Elisa and HPLC analyses of deoxynivalenol in maize and wheat

Deoxynivalenol (DON) is a part of the family of mycotoxins called trichothecenes which are produced by a number of different Fusarium mold species. The presence of DON in 25 wheat and 25 maize samples was examined by Enzyme Linked Immunosorbent Assay (ELISA) and High Performance Liquid Chromatography (HPLC) methods. The presence of DON was detected and determined in 5 (20%) maize and 6 (25%) wheat samples by both of the methods. Correlation between ELISA and HPLC results was established, with the correlation coefficients (r) of 0.9691 and 0.9735 for wheat and maize samples, respectively. The results obtained by ELISA method were significantly higher than those obtained by HPLC method. This fact can be explained by the presence of conjugated or masked mycotoxins in the samples, especially DON-3-glucoside (DON-3-Glc), which could not be determined by HPLC method due to the lack of external standards. Contrary to this, being insufficiently selective towards masked DON, ELISA method measures total DON content of a sample. According to the obtained results, ELISA can be used as a reliable screening method, but the confirmation of positive results must be done by HPLC method

Screening of mycotoxins in animal feed from the region of Vojvodina

This paper shows the results of screening of mycotoxins in animal feed originating from the region of Vojvodina. Permanent screening is needed on all levels of production and storage, as well as the use of known methods to reduce mould contamination or toxin content in feedstuffs and feed. A total of 56 representative samples were collected from feed companies from the region of Vojvodina. Samples were collected during February 2009. The collected samples included 41 samples of feedstuffs (soybean, soybean meal, soybean grits, soybean cake, maize, sunflower meal, barley, wheat feed flour, rapeseed meal, dehydrated sugar beet pulps, alfalfa meal, yeast, dried whey, fish meal, meat-bone meal) and 15 samples of complete feedingstuffs. The amounts of aflatoxins, ochratoxin A, zearalenone, fumonisin and deoxynivalenol were determined. Screening method for the analysis was done using Neogen Veratox® testing kits. The test itself is a competitive direct enzyme-linked immunosorbent assay (CD-ELISA). Mycotoxins were present in 71.4% of the samples, but the values determined were below the maximum allowed limits for both Serbian and EC reference values. Zearalenone was found with the highest incidence (57.1% of samples), followed by ochratoxin A (37.5%), fumonisin (33.9%), deoxynivalenol (14.3%) and aflatoxins (3.6%)

Left and right handedness of fermions and bosons

It is shown, by using Grassmann space to describe the internal degrees of freedom of fermions and bosons, that the Weyl like equation exists not only for massless fermions but also for massless gauge bosons. The corresponding states have well defined helicity and handedness. It is shown that spinors and gauge bosons of the same handedness only interact.Comment: 18 pages, LaTeX, no figures, typographical errors corrected and a few sentences added to clarify some issue

Catalog of Galactic Beta Cephei Stars

We present an extensive and up-to-date catalog of Galactic Beta Cephei stars. This catalog is intended to give a comprehensive overview of observational characteristics of all known Beta Cephei stars. 93 stars could be confirmed to be Beta Cephei stars. For some stars we re-analyzed published data or conducted our own analyses. 61 stars were rejected from the final Beta Cephei list, and 77 stars are suspected to be Beta Cephei stars. A list of critically selected pulsation frequencies for confirmed Beta Cephei stars is also presented. We analyze the Beta Cephei stars as a group, such as the distributions of their spectral types, projected rotational velocities, radial velocities, pulsation periods, and Galactic coordinates. We confirm that the majority of these stars are multiperiodic pulsators. We show that, besides two exceptions, the Beta Cephei stars with high pulsation amplitudes are slow rotators. We construct a theoretical HR diagram that suggests that almost all 93 Beta Cephei stars are MS objects. We discuss the observational boundaries of Beta Cephei pulsation and their physical parameters. We corroborate that the excited pulsation modes are near to the radial fundamental mode in frequency and we show that the mass distribution of the stars peaks at 12 solar masses. We point out that the theoretical instability strip of the Beta Cephei stars is filled neither at the cool nor at the hot end and attempt to explain this observation

Second generation 4,5,6,7-tetrahydrobenzo[d]thiazoles as novel DNA gyrase inhibitors

Aim: DNA gyrase and topoisomerase IV are essential bacterial enzymes, and in the fight against bacterial resistance, they are important targets for the development of novel antibacterial drugs. Results: Building from our first generation of 4,5,6,7-tetrahydrobenzo[d]thiazole-based DNA gyrase inhibitors, we designed and prepared an optimized series of analogs that show improved inhibition of DNA gyrase and topoisomerase IV from Staphylococcus aureus and Escherichia coli, with IC50 values in the nanomolar range. Importantly, these inhibitors also show improved antibacterial activity against Gram-positive strains. Conclusion: The most promising inhibitor, 29, is active against Enterococcus faecalis, Enterococcus faecium and S. aureus wild-type and resistant strains, with minimum inhibitory concentrations between 4 and 8 mu g/ml, which represents good starting point for development of novel antibacterials. Graphical abstractPeer reviewe

Discovery of Protein-Protein Interaction Inhibitors by Integrating Protein Engineering and Chemical Screening Platforms

Protein-protein interactions (PPIs) govern intracellular life, and identification of PPI inhibitors is challenging. Roadblocks in assay development stemming from weak binding affinities of natural PPIs impede progress in this field. We postulated that enhancing binding affinity of natural PPIs via protein engineering will aid assay development and hit discovery. This proof-of-principle study targets PPI between linear ubiquitin chains and NEMO UBAN domain, which activates NF-κB signaling. Using phage display, we generated ubiquitin variants that bind to the functional UBAN epitope with high affinity, act as competitive inhibitors, and structurally maintain the existing PPI interface. When utilized in assay development, variants enable generation of robust cell-based assays for chemical screening. Top compounds identified using this approach directly bind to UBAN and dampen NF-κB signaling. This study illustrates advantages of integrating protein engineering and chemical screening in hit identification, a development that we anticipate will have wide application in drug discovery

Overcoming Ostrea edulis seed production limitations to meet ecosystem restoration demands in the UN decade on restoration

The European flat oyster, Ostrea edulis, is a habitat-forming bivalve which was historically widespread throughout Europe. Following its decline due to overfishing, pollution, sedimentation, invasive species, and disease, O. edulis and its beds are now listed as a threatened and/or declining species and habitat by OSPAR. Increasing recognition of the plight of the oyster, alongside rapidly developing restoration techniques and growing interest in marine restoration, has resulted in a recent and rapid growth in habitat restoration efforts. O. edulis seed supply is currently a major bottleneck in scaling up habitat restoration efforts in Europe. O. edulis has been cultured for centuries, however, research into its culture declined following the introduction of the Pacific oyster, Crassostrea gigas to Europe in the early 1970 s. Recent efforts to renew both hatchery and pond production of O. edulis seed for habitat restoration purposes are hampered by restoration project timelines and funding typically being short, or projects not planning appropriately for the timescales required for investment, research-and-development and delivery of oyster seed by commercial producers. Furthermore, funding for restoration is intermittent, making long-term commitments between producers and restoration practitioners difficult. Long-term, strategic investment in research and production are needed to overcome these bottlenecks and meet current ambitious restoration targets across Europe

Uloga testova otpuštanja interferona gama u nadzoru nad tuberkulozom

Tuberculosis is still one of the major global public health threats. Countries with low incidence must focus on exhausting the reservoir of future cases by preventing reactivation. Therefore, it is important to identify and effectively treat those individuals who have latent tuberculosis infection and who may develop active disease. The tuberculin skin test has been the standard for detection of immune response against M. tuberculosis since the beginning of the 20th century. The new millennium has brought advancement in the diagnosis of latent tuberculosis infection. The name of the new blood test is interferon-gamma release assay (IGRA). Croatia is a middle-incidence country with a long decreasing trend and developed tuberculosis control. To reach low incidence and finally eliminate tuberculosis, its tuberculosis programme needs a more aggressive approach that would include intensive contact investigation and treatment of persons with latent tuberculosis infection. This article discusses the current uses of IGRA and its role in tuberculosis control.Tuberkuloza je i danas jedan od vodećih javnozdravstvenih problema. Zemlje s niskom incidencijom fokusiraju se na iscrpljivanje rezervoara budućih slučajeva sprječavanjem reaktivacije bolesti. To se odnosi na traženje i učinkovito liječenje infi ciranih osoba, primarno onih koje su u riziku od obolijevanja nakon infekcije. Tuberkulinski test je od početka 20. stoljeća bio standard u otkrivanju imunosnog odgovora na kontakt s Mycobacterium tuberculosis. Novo tisućljeće donijelo je određeni napredak u obliku novih testova za dijagnozu latentne tuberkulozne infekcije, krvne testove otpuštanja interferona gama. Hrvatska je zemlja srednje incidencije tuberkuloze s dugogodišnjim silaznim trendom i razvijenim protutuberkuloznim aktivnostima. U težnji prema niskoj incidenciji i u konačnici eliminaciji tuberkuloze potrebne su opsežnije aktivnosti unutar državnog programa nadzora nad tuberkulozom, uključujući intenzivnu obradu kontakata i probir na postojanje latentne tuberkulozne infekcije. Ovaj rad razmatra trenutačnu uporabu IGRE (engl. interferon - gamma release assay) i njezinu ulogu u nadzoru nad tuberkulozom

Kemijski profil sedimenata Plominskog zaljeva

Granulometric, chemical, and leaching properties of sediments dredged in the Plomin Bay (Northern Adriatic Sea, Croatia) were investigated in order to asses the risk of remobilisation of heavy metals into the water column. In total 65 samples from 65 sampling sites were taken from different sediment depths within the bay. Analysis of variance confirmed the homogeneity of granulometric and elemental composition of the investigated sediment throughout its volume. Granulometric analysis showed that all samples corresponded to a pelitic fraction (<0.063 mm). Bulk elemental mass fractions in the sediments were similar to literature data on relatively unpolluted areas of the Adriatic Sea. High sedimentation rate caused by constant infl ow of material from the Boljunčica River drainage may be responsible for low levels of heavy metals and negligible infl uence of fl y and bottom ash from a nearby disposal site on the chemical composition of the sediments. In contact with sea water only 0.29 mg kg-1 of V, 0.04 mg kg-1 of Cr, 0.07 mg kg-1 of Ni, 0.33 mg kg-1 of Cu, 0.67 mg kg-1 of Zn and 0.06 mg kg-1 of Pb could be remobilised from sediment material into the water column. However, these values increased three to ten times in case of leaching with organic acids.Granulometrijska i kemijska svojstva te mogućnost otpuštanja teških metala ispitivani su u sedimentima Plominskog zaljeva (Sjeverni Jadran, Hrvatska) u svrhu utvrđivanja rizika od remobilizacije teških metala iz sedimenta u stupac vode. Uzeto je 65 uzoraka s različitih točaka i dubina unutar zaljeva. Analizom varijance potvrđena je granulometrijska i kemijska homogenost cijelog volumena sedimenta, što upućuje na jedan prevladavajući izvor tijekom cijeloga sedimentacijskog razdoblja. Granulometrijskom analizom je utvrđeno da u svim uzorcima prevladava sitnozrnata frakcija (<0,063 mm). Koncentracije elemenata u ukupnim uzorcima sedimenata slične su literaturnim vrijednostima objavljenim za relativno onečišćena područja Jadranskog mora. Velika brzina sedimentacije uzrokovana konstantnim donosom materijala iz slijevnog područja Boljunčice vjerojatan je uzrok niskih koncentracija teških metala i slabo vidljivog utjecaja odlagališta šljake i pepela na sastav sedimenata. U kontaktu s morskom vodom moguća je remobilizacija samo 0,29 mg kg-1 V, 0,04 mg kg-1 Cr, 0,07 mg kg-1 Ni, 0,33 mg kg-1 Cu, 0,67 mg kg-1 Zn i 0,06 mg kg-1 Pb iz sedimenta u stupac morske vode. Ipak ove vrijednosti su tri do deset puta povećane u slučaju izluživanja s pomoću organskih kiselina

Validation of distinct pathogenic patterns in a cohort of membranoproliferative glomerulonephritis patients by cluster analysis

Background: A novel data-driven cluster analysis identified distinct pathogenic patterns in C3-glomerulopathies and immune complex-mediated membranoproliferative glomerulonephritis. Our aim was to replicate these observations in an independent cohort and elucidate disease pathophysiology with detailed analysis of functional complement markers. ----- Methods: A total of 92 patients with clinical, histological, complement and genetic data were involved in the study, and hierarchical cluster analysis was done by Ward method, where four clusters were generated. ----- Results: High levels of sC5b-9 (soluble membrane attack complex), low serum C3 levels and young age at onset (13 years) were characteristic for Cluster 1 with a high prevalence of likely pathogenic variations (LPVs) and C3 nephritic factor, whereas for Cluster 2-which is not reliable because of the small number of cases-strong immunoglobulin G staining, low C3 levels and high prevalence of nephritic syndrome at disease onset were observed. Low plasma sC5b-9 levels, decreased C3 levels and high prevalence of LPV and sclerotic glomeruli were present in Cluster 3, and patients with late onset of the disease (median: 39.5 years) and near-normal C3 levels in Cluster 4. A significant difference was observed in the incidence of end-stage renal disease during follow-up between the different clusters. Patients in Clusters 3-4 had worse renal survival than patients in Clusters 1-2. ----- Conclusions: Our results confirm the main findings of the original cluster analysis and indicate that the observed, distinct pathogenic patterns are replicated in our cohort. Further investigations are necessary to analyse the distinct biological and pathogenic processes in these patient groups