Ivy leaves dry extract EA 575® decreases LPS-induced IL-6 release from murine macrophages.

IL-6 plays a key role in the course of inflammatory processes as well as in the regulation of immune responses by the release of different cytokines. IL-6 is produced e.g. by macrophages recruited to the airways in response to a variety of inflammatory stimuli like allergens and respiratory viruses. Patients with inflammatory airway diseases therefore may benefit from therapies targeting the IL-6 pathway, e.g. reduction of the IL-6 release. Within this context, we tested the influence of the ivy leaves dry extract EA 575® on the LPS-induced release of IL-6 from murine macrophages (J774.2). One point seven µg/ml (5 µM) corticosterone served as positive control and was able to reduce LPS-induced IL-6 release by 46 ± 4%. EA 575® was tested in concentrations between 40 and 400 µg/ml. EA 575® decreased the LPS-induced IL-6 release in a dose-dependent manner and statistically significant by 25 ± 4%, 32 ± 4%, and 40 ± 7% in concentrations of 80, 160, and 400 µg/ml, respectively. The present data suggest an anti-inflammatory effect of EA 575® used in therapy of chronic- and acute inflammatory airway diseases accompanied with cough.

Development of a skin-friendly microemulsion for dermal allergen-specific immunotherapy.

Due to their role in immune responses, the skin dendritic cells (i.e. epidermal Langerhans cells and dermal dendritic cells) have become of great interest to researchers in the past decades. A dermal administration of allergens could target these professional antigen-presenting cells directly and build up immunotolerance. Additionally, many of the adverse side effects, which are seen in the current state of the art specific immunotherapy routes, could be avoided. Therefore, in this study a penetration enhancing microemulsion was developed and its physicochemical properties were determined under several storage conditions. The influence of different preservatives on the microemulsion stability was observed. We examined epidermal penetration of Alexa Fluor-647 labelled bee-venom phospholipase A2 (Api m 1) using the Franz diffusion cell set up and confocal laser-scanning microscopy. First results of an in-vivo Api m 1-allergic mouse model indicated the protective efficacy of dermal AIT with our newly developed microemulsion. Summarily, the developed microemulsion is a suitable, stable drug delivery system for the topical administration of proteogenic allergens into the epidermis and is able to reach dendritic cells in the skin.

Alopecia in a novel mouse model RCO3 is caused by mK6irs1 deficiency.

Reduced coat 3 (Rco3) is a new spontaneous autosomal recessive mutation with defects in hair structure and progressive alopecia. Here we describe chromosomal mapping and molecular identification of the Rco3 mutation. The murine Rco3 locus maps to a 2-Mb interval on chromosome 15 encompassing the keratin type II gene cluster. Recently, mK6irs1 was described as a type II keratin expressed in Henle's and Huxley's layer of the murine inner root sheath. Genomic sequencing revealed a 10-bp deletion in exon 1 of mK6irs1 resulting in a frameshift after 58 amino acid residues and, therefore, the absence of 422 carboxy-terminal amino acid residues containing the complete alpha-helical rod domain. Henle's and Huxley's layers show no immunoreactivity with mK6irs1-specific antibodies and the absence of intermediate filament formation in electron microscopic images. These results indicate that the expression of functional mK6irs1 is indispensable for intermediate filament formation in the inner root sheath and highlights the importance of the keratinization of the inner root sheath in the normal formation of the hair shaft.

Pharmacological properties of naturally occurring variants of the human norepinephrine transporter.

The human norepinephrine transporter (hNET) gene has five sequence polymorphisms that predict amino acid substitutions in the transporter protein: Val69Ile, Thr99Ile, Val245Ile, Val449Ile, and Gly478Ser. In order to functionally characterize the naturally occurring transporter variants, we used site-directed mutagenesis to establish the hNET variants and compared some basic pharmacological properties (uptake of norepinephrine and its inhibition by the tricyclic antidepressant desipramine) in COS-7 cells transiently expressing variant hNETs and wild-type hNET. None of the hNET variants displayed changes in the potency (Ki) of desipramine for inhibition of norepinephrine uptake. Furthermore, variants Val69Ile, Thr99Ile, ValZ45Ile, and Val449Ile did not affect kinetic constants (Km, Vmax) of norepinephrine uptake. However, COS-7 cells expressing the hNET variant Gly478Ser displayed an approximately four-fold increase in the Km for norepinephrine, while the Vmax was unaffected. The increase in the Km, which is equivalent to a four-fold reduction in the affinity of the variant hNET for its natural substrate norepinephrine, indicates that the glycine in position 478 is part of a substrate recognition domain. The reduced clearance of released norepinephrine by reuptake through the Gly478Ser variant might cause an increase in the synaptic and the circulating concentration of norepinephrine. Elevated norepinephrine concentrations have been associated with human diseases and it will be interesting to explore a possible contribution by the Gly478Ser variant to certain disease states.

[Treatment of chronic bronchitis with ivy leaf special extract--multicenter post-marketing surveillance study in 1,350 patients]. / Behandlung chronischer Bronchitis mit einem Spezialextrakt aus Efeublättern - multizentrische Anwendungsbeobachtung mit 1350 Patienten.

BACKGROUND AND OBJECTIVES: The changes of clinical symptoms and the tolerability of Prospan((R)) acute Effervescent Cough Tablets were investigated in a multicenter, prospective post-marketing surveillance (PMS) study focusing on patients with chronic bronchitis. PATIENTS AND METHODS: The study included 1,350 male and female patients aged 4 years and above who were treated in one of 135 participating medical practices and who suffered from chronic bronchitis (with or without airway obstruction). During a scheduled observational period of 4 weeks, the patients had to take 1(1/2) or 2 tablets per day (depending on their age), according to the manufacturer's dosing recommendations, corresponding to 97.5 or 130 mg of dried ivy leaf extract (about 585- 780 mg of drug). Treatment success was assessed by observing the changes in the direct symptoms of chronic bronchitis between the baseline examination and the end of treatment. Safety was evaluated by analyzing adverse events. RESULTS: In comparison to baseline, the following percentages of patients showed improved symptoms or were cured at treatment end: cough 92.2%; expectoration 94.2%; dyspnea 83.1%; respiratory pain 86.9%. In each of the four symptoms at least 38% of the initially affected patients were completely free of complaints. Three patients (0.2%) experienced adverse events in which a causal relationship to the drug under investigation could not be excluded (2x eructation, 1x nausea). CONCLUSIONS: Considering the favorable changes in all investigated clinical symptoms as well as the excellent tolerability in children and adults, the ivy leaf extract preparation Prospan acute Effervescent Cough Tablets can be considered as a therapeutic option in alleviating the symptoms of chronic bronchitis.

Development and validation of an alternative disturbed skin model by mechanical abrasion to study drug penetration.

Pharmaceuticals and cosmetics for dermal application are usually tested on healthy skin, although the primary permeation barrier, the stratum corneum, is often impaired by skin diseases or small skin lesions, especially on the hands. These skin conditions can considerably influence the permeation of chemicals and drugs. Furthermore, risk assessment for example of nanoparticles should be performed under various skin conditions to reflect the true circumstances. Therefore, an alternative and reproducible method for a high throughput of skin samples with impaired skin barrier was developed and verified by skin permeation studies (25 h) of caffeine, sorbic acid and testosterone compared to healthy (untreated) and tape-stripped skin. Skin barrier disruption was controlled by TEWL measurement. Skin permeation of the three substances was increased in tape-stripped and abraded skin compared to untreated skin due to the reduced barrier integrity. Enhancement of drug uptake was highest for the most hydrophilic substance, caffeine, followed by sorbic acid and lipophilic testosterone. No significant difference in drug uptake studies was observed between the new abrasion method with an aluminum-coated sponge and the tape-stripping method. The obtained results demonstrate that this abrasion method is an alternative way to achieve a disturbed skin barrier for drug and chemical uptake studies.

Development of multiple W/O/W emulsions as dermal carrier system for oligonucleotides: effect of additives on emulsion stability.

Multiple water-in-oil-in-water (W/O/W) emulsions are of major interest as potential skin delivery systems for water-soluble drugs like oligonucleotides due to their distinct encapsulation properties. However, multiple emulsions are highly sensitive in terms of variations of the individual components. The presence of osmotic active ingredients in the inner water phase is crucial for the generation of stable multiple emulsions. In order to stabilize the emulsions the influence of NaCl, MgSO(4), glucose and glycine and two cellulose derivatives was investigated. Briefly, multiple W/O/W emulsions using Span 80 as a lipophilic emulsifier and different hydrophilic emulsifiers (PEG-40/50 stearate, steareth-20 and polysorbate 80) were prepared. Stability of the emulsions was analyzed over a period of time using rheological measurements, droplet size observations and conductivity analysis. In this study we show that additives strongly influence the properties stability of multiple emulsions. By increasing the concentration of the osmotic active ingredients, smaller multiple droplets are formed and the viscosity is significantly increased. The thickening agents resulted in a slightly improved stability. The most promising emulsions were chosen and further evaluated for their suitability and compatibility to incorporate a DNAzyme oligonucleotide as active pharmaceutical ingredient.

[Validity of clinical trials: are there differences between conventional and complementary alternative medicine?]. / Studienvalidität: Gibt es Unterschiede zwischen Schul- und Komplementärmedizin?

So far there has been no consensus on the criteria which confirm the validity of scientific contributions in conventional medicine (CM) and complementary/ alternative medicine (CAM). An interdisciplinary group of experts from various disciplines within each of the areas of medicine held six well-documented sessions in an effort to reach a consensus. The group agreed that the methods to confirm the validity of clinical trials are identical in CM and CAM. There are differences in research strategies and there may also be differences in interpreting the results, depending on the concept of medicine.

Influence of hydrophilic surfactants on the properties of multiple W/O/W emulsions.

Multiple W/O/W emulsions for topical application using Span 80 as a lipophilic emulsifier were prepared. Several hydrophilic emulsifiers were tested in respect of their suitability for the preparation of multiple emulsions. In addition, the effect of different oil-phase compositions on emulsion stability was investigated. The physicochemical parameters of the formulations were characterized and their long-term stability was evaluated by means of rheological measurements, droplet size observations and conductivity analysis. As discovered, the modification of an oil-phase composition results in a decrease in the diffusion coefficient of water and water-soluble substances and, consequently, in enhanced stability. The influence of the release of electrolytes from the inner to the outer water phase on the emulsion stability behaviour was investigated. It was found, that the effect of the hydrophilic emulsifiers on the formulation properties is related not only to its HLB value, but rather to its chemical composition. As a result, polyethoxylated ethers of fatty alcohols (C=16-18) with HLBs between 15.3 and 16.2 appear to be the most suitable ones for creating stable formulations.

The human desipramine-sensitive noradrenaline transporter and the importance of defined amino acids for its function.

1. This article gives a short overview of the physiology, pharmacology and the molecular biology of the human Na+/Cl(-)-dependent noradrenaline transporter (hNAT) and its gene. 2. Furthermore, naturally occurring variants of the hNAT are described and new results obtained through site-directed mutagenesis of the hNAT are presented, which increase our understanding about structural domains and amino acids critically involved in substrate, cosubstrate and inhibitor binding to the hNAT.

Studies with clonidine (dixarit) in menopausal women.

Eleven women with menopausal symptoms were treated with 150 micrograms Clonidine (dixarit) daily. Before and during therapy, plasma estradiol-17 beta, LH, FSH and prolactin levels were measured by specific radio-immunoassays. In addition, each patient recorded the number of hot flushes a day and symptoms were monitored by the Kupperman-index. A highly significant fall in the number of hot flushes by day and night occurred during therapy (p less than 0.001) and the Kupperman-index similarly improved. Significant pulse and blood pressure changes were not noted and plasma hormone concentrations remained unaltered. Medication was well tolerated. Clonidine therapy would appear to be the treatment of choice for menopausal symptoms if estrogens are contraindicated.

Pharmacokinetic and -dynamic investigations on HL60 cells with a new protoporphyrin dimethyl ester hematoporphyrin dimer.

Pharmacokinetic and -dynamic studies using a novel porphyrin dimer were performed in human line HL60 promyelocytic leukemia cells. Uptake of the dimer into leukemic cells was observed to occur at substantially lower concentrations in comparison to a previously described monomeric dihematoporphyrin ether-free hematoporphyrin derivative. Both dimer and monomer derivatives could be demonstrated to inhibit cell growth, with no remarkable quantitative differences being found in cell proliferation studies regarding [3H]-thymidine incorporation and assay for colony formation. Structurally, the new compound represents an unsymmetrically substituted diethyl ether having protoporphyrin dimethyl ester and hematoporphyrin as substituents. For this reason the compound was designated as protoporphyrin dimethyl ester hematoporphyrin ether.

The dominant alopecia phenotypes Bareskin, Rex-denuded, and Reduced Coat 2 are caused by mutations in gasdermin 3.

Reduced Coat 2 (Rco2) is an ENU-induced mutation affecting hair follicle morphogenesis by an abnormal and protracted catagen. We describe chromosomal mapping and molecular identification of the autosomal dominant Rco2 mutation. The Rco2 critical region on mouse chromosome 11 encompasses the alopecia loci, Bareskin (Bsk), Rex-denuded (Re(den)), Recombination induced mutation 3 (Rim3), and Defolliculated (Dfl). Recently, the gasdermin (Gsdm) gene was described as predominantly expressed in skin and gastric tissues. We provide evidence for a murine-specific gene cluster consisting of Gsdm and two closely related genes which we designate as Gsdm2 and Gsdm3. We show that Gsdm3 reflects a mutation hotspot and that Gsdm3 mutations cause alopecia in Rco2, Re(den), and Bsk mice. We infer a role of Gsdm3 during the catagen to telogen transition at the end of hair follicle morphogenesis and the formation of hair follicle-associated sebaceous glands.