Viscosity reduction of isotonic solutions of the photosensitizer TPCS2a by cyclodextrin complexation

Meso-tetraphenyl chlorin disulphonate (TPCS_2a) is a photosensitizer (PS) particularly developed and patented for use in the technology of photochemical internalization (PCI) against cancer. TPCS_2a is known to aggregate in aqueous media even at low concentrations (≥0.1?µM) and to form a high-viscosity network at clinically relevant concentrations (mM). The aim of this work was to evaluate the effect of two hydroxypropylated cyclodextrin derivatives of beta and gamma type, respectively i.e. HPβCD and HPγCD, on the aggregation and solubilization of TPCS_2a in isotonic solutions. Samples containing micromolar concentrations of TPCS_2a were studied spectrophotometrically, while samples containing a clinical relevant concentration (10?mM?=?9?mg/ml) of TPCS_2a were evaluated by dynamic viscosity measurements. HPβCD was determined to be a more suitable solubilizer of TPCS_2a than HPγCD in aqueous media both in the absence and presence of salt. The complexation stoichiometry between TPCS_2a/HPβCD at micromolar to millimolar concentrations of TPCS_2a was determined to be 1:3 and 1:2 in the absence and presence of isotonic NaCl, respectively. The network of TPCS_2a (10?mM) was broken down in the presence of 3% w/v (=?20?mM) HPβCD, i.e. a 1:2 molar ratio between TPCS_2a and the cyclodextrin. Formation of the inclusion complex resulted in low viscosity samples both in water and in the presence of isotonic NaCl or phosphate buffered saline (PBS) at 25?°C and 37?°C.     

Using the past to indicate the possible presence of relics in the present-day landscape: the Western Front of the Great War in Belgium

The First World War (WWI) had a notable influence on the landscape at the former Western Front in Belgium. Research on a landscape scale is necessary to understand the destructive and constructive impact of the war and its consequences for the post-war landscape in a holistic manner. This paper focuses on the trajectory and impact analysis of three study areas with contrasting landscape types and aims to indicate possible preserved military relics today. Therefore, landscape changes are studied using historical aerial photographs (WWI and WWII) and contemporary orthophotos. A military landscape characterisation is made based on land use/land cover and linear structures that were mapped throughout time. Specific landscape trajectories could be designated as areas with a possible preservation of WWI heritage, with a related impact degree. The results are useful for sustainable heritage management and for further interdisciplinary research on WWI-heritage, by providing a broad knowledge of the area.     

Classification of Amyloidosis by Model-Assisted Mass Spectrometry-Based Proteomics

Amyloidosis is a rare disease caused by the misfolding and extracellular aggregation of proteins as insoluble fibrillary deposits localized either in specific organs or systemically throughout the body. The organ targeted and the disease progression and outcome is highly dependent on the specific fibril-forming protein, and its accurate identification is essential to the choice of treatment. Mass spectrometry-based proteomics has become the method of choice for the identification of the amyloidogenic protein. Regrettably, this identification relies on manual and subjective interpretation of mass spectrometry data by an expert, which is undesirable and may bias diagnosis. To circumvent this, we developed a statistical model-assisted method for the unbiased identification of amyloid-containing biopsies and amyloidosis subtyping. Based on data from mass spectrometric analysis of amyloid-containing biopsies and corresponding controls. A Boruta method applied on a random forest classifier was applied to proteomics data obtained from the mass spectrometric analysis of 75 laser dissected Congo Red positive amyloid-containing biopsies and 78 Congo Red negative biopsies to identify novel “amyloid signature” proteins that included clusterin, fibulin-1, vitronectin complement component C9 and also three collagen proteins, as well as the well-known amyloid signature proteins apolipoprotein E, apolipoprotein A4, and serum amyloid P. A SVM learning algorithm were trained on the mass spectrometry data from the analysis of the 75 amyloid-containing biopsies and 78 amyloid-negative control biopsies. The trained algorithm performed superior in the discrimination of amyloid-containing biopsies from controls, with an accuracy of 1.0 when applied to a blinded mass spectrometry validation data set of 103 prospectively collected amyloid-containing biopsies. Moreover, our method successfully classified amyloidosis patients according to the subtype in 102 out of 103 blinded cases. Collectively, our model-assisted approach identified novel amyloid-associated proteins and demonstrated the use of mass spectrometry-based data in clinical diagnostics of disease by the unbiased and reliable model-assisted classification of amyloid deposits and of the specific amyloid subtype.     

Mucoadhesive Electrospun Nanofiber-Based Hybrid System with Controlled and Unidirectional Release of Desmopressin

The sublingual mucosa is an attractive route for drug delivery, although challenged by a continuous flow of saliva that leads to a loss of drug by swallowing. It is of great benefit that drugs absorbed across the sublingual mucosa avoid exposure to the harsh environment of the gastro-intestinal lumen; this is especially beneficial for drugs of low physicochemical stability such as therapeutic peptides. In this study, a two-layered hybrid drug delivery system was developed for the sublingual delivery of the therapeutic peptide desmopressin. It consisted of peptide-loaded mucoadhesive electrospun chitosan/polyethylene oxide-based nanofibers (mean diameter of 183 ± 20 nm) and a saliva-repelling backing film to promote unidirectional release towards the mucosa. Desmopressin was released from the nanofiber-based hybrid system (approximately 80% of the loaded peptide was released within 45 min) in a unidirectional manner in vitro. Importantly, the nanofiber–film hybrid system protected the peptide from wash-out, as demonstrated in an ex vivo flow retention model with porcine sublingual mucosal tissue. Approximately 90% of the loaded desmopressin was retained at the surface of the ex vivo porcine sublingual mucosa after 15 min of exposure to flow rates representing salivary flow.     

Reduction of immunoreactivity of bovine beta-lactoglobulin upon combined physical and proteolytic treatment

Bovine beta-lactoglobulin was hydrolyzed with trypsin or chymotrypsin before, during and after treatment at 600 MPa and pH 6.8 for 10 min at 30, 37 and 44 degrees C. The extent of beta-lactoglobulin hydrolysis under pressure was noticeably higher than at atmospheric pressure, particularly when chymotrypsin was used. Addition of proteases at ambient pressure to previously pressure-treated beta-lactoglobulin gave only a modest increase in proteolysis with respect to the untreated protein. Products of enzyme hydrolysis under pressure were separated by reverse-phase HPLC, and were found to be different from those obtained at atmospheric pressure when chymotrypsin was used. The residual immunochemical reactivity of the products of combined pressure-enzyme treatment was assessed on the unresolved hydrolysates by ELISA tests using polyclonal and monoclonal antibodies, and on individual hydrolytic fractions by Western Blotting using sera of paediatric patients allergic to whey proteins in cow milk. The immunoreactivity of the whole hydrolysates was related to their content of residual intact beta-lactoglobulin, and no immunochemical reactivity was found for all the products of chymotrypsin hydrolysis under pressure. The results indicate that chymotrypsin effectively hydrolysed hydrophobic regions of beta-lactoglobulin that were transiently exposed during the pressure treatments and that were not accessible in the native protein or in the protein that had been previously pressure treated.     

Applications of real number theorem proving in PVS

Real number theorem proving has many uses, particularly for verification of safety critical systems and systems for which design errors may be costly. We discuss a chain of developments building on real number theorem proving in PVS. This leads from the verification of aspects of an air traffic control system, through work on the integration of computer algebra and automated theorem proving to a new tool, NRV, first presented here that builds on the capabilities of Maple and PVS to provide a verified and automatic analysis of Nichols plots. This automates a standard technique used by control engineers and greatly improves assurance compared with the traditional method of visual inspection of the Nichols plots.     

Studies on curcumin and curcuminoids XVII. Variation in the content of curcuminoids inCurcuma longa L. from Nenal during one season

Summary The content of curcuminoids inCurcuma longa L. before, during and after a normal harvesting period was investigated. The plant material was grown in the Kathmandu area. Bis-demethoxycurcumin was found to be the main constituent. No significant change in the curcuminoid content was observed during a 17-week sampling period.

---

Studien über Curcumin und Curcuminoide. XVII. Veränderung des Gehaltes an Curcuminoiden inCurcuma longa L. (Zingiberaceae) aus Nepal warend der Ernte

Zusammenfassung Der Gehalt an Curcuminoiden inCurcuma longa L. (Zingiberaceae) wurde vor, während und nach der normalen Erntezeit untersucht. Das Pflanzenmaterial stammt aus dim Kathmandu-Tal in Nepal. Der Hauptinhaltsstoff ist Bisdemethoxycurcumin. Im Laufe von 17 Wochen konnten keine signifikante Schwankungen observiert werden.

     

A GC–MS-based metabonomic investigation of blood serum from irritable bowel syndrome patients undergoing intervention with acidified milk products

In the present study, the use of gas chromatography mass spectrometry (GC–MS)-based metabonomics to characterize blood serum in an intervention study of patients suffering from the common gastrointestinal disorder irritable bowel syndrome (IBS) was investigated. The patients included in the study consumed an acidified milk product with (n = 30) or without probiotics (n = 31) (Lactobacillus paracasei F19, Lactobacillus acidophilus LA-5 and Bifidobacterium lactis BB-12) for an 8-week period, and blood serum samples were collected before and after the intervention. Acidified milk is commonly used as a delivering vector for probiotics in commercial consumer settings. The serum samples were extracted and derivatized using N-Methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA), and GC–MS analysis was carried out. Multivariate data analysis including principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), and S-plot was applied on the obtained GC–MS data, which revealed higher serum lactate, glutamine, proline creatinine/creatine, and aspartic acid levels and lower serum glucose levels after the intervention period for both treatment groups. Consequently, the present study indicated an effect of acidified milk consumption on the plasma metabolite profile, which was independent of a concomitant intake of probiotics. In addition, the present study demonstrates that GC–MS is a useful analytical technique for metabonomics studies of blood serum.