NmPin from the marine thaumarchaeote Nitrosopumilus maritimus is an active membrane associated prolyl isomerase.

We cordially thank Alma Rute for excellent technical assistance and the DFG (GRK 1431-1 and 1431-2) for financial support (PB). We thank the Microscope and Histology Facility of the University of Aberdeen for providing their equipment. We also thank Jacob Hargreaves for proofreading the manuscript.Peer reviewedPublisher PD

Design and Characterization of Surface‐Crosslinked Gelatin Nanoparticles for the Delivery of Hydrophilic Macromolecular Drugs

For nanotechnology enabled delivery of hydrophilic protein‐based drugs, several polymer‐based carrier systems have been used in the past to protect the sensitive load and to facilitate cellular uptake and crossing of biological barriers. This study uses gelatin, a natural and biodegradable macromolecule, as carrier material which is approved for several applications. Nanoprecipitation is used to form nanoparticles and to maintain the physicochemical integrity of gelatin, hydrophilic crosslinkers, e.g., paraformaldehyde, glutaraldehyde, carbodiimide, and transglutaminase are employed. However, these crosslinkers diffuse homogenously into the carrier matrix also crosslinking the polymeric matrix with the entrapped protein‐based molecules thus rendering it inactive. Hence a hydrophobic zero‐length crosslinker, diisopropylcarbodiimide, is applied to avoid diffusion into the particles. This will provide an opportunity to encapsulate protein‐based drugs in the non‐crosslinked matrix. The hypothesis of surface crosslinking is proven by the extent of crosslinking and more importantly by encapsulation and the release of lysozyme as a model hydrophilic protein. Furthermore, essential process parameters are evaluated such as crosslinker concentration, crosslinking time and crosslinking reaction temperature with regard to the effect on particle size, size distribution and zeta‐potential of gelatin nanoparticles. The optimum formulation results in the production of gelatin nanoparticles with 200‐300 nm and a polydispersity index < 0.2

Production of oceanic nitrous oxide by ammonia-oxidizing archaea

The recent finding that microbial ammonia oxidation in the ocean is performed by archaea to a greater extent than by bacteria has drastically changed the view on oceanic nitrification. The numerical dominance of archaeal ammonia-oxidizers (AOA) over their bacterial counterparts (AOB) in large parts of the ocean leads to the hypothesis that AOA rather than AOB could be the key organisms for the oceanic production of the strong greenhouse gas nitrous oxide (N2O) that occurs as a by-product of nitrification. Very recently, enrichment cultures of marine ammonia-oxidizing archaea have been reported to produce N2O. Here, we demonstrate that archaeal ammonia monooxygenase genes (amoA) were detectable throughout the water column of the eastern tropical North Atlantic (ETNA) and eastern tropical South Pacific (ETSP) Oceans. Particularly in the ETNA, comparable patterns of abundance and expression of archaeal amoA genes and N2O co-occurred in the oxygen minimum, whereas the abundances of bacterial amoA genes were negligible. Moreover, selective inhibition of archaea in seawater incubations from the ETNA decreased the N2O production significantly. In studies with the only cultivated marine archaeal ammonia-oxidizer Nitrosopumilus maritimus SCM1, we provide the first direct evidence for N2O production in a pure culture of AOA, excluding the involvement of other microorganisms as possibly present in enrichments. N. maritimus showed high N2O production rates under low oxygen concentrations comparable to concentrations existing in the oxycline of the ETNA, whereas the N2O production from two AOB cultures was comparably low under similar conditions. Based on our findings, we hypothesize that the production of N2O in tropical ocean areas results mainly from archaeal nitrification and will be affected by the predicted decrease in dissolved oxygen in the ocean

Effect of physical stimuli on hair follicle deposition of clobetasol-loaded Lipid Nanocarriers

Clobetasol propionate (CLO) is a potent glucocorticoid used to treat inflammation-based skin, scalp, and hair disorders. In such conditions, hair follicles (HF) are not only the target site but can also act as drug reservoirs when certain formulations are topically applied. Recently, we have demonstrated nanostructured lipid carriers (NLC) containing CLO presenting epidermal-targeting potential. Here, the focus was evaluating the HF uptake provided by such nanoparticles in comparison to a commercial cream and investigating the influence of different physical stimuli [i.e., infrared (IR) irradiation (with and without metallic nanoparticles-MNP), ultrasound (US) (with and without vibration) and mechanical massage] on their follicular targeting potential. Nanosystems presented sizes around 180 nm (PdI < 0.2) and negative zeta potential. The formulation did not alter skin water loss measurements and was stable for at least 30 days at 5 °C. Nanoparticles released the drug in a sustained fashion for more than 3 days and increased passively about 40 times CLO follicular uptake compared to the commercial cream. Confocal images confirmed the enhanced follicular delivery. On the one hand, NLC application followed by IR for heat generation showed no benefit in terms of HF targeting even at higher temperatures generated by metallic nanoparticle heating. On the other hand, upon US treatment, CLO retention was significantly increased in deeper skin layers. The addition of mechanical vibration to the US treatment led to higher follicular accumulation compared to passive exposure to NLC without stimuli. However, from all evaluated stimuli, manual massage presented the highest follicular targeting potential, driving more than double the amount of CLO into the HF than NLC passive application. In conclusion, NLC showed great potential for delivering CLO to HF, and a simple massage was capable of doubling follicular retention

Carbon isotope fractionation by the marine ammonia-oxidizing archaeon Nitrosopumilus maritimus

Ammonia-oxidizing archaea (AOA) are abundant and widely distributed microorganisms in aquatic and terrestrial habitats. By catalyzing the first and rate limiting step in nitrification, these chemolithoautotrophs play a significant role in the global nitrogen cycle and contribute to primary production. Here, the carbon isotopic fractionation relative to inorganic carbon source was determined for bulk biomass, biphytanes and polar lipid bound sugars of a marine AOA pure culture. Bulk biomass and biphytanes from Nitrosopumilus maritimus showed identical carbon isotope fractionation (εDIC/bulk and εDIC/byphytanes) of ca. −20‰. The glycoside head groups were mainly glucose, mannose and inositol, and exhibited different carbon isotopic composition. In general, these monosaccharides were enriched in 13C (ε −6.1‰ to −13.8‰) relative to bulk biomass and biphytanes. The fact that the carbon isotope composition of the biphytanes reflected that of the bulk biomass of N. maritimus suggests that the depletion of 13C in both biomass and biphytanes resulted mainly from the carbon isotope discrimination by the bicarbonate-fixing enzyme in the autotrophic hydroxypropionate/hydroxybutyrate cycle. Our results further revealed that lipid compounds represent suitable biomarkers for determining δ13C values of archaeal ammonia oxidizers without biosynthetic correction

Identity and abundance of active sulfate-reducing bacteria in deep tidal flat sediments determined by directed cultivation and CARD-FISH analysis

The identity and abundance of potentially active sulfate-reducing bacteria (SRB) in several metre deep sediments of a tidal sand flat in the German Wadden Sea were assessed by directed cultivation and cultivation-independent CARD-FISH analysis (catalysed reporter deposition fluorescence in situ hybridization). Presumably abundant SRB from different sediment layers between 0.5 and 4 m depth were selectively enriched in up to million-fold diluted cultures supplemented with lactate, acetate or hydrogen. Partial 16S rRNA gene sequences obtained from highest dilution steps showing sulfide formation indicated growth of deltaproteobacterial SRB belonging to the Desulfobulbaceae and the Desulfobacteraceae as well as of members of the Firmicutes. Subsequent isolation resulted in 10 novel phylotypes of both litho- and organotrophic sulfate-reducing Deltaproteobacteria. Molecular pre-screening identified six isolates as members of the Desulfobulbaceae, sharing highest identities with either candidatus 'Desulfobacterium corrodens' (95-97%) or Desulfobacterium catecholicum (98%), and four isolates as members of Desulfobacteraceae, being related to either Desulfobacter psychrotolerans (98%) or Desulfobacula phenolica (95-97%). Relatives of D. phenolica were exlusively isolated from 50 and 100 cm deep sediments with 10 and 2 mM of pore water sulfate respectively. In contrast, relatives of D. corrodens, D. psychrotolerans and D. catecholicum were also obtained from layers deeper than 100 cm and with less than 2 mM sulfate. The high in situ abundance of members of both families in sediment layers beneath 50 cm could be confirmed via CARD-FISH analysis performed with a set of six SRB-specific oligonucleotide probes. Moreover, SRB represented a numerically significant fraction of the microbial community throughout the sediment (up to 7%) and reached even higher cell numbers in deep, sulfate-poor layers than in the sulfate-rich surface sediment. This relatively large community size of potentially active SRB in deep sandy sediments might on the one hand be a result of their syntrophic association with other anaerobes. Our results furthermore support the hypothesis that enhanced advective pore water transport might supply nutrients to microbial communities in deep sandy sediments and point to their so far unrecognized contribution to biogeochemical processes in Wadden Sea sediments

Carbon recycling efficiency and phosphate turnover by marine nitrifying archaea

Thaumarchaeotal nitrifiers are among the most abundant organisms in the ocean, but still unknown is the carbon (C) yield from nitrification and the coupling of these fluxes to phosphorus (P) turnover and release of metabolites from the cell. Using a dual radiotracer approach, we found that Nitrosopumilus maritimus fixed roughly 0.3 mol C, assimilated 2 mmol P, and released ca. 10(-2) mol C and 10(-5) mol P as dissolved organics (DOC and DOP) per mole ammonia respired. Phosphate turnover may influence assimilation fluxes by nitrifiers in the euphotic zone, which parallel those of the dark ocean. Collectively, marine nitrifiers assimilate up to 2 Pg C year(-1) and 0.05 Pg P year(-1) and thereby recycle roughly 5% of mineralized C and P into marine biomass. Release of roughly 50 Tg DOC and 0.2 Tg DOP by thaumarchaea each year represents a small but fresh input of reduced substrates throughout the ocean

Ernaehrungswirtschaft im Oblast Kaliningrad (Gebiet Koenigsberg) in der Transformation: Produktions- und Marktprobleme

Summary in EnglishAvailable from Bibliothek des Instituts fuer Weltwirtschaft, ZBW, Duesternbrook Weg 120, D-24105 Kiel C 192832 / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekSIGLEDEGerman

Additional file 1: Figure S1. of NmPin from the marine thaumarchaeote Nitrosopumilus maritimus is an active membrane associated prolyl isomerase

Sequence alignment of NmPin from N. maritimus with various homologues from other phyla to investigate a potential conservation of the positively charged lysine patch of NmPin. Representatives were found by BLAST search. Each group was separately aligned to NmPin (green). Lysines of the patch (K5, K7, K31, K34, K37, K47, K48, K90) are labelled in dark blue and Arg or positively charged residues which are in close proximity to the conserved position are labelled in light blue. The positively charged patch on the surface of NmPin might be conserved also in other members of the TACK phylum. (PDF 464 kb