Technologies for Single-Cell Isolation

The handling of single cells is of great importance in applications such as cell line development or single-cell analysis, e.g., for cancer research or for emerging diagnostic methods. This review provides an overview of technologies that are currently used or in development to isolate single cells for subsequent single-cell analysis. Data from a dedicated online market survey conducted to identify the most relevant technologies, presented here for the first time, shows that FACS (fluorescence activated cell sorting) respectively Flow cytometry (33% usage), laser microdissection (17%), manual cell picking (17%), random seeding/dilution (15%), and microfluidics/lab-on-a-chip devices (12%) are currently the most frequently used technologies. These most prominent technologies are described in detail and key performance factors are discussed. The survey data indicates a further increasing interest in single-cell isolation tools for the coming years. Additionally, a worldwide patent search was performed to screen for emerging technologies that might become relevant in the future. In total 179 patents were found, out of which 25 were evaluated by screening the title and abstract to be relevant to the field.     

Biocatalytic Asymmetric Phosphorylation Catalyzed by Recombinant Glycerate‐2‐Kinase

The efficient synthesis of pure d ‐glycerate‐2‐phosphate is of great interest due to its importance as an enzyme substrate and metabolite. Therefore, we investigated a straightforward one‐step biocatalytic phosphorylation of glyceric acid. Glycerate‐2‐kinase from Thermotoga maritima was expressed in Escherichia coli, allowing easy purification. The selective glycerate‐2‐kinase‐catalyzed phosphorylation was followed by ³¹P NMR and showed excellent enantioselectivity towards phosphorylation of the d ‐enantiomer of glyceric acid. This straightforward phosphorylation reaction and subsequent product isolation enabled the preparation of enantiomerically pure d ‐glycerate 2‐phosphate. This phosphorylation reaction, using recombinant glycerate‐2‐kinase, yielded d ‐glycerate 2‐phosphate in fewer reaction steps and with higher purity than chemical routes.     

A Conjugate of an Anti‐Epidermal Growth Factor Receptor (EGFR) VHH and a Cell‐Penetrating Peptide Drives Receptor Internalization and Blocks EGFR Activation

Overexpression of (mutated) receptor tyrosine kinases is a characteristic of many aggressive tumors, and induction of receptor uptake has long been recognized as a therapeutic modality. A conjugate of a synthetically produced cell‐penetrating peptide (CPP), corresponding to amino acids 38–59 of human lactoferrin, and the recombinant llama single‐domain antibody (VHH) 7D12, which binds the human epidermal growth factor receptor (EGFR), was generated by sortase A mediated transpeptidation. The conjugate blocks EGF‐mediated EGFR activation with higher efficacy than that of both modalities alone; a phenomenon that is caused by both effective receptor blockade and internalization. Thus, the VHH–CPP conjugate shows a combination of activities that implement a highly powerful new design principle to block receptor activation by its clearance from the cell surface.     

Effect of solvent type on preparation of ethyl cellulose microparticles by solvent evaporation method with double emulsion system using focused beam reflectance measurement

The purpose of this study was to investigate the effect of solvent type on the solidification rate of ethyl cellulose (EC) microparticles and particle size/distribution of emulsion droplets/hardened microparticles during the solvent evaporation process using focused beam reflectance measurement (FBRM). EC microparticles were prepared with a water‐in‐oil‐in‐water solvent evaporation method using various solvents, including dichloromethane, dichloromethane–methanol (1:1), ethyl acetate and chloroform. The particle size/distribution of the emulsion droplets/hardened microparticles was monitored using FBRM. The morphology of EC microparticles was characterized using scanning electron microscopy (SEM). The transformation of the emulsion droplets into solid microparticles for all solvents occurred within the first 10–90 min. The square weighted mean chord length of EC microparticles prepared using chloroform was smallest, but the chord count was not the highest. The chord length distribution (CLD) measured by FBRM showed that a larger mean particle size gave longer CLD and a lower peak of particle number. SEM data revealed that the morphology of microparticles was influenced by the type of solvent. FBRM can be employed for online monitoring of the shift in the microparticle CLD and detect transformation of emulsion droplets into solid microparticles during the solvent evaporation process. The microparticle CLD and transformation process were strongly influenced by solvent type. © 2017 Society of Chemical Industry     

Steam Drying - Modelling and Applications

ABSTRACT

The concept of steam drying originates from the mid of the last century. However, a broad industrial acceptance of the technique has so far not taken place. The paper deals with modelling the steam drying process and applications of steam drying with in certain industrial sectors where the technique has been deemed to hove special opponunities.

In the modelling scction the mass and heat transfer proceases are described along with equilibrium, capillarity and sorption phenomena occurring in porous materials during the steam drying process. In addition existing models in the literslure are presented.

The applications discussed involve drying of fuels with high moisture contcna, cattle feed exemplified by sugar beet pulp. lumber. paper pulp. paper and sludges.

Steam drying is compared to flue gas drying of biofuels prior to combustion in a boiler. With reference to a current insrallation in Sweden. the exergy losses. as manifested by loss of co-generation cupacity. are discussed. The energy saving potentid when using steam drying of sugar beet pulp as compared to other possible plant configurations is demonstrated.

Mechanical vapour recompression applied to steam drying is analysed with reference to reponed dau from industriul plsnts. Finally. environmcntul advantages when using steam drying are presented.     

Dissolution,Transport and Reaction at a DICY/DGEBA Interface

The curing of an epoxy consisting of the solid hardener dicyandiamide (DICY) and the resin diglycidyl ether of bisphenol A (DGEBA) is studied in a system consisting of a tablet of DICY embedded in liquid DGEBA. Dissolution of DICY within the liquid DGEBA in combination with the transport of dissolved DICY from the tablet border into DGEBA and the chemical reaction of both reactants is studied by scanning Brillouin microscopy and infrared spectroscopy. Scanning Brillouin microscopy demonstrates the spatial and temporal evolution of the static and dynamic hypersonic properties in the course of curing in the vicinity of the DICY tablet. Infrared spectroscopy performed on epoxy pieces extracted from the final sample at different distances from the tablet surface give information about the spatial evolution of the curing process. The results achieved by both techniques are finally combined to yield a better understanding of the curing of DICY-based epoxies, which transform upon curing from strongly heterogeneous systems towards increasingly homogeneous systems.     

Thermal Degradation Kinetics of Carotenoids in Palm Oil

In the present work, a detailed study is performed for carotene thermal degradation in palm oil at four temperatures ranging from 170 to 230 °C. The heating process was carried out with injection of nitrogen, and the samples were collected every 20 min during a total heating period of 140 min. HPLC analysis was conducted to monitor the carotenoids and tocols variations over the heating time at each temperature. The experimental data were then compared to literature data concerning carotenoids thermal degradation. The thermal degradation kinetics of carotenoids in palm oil followed an order superior to 1. The dependence of constant rates with temperature obeyed the Arrhenius relationship. The activation energy for the carotenoids thermal degradation in palm oil was found to be 109.4 kJ/mol.     

Enzymatic hydrolysis of polyester based coatings

The potential of two hydrolytic enzymes, namely a lipase from Thermomyces lanuginosus (TlL) and a cutinase from Humicola insolens (HiC) for hydrolysis of the phthalic acid backbone based polyester coatings was assessed. Two phthalic acid/trimethylolpropane based model substrates resembling the structure of the polyester backbone of coatings were synthesized. Out of both enzymes, only the cutinase was able to hydrolyze both model substrates while the larger substrate was hydrolyzed at a lower rate. The cutinase was also able to hydrolyze a coating (alkyd resin) both in suspension and as dried film. LC–MS analysis of the hydrolysis products released from the coating revealed the presence of oleic acid, its monoglyceride, phthalic acid and 2-((3-((2-((2,3-dihydroxypropoxy)carbonyl)benzoyl)oxy)-2-hydroxypropoxy)carbonyl)benzoic acid. These results indicate that the enzyme was able to hydrolyze the polyester backbone as well as to release fatty acid side chains. Consequently, enzymatic hydrolysis has a potential for the removal of coatings, their recycling or their functionalization.     

Dendrimers or Nanoparticles as Supports for the Design of Efficient and Recoverable Organocatalysts?

The Jørgensen–Hayashi catalyst [(S)‐α,α‐diphenylprolinol trimethylsilyl ether] was grafted onto the surface of two different supports: phosphorus dendrimers (generations 1 to 3) and magnetic, polymer‐coated cobalt/carbon (Co/C) nanobeads. These new supported catalysts displayed high activities and selectivities in the Michael additions of a wide range of aldehydes to different nitroolefins. Moreover, the dendrimer of the third generation displayed excellent recycling abilities since it could be recovered and reused in 7 consecutive runs without loss of activity.