Production of Radiobromide: new Nickel Selenide target and optimized separation by dry distillation

Introduction Radioisotopes of bromine are of special interest for nuclear medical applications. The positron emitting isotopes 75Br (T½ = 1.6 h; β+ = 75.5 %) and 76Br (T½ = 16.2 h; β+ = 57 %) have suitable decay properties for molecular imaging with PET, while the Auger electron emitters 77Br (T½ = 57.0 h) and 80mBr (T½ = 4.4 h) as well as the β−-emitter 82Br (T½ = 35.3 h) are useful for internal radiotherapy. 77Br is additionally suited for SPECT. The isotopes 75Br, 76Br and 77Br are usually produced at a cyclotron either by 3He and α-particle induced reactions on natural arsenic or by proton and deuteron induced reactions on enriched selenium isotopes [1]. As target mate-rials for the latter two reactions, earlier ele-mental selenium [2] and selenides of Cu, Ag, Mn, Mo, Cr, Ti, Pb and Sn were investigated [cf. 3–7]. Besides several wet chemical separation techniques the dry distillation of bromine from the irradiated targets was investigated, too [cf. 2, 4, 5]. However, the method needs further development. Nickel selenide was investigated as a promising target to withstand high beam currents, and the dry distillation technique for the isolation of n.c.a. radiobromine from the target was optimized. Material and Methods Crystalline Nickel-(II) selenide (0.3–0.5 g) was melted into a 0.5 mm deep cavity of a 1 mm thick Ni plate covered with a Ni grid. NiSe has a melting point of 959 °C. For development of targeting and the chemical separation, natural target material was used. Irradiations of NiSe were usually performed with protons of 17 MeV using a slanting water cooled target holder at the cyclotron BC1710 [8]. For radiochemical studies a beam current of 3 µA and a beam time of about 1 h were appropriate. To separate the produced no-carrier-added (n.c.a.) radiobromine from the target material a dry distillation method was chosen. The apparatus was developed on the basis of a dry distillation method for iodine [cf. 9,10] and optimized to obtain the bromine as n.c.a. [*Br]bromide in a small volume of sodium hydroxide solution. Changing different components of the apparatus, the dead volume could be minimized and an almost constant argon flow as carrier medium was realized. Various capillaries of platinum, stainless steel and quartz glass with different diameters and lengths were tested to trap the radiobromine. Results and Conclusion Nickel selenide proved successful as target material for the production of radiobromine by proton irradiation with 17 MeV protons. The target was tested so far only at beam currents up to 10 µA, but further investigations are ongoing. The optimized dry distillation procedure allows trapping of 80–90 % of the produced radiobromine in a capillary. For this purpose quartz glass capillaries proved to be most suitable. After rinsing the capillary with 0.1 M NaOH solution the activity can be nearly completely obtained in less than 100 µL solution as [*Br]bromide immediately useable for radiosynthesis. So, the overall separation yield was estimated to 81 ± 5 %. The radionuclidic composition and activity of the separated radiobromide was measured by γ-ray spectrometry. Due to the use of natural selenium the determination of the isotopic purity was not meaningful, but it could be shown that the radiobromine was free from other radioisotopes co-produced in the target material and the backing. The radiochemical purity as well as the specific activity were determined by radio ionchromatography. Further experiments using NiSe produced from nickel and enriched selenium are to be per-formed. The isotopic purity of the produced respective radiobromide, the production yield at high beam currents and the reusability of the target material have to be studied

A European reference collection of rose varieties : final report

An integrated pilot database was constructed containing administrative, morphological and molecular data as well as pictures of each variety. In spite of some encountered difficulties, it was demonstrated that two laboratories can produce substantially equivalent data and that the molecular data produced is useful as a tool for managing reference collections, prescreening and quality assuranc

BSA Hydrogel Beads Functionalized with a Specific Aptamer Library for Capturing Pseudomonas aeruginosa in Serum and Blood

Systemic blood stream infections are a major threat to human health and are dramatically increasing worldwide. Pseudomonas aeruginosa is a WHO-alerted multi-resistant pathogen of extreme importance as a cause of sepsis. Septicemia patients have significantly increased survival chances if sepsis is diagnosed in the early stages. Affinity materials can not only represent attractive tools for specific diagnostics of pathogens in the blood but can prospectively also serve as the technical foundation of therapeutic filtration devices. Based on the recently developed aptamers directed against P. aeruginosa, we here present aptamer-functionalized beads for specific binding of this pathogen in blood samples. These aptamer capture beads (ACBs) are manufactured by crosslinking bovine serum albumin (BSA) in an emulsion and subsequent functionalization with the amino-modified aptamers on the bead surface using the thiol- and amino-reactive bispecific crosslinker PEG(4)-SPDP. Specific and quantitative binding of P. aeruginosa as the dedicated target of the ACBs was demonstrated in serum and blood. These initial but promising results may open new routes for the development of ACBs as a platform technology for fast and reliable diagnosis of bloodstream infections and, in the long term, blood filtration techniques in the fight against sepsis

FluCell-SELEX Aptamers as Specific Binding Molecules for Diagnostics of the Health Relevant Gut Bacterium Akkermansia muciniphila

Based on their unique properties, oligonucleotide aptamers have been named a gift of biological chemistry to life science. We report the development of DNA aptamers as the first high-affinity binding molecules available for fast and rapid labeling of the human gut bacterium Akkermansia muciniphila with a certain impact on Alzheimer´s disease. Fast and reliable analyses of the composition of microbiomes is an emerging field in microbiology. We describe the molecular evolution and biochemical characterization of a specific aptamer library by a FluCell-SELEX and the characterization of specific molecules from the library by bioinformatics. The aptamer AKK13.1 exerted universal applicability in different analysis techniques in modern microbiology, including fluorimetry, confocal laser scanning microscopy and flow cytometry. It was also functional as a specific binding entity hybridized to anchor primers chemically coupled via acrydite-modification to the surface of a polyacrylamide-hydrogel, which can be prototypically used for the construction of affinity surfaces in sensor chips. Together, the performance and methodological flexibility of the aptamers presented here may open new routes not only to develop novel Akkermansia-specific assays for clinical microbiology and the analyses of human stool samples but may also be an excellent starting point for the construction of novel electronic biosensors

Group B streptococcal colonization in elderly women.

BACKGROUND In non-pregnant adults, the incidence of invasive Group B Streptococcus (GBS) disease is continuously increasing. Elderly and immunocompromised persons are at increased risk of infection. GBS commonly colonizes the vaginal tract, though data on colonization in the elderly are scarce. It is unknown whether the prevalence of GBS colonization is increasing in parallel to the observed rise of invasive infection. We conducted a three-year (2017-2019) prospective observational cross-sectional study in two teaching hospitals in Switzerland to determine the rate of GBS vaginal colonization in women over 60 years and i) to compare the proportions of known risk factors associated with invasive GBS diseases in colonized versus non-colonized women and ii) to evaluate the presence of GBS clusters with specific phenotypic and genotypic patterns in this population. METHODS GBS screening was performed by using vaginal swabs collected during routine examination from women willing to participate in the study and to complete a questionnaire for risk factors. Isolates were characterized for antibiotic resistance profile, serotype and sequence type (ST). RESULTS The GBS positivity rate in the elderly was 17% (44/255 positive samples), and similar to the one previously reported in pregnant women (around 20%). We could not find any association between participants' characteristics, previously published risk factors and GBS colonization. All strains were susceptible to penicillin, 22% (8/36) were not susceptible to erythromycin, 14% (5/36) were not susceptible to clindamycin and 8% (3/36) showed inducible clindamycin resistance. Both M and L phenotypes were each detected in one isolate. The most prevalent serotypes were III (33%, 12/36) and V (31%, 11/36). ST1 and ST19 accounted for 11% of isolates each (4/36); ST175 for 8% (3/36); and ST23, ST249 and ST297 for 6% each (2/36). Significantly higher rates of resistance to macrolides and clindamycin were associated with the ST1 genetic background of ST1. CONCLUSIONS Our findings indicate a similar colonization rate for pregnant and elderly women. TRIAL REGISTRATION Current Controlled Trial ISRCTN15468519 ; 06/01/2017

A Polyclonal SELEX Aptamer Library Allows Differentiation of Candida albicans, C. auris and C. parapsilosis Cells from Human Dermal Fibroblasts

Easy and reliable identification of pathogenic species such as yeasts, emerging as problematic microbes originating from the genus Candida, is a task in the management and treatment of infections, especially in hospitals and other healthcare environments. Aptamers are seizing an already indispensable role in different sensing applications as binding entities with almost arbitrarily tunable specificities and optimizable affinities. Here, we describe a polyclonal SELEX library that not only can specifically recognize and fluorescently label Candida cells, but is also capable to differentiate C. albicans, C. auris and C. parapsilosis cells in flow-cytometry, fluorometric microtiter plate assays and fluorescence microscopy from human cells, exemplified here by human dermal fibroblasts. This offers the opportunity to develop diagnostic tools based on this library. Moreover, these specific and robust affinity molecules could also serve in the future as potent binding entities on biomaterials and as constituents of technical devices and will thus open avenues for the development of cost-effective and easily accessible next generations of electronic biosensors in clinical diagnostics and novel materials for the specific removal of pathogenic cells from human bio-samples

Albumin Microspheres as “Trans-ferry-beads” for Easy Cell Passaging in Cell Culture Technology

Protein hydrogels represent ideal materials for advanced cell culture applications, including 3D-cultivation of even fastidious cells. Key properties of fully functional and, at the same time, economically successful cell culture materials are excellent biocompatibility and advanced fabrication processes allowing their easy production even on a large scale based on affordable compounds. Chemical crosslinking of bovine serum albumin (BSA) with N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) in a water-in-oil emulsion with isoparaffinic oil as the continuous phase and sorbitan monooleate as surfactant generates micro-meter-scale spherical particles. They allow a significant simplification of an indispensable and laborious step in traditional cell culture workflows. This cell passaging (or splitting) to fresh culture vessels/flasks conventionally requires harsh trypsinization, which can be omitted by using the “trans-ferry-beads” presented here. When added to different pre-cultivated adherent cell lines, the beads are efficiently boarded by cells as passengers and can be easily transferred afterward for the embarkment of novel flasks. After this procedure, cells are perfectly viable and show normal growth behavior. Thus, the trans-ferry-beads not only may become extremely affordable as a final product but also may generally replace trypsinization in conventional cell culture, thereby opening new routes for the establishment of optimized and resource-efficient workflows in biological and medical cell culture laboratories