolved the ability to utilize defence compounds synthesized and released by neighbouring heterospecific plants. In two experiments, we incubated clover (Trifolium repens L.) seedlings with individual benzoxazinoid (BX) compounds (2,4-dihydroxy-1,4-benzoxazin-3-one, 2-hydroxy-1,4-benzoxazin-3-one, benzoxazolinone, and 6-methoxy- benzoxazolin-2-one), a group of bioactive compounds produced by cereals, to allow clover BX uptake. Subsequently, we transplanted the seedlings into soil and quantified BX root and shoot content and invasion of root-knot nematodes in clover roots up to 8 weeks after transplantation. We show that clover root uptake of BXs substantially enhanced clover's resistance against the root-knot nematode Meloidogyne incognita. This effect lasted up to 6 weeks after the clover roots were exposed to the BXs. BXs were absorbed by clover roots, and then translocated to the shoots. As a result of clover metabolization, we detected the parent BXs and a range of their transformation products in the roots and shoots. Based on these novel findings, we envisage that co-cultivation of crop species with complementary and transferable chemical defence systems can add to plant protection. [Display omitted] • Clover roots take up and metabolize cereal benzoxazinoids (BXs). • BXs are translocated into aboveground clover tissues. • BX transformation products are a result of enzymatic reactions in clover. • BXs uptake enhanced clover's resistance against root-knot nematodes. [ABSTRACT FROM AUTHOR]

In 2008–2009 a tick-borne encephalitis virus (TBEV) micro-focus was detected in Northern Zealand, Denmark. No new cases of TBE with an epidemiological link to Northern Zealand has been reported since. Here we undertook to investigate Ixodes ricinus ticks from this endemic micro-focus in 2016 and 2017. In addition to TBEV, I. ricinus ticks may host other pathogens that include Borrelia spp., Babesia spp., Rickettsia spp. and Neoehrlichia mikurensis , together with various endosymbiont microorganisms. To detect multiple organisms we used a metagenomics PanVirus microarray and next-generation sequencing to examine the persistence and evolution of other emerging viruses, bacteria and parasites. Here we report the rise and fall of the Danish TBEV micro-focus in Northern Zealand. However, we identify for the first time in Danish I. ricinus ticks the presence of Uukuniemi virus in addition to a tick-borne phlebovirus and a range of bacteria. [ABSTRACT FROM AUTHOR]

Weed suppressive potential of 33 winter wheat, 24 winter rye and 11 winter triticale cultivars recently introduced to the Scandinavian market was investigated. Competitive traits recorded were early vigor, leaf area index and crop height. Allelopathic potential was assessed by analyzing plant root and shoot material by LC-MS/MS for their content of 12 phytotoxic metabolites of the chemical group of benzoxazinoids (BX). Total BX content was highest in rye, followed by triticale and wheat. Benzoxazinoid composition varied between species with non-methoxy substituted BX dominating in rye and methoxy substituted BX dominating in wheat and triticale. Principal component analysis exhibited a clear relationship of allelopathic and competitive traits, together explaining 62% of the variance in the data set. This result underlines the need for further investigations of the relative contribution of allelopathy and competition to weed suppression in future studies. [ABSTRACT FROM AUTHOR]

Abstract: Therapeutic immunization of HIV-1-infected individuals with or without anti-retroviral therapy is a new promising disease prevention. To induce a new cytotoxic TCD8 lymphocyte (CTL) immunity during chronic HIV-1 infection 15 infrequently targeted but conserved HLA-supertype binding CTL epitopes from Gag, Pol, Nef, Env, Vpu and Vif were identified. The 15 TCD8 and three TCD4 helper peptides were GMP synthesised and formulated with a new adjuvant CAF01 which is a synthetic two-component liposomic adjuvant comprising the quaternary ammonium dimethyl-dioctadecyl-ammonium (DDA) and the immune modulator trehalose 6,6′-dibehenate (TDB). Using IFN-γ ELISPOT assay, T-cell immune induction by the vaccine was found to both CD4 and CD8 T-cell restricted peptides in HLA-A2 transgenic mice. Comprehensive toxicity studies of the CAF01 adjuvant-alone and together with different vaccines showed that CAF01 when tested at human dose levels was safe and well tolerated with only local inflammation at the site of injection and no systemic reactions. No pharmacological safety issues were observed in Beagle dogs. The HIV-1 vaccine toxicity study in the Göttingen Minipig® showed no systemic toxicity from five repetitive i.m. injections, each with a 2-week interval, of either the 18 HIV-1 peptide antigen solution (AFO18) or the AFO18–CAF01, in which the 18 HIV-1 peptides were formulated with the CAF01 adjuvant. Distinct inflammatory responses were observed in the injected muscles of the AFO18–CAF01 vaccine treated animals as a result of the immune stimulating effect of the adjuvant on the vaccine. The results of the toxicity studies provide optimism for phase I clinical trials evaluating the therapeutic HIV-1 T-cell vaccination approach using multiple subdominant minimal epitope peptides applying the novel cationic adjuvant CAF01. [Copyright &y& Elsevier]

Abstract: Immunotherapies based on reinfusion of autologous cells incubated ex vivo with peptides reconstituted in toxic solvents, such as DMSO, are now performed on a routine basis. However, the toxic effects of the most common solvent used, DMSO, on T cell responses from human PBMCs, have not previously been evaluated in detail. Here, in preparation for a first-in-man human phase I vaccine trial comprising reinfusion of autologous HIV peptide-pulsed PBMCs, human PBMCs from healthy and HIV-infected donors were exposed in vitro to a range of DMSO concentrations, and for a range of time periods. Polychromatic flow cytometry was used to evaluate the influence of DMSO on functional T cell responses. We report that high concentrations of up to 10% of DMSO for 1hour do not affect the cell viability, the magnitude or the functional profile of CD4+ and CD8+ T cell responses, regardless of antigen specificity and HLA class I restriction. In contrast, >2% for >2hours compromises these responses. These data are relevant in the design of immunotherapies based on pulsing a large number of peptides onto antigen presenting cells prior to reinfusion. [Copyright &y& Elsevier]

Cyclic hydroxamic acids and lactams are allelochemicals present in the common agricultural crops wheat, rye, and maize. The hydroxamic acids are mainly present in the plants as glucosides. Upon injury or insect attack or when exuded to the soil environment, the hydroxamic acids occur in their unstable agluconic form. In the first step in the transformation of hydroxamic acids, benzoxazolinones are formed spontaneously. It is necessary to elucidate the further microbial transformation of these compounds in the soil environment for a purposeful exploitation of the allelopathic properties of wheat, rye, and maize. In the present paper, the existing knowledge on microbial transformation products of benzoxazolin-2-one (BOA), 6-methoxy-benzoxazolin-2-one (MBOA), and 2-hydroxy-1,4-benzoxazin-3-one (HBOA) was reviewed. Three main groups of transformation products were identified: aminophenoxazinones, malonamic acids, and acetamides. Future research needs concerning the transformation of these chemicals in soil are discussed, when their properties for suppressing weeds and soil-borne diseases are going to be exploited. [Copyright &y& Elsevier]

The atmospheric CO 2 level is rising, and the consequent climate change is causing an increase in drought events. Furthermore, the CO 2 level is known to induce changes in the physiological responses to stress in plants. Exogenous melatonin is suggested to play roles in the response of plants to abiotic stresses, including drought. We investigated physiological drought stress responses at ambient and elevated CO 2 levels (aCO 2 and eCO 2) of melatonin-treated and untreated tomato plants, aiming to link effects of water use efficiency of photosynthesis at (WUE Leaf) and stomatal conductance (g s) with the hormonal regulation of stomata. Tomatoes grown at eCO 2 had reduced water use of both irrigated and drought stressed plants during the progression of drought at the whole plant level. This was also reflected in a CO 2 -affected increase in WUE Leaf at eCO 2 across irrigated and drought-stressed plants. These CO 2 -induced effects were mediated through stomatal closing and reductions in stomatal pore area rather than stomatal density or size. Abscisic acid (ABA) and its conjugated form, ABA glucose ester (ABA-GE), increased at drought stress in aCO 2 , while only ABA-GE increased at eCO 2. Contrary, salicylic acid (SA) increased to a greater magnitude at drought stress in eCO 2 than aCO 2. Melatonin treatment showed no effects on the stomatal regulation. Our findings imply that eCO 2 changes in the balance of hormonal effectors in stomatal regulation during drought, shifting from it ABA to SA regulation, suggesting to consider stomatal reactions at eCO 2 in a perspective of a hormonal interplay rather than only ABA. • ECO 2 reduced water use of both irrigated and drought stressed plants. • ABA increased in tomato in response to drought at aCO 2 , but not at eCO 2. • Salicylic acid increased in drought stress to a larger extend at eCO2 than at aCO2. • Exogenous melatonin had no effect on stomatal regulation or associated hormones. [ABSTRACT FROM AUTHOR]

The SARS-CoV-2 pandemic revealed the critical shortfalls of global vaccine availability for emergent pathogens and the need for exploring additional vaccine platforms with rapid update potential in response to new variants. Thus, it remains essential, for the present evolving SARS-CoV-2/Covid-19 and future pandemics, to continuously develop and characterize new and different vaccine platforms. Here, we describe an expression-optimized DNA vaccine candidate based on the SARS-CoV-2 spike protein of the Beta variant (B.1.351), pNTC-Spike.351, and, in animal models, compare its immunogenicity with a similar DNA vaccine encoding the ancestral index strain spike protein, pNTC-Spike. Both DNA vaccines induced neutralizing antibodies and a Th1 biased immune response. In contrast to the index-specific vaccine, the Beta-specific DNA vaccine induced antibodies in mice and rabbits that, even at low levels, efficiently neutralize the otherwise antibody resistant Beta variant. It similarly neutralized unrelated variants bearing the neutralization resistant E484K spike mutation. Intensive priming using two vaccinations with pNTC-Spike and a single booster immunization with the pNTC-Spike.351 induced a more robust neutralizing antibody response with comparable magnitude against different variants of concern. Thus, DNA vaccine technology with heterologous spike protein prime-boost should be explored further using the Beta derived pNTC-Spike.351 to broaden neutralizing antibody responses against emerging variants of concern. [ABSTRACT FROM AUTHOR]

Hairy roots induced by Agrobacterium rhizogenes are well established models to study the metabolism of xenobiotics in plants for phytoremediation purposes. However, the model requires special skills and resources for growing and is a time-consuming process. The roots induction process alters the genetic construct of a plant and is known to express genes that are normally absent from the non-transgenic plants. In this study, we propose and establish a non-transgenic maize root model to study xenobiotic metabolism in plants for phytoremediation purpose using azoxystrobin as a xenobiotic compound. Maize roots were grown aseptically in Murashige and Skoog medium for two weeks and were incubated in 100 μM azoxystrobin solution. Azoxystrobin was taken up by the roots to the highest concentration within 15 min of treatment and its phase I metabolites were also detected at the same time. Conjugated metabolites of azoxystrobin were detected and their identities were confirmed by enzymatic and mass spectrometric methods. Further, azoxystrobin metabolites identified in maize root culture were compared against azoxystrobin metabolites in azoxystrobin sprayed lettuce grown in green house. A very close similarity between metabolites identified in maize root culture and lettuce plant was obtained. The results from this study establish that non-transgenic maize roots can be used for xenobiotic metabolism studies instead of genetically transformed hairy roots due to the ease of growing and handling. [ABSTRACT FROM AUTHOR]

Estimates of immunity and severity for the SARS-CoV-2 omicron subvariant BA.5 are important to assess the public health impact associated with its rapid global spread despite vaccination. We estimated natural and vaccine immunity and severity of BA.5 relative to BA.2 in Denmark, a country with high mRNA-vaccination coverage and free-of-charge RT-PCR testing. This nation-wide population-based study in Denmark included residents aged 18 years or older who had taken an RT-PCR test between 10 April and 30 June, 2022 (ie, the outcome period), and who the national COVID-19 surveillance system identified as having information since February 2020 on RT-PCR tests, whole-genome sequencing, vaccinations, and hospitalisation with a positive RT-PCR test and COVID-19 as the main diagnosis. First, we used a case–control design, in which cases were people infected with BA.5 or BA.2 during the outcome period and controls were people who tested negative for SARS-CoV-2 infection during the outcome period. We calculated the protection provided by a previous PCR-confirmed omicron infection against BA.5 and BA.2 infection and hospitalisation among triple-vaccinated individuals. Second, we compared vaccination status in people infected with BA.5 versus BA.2 and estimated relative vaccine protection against each subvariant. Third, we compared rates of hospitalisation for COVID-19 among people infected with BA.5 versus BA.2. We estimated effects using logistic regression with adjustment for sex, age, region, PCR-test date, comorbidity and, as appropriate, vaccination and previous infection status. A total of 210 (2·4%) of 8678 of BA.5 cases, 192 (0·7%) of 29 292 of BA.2 cases, and 33 972 (19·0%) of 178 669 PCR-negative controls previously had an omicron infection, which was estimated in the adjusted analyses to offer 92·7% (95% CI 91·6–93·7) protection against BA.5 infection and 97·1% (96·6–97·5) protection against BA.2 infection. We found similarly high amounts of protection against hospitalisation owing to infection with BA.5 (96·4% [95% CI 74·2–99·5]) and BA.2 (91·2% [76·3–96·7]). Vaccine coverage (three mRNA doses vs none) was 9307 (94·2%) of 9878 among BA.5 cases and 30 581 (94·8%) of 32 272 among BA.2 cases, although in the adjusted analysis, there was a trend towards slightly higher vaccination coverage among BA.5 cases than BA.2 cases (OR 1·18 [95% CI 0·99–1·42]; p=0·064), possibly suggesting marginally poorer vaccine protection against BA.5. The rate of hospitalisation due to COVID-19 was higher among the BA.5 cases (210 [1·9%] of 11 314) than among the BA.2 cases (514 [1·4%] of 36 805), with an OR of 1·34 (95% CI 1·14–1·57) and an adjusted OR of 1·69 (95% CI 1·22–2·33), despite low and stable COVID-19 hospitalisation numbers during the study period. The study provides evidence that a previous omicron infection in triple-vaccinated individuals provides high amounts of protection against BA.5 and BA.2 infections. However, protection estimates greater than 90% might be too high if individuals with a previous infection were more likely than those without one to come forward for a test for reasons other than suspicion of COVID-19. Our analysis also showed that vaccine protection against BA.5 infection was similar to, or slightly weaker than, protection against BA.2 infection. Finally, there was evidence that BA.5 infections were associated with an increased risk of hospitalisation compared with BA.2 infections. There was no funding source for this study. [ABSTRACT FROM AUTHOR]

Background: The more infectious SARS-CoV-2 lineage B.1.1.7 rapidly spread in Europe after December, 2020, and a concern that B.1.1.7 could cause more severe disease has been raised. Taking advantage of Denmark's high RT-PCR testing and whole genome sequencing capacities, we used national health register data to assess the risk of COVID-19 hospitalisation in individuals infected with B.1.1.7 compared with those with other SARS-CoV-2 lineages.Methods: We did an observational cohort study of all SARS-CoV-2-positive cases confirmed by RT-PCR in Denmark, sampled between Jan 1 and March 24, 2021, with 14 days of follow-up for COVID-19 hospitalisation. Cases were identified in the national COVID-19 surveillance system database, which includes data from the Danish Microbiology Database (RT-PCR test results), the Danish COVID-19 Genome Consortium, the National Patient Registry, the Civil Registration System, as well as other nationwide registers. Among all cases, COVID-19 hospitalisation was defined as first admission lasting longer than 12 h within 14 days of a sample with a positive RT-PCR result. The study population and main analysis were restricted to the proportion of cases with viral genome data. We calculated the risk ratio (RR) of admission according to infection with B.1.1.7 versus other co-existing lineages with a Poisson regression model with robust SEs, adjusted a priori for sex, age, calendar time, region, and comorbidities. The contribution of each covariate to confounding of the crude RR was evaluated afterwards by a stepwise forward inclusion.Findings: Between Jan 1 and March 24, 2021, 50 958 individuals with a positive SARS-CoV-2 test and at least 14 days of follow-up for hospitalisation were identified; 30 572 (60·0%) had genome data, of whom 10 544 (34·5%) were infected with B.1.1.7. 1944 (6·4%) individuals had a COVID-19 hospitalisation and of these, 571 (29·4%) had a B.1.1.7 infection and 1373 (70·6%) had an infection with other SARS-CoV-2 lineages. Although the overall number of hospitalisations decreased during the study period, the proportion of individuals infected with B.1.1.7 increased from 3·5% to 92·1% per week. B.1.1.7 was associated with a crude RR of hospital admission of 0·79 (95% CI 0·72-0·87; p<0·0001) and an adjusted RR of 1·42 (95% CI 1·25-1·60; p<0·0001). The adjusted RR was increased in all strata of age and calendar period-the two covariates with the largest contribution to confounding of the crude RR.Interpretation: Infection with SARS-CoV-2 lineage B.1.1.7 was associated with an increased risk of hospitalisation compared with that of other lineages in an analysis adjusted for covariates. The overall effect on hospitalisations in Denmark was lessened due to a strict lockdown, but our findings could support hospital preparedness and modelling of the projected impact of the epidemic in countries with uncontrolled spread of B.1.1.7.Funding: None. [ABSTRACT FROM AUTHOR]

This study describes the protective efficacy of a novel influenza plasmid DNA vaccine in the ferret challenge model. The rationally designed polyvalent influenza DNA vaccine encodes haemagglutinin and neuraminidase proteins derived from less glycosylated pandemic H1N1 (2009) and H3N2 (1968) virus strains as well as the nucleoprotein (NP) and matrix proteins (M1 and M2) from a different pandemic H1N1 (1918) strain. Needle-free intradermal immunisation with the influenza DNA vaccine protected ferrets against homologous challenge with an H1N1pdm09 virus strain, demonstrated by restriction of viral replication to the upper respiratory tract and reduced duration of viral shedding post-challenge. Breadth of protection was demonstrated in two heterologous efficacy experiments in which animals immunised with the influenza DNA vaccine were protected against challenge with a highly pathogenic avian influenza H5N1 virus strain with reproducible survival and clinical outcomes. [ABSTRACT FROM AUTHOR]

Secondary metabolites secreted by microbes and plants act as mediators in plant-microbe interactions including nutrient uptake. However, until now very little is known about their role in nutrient assimilation, particularly amino acids, which are important compounds due to their high N content. Here we show that the addition of flavonoid secondary metabolites, derived from clover, to soil changed the bacterial diversity, enhanced the flux of asparagine, and increased the pools of glutamine/glutamate in the soil. This indicates that flavonoids are functionally important qualitative and quantitative components of clover root exudates. Furthermore, the addition of microbial secondary metabolites negatively affected clover uptake of asparagine and plant performance, which demonstrates that microbial competition for nutrients may have multiple physiological targets in the plant. Finally, the detection of intact asparagine in clover roots confirms that amino acid uptake is significant to the plant in agricultural soil. In conclusion, amino-acid flow in the clover rhizosphere can be modified by the effects of clover-derived flavonoids on the bacterial community structure, which affects the flux and pools of amino acids; microbial secondary metabolites, which reduce clover uptake of asparagine; and direct recapture of amino acids by clover. [ABSTRACT FROM AUTHOR]

Benzoxazinoids are secondary metabolites with plant defense properties and possible health-promoting effects in humans. In this study, the transcriptional activity of ScBx genes ( ScBx1-ScBx5 ; ScBx6-like ), involved in benzoxazinoid biosynthesis, was analyzed during germination and early seedling development in rye. Our results showed that ScBx genes had highest levels of expression at 24–30 h after germination, followed by a decrease at later stages. For ScBx1-ScBx5 genes expression was higher in shoots compared with root tissues and vice versa for ScBx6-like gene transcripts. Moreover, methylated forms of benzoxazinoids accumulated in roots rather than in shoots during seedling development, in particular reaching high levels of HMBOA-glc in roots. Chemical profiles of benzoxazinoid accumulation in the developing seedling reflected the combined effects of de novo biosynthesis of the compounds as well as the turnover of compounds either pre-stored in the embryo or de novo biosynthesized. Bioinformatic analysis, together with the differential distribution of ScBx6-like transcripts in root and shoot tissues, suggested the presence of a ZmBx6 homolog encoding a 2-oxoglutarate dependent dehydrogenase in rye. The ScBx6-like cDNA was expressed in E. coli for functional characterization in vitro . LC-MS/MS analysis showed that the purified enzyme was responsible for the oxidation of DIBOA-glc into TRIBOA-glc, strongly suggesting the ScBX6-like enzyme in rye to be a functional ortholog of maize ZmBX6. [ABSTRACT FROM AUTHOR]

Background Pigs are natural hosts for influenza A viruses, and the infection is widely prevalent in swine herds throughout the world. Current commercial influenza vaccines for pigs induce a narrow immune response and are not very effective against antigenically diverse viruses. To control influenza in pigs, the development of more effective swine influenza vaccines inducing broader cross-protective immune responses is needed. Previously, we have shown that a polyvalent influenza DNA vaccine using vectors containing antibiotic resistance genes induced a broadly protective immune response in pigs and ferrets using intradermal injection followed by electroporation. However, this vaccination approach is not practical in large swine herds, and DNA vaccine vectors containing antibiotic resistance genes are undesirable. Objectives To investigate the immunogenicity of an optimized version of our preceding polyvalent DNA vaccine, characterized by a next-generation expression vector without antibiotic resistance markers and delivered by a convenient needle-free intradermal application approach. Methods The humoral and cellular immune responses induced by three different doses of the optimized DNA vaccine were evaluated in groups of five to six pigs. The DNA vaccine consisted of six selected influenza genes of pandemic origin, including internally expressed matrix and nucleoprotein and externally expressed hemagglutinin and neuraminidase. Results Needle-free vaccination of growing pigs with the optimized DNA vaccine resulted in specific, dose-dependent immunity down to the lowest dose (200 μg DNA/vaccination). Both the antibody-mediated and the recall lymphocyte immune responses demonstrated high reactivity against vaccine-specific strains and cross-reactivity to vaccine-heterologous strains. Conclusion The results suggest that polyvalent DNA influenza vaccination may provide a strong tool for broad protection against swine influenza strains threatening animal as well as public health. In addition, the needle-free administration technique used for this DNA vaccine will provide an easy and practical approach for the large-scale vaccination of swine. [ABSTRACT FROM AUTHOR]

We have used a metagenomic microarray to detect genomic RNA from human pegivirus in serum and cerebrospinal fluid from a patient suffering from severe encephalitis. No other pathogen was detected. HPgV in cerebrospinal fluid during encephalitis has never been reported before and its prevalence in cerebrospinal fluid needs further investigation. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB) is a devastating disease of wheat ( Triticum aestivum L.) caused by a mycotoxigenic fungus Fusarium graminearum resulting in significantly decreased yields and accumulation of toxic trichothecenes in grains. We tested 7 major secondary metabolites from wheat for their effect on trichothecene production in liquid cultures of F. graminearum producing trichothecene 15-acetyldeoxynivalenol (15-ADON). 2,4-Dihydroxy-7-methoxy-2 H -1,4-benzoxazin-3(4 H )-one (DIMBOA) benzoxazinoid completely abolished toxin production without any apparent effect on fungal growth. DIMBOA strongly affected the expression of Tri6 , encoding a major transcriptional regulator of several genes of the trichothecene biosynthesis pathway. DIMBOA also repressed expression of Tri5 , encoding trichodiene synthase, the first enzyme in the trichothecene biosynthesis pathway. Thus, DIMBOA could play an important role against the accumulation of trichothecenes in wheat grain. Breeding or engineering of wheat with increased levels of benzoxazinoids could provide varieties with increased resistance against trichothecene contamination of grain and lower susceptibility to FHB. [ABSTRACT FROM AUTHOR]

The use of neonicotinoids as an insecticide group in Ghana has been quite significant particularly in cocoa production. The high usage has been mainly as a result of a government policy of free insecticide spraying on cocoa farms, in an effort to curb declining yields caused by pests and diseases and to prevent the use of unapproved or banned insecticides on cocoa farms. However the scale of cocoa farming, the frequency and intensity of usage coupled with the mode of application may result in large physical volumes of insecticides in the environment. This makes the knowledge of the concentration and fate of neonicotinoids in the environment extremely important. The present study was aimed at assessing the levels of five major neonicotinoids in soils from cocoa farmlands in Ghana. Extraction and cleanup of analytes were performed by use of a method based on the original QuEChERS procedure after optimizing salts, sorbents and instrumental conditions. Analyte extraction with NaCl and MgSO 4 in acidified acetonitrile followed by cleanup with primary secondary amine (PSA) presented the optimum conditions for extraction. Quantification was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionization (ESI). Validation of the procedure showed average recoveries ranging from 72.0 to 104.8% for all analytes at all fortification levels with relative standard deviation (RSD) ≤ 15.0. Limits of quantitation were < 10 μg kg − 1 for all neonicotinoids studied. The results obtained from the analysis of 52 samples from cocoa farms revealed imidacloprid as the predominant neonicotinoid with concentrations ranging from 4.3 to 251.4 μg kg − 1 in > 50% of samples analyzed. [ABSTRACT FROM AUTHOR]

Highlights: [•] High amounts of benzoxazinoids are present in germ of wheat and rye. [•] Dihexoses of DIBOA and HBOA are prominent compounds in wheat and rye grains. [•] Soaking of pearled rye transforms DIBOA-glc-hexose into DIBOA-glc more than boiling. [•] Considerable amounts of benzoxazinoids are leached into water used for cooking. [•] Benzoxazinoid metabolism is different among rye flakes and pearled rye upon cooking. [ABSTRACT FROM AUTHOR]

Abstract: In line of the development of a food composition database for Bangladesh, 10 minor indigenous fruits were analysed for their nutrient composition comprising ascorbic acid, carotenoids and mineral values. Nutrient data obtained have been compared with published data reported in different literatures, book and United States Department of Agriculture-National Nutrient Database for Standard Reference. Ascorbic acid was highest in Wood apple and lowest in Roselle. Monkey jack contained the highest amount of carotenoids, zinc and copper. Content of calcium, magnesium and phosphorous were found highest in Antidesma velutinum. Potassium was the highest in Wood apple followed by in Moneky jack. It was noted that most of the minor fruits have much higher amount of ascorbic acid than the national fruit – Jack fruit ripe, the king fruit – Mango ripe of Bangladesh and exotic fruits – Apple and Grapes. The nutrient values of these minor fruits would make awareness among the people for their mass consumption for healthy life and to grow more minor fruit trees from extinction in order to maintain biodiversity. [Copyright &y& Elsevier]

Highlights: [•] Influenza A(H1N1)pdm09 DNA vaccine protects pigs against heterologous challenge. [•] DNA vaccine based on pandemic 1918 and 2009 genes induces cross-protective immunity. [•] DNA vaccine is highly efficient in influenza virus clearance. [•] DNA vaccine induces immunity and protection in a higher animal model, the pig. [•] Protection against influenza is not merely dependent on HI antibodies [ABSTRACT FROM AUTHOR]

Clin Microbiol Infect 2012; 18: 171-176 Abstract Dengue fever (DF) remains one of the most important emerging infectious diseases. Whereas DF is well recognized in endemic countries, there are indications that the disease is underdiagnosed among travellers to endemic regions. Here, we present the first descriptive survey on cases of travel-acquired DF imported to Denmark diagnosed at the national reference laboratory for dengue virus diagnostics during a 9-year period. In our study, 16 - 46 travel-acquired dengue virus infections were diagnosed per year. DF is mainly imported by adults, mostly men, returning from Southeast Asian countries. The minimum incidence of dengue virus infection among Danish travellers is estimated to be 4.9 per 100 000 travellers. Our results confirm and expand studies from other European countries, and underline the importance of surveillance based on relevant diagnostic analyses. [ABSTRACT FROM AUTHOR]

Abstract: In a cultivation experiment 18 different Amaranthus genotypes were cultivated in parallel in Argentina, Mexico, Spain and two different locations in the Czech Republic. The ripe seeds were analysed for their content of 11 polyphenols and the variations among genotype, species and location were analysed by principal component analysis (PCA). The flavonoid, rutin, exhibited large variations with varying environmental conditions whereas the flavonoid, nicotiflorin, was affected less. Amaranthus hypochondriacus displayed the most stable content of polyphenols with a high end content of flavonoids. The variations between location/environmental condition were primarily described by the variations in the content of p-coumaric acid and protocatechuic acid in the seed samples. [Copyright &y& Elsevier]

Abstract: Bread is consumed in large quantities all over the world, and rye bread is especially popular throughout the Nordic countries. Wholemeal bread is highly recommended as a basic ingredient in daily food, because wholemeal food products generally promote good health due to their vitamin, mineral and fibre content. The literature suggests that wholemeal products have other health-promoting effects even if the ingredients responsible have not been identified. Benzoxazinoids are a group of natural products that have not previously been reported in mature grains. Here, we report for the first time the identity and quantity of 10 compounds of the benzoxazinoid family in mature grains, hydrothermally processed grains of durum wheat (Triticum durum, cv. Kamut), a commercial variety of rye (Secale cereale cv. Picasso) and an old Nordic rye landrace (S. cereale, Svedjerug), as well as in bread baked with flour milled from those grains. Concentrations of the 10 benzoxanoids were determined using LC–MS/MS and ranged from 0 to 348nmolg−1 for conventional flour, to 772–1177nmolg−1 in bread baked with flour from hydrothermally processed grains and to 3116–5570nmolg−1 in flour from hydrothermally processed grains. Benzoxazinoids possess documented physiological effects, and research into the importance of these compounds in the daily diet is therefore needed. Ongoing studies in our lab on the uptake and transformation of benzoxazinoids in mammals will be reported in the near future. [Copyright &y& Elsevier]

Abstract: The New Zealand white rabbit model (Oryctolagus cuniculus) is widely used to test whether HIV vaccine candidates elicit systemic antibody responses; however, its use in mucosal immunology has not been fully exploited due to the difficulty in collecting mucosal specimens longitudinally and reproducibly. Here we describe feasible and non-feasible methods to collect vaginal and nasal specimens from nulliparous rabbits. Non-feasible methods were those resulting in poor reproducibility and considerable animal twitching during sampling, whereas feasible methods resulted in no animal twitching and potential for sampling reproducibility. Standard operating procedures (SOPs) were implemented to collect vaginal swabs yielding total IgA titres ranging from 12,500 to 312,500. Intranasal immunisation with a naked DNA vaccine encoding HIV gp140 elicited HIV envelope-specific IgA detectable in nasal but not in vaginal secretions. Our methods provide an alternative to reliably assess pre- and post-vaccination mucosal antibody titres longitudinally in rabbits as part of mucosal HIV vaccine immunogenicity studies. [ABSTRACT FROM AUTHOR]

Abstract: Fusarium Head Blight (FHB) is a destructive disease that affects the grain yield and quality of cereals. The relationship between the natural defense chemicals benzoxazinoids and the FHB resistance of field grown winter wheat varieties was investigated. FHB resistance was assessed by the inoculation of wheat ears with mixtures of Fusarium avenaceum, Fusarium culmorum, Fusarium graminearum, and Microdochium nivale. The benzoxazinoids detected in the highest concentration were 2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one (3.7–9.4μmol/kgDW) and 2-hydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one (HMBOA, 2.0–11μmol/kgDW). The cultivars most susceptible to FHB were cvs. Hanseat, Asketis, and Ritmo, while cvs. Petrus, Terra, and Hattrick showed high resistance. 2-O-β-d-Glucopyranosyloxy-4,7-dimethoxy-(2H)-1,4-benzoxazin-3(4H)-one (HDMBOA-glc) and 2-O-β-d-glucopyranosyloxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one (HMBOA-glc) were detected. HMBOA-glc was found in higher concentrations than 2-O-β-d-glucopyranosyloxy-2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one (DIMBOA-glc). Principal component analyses demonstrated correlation between the susceptibility to FHB and the concentrations of DIMBOA-glc, HMBOA-glc, HMBOA, 2-O-β-d-glucopyranosyloxy-4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one (DIBOA-glc), 2-O-β-d-glucopyranosyloxy-1,4-benzoxazin-3(4H)-one, and 2-O-β-d-glucopyranosyloxy-4-dihydroxy-(2H)-7,8-dimethoxy-1,4-benzoxazin-3(4H)-one (DIM2BOA-glc). [Copyright &y& Elsevier]

Abstract: A large number of human tumor-associated antigen-derived peptides have been identified that are recognized by CTLs in a MHC-I restricted fashion. The apoptosis inhibitory protein Bcl2 is overexpressed in many human cancers as part of their neoplastic phenotype. Since inhibition or loss of Bcl2 expression might impair tumor growth and survival, this protein may serve as a rational target for vaccine-induced CTL responses. By Western blot technique, we screened a panel of established human tumor cell lines for proteins involved in the apoptotic process. Two of eight tumor cell lines, a B lymphoma (Loukes) and a colon carcinoma (CCL220) cell line showed increased Bcl2 protein expression whereas the majority of tumor cell lines expressed proapoptotic proteins. Neither fibroblasts nor peripheral blood mononuclear cells showed Bcl2 expression. An HLA-A*0201 restricted CTL epitope was deduced in silica from the amino acid sequence of the Bcl2 protein and its binding affinity for HLA-A*0201 was confirmed using a biochemical binding assay. We here demonstrate that the 9-mer peptide Bcl285–93 induces specific CTL reactivity in immunized C57-A2Kb or -A2Db tg mice. These Bcl285–93 specific CTLs react with and lyse Bcl2-expressing human colon carcinoma CCL220 cells which have been transfected with a chimeric HLA-A*0201/H2-Kb DNA construct similar to that expressed in the transgenic mice. Based on these observations, we suggest that Bcl285–93 may be a target for immune therapy. [Copyright &y& Elsevier]

Abstract: We full genome characterised the newly discovered avian influenza virus H5N7 subtype combination isolated from a stock of Danish game ducks to investigate the composition of the genome and possible features of high pathogenicity. It was found that the haemagglutinin and the acidic polymerase genes were closely related to a low pathogenic H5 strain (A/Duck/Denmark/65047/04 H5N2). The neuraminidase and the non-structural genes were closely related to the highly pathogenic H7N7 strains from The Netherlands 2003. The basic polymerase genes 1 and 2 were shared between the Danish H5N7 and H5N2 and the H7N7 from The Netherlands. The nucleoprotein and the matrix genes were closely related to H6 strains. Thus, the new H5N7 subtype share genes with H5, H7 and H6 subtypes and possesses internal genes originating from highly pathogenic strains. The findings emphasize the need for surveillance presumed low pathogenic avian influenza A viruses. [Copyright &y& Elsevier]

Abstract: Three degradation experiments were performed to examine the formation of transformation products from 2-benzoxazolinone (BOA) in different soil types and concentrations. In two experiments using BOA in low concentration (400μgkg−1) only one unidentified transformation product was found, whereas in the degradation experiment in high concentration (400mgkg−1) several metabolites occurred. Two of these metabolites, 2-amino-(3H)-phenoxazin-3-one (APO); and 2-acetylamino-(3H)-phenoxazin-3-one (AAPO) were synthesized to prove their identity. This is the first time that the successive formation of these types of compounds from BOA is shown in soil. A number of other APO related transformation products were detected and provisionally characterized. The formation of APO, which is a much more biologically active compound than BOA, and the concurrent formation of a number of other APO-related metabolites in soil underline the importance of performing transformation studies as part of the evaluation of the effect of allelochemicals on weeds and soil-borne diseases. [Copyright &y& Elsevier]

Abstract: During the past years increasing incidences of influenza A zoonosis have made it of uppermost importance to possess methods for rapid and precise identification and characterisation of influenza A viruses. We present here a convenient one-step RT-PCR method that will amplify full-length haemagglutinin (HA) and neuraminidase (NA) directly from clinical samples and from all known subtypes of influenza A. We applied the method on samples collected in September 2003 from a Danish flock of mallards with general health problems and by this a previously undescribed influenza A subtype combination, H5N7, was identified. The HA gene showed great sequence similarity to the highly pathogenic avian influenza A virus (HPAIV) A/Chicken/Italy/312/97 (H5N2); however, the cleavage site sequence between HA1 and HA2 had a motif typical for low pathogenic avian influenza viruses (LPAIV). The full-length NA sequence was most closely related to the HPAIV A/Chicken/Netherlands/01/03 (H7N7) that infected chickens and humans in the Netherlands in 2003. Ten persons with direct or indirect contact with the Danish mallard ducks showed signs of influenza-like illness 2–3 days following the killing of the ducks, but no evidence of influence infections was detected. To our knowledge this is the first report of an H5N7 influenza A virus. [Copyright &y& Elsevier]

[Display omitted] • Dietary phytochemicals affect the metabolism of pesticides in honey bees. • Dietary Phacelia and Reseda pollen reduce the level of imidacloprid in honey bees. • Dietary Reseda pollen diminishes the concentration of tebuconazole in honey bees. • Metabolomics is a promising tool to explore the impact of phytochemicals in bees. The losses of honey bee colonies and declines of other insect pollinators have been associated with negative effects of pesticides. Honey bees as well as other pollinators are nectar and pollen foragers and thus are exposed to an extensive range of phytochemicals. Understanding the synergistic, additive, and antagonistic effects of plant secondary metabolites and pesticides in honey bees may help to protect honey bee colonies against agrochemicals. In this study, we used untargeted metabolomics to investigate the impact of dietary phytochemical composition on the residual concentration of three pesticides: imidacloprid, tau-fluvalinate and tebuconazole in exposed honey bees. Honey bees were given different diets based on pollen or nectar from four plants: Reseda odorata , Borago officinalis , Phacelia tanacetifolia , and Trifolium repens for two days. Thereafter, they were orally exposed to 10 ng/bee imidacloprid or contact-exposed to 0.9 μg/bee tau-fluvalinate or 5 μg/bee tebuconazole. After 1 h of oral exposure or 24 h of contact exposure, the honey bees were anaesthetised with CO 2 , sacrificed by freezing, extracted with a validated QuEChERS method, and residual pesticide concentrations were determined by LC-QTRAP-MS/MS. The phytochemical composition in the given diets were profiled with an UHPLC-Q Exactive-MS/MS. The results revealed that the dietary phytochemical composition has a noteworthy influence on the concentration of residual pesticides in honey bees. The correlation coefficient analysis demonstrated that flavonoids have a reducing effect on the residual concentration of imidacloprid and tau-fluvalinate in honey bees. The results also highlighted that exposure to imidacloprid impaired the metabolism of sugars in honey bees. Exploiting flavonoid-rich plants may protect honey bees against pesticides and hold promise as forage plants in future beekeeping. [ABSTRACT FROM AUTHOR]

Honey bees are important pollinators and are subject to numerous stressors, such as changing floral resources, parasites, and agrochemical exposure. Pesticide exposure has been linked to the decline in the global honey bee population. We have limited knowledge of the metabolic pathways and synergistic effects of xenobiotics in bees. Quercetin is one of the most abundant phytochemicals in plants and is therefore abundant in the honey bee diet. Quercetin can upregulate the detoxification system in honey bees; however, it is still unknown to what extent quercetin ingestion can reduce the content of absorbed pesticides. In this study, we investigated the effect of dietary quercetin on the contents of three pesticides in honey bees: imidacloprid (insecticide), tebuconazole (fungicide), and tau-fluvalinate (insecticide and acaricide). Bees were divided into two main groups and fed either quercetin-sucrose paste or only sucrose for 72 h. Thereafter, they were orally exposed to ∼10 ng/bee imidacloprid or contact-exposed to ∼0.9 μg/bee tau-fluvalinate or ∼5.2 μg/bee tebuconazole. After 1 h of oral exposure or 24 h of contact exposure, the bees were anaesthetised with CO 2 , sacrificed by freezing, and extracted with a validated QuEChERS method. Subsequently, the concentrations of the three pesticides and quercetin in the bees were determined with a triple quadrupole tandem mass spectrometer coupled to an HPLC system. No significant effect on the concentration of tebuconazole or tau-fluvalinate was observed in bees fed quercetin. Intake of quercetin led to a reduction in the concentration of imidacloprid in honey bees. Quercetin-rich plants may be exploited in future beekeeping. Image 1 • Honey bees orally exposed to imidacloprid exhibit reduced metabolism of quercetin. • Dietary quercetin reduces the concentration of imidacloprid absorbed in honey bees. • Dietary quercetin does not affect the concentration of tebuconazole or tau-fluvalinate in contact-exposed honey bees. [ABSTRACT FROM AUTHOR]