Topical delivery of interleukin‐13 antisense oligonucleotides with cationic elastic liposome for the treatment of atopic dermatitis

Background

Interleukin (IL)‐13, overproduced in the skin of atopic dermatitis (AD), has been shown to play an essential role in the pathogenesis of the disease. Thus, inhibition of IL‐13 production should provide a key step to alleviate disease conditions of the atopic skin. In the present study, IL‐13 antisense oligonucleotide (ASO) was designed and formulated with cationic elastic liposome (cEL) to improve transdermal delivery.

Methods

ASOs were generated against murine IL‐13 mRNA (+4 to + 23) and complexed with cEL. Physicochemical properties of IL‐13 ASO/cEL complex were examined by DNA retardation and DNase I protection assay. An in vitro inhibition study was performed in T‐helper 2 (Th2) cells and cytotoxicity was tested by the XTT assay. The in vivo effect of IL‐13 ASO/cEL complex was tested in a murine model of AD.

Results

In vitro, the IL‐13 ASO/cEL complex showed dose‐ and ratio‐dependent inhibition of IL‐13 secretion in Th2 cells. At the IL‐13 ASO/cEL ratio of 6, maximum inhibition of IL‐13 secretion was observed. When applied to the ovalbumin‐sensitized murine model of AD, topically administered IL‐13 ASO/cEL complex dramatically suppressed IL‐13 production (by up to 70% of the control) in the affected skin region. In addition, the levels of IL‐4 and IL‐5 were also significantly reduced. Moreover, IL‐13 ASO/cEL‐treated AD mice showed reduced infiltration of inflammatory cells into the epidermal and dermal areas, with concomitant reduction of skin thickness.

Conclusions

These data suggests that IL‐13 ASO/cEL complex can provide a potential therapeutic tool for the treatment of AD and also be applied to other immune diseases associated with the production of Il‐13. Copyright © 2008 John Wiley & Sons, Ltd.     

Long-term effects of graduated compression stockings on cardiorespiratory performance

The use of graduated compression stockings (GCS) in sport has been increasing in the last years due to their potential positive effects for athletes. However, there is little evidence to support whether these types of garments actually improve cardiorespiratory performance. The aim of this study was to examine the cardiorespiratory responses of GCS during running after three weeks of regular use. Twenty recreational runners performed three tests on different days: test 1) – a 5-min maximal effort run in order to determine the participants’ maximal aerobic speed; and tests 2) and 3) – a fatigue running test of 30 minutes at 80% of their maximal aerobic speed with either GCS or PLACEBO stockings at random. Cardiorespiratory parameters (minute ventilation, heart rate, relative oxygen consumption, relative carbon dioxide production, ventilatory equivalents for oxygen and carbon dioxide, and oxygen pulse) were measured. Before each test in the laboratory, the participants trained with the randomly assigned stockings (GCS or PLACEBO) for three weeks. No significant differences between GCS and PLACEBO were found in any of the cardiorespiratory parameters. In conclusion, the present study provides evidence that running with GCS for three weeks does not influence cardiorespiratory parameters in recreational runners.     

Enhancing operational stability and exhibition of enzyme activity by removing water in the immobilized lipase‐catalyzed production of erythorbyl laurate

Erythorbyl laurate was continuously synthesized by esterification in a packed‐bed enzyme reactor with immobilized lipase from Candida antarctica. Response surface methodology based on a five‐level three‐factor central composite design was adopted to optimize conditions for the enzymatic esterification. The reaction variables, such as reaction temperature (10–70°C), substrate molar ratio ([lauric acid]/[erythorbic acid], 5–15), and residence time (8–40 min) were evaluated and their optimum conditions were found to be 56.2°C, 14.3, and 24.2 min, respectively. Under the optimum conditions, the molar conversion yield was 83.4%, which was not significantly different (P < 0.05) from the value predicted (84.4%). Especially, continuous water removal by adsorption on an ion‐exchange resin in a packed‐bed enzyme reactor improved operational stability, resulting in prolongation of half‐life (2.02 times longer compared to the control without water‐removal system). Furthermore, in the case of batch‐type reactor, it exhibited significant increase in initial velocity of molar conversion from 1.58% to 2.04%/min. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:882–889, 2013     

Sensitive fluorescence in situ hybridization signal detection in maize using directly labeled probes produced by high concentration DNA polymerase nick translation

The signal produced by fluorescence in situ hybridization (FISH) often is inconsistent among cells and sensitivity is low. Small DNA targets on the chromatin are difficult to detect. We report here an improved nick translation procedure for Texas red and Alexa Fluor 488 direct labeling of FISH probes. Brighter probes can be obtained by adding excess DNA polymerase I. Using such probes, a 30 kb yeast transgene, and the rp1, rp3 and zein multigene clusters were clearly detected.     

The Symbiotic Anthozoan: A Physiological Chimera between Alga and Animal

The symbiotic life style involves mutual ecological, physiological,structural, and molecular adaptations between the partners.In the symbiotic association between anthozoans and photosyntheticdinoflagellates (Symbiodinium spp., also called zooxanthellae),the presence of the endosymbiont in the animal cells has constrainedthe host in several ways. It adopts behaviors that optimizephotosynthesis of the zooxanthellae. The animal partner hashad to evolve the ability to absorb and concentrate dissolvedinorganic carbon from seawater in order to supply the symbiont'sphotosynthesis. Exposing itself to sunlight to illuminate itssymbionts sufficiently also subjects the host to damaging solarultraviolet radiation. Protection against this is provided bybiochemical sunscreens, including mycosporine-like amino acids,themselves produced by the symbiont and translocated to thehost. Moreover, to protect itself against oxygen produced duringalgal photosynthesis, the cnidarian host has developed certainantioxidant defenses that are unique among animals. Finally,living in nutrient-poor waters, the animal partner has developedseveral mechanisms for nitrogen assimilation and conservationsuch as the ability to absorb inorganic nitrogen, highly unusualfor a metazoan. These facts suggest a parallel evolution ofsymbiotic cnidarians and plants, in which the animal host hasadopted characteristics usually associated with phototrophicorganisms.     

Proof-of-concept human laboratory study for protracted abstinence in alcohol dependence: effects of gabapentin

There is a need for safe medications that can effectively support recovery by treating symptoms of protracted abstinence that may precipitate relapse in alcoholics, e.g. craving and disturbances in sleep and mood. This proof-of-concept study reports on the effectiveness of gabapentin 1200 mg for attenuating these symptoms in a non-treatment-seeking sample of cue-reactive, alcohol-dependent individuals. Subjects were 33 paid volunteers with current Diagnostic and Statistical Manual of Mental Disorders-IV alcohol dependence and a strength of craving rating 1 SD or greater for alcohol than water cues. Subjects were randomly assigned to gabapentin or placebo for 1 week and then participated in a within-subjects trial where each was exposed to standardized sets of pleasant, neutral and unpleasant visual stimuli followed by alcohol or water cues. Gabapentin was associated with significantly greater reductions than placebo on several measures of subjective craving for alcohol as well as for affectively evoked craving. Gabapentin was also associated with significant improvement on several measures of sleep quality. Side effects were minimal, and gabapentin effects were not found to resemble any major classes of abused drugs. Results suggest that gabapentin may be effective for treating the protracted abstinence phase in alcohol dependence and that a randomized clinical trial would be an appropriate next step. The study also suggests the value of cue-reactivity studies as proof-of-concept screens for potential antirelapse drugs.