Whey Protein/Polysaccharide-Stabilized Emulsions: Effect of Polymer Type and pH on Release and Topical Delivery of Salicylic Acid

Emulsions are widely used as topical formulations in the pharmaceutical and cosmetic industries. They are thermodynamically unstable and require emulsifiers for stabilization. Studies have indicated that emulsifiers could affect topical delivery of actives, and this study was therefore designed to investigate the effects of different polymers, applied as emulsifiers, as well as the effects of pH on the release and topical delivery of the active. O/w emulsions were prepared by the layer-by-layer technique, with whey protein forming the first layer around the oil droplets, while either chitosan or carrageenan was subsequently adsorbed to the protein at the interface. Additionally, the emulsions were prepared at three different pH values to introduce different charges to the polymers. The active ingredient, salicylic acid, was incorporated into the oil phase of the emulsions. Physical characterization of the resulting formulations, i.e., droplet size, zeta potential, stability, and turbidity in the water phase, was performed. Release studies were conducted, after which skin absorption studies were performed on the five most stable emulsions, by using Franz type diffusion cells and utilizing human, abdominal skin membranes. It was found that an increase in emulsion droplet charge could negatively affect the release of salicylic acid from these formulations. Contrary, positively charged emulsion droplets were found to enhance dermal and transdermal delivery of salicylic acid from emulsions. It was hypothesized that electrostatic complex formation between the emulsifier and salicylic acid could affect its release, whereas electrostatic interaction between the emulsion droplets and skin could influence dermal/transdermal delivery of the active.     

Carbohydrate microarrays reveal sulphation as a modulator of siglec binding

Siglecs are receptors on cells of the immune, haemopoietic, and nervous systems that recognize sialyl-glycans with differing preferences for sialic acid linkage and oligosaccharide backbone sequence. We investigate here siglec binding using microarrays of Lewis(x) (Le(x))- and 3'-sialyl-Le(x)-related probes with different sulphation patterns. These include sulphation at position 3 of the terminal galactose of Le(x), position 6 of the galactose of Le(x) and sialyl-Le(x), position 6 of N-acetylglucosamine of Le(x) and sialyl-Le(x), or both positions of sialyl-Le(x). Recombinant soluble forms of five siglecs have been investigated: human Siglec-7, -8, -9, and murine Siglec-F and CD22 (Siglec-2). Each siglec has a different binding pattern. Unlike two C-type lectins of leukocytes, L-selectin and Langerin, which also bind to sulphated analogues of sialyl-Le(x), the siglecs do not give detectable binding signals with sulphated analogues that are lacking sialic acid. The sulphate groups modulate, however, positively or negatively the siglec binding intensities to the sialyl-Le(x) sequence.     

Comparative kinetic studies of the dealkylation reaction and steroid hydroxylations in various oxygen and carbon monoxide:oxygen atmospheres

The time-course kinetics of the cytochrome P-450-catalyzed dealkylations of the exogenous compounds benzphetamine, ethylmorphine, codeine, and 7-ethoxycoumarin were compared to the hydroxylation of the endogenous compound testosterone. Using liver microsomes from phenobarbital-induced rats, the time course of the demethylations of ethylmorphine, codeine, and especially benzphetamine was characterized by a fast initial phase of enzymatic activity and then a steady decline in the rate throughout the remainder of the reaction. In contrast, under the same experimental conditions, both the dealkylation of 7-ethoxycoumarin and the hydroxylation of testosterone showed no initial fast phase of activity and a constant rate of product formation for most of the remainder of the time course. The difference also held for the carbon monoxide inhibition studies in which the degree of inhibition of the demethylation reactions by a variety of CO:O₂ mixtures was time dependent, in contrast to the constant, time-independent degree of CO inhibition of the other two reactions. The kinetics of the demethylation reactions could not be explained by enzyme destruction, back reaction, or product adduct formation and were further confirmed by measurements of the rate of O₂ utilization and NADPH oxidation. The complexity of the demethylation reaction should be taken into consideration in any detailed studies of the monooxygenation reaction system.     

Thalictricoside, a new phenolic compound from Thalictrum orientale

From the underground parts of Thalictrum orientale Boiss., a new phenolic compound 1 was isolated in addition to one known cyanoglycoside, lithospermoside (2). For the structure elucidation of all compounds, 1D- and 2D-NMR techniques (DEPT, COSY, HMBC, HSQC) and MS (HR-MALDI) were used. The structure of the new compound was established as 2-(4'-hydroxyphenyl)-nitroethane-4'-O-[beta-xylopyranosyl-(1 --> 6)-beta-glucopyranoside] (1).     

EVOLUTIONARILY STABLE SEX RATIOS AND MUTATION LOAD

Frequency‐dependent selection should drive dioecious populations toward a 1:1 sex ratio, but biased sex ratios are widespread, especially among plants with sex chromosomes. Here, we develop population genetic models to investigate the relationships between evolutionarily stable sex ratios, haploid selection, and deleterious mutation load. We confirm that when haploid selection acts only on the relative fitness of X‐ and Y‐bearing pollen and the sex ratio is controlled by the maternal genotype, seed sex ratios evolve toward 1:1. When we also consider haploid selection acting on deleterious mutations, however, we find that biased sex ratios can be stably maintained, reflecting a balance between the advantages of purging deleterious mutations via haploid selection, and the disadvantages of haploid selection on the sex ratio. Our results provide a plausible evolutionary explanation for biased sex ratios in dioecious plants, given the extensive gene expression that occurs across plant genomes at the haploid stage.     

The HIF-1 response to simulated ischemia in mouse skeletal muscle cells neither enhances glycolysis nor prevents myotube cell death

Hypoxia-inducible factor (HIF) plays an important role in regulating gene expression in response to ischemia. Although activation of HIF-1 in muscle tissue was found during ischemia in vivo, the meaning and mechanisms in isolated cells are still incompletely understood. We studied activation of HIF-1 in skeletal muscle cells cultured in either their undifferentiated myoblast state or differentiated into myotubes. HIF-1 was activated in myoblasts and myotubes by hypoxia and simulated ischemia. Induction of adrenomedullin mRNA and, to a lesser extent, VEGF mRNA correlated well with the induction of HIF-1alpha protein in both cell types. Enzymes of glycolysis-like lactate dehydrogenase and pyruvate kinase showed upregulation of their mRNA only under hypoxic conditions but not during simulated ischemia. Phosphofructokinase mRNA showed no significant upregulation at all. Although HIF-1 was activated in myotubes during simulated ischemia, myotubes died preceded by a loss of ATP. Myoblasts survived simulated ischemia with no decrease in ATP or ATP turnover. Furthermore, pharmacological inhibition of HIF-1 hydroxylases by dimethyloxalylglycine (DMOG) increased HIF-1alpha accumulation and significantly upregulated the expression of adrenomedullin, VEGF, lactate dehydrogenase, and pyruvate kinase in myoblasts and myotubes. However, DMOG provided no protection from cell death. Our data indicate that HIF-1, although activated in myotubes during simulated ischemia, cannot protect against the loss of ATP and cell viability. In contrast, myoblasts survive ischemia and thus may play an important role during regeneration and HIF-1-induced revascularization.     

Zur Kenntnis der Mitose. XIII

Zusammenfassung In Ergänzung zu allen bisherigen Befunden über die von W. v. Möllendorff aufgefundene spezifische Mitosenstörung sind weitere Substanzen auf ihre Mitosenschädlichkeit in Kulturen von Kaninchenfibrocyten geprüft worden. Es gelangten zunächst Cholesterin und Cholestenon als zwei Steroide zur Untersuchung, denen keine geschlechtsspezifische Wirkung zugeschrieben werden kann. Beide haben sich als mitosenunwirksam erwiesen. Diese Tatsache ist von besonderer Bedeutung. Sie stimmt erstens mit A. H. Roffos Angaben überein, welche das Cholesterin von der tumorigenen Seite her betrachten und an und für sich als unschädlich bezeichnen. Nur tiefgreifende chemische Veränderungen, z. B. durch Oxydation, vermögen die harmlose Substanz in eine carcinogene umzuwandeln. Zweitens muß darauf aufmerksam gemacht werden, daß sowohl Cholesterin als auch Cholestenon mit je einer Doppelbindung im Phenanthrenkern ungesättigte Substanzen darstellen, die trotzdem unwirksam sind. Somit darf der Befund, den W. v. Möllendorff für die männlichen Steroide geltend gemacht hat, nicht verallgemeinert werden. Von Interesse wäre die Beantwortung der Frage, ob stärker dehydrierte Cholesterinpräparate die Wachstumsschädigung bewirken können.Die Ergebnisse mit Stilböstrol passen sehr gut in das Bild, welches aus anderen Forschungen über die Wirksamkeit dieses Präparates gewonnen wurde. Übertrifft das Stilböstrol die natürlichen Hormone schon in seiner hormonalen Wirkung, so zeigt es auch im Tierversuch (bei krebsbereiten Mäusestämmen) eine stärkere tumorigene Potenz. In unseren Versuchen ruft es ebenso in entschieden überlegener Weise die spezifische Wachstumsstörung hervor, zu welcher sich zusätzlich noch eine weitere Giftwirkung hinzugesellt. Diese zweite Giftwirkung greift tief in den eigentlichen Ablaufmechanismus der Mitose ein und beginnt bereits schon in der frühen Metaphase. Dadurch gestaltet sich der Vergiftungstypus des Stilböstrols umfangreicher und ist schwerer zu analysieren als der Störungstyp bei den Steroiden.Endlich sind auch einige cis- und trans-Formen sonst gleich konstituierter Steroide untersucht worden. Die Wachstumsstörung ist nachweisbar unabhängig von der Baueigentümlichkeit der Substanzen. Oben haben wir schon erwähnt, daß dieser Umstand die bereits von W. v. Möllendorff betonte Tatsache unterstreicht, daß die Wachstumsstörung eine hormonunspezifische Nebenwirkung ist.     

Beitrag zum funktionellen Bau des hyalinen Knorpels (auf Grund von Untersuchungen im polarisierten Lichte)

Zusammenfassung Es wurde der Verlauf der interterritorialen kollagenen Fibrillenzüge in verschiedenen Knorpeln des erwachsenen Menschen, einiger Säugetiere (Pferd, Rind, Hund, Kaninchen) und eines Haifisches mittels des Polarisationsmikroskopes untersucht.Alle untersuchten Knorpel zeigen das gleiche Konstruktionsprinzip: zwei Systeme von S-förmigen Fibrillenzügen (vgl. Schema Abb. 1b, S. 282 und Abb. 4a, S. 288), welche, sich unter einem Winkel von durchschnittlich 30–45° kreuzend, den Knorpel auf Quer- und Längsschnitten schräg durchziehen und beiderseits in das Perichondrium einbiegen.Dieses Verhalten entspricht demjenigen, welches zuerst von H. Bormuth für verschiedene Selachierknorpel beschrieben worden ist. Im übrigen werden die Ansichten verschiedener Autoren über den funktionellen Bau des Knorpels diskutiert, und besonders wird auf die Arbeiten von A. Benninghoff eingetreten.Das architektonische Prinzip mit den schräg verlaufenden, sich überkreuzenden Fibrillensystemen ist vielseitiger als das ältere Schema mit den senkrecht zur Oberfläche angeordneten Fibrillenzügen; zudem vermittelt es mit einem geringeren Materialaufwand eine größere Festigkeit.     

Distance models as a tool for modelling detection probability and density of native bumblebees

Effective monitoring of native bee populations requires accurate estimates of population size and relative abundance among habitats. Current bee survey methods, such as netting or pan trapping, may be adequate for a variety of study objectives but are limited by a failure to account for imperfect detection. Biases due to imperfect detection could result in inaccurate abundance estimates or erroneous insights about the response of bees to different environments. To gauge the potential biases of currently employed survey methods, we compared abundance estimates of bumblebees (Bombus spp.) derived from hierarchical distance sampling models (HDS) to bumblebee counts collected from fixed‐area net surveys (“net counts”) and fixed‐width transect counts (“transect counts”) at 47 early‐successional forest patches in Pennsylvania. Our HDS models indicated that detection probabilities of Bombus spp. were imperfect and varied with survey‐ and site‐covariates. Despite being conspicuous, Bombus spp. were not reliably detected beyond 5 m. Habitat associations of Bombus spp. density were similar across methods, but the strength of association with shrub cover differed between HDS and net counts. Additionally, net counts suggested sites with more grass hosted higher Bombus spp. densities whereas HDS suggested that grass cover was associated with higher detection probability but not Bombus spp. density. Density estimates generated from net counts and transect counts were 80%–89% lower than estimates generated from distance sampling. Our findings suggest that distance modelling provides a reliable method to assess Bombus spp. density and habitat associations, while accounting for imperfect detection caused by distance from observer, vegetation structure, and survey covariates. However, detection/non‐detection data collected via point‐counts, line‐transects and distance sampling for Bombus spp. are unlikely to yield species‐specific density estimates unless individuals can be identified by sight, without capture. Our results will be useful for informing the design of monitoring programs for Bombus spp. and other pollinators.