Direct-Push-Technologien – Effiziente Untersuchungsmethoden für die Untergrunderkundung

About ten years ago, Direct Push technologies were introduced in Germany as a “new” tool for subsurface characterization. Ever since, they have been frequently used in various fields of site investigations. However, despite over ten years of experience, their application potential is often perceived with scepticism. Closer consideration of the range of applications of this method shows that the technology is indeed being increasingly used, even though it does not seem to be completely accepted in the “consulting world”. This is surprising as Direct Push is already used in Germany in its simplest form—i.e. by percussion coring—on a routine basis. Nonetheless, the full potential of Direct Push has not yet been exploited. This article aims to provide a comprehensive overview on the state of the art of this technology and its various application potentials and limitations.     

Selective Enzymatic Transformation to Aldehydes in vivo by Fungal Carboxylate Reductase from Neurospora crassa

The enzymatic reduction of carboxylic acids is in its infancy with only a handful of biocatalysts available to this end. We have increased the spectrum of carboxylate‐reducing enzymes (CARs) with the sequence of a fungal CAR from Neurospora crassa OR74A (NcCAR). NcCAR was efficiently expressed in E. coli using an autoinduction protocol at low temperature. It was purified and characterized in vitro, revealing a broad substrate acceptance, a pH optimum at pH 5.5–6.0, a T_m of 45 °C and inhibition by the co‐product pyrophosphate which can be alleviated by the addition of pyrophosphatase. The synthetic utility of NcCAR was demonstrated in a whole‐cell biotransformation using the Escherichia coli K‐12 MG1655 RARE strain in order to suppress overreduction to undesired alcohol. The fragrance compound piperonal was prepared from piperonylic acid (30 mM) on gram scale in 92 % isolated yield in >98% purity. This corresponds to a productivity of 1.5 g/L/h.

     

Diffusion-weighted echo planar MR imaging of the neck at 3 T using integrated shimming: comparison of MR sequence techniques for reducing artifacts caused by magnetic-field inhomogeneities

Objective

Our objective was to compare available techniques reducing artifacts in echo planar imaging (EPI)-based diffusion-weighed magnetic resonance imaging MRI (DWI) of the neck at 3 Tesla caused by B0-field inhomogeneities.

Materials and methods

A cylindrical fat–water phantom was equipped with a Maxwell coil allowing for additional linear B0-field variations in z-direction. The effect of increasing strength of this superimposed gradient on image quality was observed using a standard single-shot EPI-based DWI sequence (sEPI), a zoomed single-shot EPI sequence (zEPI), a readout-segmented EPI sequence (rsEPI), and an sEPI sequence with integrated dynamic shimming (intEPI) on a 3-Tesla system. Additionally, ten volunteers were examined over the neck region using these techniques. Image quality was assessed by two radiologists. Scan durations were recorded.

Results

With increasing strength of the external gradient, marked distortions, signal loss, and failure of fat suppression were observed using sEPI, zEPI, and rsEPI. These artifacts were markedly reduced using intEPI. Significantly better in vivo image quality was also observed using intEPI compared with the other techniques. Scan time of intEPI was similar to sEPI and zEPI and shorter than rsEPI.

Conclusion

The use of integrated 2D shim and frequency adjustment for EPI-based DWI results in a significant improvement in image quality of the head/neck region at 3 Tesla. Combining integrated shimming with rsEPI or zEPI can be expected to provide additional improvements.

     

Structure-cytotoxicity studies on alkyl lysophospholipids and some analogs in leukemic blasts of human origin in vitro

Eleven lipids have been tested for cytotoxic (trypan blue dye exclusion) activity in cells from eight freshly explanted human leukemias in vitro. 4-Aminomethyl-1-[2,3-(di-N-decyloxy)N-propyl]-4-phenylpiperidine (CP-46,665), 1-mercapto-hexadecyl-2-methoxymethyl-rac-glycero-3-phosphocholine (BM 41.440), the 2-acetamide analog of platelet-activating factor (PAF) and 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH₃) were found among the most active compounds. 2-Lysophosphatidylcholine (2-LPC) showed the lowest activity. However, in addition there was variation among the results regarding the activity of the 1-octadecyl-rac-glycero-3-phosphocholine (ET-18-OH) and its D- and L-forms, but a significantly higher cytotoxic activity of D-ET-18-OH compared with L-ET-18-OH on the basis of 2-LPC as control after an incubation time of 48 hr. We conclude that with the limited number of structures available, this type of study is not sufficient to yield further information about the mode of the accumulation and toxicity of this type of lipids.     

Break on through to the other side-biophysics and cell biology shed light on cell-penetrating peptides

Cell-penetrating peptides (CPPs) have become widely used vectors for the cellular import of molecules in basic and applied biomedical research. Despite the broad acceptance of these molecules as molecular carriers, the details of the mode of cellular internalization and membrane permeation remain elusive. Within the last two years endocytosis has been demonstrated to be a route of uptake shared by several CPPs. These findings had a significant impact on CPP research. State-of-the-art cell biology is now required to advance the understanding of the intracellular fate of the CPP and cargo molecules. Owing to their presumed ability to cross lipid bilayers, CPPs also represent highly interesting objects of biophysical research. Numerous studies have investigated structure-activity relationships of CPPs with respect to their ability to bind to a lipid bilayer or to cross this barrier. Endocytosis route only relocates the membrane permeation from the cell surface to endocytic compartments. Therefore, biophysical experiments are key to a mechanistic molecular understanding of the cellular uptake of CPPs. However, biophysical investigations have to consider the molecular environment encountered by a peptide inside and outside a cell. In this contribution we will review biophysical and cell-biology data obtained for several prominent CPPs. Furthermore, we will summarize recent findings on the cell-penetrating characteristics of antimicrobial peptides and the antimicrobial properties of CPPs. Peptides of both groups have overlapping characteristics. Therefore, both fields may greatly benefit from each other. The review will conclude with a perspective of how biophysics and cell biology may synergize even more efficiently in the future.     

Efficient Synthesis of Highly Active Phospha‐Isosteres of the Influenza Neuraminidase Inhibitor Oseltamivir

With a Hunsdiecker–Barton iododecarboxylation strategy, we converted the carboxylate group of the oseltamivir precursor into exemplary phosphonate monoesters. In all cases, K_i values towards influenza virus sialidase remained in the sub‐nanomolar range. We have thus made valuable structural space available for the design of novel oseltamivir‐based tools for influenza virus research.

     

Effects of hexadecylphosphocholine on protein kinase C and TPA-induced differentiation of HL60 cells

Several structural analogs of alkylphosphocholine (APC) were studied for their effects on protein kinase C (PKC) and 12-O-tetradecanoylphorbol-13-acetate (TPA) elicited biochemical and cellular events in HL60 cells. Hexadecylphosphocholine (He-PC₂), the APC prototype, inhibited PKC competitively with respect to phosphatidylserine and noncompetitively with respect to CaCl₂, both with an apparent K_i of about 15 μM. Inhibition of PKC by He-PC₂ was selective, since cyclic AMP dependent protein kinase and Ca²⁺/calmodulin dependent protein kinase II were relatively unaffected. He-PC₂ inhibited TPA-induced depletion of PKC and TPA-stimulated phosphorylation of cellular proteins in HL60 cells. TPA-induced differentiation of HL60 cells was also inhibited by He-PC₂, and this inhibition was synergistic or additive to the effects of 1-(5-isoquinoline-sulfonyl)-2-methylpiperazine (H-7), a PKC inhibitor. The present findings are consistent with the hypothesis that inhibition of PKC might be related, in part, to the antineoplastic effect of He-PC₂ and ether lipid analogs such as ET-18-OCH₃ (1-octadecyl-2-methyl-glycero-3-phosphocholine).     

Choosing heat sinks for cooling in tropical climates

Efforts to limit CO₂emissions from buildings in the tropics either focus on reducing energy demand, i.e., air-conditioning, or on replacing fossil with renewable sources. The link between energy demand and supply is often overlooked, especially the effect of the temperature lift of air-conditioning systems on energy consumption. But while heat and humidity gains define energy demand, operating temperatures of the system define the necessary energy input. We aim to transfer our experience of using the LowEx paradigm for heat pump systems in moderate climates to the tropical climate of Singapore. In this paper, we took a systematic overview of a range of heat sinks, to which we refer to as anergy sinks. We analysed their thermal properties and their effect on the performance of air-conditioning systems, expressed as COP. The predominantly used dry air-cooled condenser units performed worst, especially when subject to a stack effect in (semi-)confined spaces. The performance is highest for cooling towers using the wet bulb temperature followed by water body based anergy sinks and the soil. The wide spread of results confirms that the heat rejection temperature is a decisive factor for the performance of the overall cooling system and the input of primary energy.     

Cytotoxic effects of ether lipids and derivatives in human nonneoplastic bone marrow cells and leukemic cells in vitro

The effects of 2-lysophosphatidylcholine (2-LPC), the alkyl lysophospholipid derivatives (ALP) 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH₃) and 1-O-hexadecyl-sn-glycero-3-phospho-trimethyl-ammonio-hexanol, the 2-acetamide analog of platelet-activating factor (PAF) 1-O-octadecyl-2-acetamide-sn-glycero-3-phosphocholine, the thioether lysophospholipid derivative (TLP) BM 41.440 and the ether-linked lipoidal amine CP-46,665 on tritiated thymidine uptake and trypan blue dye exclusion were tested in vitro in various freshly explanted cell samples from human nonneoplastic bone marrow and human leukemias. In both assay systems, a dose range of 1–20 μg/ml of the compounds was tested after 24, 48 and 72 hr of coincubation with the cells. The trypan blue dye exclusion revealed statistically significant preferential cytotoxicity in leukemic cells for three compounds with the order of quantitative selectiveness: ET-18-OCH₃>BM41.440>2-acetamide analog of PAF. CP-46,665 was the most toxic compound, but did not reveal significant differences between nonneoplastic bone marrow and leukemic cells when added in concentrations greater than 1 μg/ml. The trimethyl-ammoniohexanol compound showed only minor activity in the majority of tests, when added at concentrations <20 μg/ml. 2-LPC was rather ineffective. The tritiated thymidine uptake showed only preferential antiproliferative effects towards leukemic cells of ET-18-OCH₃ and, sometimes, within the dose time frame tested of BM 41.440. All compounds tested except 2-LPC and the trimethyl-ammonio-hexanol compound were active also in this assay (inhibition of uptake>50% of the controls). Based on these results, ET-18-OCH₃ and BM 41.440 are recommended for experimental bone marrow purging.     

Degenerate Quantum Gases in Microgravity

Clouds of ultra-cold atoms and especially Bose–Einstein condensates (BEC) provide a source for coherent matter-waves in numerous earth bound experiments. Analogous to optical interferometry, matter-wave interferometers can be used for precision measurements allowing for a sensitivity orders of magnitude above their optical counterparts. However, in some respects the presence of gravitational forces in the lab limits experimental possibilities. In this article, we report about a compact and robust experiment generating Bose–Einstein condensates in the drop tower facility in Bremen, Germany. We also present the progress of building the succeeding experiment in which a two species atom interferometer will be implemented to test the weak equivalence principle with quantum matter.