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

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

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

Effects of Temperature–Climate Patterns on the Production of Some Competitive Species on Grounds of Modelling

Climate change has serious effects on the setting up and the operation of natural ecosystems. Small increase in temperature could cause rise in the amount of some species or potential disappearance of others. During our researches, the dispersion of the species and biomass production of a theoretical ecosystem were examined on the effect of the temperature–climate change. The answers of the ecosystems which are given to the climate change could be described by means of global climate modelling and dynamic vegetation models. The examination of the operation of the ecosystems is only possible in huge centres on supercomputers because of the number and the complexity of the calculation. The number of the calculation could be decreased to the level of a PC by considering the temperature and the reproduction during modelling a theoretical ecosystem, and several important theoretical questions could be answered

Remodeling of the Streptococcus agalactiae Transcriptome in Response to Growth Temperature

BACKGROUND: To act as a commensal bacterium and a pathogen in humans and animals, Streptococcus agalactiae (group B streptococcus, GBS) must be able to monitor and adapt to different environmental conditions. Temperature variation is a one of the most commonly encountered variables. METHODOLOGY/PRINCIPAL FINDINGS: To understand the extent to which GBS modify gene expression in response to temperatures encountered in the various hosts, we conducted a whole genome transcriptome analysis of organisms grown at 30 degrees C and 40 degrees C. We identified extensive transcriptome remodeling at various stages of growth, especially in the stationary phase (significant transcript changes occurred for 25% of the genes). A large proportion of genes involved in metabolism was up-regulated at 30 degrees C in stationary phase. Conversely, genes up-regulated at 40 degrees C relative to 30 degrees C include those encoding virulence factors such as hemolysins and extracellular secreted proteins with LPXTG motifs. Over-expression of hemolysins was linked to larger zones of hemolysis and enhanced hemolytic activity at 40 degrees C. A key theme identified by our study was that genes involved in purine metabolism and iron acquisition were significantly up-regulated at 40 degrees C. CONCLUSION/SIGNIFICANCE: Growth of GBS in vitro at different temperatures resulted in extensive remodeling of the transcriptome, including genes encoding proven and putative virulence genes. The data provide extensive new leads for molecular pathogenesis research

Extensive Adaptive Changes Occur in the Transcriptome of Streptococcus agalactiae (Group B Streptococcus) in Response to Incubation with Human Blood

To enhance understanding of how Streptococcus agalactiae (group B streptococcus, GBS) adapts during invasive infection, we performed a whole-genome transcriptome analysis after incubation with whole human blood. Global changes occurred in the GBS transcriptome rapidly in response to blood contact following shift from growth in a rich laboratory medium. Most (83%) of the significantly altered transcripts were down-regulated after 30 minutes of incubation in blood, and all functional categories of genes were abundantly represented. We observed complex dynamic changes in the expression of transcriptional regulators and stress response genes that allow GBS to rapidly adapt to blood. The transcripts of relatively few proven virulence genes were up-regulated during the first 90 minutes. However, a key discovery was that genes encoding proteins involved in interaction with the host coagulation/fibrinolysis system and bacterial-host interactions were rapidly up-regulated. Extensive transcript changes also occurred for genes involved in carbohydrate metabolism, including multi-functional proteins and regulators putatively involved in pathogenesis. Finally, we discovered that an incubation temperature closer to that occurring in patients with severe infection and high fever (40°C) induced additional differences in the GBS transcriptome relative to normal body temperature (37°C). Taken together, the data provide extensive new information about transcriptional adaptation of GBS exposed to human blood, a crucial step during GBS pathogenesis in invasive diseases, and identify many new leads for molecular pathogenesis research

Dual Role for Pilus in Adherence to Epithelial Cells and Biofilm Formation in Streptococcus agalactiae

Streptococcus agalactiae is a common human commensal and a major life-threatening pathogen in neonates. Adherence to host epithelial cells is the first critical step of the infectious process. Pili have been observed on the surface of several gram-positive bacteria including S. agalactiae. We previously characterized the pilus-encoding operon gbs1479-1474 in strain NEM316. This pilus is composed of three structural subunit proteins: Gbs1478 (PilA), Gbs1477 (PilB), and Gbs1474 (PilC), and its assembly involves two class C sortases (SrtC3 and SrtC4). PilB, the bona fide pilin, is the major component; PilA, the pilus associated adhesin, and PilC, are both accessory proteins incorporated into the pilus backbone. We first addressed the role of the housekeeping sortase A in pilus biogenesis and showed that it is essential for the covalent anchoring of the pilus fiber to the peptidoglycan. We next aimed at understanding the role of the pilus fiber in bacterial adherence and at resolving the paradox of an adhesive but dispensable pilus. Combining immunoblotting and electron microscopy analyses, we showed that the PilB fiber is essential for efficient PilA display on the surface of the capsulated strain NEM316. We then demonstrated that pilus integrity becomes critical for adherence to respiratory epithelial cells under flow-conditions mimicking an in vivo situation and revealing the limitations of the commonly used static adherence model. Interestingly, PilA exhibits a von Willebrand adhesion domain (VWA) found in many extracellular eucaryotic proteins. We show here that the VWA domain of PilA is essential for its adhesive function, demonstrating for the first time the functionality of a prokaryotic VWA homolog. Furthermore, the auto aggregative phenotype of NEM316 observed in standing liquid culture was strongly reduced in all three individual pilus mutants. S. agalactiae strain NEM316 was able to form biofilm in microtiter plate and, strikingly, the PilA and PilB mutants were strongly impaired in biofilm formation. Surprisingly, the VWA domain involved in adherence to epithelial cells was not required for biofilm formation

Effects of vegetation structure and human impact on understory honey plant richness: implications for pollinator visitation

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.Background Though the biomass of floral vegetation in understory plant communities in a forested ecosystem only accounts for less than 1% of the total biomass of a forest, they contain most of the floral resources of a forest. The diversity of understory honey plants determines visitation rate of pollinators such as honey bee (Apis mellifera) as they provide rich food resources. Since the flower visitation and foraging activity of pollinators lead to the provision of pollination service, it also means the enhancement of plant-pollinator relationship. Therefore, an appropriate management scheme for understory vegetation is essential in order to conserve pollinator population that is decreasing due to habitat destruction and disease infection. This research examined the diversity of understory honey plant and studied how it is related to environmental variables such as (1) canopy density, (2) horizontal heterogeneity of canopy surface height, (3) slope gradient, and (4) distance from roads. Vegetation survey data of 39 plots of mixed forests in Chuncheon, Korea, were used, and possible management practices for understory vegetation were suggested. Results This study found that 113 species among 141 species of honey plant of the forests were classified as understory vegetation. Also, the understory honey plant diversity is significantly positively correlated with distance from the nearest road and horizontal heterogeneity of canopy surface height and negatively correlated with canopy density. Conclusions The diversity of understory honey plant vegetation is correlated to vegetation structure and human impact. In order to enhance the diversity of understory honey plant, management of density and height of canopy is necessary. This study suggests that improved diversity of canopy cover through thinning of overstory vegetation can increase the diversity of understory honey plant species