Untersuchungen über Phenthiazinderivate III: Über die Synthese von 10-(Piperazinoalkyl)-phenthiazinen

Zusammenfassung Zum Zwecke einer pharmakologischen Untersuchung wurden weitere 10-(Piperazinoalkyl)-phenthiazine und deren Hydrochloride hergestellt. Anders als bei der Veröffentlichung I dieser Reihe erfolgte die Synthese in vorliegender Arbeit durch Umsetzung von 10-(Haloalkyl)-phenthiazinen mit N-Alkyl-,N-Aryl-und N-Aralkylpiperazinen.     

Untersuchungen über Phenthiazinderivate IV: Über die Synthese von 10-(Piperazinoacyl)-phenthiazinen

Zusammenfassung Die Kondensation von 10-(-Chlorpropionyl)-phenthiazin und 10-(-Bromcaproyl)-phenthiazin mit N-monosubstituierten Piperazinbasen gab acht 10-(Piperazinoacyl)-phenthiazine, deren Dihydrochloride der pharmakologischen Untersuchung zugeführt wurden. An einem dieser Derivate wurde versucht, die Acylgruppe mit LiAlH₄ zu reduzieren.     

Stability evaluation of morphine,hydromorphone, metamizole and esketamine containing analgesic mixtures applied for patient-controlled analgesia in hospice and palliative care

In this study, different injection solutions containing opioid and nonopioid compounds used for patient-controlled analgesia in hospice and palliative care were evaluated in terms of analyte stability. Investigated injection solutions contained different combinations of morphine, hydromorphone, metamizole and esketamine. For the practical implementation, samples from infusion pumps were daily drawn over a period of 7 days at 22 and 37°C. Quantitative measurements were performed on a high-performance liquid chromatography system with ultraviolet detection applying a validated analytical method. All compounds apart from morphine showed no evident changes in concentration. However, a significant loss of morphine was observed for injection mixtures containing both morphine and metamizole at 37°C. After 7 days, only 72% of the initially measured morphine concentration was measured in the binary and 77% in the ternary mixture. Furthermore, an additional compound was detected that could represent the morphine-metamizole-adduct, “metamorphine”. Based on these results, a significantly reduced morphine concentration must be expected after only 3 days if an injection solution mixture containing both morphine and metamizole is administered to a patient at 37°C. Since the analgesic effects of morphine–metamizole adducts have not yet been thoroughly investigated, further clinical studies are necessary before accurate conclusions can be drawn in this regard.     

Cellular interaction of different forms of aluminum nanoparticles in rat alveolar macrophages

Nanomaterials, with dimensions in the 1-100 nm range, possess numerous potential benefits to society. However, there is little characterization of their effects on biological systems, either within the environment or on human health. The present study examines cellular interaction of aluminum oxide and aluminum nanomaterials, including their effect on cell viability and cell phagocytosis, with reference to particle size and the particle's chemical composition. Experiments were performed to characterize initial in vitro cellular effects of rat alveolar macrophages (NR8383) after exposure to aluminum oxide nanoparticles (Al2O3-NP at 30 and 40 nm) and aluminum metal nanoparticles containing a 2-3 nm oxide coat (Al-NP at 50, 80, and 120 nm). Characterization of the nanomaterials, both as received and in situ, was performed using transmission electron microscopy (TEM), dynamic light scattering (DLS), laser Doppler velocimetry (LDV), and/or CytoViva150 Ultra Resolution Imaging (URI)). Particles showed significant agglomeration in cell exposure media using DLS and the URI as compared to primary particle size in TEM. Cell viability assay results indicate a marginal effect on macrophage viability after exposure to Al2O3-NP at doses of 100 microg/mL for 24 h continuous exposure. Al-NP produced significantly reduced viability after 24 h of continuous exposure with doses from 100 to 250 microg/mL. Cell phagocytotic ability was significantly hindered by exposure to 50, 80, or 120 nm Al-NP at 25 microg/mL for 24 h, but the same concentration (25 microg/mL) had no significant effect on the cellular viability. However, no significant effect on phagocytosis was observed with Al2O3-NP. In summary, these results show that Al-NP exhibit greater toxicity and more significantly diminish the phagocytotic ability of macrophages after 24 h of exposure when compared to Al2O3-NP.     

Liquid chromatography-electrochemical detection of ferro- and ferricytochrome c at a chemically modified gold electrode

Chemically modified electrodes, constructed by adsorption of 4,4'-dithiodipyridine onto a polyvinylferrocene-treated gold surface, were employed for the amperometric detection of cytochrome c following size-exclusion chromatography. The electrode response was nearly reversible, permitting quantitation both of the ferro-form of the protein at +0.15 V vs. Ag/AgCl and of the ferri-form at -0.15 V. The limit of detection for the reduced species was 3 pmol injected, and the response was linear over three orders of magnitude. Using the chemically modified electrode approach, cytochrome c monitoring was sufficiently selective that the compound could be determined in human plasma pretreated only by dilution and particulate filtration.     

A Pseudo MS<Superscript>3</Superscript> Approach for Identification of Disulfide-Bonded Proteins: Uncommon Product Ions and Database Search

It has previously been reported that disulfide and backbone bonds of native intact proteins can be concurrently cleaved using electrospray ionization (ESI) and collision-induced dissociation (CID) tandem mass spectrometry (MS/MS). However, the cleavages of disulfide bonds result in different cysteine modifications in product ions, making it difficult to identify the disulfide-bonded proteins via database search. To solve this identification problem, we have developed a pseudo MS³ approach by combining nozzle-skimmer dissociation (NSD) and CID on a quadrupole time-of-flight (Q-TOF) mass spectrometer using chicken lysozyme as a model. Although many of the product ions were similar to those typically seen in MS/MS spectra of enzymatically derived peptides, additional uncommon product ions were detected including c_i-1 ions (the i^th residue being aspartic acid, arginine, lysine and dehydroalanine) as well as those from a scrambled sequence. The formation of these uncommon types of product ions, likely caused by the lack of mobile protons, were proposed to involve bond rearrangements via a six-membered ring transition state and/or salt bridge(s). A search of 20 pseudo MS³ spectra against the Gallus gallus (chicken) database using Batch-Tag, a program originally designed for bottom up MS/MS analysis, identified chicken lysozyme as the only hit with the expectation values less than 0.02 for 12 of the spectra. The pseudo MS³ approach may help to identify disulfide-bonded proteins and determine the associated post-translational modifications (PTMs); the confidence in the identification may be improved by incorporating the fragmentation characteristics into currently available search programs.     

Untersuchungen über Phenthiazinderivate II: Über die Synthese von 10-(Piperazinoacetyl)-phenthiazinen

Zusammenfassung Um weitere Beiträge zur Untersuchung der pharmakologischen Eigenschaften von Phenthiazinen, die als basischen Substituenten in Stellung 10 den Piperazinring enthalten, zu ermöglichen, wurden Verbindungen hergestellt, bei denen die Verknüpfung der beiden Ringsysteme durch die –CO–CH₂–Gruppe erfolgt.     

Scaling behavior of the Hall coefficient of Si:P at the metal-insulator transition

Measurements of the Hall coefficient R _H (T, B) of Si:P with P concentration N between 3.54 and 7.0·10¹⁸ cm^?3 are reported for the temperature range 0.04 K ≤ T ≤ 4K and in magnetic fields up to 7 T. Even far above the metal-insulator transition (MIT), a sign change of the temperature coefficient similar to the behavior of the conductivity σ(T) in moderate fields is not observed in R _H (T). Field and temperature dependence of R _H both increase as the MIT (at the critical concentration N _c = 3.52 · 10¹⁸ cm^?3) is approached. A careful extrapolation to T → 0 and B → 0 indicates that R _H ^?1 scales to zero as R _H ^?1 ～| N ? N _c ^μH with μ _H = (0.44 ± 0.04) in agreement with previous results.     

Unique cellular interaction of silver nanoparticles: size-dependent generation of reactive oxygen species

The rapid advancement of nanotechnology has created a vast array of engineered nanomaterials (ENMs) which have unique physical (size, shape, crystallinity, surface charge) and chemical (surface coating, elemental composition and solubility) attributes. These physicochemical properties of ENMs can produce chemical conditions to induce a pro-oxidant environment in the cells, causing an imbalanced cellular energy system dependent on redox potential and thereby leading to adverse biological consequences, ranging from the initiation of inflammatory pathways through to cell death. The present study was designed to evaluate size-dependent cellular interactions of known biologically active silver nanoparticles (NPs, Ag-15 nm, Ag-30 nm, and Ag-55 nm). Alveolar macrophages provide the first defense and were studied for their potential role in initiating oxidative stress. Cell exposure produced morphologically abnormal sizes and adherence characteristics with significant NP uptake at high doses after 24 h. Toxicity evaluations using mitochondrial and cell membrane viability along with reactive oxygen species (ROS) were performed. After 24 h of exposure, viability metrics significantly decreased with increasing dose (10-75 microg/mL) of Ag-15 nm and Ag-30 nm NPs. A more than 10-fold increase of ROS levels in cells exposed to 50 microg/mL Ag-15 nm suggests that the cytotoxicity of Ag-15 nm is likely to be mediated through oxidative stress. In addition, activation of the release of traditional inflammatory mediators were examined by measuring levels of cytokines/chemokines, including tumor necrosis factor (TNF-alpha), macrophage inhibitory protein (MIP-2), and interleukin-6 (IL-6), released into the culture media. After 24 h of exposure to Ag-15 nm nanoparticles, a significant inflammatory response was observed by the release of TNF-alpha, MIP-2, and IL-1beta. However, there was no detectable level of IL-6 upon exposure to silver nanoparticles. In summary, a size-dependent toxicity was produced by silver nanoparticles, and one predominant mechanism of toxicity was found to be largely mediated through oxidative stress.     

Automation and Standardization—A Coupled Approach towards Reproducible Sample Preparation Protocols for Nanomaterial Analysis

Whereas the characterization of nanomaterials using different analytical techniques is often highly automated and standardized, the sample preparation that precedes it causes a bottleneck in nanomaterial analysis as it is performed manually. Usually, this pretreatment depends on the skills and experience of the analysts. Furthermore, adequate reporting of the sample preparation is often missing. In this overview, some solutions for techniques widely used in nano-analytics to overcome this problem are discussed. Two examples of sample preparation optimization by automation are presented, which demonstrate that this approach is leading to increased analytical confidence. Our first example is motivated by the need to exclude human bias and focuses on the development of automation in sample introduction. To this end, a robotic system has been developed, which can prepare stable and homogeneous nanomaterial suspensions amenable to a variety of well-established analytical methods, such as dynamic light scattering (DLS), small-angle X-ray scattering (SAXS), field-flow fractionation (FFF) or single-particle inductively coupled mass spectrometry (sp-ICP-MS). Our second example addresses biological samples, such as cells exposed to nanomaterials, which are still challenging for reliable analysis. An air–liquid interface has been developed for the exposure of biological samples to nanomaterial-containing aerosols. The system exposes transmission electron microscopy (TEM) grids under reproducible conditions, whilst also allowing characterization of aerosol composition with mass spectrometry. Such an approach enables correlative measurements combining biological with physicochemical analysis. These case studies demonstrate that standardization and automation of sample preparation setups, combined with appropriate measurement processes and data reduction are crucial steps towards more reliable and reproducible data.