Rapid and sensitive method for the determination of propylene oxide in cigarette mainstream smoke by gas chromatography-mass spectrometry

A rapid method using gas chromatography-mass spectrometry for the analysis of propylene oxide in cigarette mainstream smoke is reported. Validation data show the method, which requires a minimum of sample preparation, to be selective, sensitive, reliable, and robust. Propylene oxide is found in the University of Kentucky Reference Cigarettes 1R4F and 2R4F at concentrations of 0.93 and 0.65 microg/cigarette, respectively, with a quantitation limit of 0.135 microg/cigarette.     

Novel Inhibitors of Nicotinamide-N-Methyltransferase for the Treatment of Metabolic Disorders

Nicotinamide-N-methyltransferase (NNMT) is a cytosolic enzyme catalyzing the transfer of a methyl group from S-adenosyl-methionine (SAM) to nicotinamide (Nam). It is expressed in many tissues including the liver, adipose tissue, and skeletal muscle. Its expression in several cancer cell lines has been widely discussed in the literature, and recent work established a link between NNMT expression and metabolic diseases. Here we describe our approach to identify potent small molecule inhibitors of NNMT featuring different binding modes as elucidated by X-ray crystallographic studies.     

Starch Granule Size and Morphology of Arabidopsis thaliana Starch-Related Mutants Analyzed during Diurnal Rhythm and Development

Transitory starch plays a central role in the life cycle of plants. Many aspects of this important metabolism remain unknown; however, starch granules provide insight into this persistent metabolic process. Therefore, monitoring alterations in starch granules with high temporal resolution provides one significant avenue to improve understanding. Here, a previously established method that combines LCSM and safranin-O staining for in vivo imaging of transitory starch granules in leaves of Arabidopsis thaliana was employed to demonstrate, for the first time, the alterations in starch granule size and morphology that occur both throughout the day and during leaf aging. Several starch-related mutants were included, which revealed differences among the generated granules. In ptst2 and sex1-8, the starch granules in old leaves were much larger than those in young leaves; however, the typical flattened discoid morphology was maintained. In ss4 and dpe2/phs1/ss4, the morphology of starch granules in young leaves was altered, with a more rounded shape observed. With leaf development, the starch granules became spherical exclusively in dpe2/phs1/ss4. Thus, the presented data provide new insights to contribute to the understanding of starch granule morphogenesis.     

Surface engineering the cellular microenvironment via patterning and gradients

Cell organization, proliferation, and differentiation are impacted by diverse cues present in the cellular microenvironment. As a result, the surface of a material plays an important role in cellular function. Synthetic surfaces may be augmented by physical as well as chemical means. In particular, patterning and interfacial gradients may be utilized to mitigate the cellular response. Patterning is advantageous as it affords control over a range of feature sizes from several nanometers to millimeters. Gradients exist in vivo, for instance in stem cell niches, and the ability to create interfacial gradients in vitro can provide valuable insights into the influence of a series of minute surface changes on a single sample. This review focuses on fabrication methods for generating micro‐ and nanoscale surface patterns as well as interfacial gradients, the impact of these surface modifications on the cellular response, and the advantages and challenges of these surfaces in in vitro applications. © 2013 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys., 2013     

A solid phase linker strategy for the direct synthesis of EDANS-labelled peptide substrates

A novel linker strategy for the efficient synthesis of peptides C-terminally labelled with the EDANS fluorophore is described. Using this support, FRET peptide substrates bearing EDANS/Dabcyl fluorescent donor/acceptor groups can be readily prepared using standard Fmoc solid phase methods.     

Mechanistic considerations on contact-active antimicrobial surfaces with controlled functional group densities

A series of N-alkyl-N,N-dimethyldeoxyammonium celluloses is synthesized by converting tosyl celluloses with DBA and DDA, respectively. Surface coatings with these water-insoluble derivatives contain well-defined densities of quaternary ammonium functions and nonactive hydrophobic and hydrophilic groups. It is shown that the antimicrobial activity of such surfaces against S. aureus requires a delicate balance between DDA, BDA, and hydrophobic groups. A mechanism is proposed that involves the selective adhesion of anionic phospholipids from the bacterial cell membrane. This so-called phospholipid sponge effect is supported by the fact that all coatings could be deactivated by treatment with SDS or negatively charged phospholipids, but not with neutral phospholipids.     

Polyhydroxybutyrate production from carbon dioxide by cyanobacteria

Genetic characterization and enhancement of polyhydroxybutyrate (PHB) accumulation in cyanobacteria were investigated for efficient PHB production from CO₂. The genome DNAs in the PHB-accumulating strains Synechococcus sp. MA19 and Spirulina platensis NIES46 retained the highly homologous region to phaC of Synechocystis PCC6803, whereas low homology was detected in the nonaccumulating strains Synechococcus sp. PCC7942 and Anabaenacylindrica NIES19. Synechococcus sp. MA19, which accumulates PHB up to 30% of dry cell weight from CO₂ as the sole carbon source, was mutated by insertion of transposon Tn5 to enhance the PHB accumulation. Genetic and physiological analysis of the mutant indicated that decreased phosphotransacetylase activity could trigger an increase of acetyl coenzyme A leading to enhancement of PHB accumulation. PHB synthase in Synechococcus sp. MA19 was probably attached to thylakoid membrane since PHB granules were associated with pigments. A genetically engineered cyanobacteria retaining soluble PHB synthase from Ralstonia eutropha accumulated pigment-free PHB granules, which is an advantage for the purification of PHB.     

Full Thermal Switching of Enzymes by Thermoresponsive Poly(2-oxazoline)-Based Enzyme Inhibitors

Controlling the activity of enzymes is an important feature for many processes in medicine, bioanalytics, and biotechnology. So far, it has not been possible to fully switch biocatalysts on and off by thermoresponsive enzyme inhibitors. Herein, we present poly(2-oxazoline)s with iminodiacetic acid end groups (POx-IDA) that are lower critical solution temperature (LCST) polymers and thus thermosensitive. They are capable of reversibly inhibiting the activity of horse radish peroxidase and laccase by more than 99 %. Increasing the temperature makes the POx-IDA precipitate, which leads to 100 % recovery of the enzyme activity. This switching cycle is fully reversible. The LCST of the POx-IDA can be tuned by varying the polymer composition to generate a wide range of switching windows.