Simple model for the growth behaviour of mixed lecithin-bile salt micelles

Mixed lecithin-bile salt micelles are known to have a cylindrical or worm-like structure. We investigated their shape, length, flexibility and cross-sectional structure using small-angle neutron scattering (SANS). A broad range of sample compositions was studied varying both the total amphiphile concentration and the molar ratio of bile salt (sodium taurochenodeoxycholate, NaTCDC) to lecithin (egg yolk phosphatidylcholine, EYL). The length of the micelles was quantitatively linked to the micellar composition by introducing a simple model. The model takes into account the partitioning of lecithin and bile salt between the bulk, cylindrical parts and the end caps of the micelles. The model also sheds light on the organization of the micelles, both in their cylindrical regions and end caps.     

Droplet formation via flow-through microdevices in Raman and surface enhanced Raman spectroscopy--concepts and applications

This review outlines concepts and applications of droplet formation via flow-through microdevices in Raman and surface enhanced Raman spectroscopy (SERS) as well as the advantages of the approach. Even though the droplet-based flow-through technique is utilized in various fields, the review focuses on implementing droplet-based fluidic systems in Raman and SERS as these highly specific detection methods are of major interest in the field of analytics. With the combination of Raman or SERS with droplet-based fluidics, it is expected to achieve novel opportunities for analytics. Besides the approach of using droplet-based microfluidic devices as a detection platform, the unique properties of flow-through systems for the formation of droplets are capitalized to produce SERS active substrates and to accomplish uniform sample preparation. Within this contribution, previous reported applications on droplet-based flow-through Raman and SERS approaches and the additional benefit with regard to the importance in the field of analytics are considered.     

Bioaffinity magnetic reactor for peptide digestion followed by analysis using bottom-up shotgun proteomics strategy

We report an efficient and streamlined way to improve the analysis and identification of peptides and proteins in complex mixtures of soluble proteins, cell lysates, etc. By using the shotgun proteomics methodology combined with bioaffinity purification we can remove or minimize the interference contamination of a complex tryptic digest and so avoid the time-consuming separation steps before the final MS analysis. We have proved that by means of enzymatic fragmentation (endoproteinases with Arg-C or/and Lys-C specificity) connected with the isolation of specific peptides we can obtain a simplified peptide mixture for easier identification of the entire protein. A new bioaffinity sorbent was developed for this purpose. Anhydrotrypsin (AHT), an inactive form of trypsin with an affinity for peptides with arginine (Arg) or lysine (Lys) at the C-terminus, was immobilized onto micro/nanoparticles with superparamagnetic properties (silica magnetite particles (SiMAG)-Carboxyl, Chemicell, Germany). This AHT carrier with a determined binding capacity (26.8 nmol/mg of carrier) was tested with a model peptide, human neurotensin, and the resulting MS spectra confirmed the validity of this approach.     

Synthesis and characterisation of helical beta-peptide architectures that contain (S)-beta3-HDOPA(Crown Ether) derivatives

A new set of beta-amino acids that carry various crown ether receptors on their side chains of the general formula (S)-beta(3)-HDOPA(crown ether) (HDOPA: homo-3,4-dihydroxyphenylalanine; (crown ether): [15]crown-5 ([15-C-5]), [18]crown-6 ([18-C-6]), [21]crown-7 ([21-C-7]), 1,2-Benzo-[24]crown-8 ([Benzo-24-C-8]) and (R)-Binol-[20]crown-6 ([(R)-Binol-20-C-6])) was prepared. Peptides that are based on these new crowned beta-amino acids combined with (1S,2S)-ACHC (2-aminocyclohexanecarboxylic acid), which is known to be a potent 3(14)-helix inducer, to the hexamer level, with two crowned residues at the i and i+3 positions of the main-chain, were synthesized in solution by stepwise coupling using Boc-N(alpha)-protection (Boc: tert-butoxycarbonyl) and the EDC/HOAt C-activation method. Their conformational analysis was performed by using FTIR absorption, NMR and CD spectroscopy techniques. Our results are in full agreement with a 3(14)-helix conformation.     

Facile access to the pnictocenium ions [Cp*ECl]+ (E = P, As) and [(Cp*)2P]+: chloride ion affinity of Al(OR(F))3

The pnictocenium salts [Cp*PCl]⁺[μCl]^? ( 1 a ), [Cp*PCl]⁺[ClAl(OR^F)₃]^? ( 1 b ), [Cp*AsCl]⁺[ClAl(OR^F)₃]^? ( 2 ), and [(Cp*)₂P]⁺[μCl]^? ( 3 ), in which Cp*=Me₅C₅, μCl=(^FRO)₃Al? Cl? Al(OR^F)₃, and OR^F=OC(CF₃)₃, were prepared by halide abstraction from the respective halopnictines with the Lewis superacid PhF→Al(OR^F)₃. 1 The X‐ray crystal structures of 1 a , 2 , and 3 established that in the half as well as in the sandwich cations the Cp* rings are attached in an η²‐fashion. By using one or two equivalents of the Lewis acid, the two new weakly coordinating anions [μCl]^? and [ClAl(OR^F)₃]^? resulted. They also stabilize the highly reactive cations in PhF or 1,2‐F₂C₆H₄ solution at room temperature. The chloride ion affinities (CIAs) of a range of classical strong Lewis acids were also investigated. The calculations are based on a set of isodesmic BP86/SV(P) reactions and a non‐isodesmic reference reaction assessed at the G3MP2 level.     

Photoelectron spectroscopy of HC4N-

We report the 364-nm photoelectron spectrum of HC(4)N(-). We observe electron photodetachment from the bent X(2)A" state of HC(4)N(-) to both the near-linear X(3)A" and the bent ? (1)A' states of neutral HC(4)N. We observe an extended, unresolved vibrational progression corresponding to X(3)A" ← X(2)A" photodetachment, and we measure the electron affinity (EA) of the X(3)A" state of HC(4)N to be 2.05(8) eV. Photodetachment to the bent ? (1)A' state results in a single intense origin peak at a binding energy of 2.809(4) eV, from which we determine the singlet-triplet splitting (ΔE(ST)) of HC(4)N: 0.76(8) eV. For comparison and to aid in the interpretation of the HC(4)N(-) spectrum, we also report the 364-nm photoelectron spectra of HCCN(-) and DCCN(-). Improved signal-to-noise over the previous HCCN(-) and DCCN(-) photoelectron spectra allows for a more precise determination of the EAs and ΔE(ST)s of HCCN and DCCN. The EAs of HCCN and DCCN are measured to be 2.001(15) eV and 1.998(15) eV, respectively; ΔE(ST)(HCCN) is 0.510(15) eV and ΔE(ST)(DCCN) is 0.508(15) eV. These results are discussed in the context of other organic carbene chains.     

Reagent Precoated Targets for Rapid In-Tissue Derivatization of the Anti-Tuberculosis Drug Isoniazid Followed by MALDI Imaging Mass Spectrometry

Isoniazid (INH) is an important component of front-line anti-tuberculosis therapy with good serum pharmacokinetics but unknown ability to penetrate tuberculous lesions. However, endogenous background interferences hinder our ability to directly analyze INH in tissues. Chemical derivatization has been successfully used to measure isoniazid directly from tissue samples using matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS). MALDI targets were pretreated with trans-cinnamaldehyde (CA) prior to mounting tissue slices. Isoniazid present in the tissues was efficiently derivatized and the INH-CA product measured by MS/MS. Precoating of MALDI targets allows the tissues to be directly thaw-mounted and derivatized, thus simplifying the preparation. A time-course series of tissues from tuberculosis infected/INH dosed animals were assayed and the MALDI MS/MS response correlates well with the amount of INH determined to be in the tissues by high-performance liquid chromatography (HPLC)-MS/MS.     

THE EFFECT OF DIFFERENTIATION ON PHOTOSENSITIZER UPTAKE BY HL60 CELLS

The capability of human promyelocytic leukemia cells HL60 to be induced to differentiate to various stages along the monocytic or myelocytic pathway was exploited for investigation of the uptake of selected photo-sensitizers by diverse types of cells of the same origin. The results showed that there was no substantial difference in photofrin uptake between noninduced HL60 cells, immature monocytes, immature neutrophils and cells differentiated along the eosinophilic pathway. In contrast, HL60 cells differentiated into macrophages (HL609) exhibited markedly increased photofrin uptake, which was further enhanced by their pretreatment with bacterial lipopolysaccharide. Similar results were obtained with other photosensitizers tested: di-and tetrasulfonated aluminum phthalocyanines (AIPcS₂ and AIPcS₄), tetrasulfonated zinc phthalocyanine (ZnPcS₄), tetraphenylporphine tetrasulfonate (TPPS₄) and benzoporphyrin derivative monoacid (BPD). Despite marked differences in the state of self-aggregation and other chemical properties of these compounds, the degree of their preferential uptake by HL60 PH cells showed very little variation. In a typical experiment, the uptake of these photosensitizers by HL60 PH cells was four to five times higher than the uptake by noninduced HL60 cells. In addition to the fluorometric assay employed in most of the experiments, cellular concentration of AlPcS₄ was determined by measurement of elementary aluminum using atomic absorption spectroscopy.     

GC/MS determination of the mutagenic heterocyclic amines MeIQx and DiMeIQx in cooking fumes

Summary 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx) and 2-amino-3,7,8-trimethylimidazo-[4,5-f]quinoxaline (7,8-DiMeIQx) are heterocyclic aromatic amines formed during cooking of meat. The release of these mutagenic and possibly carcinogenic compounds into the ambient air was investigated. Minced beef/pork (1440 g) was fried at 210°C in a cast-iron frying pan. The cooking fumes were led through a glass funnel into a sampler which consisted of two glass fibre filters and an XAD-2 sorbent tube. MeIQx and DiMeIQx were analysed as 3,5-bistrifluoromethylbenzyl derivatives by GC/MS using electron ionisation in selected ion monitoring mode. The results showed that MeIQx and DiMeIQx are volatilised during frying. 0.014 ng/g of MeIQx and 0.007 ng/g of DiMeIQx were found on the filters and the sorbent tube in an air sample of 721. The proportions of 4,8- and 7,8-isomers of DiMeIQx were not determined. The method proved to be sufficiently sensitive (detection limit 0.1 ng/sample for MeIQx) and specific for workplace measurements.