Matrix metalloproteinase landscape in the imiquimod-induced skin inflammation mouse model.

Inflammation and autoimmunity are known as central processes in many skin diseases, including psoriasis. It is therefore important to develop pre-clinical models that describe disease-related aspects to enable testing of pharmaceutical drug candidates and formulations. A widely accepted pre-clinical model of psoriasis is the imiquimod (IMQ)-induced skin inflammation mouse model, where topically applied IMQ provokes local skin inflammation. In this study, we investigated the abundance of a subset of matrix metalloproteinases (MMPs) in skin from mice with IMQ-induced skin inflammation and skin from naïve mice using targeted proteomics. Our findings reveal a significant increase in the abundance of MMP-2, MMP-7, MMP-8, and MMP-13 after treatment with IMQ compared to the control skin, while MMP-3, MMP-9, and MMP-10 were exclusively detected in the IMQ-treated skin. The increased abundance and broader representation of MMPs in the IMQ-treated skin provide valuable insight into the pathophysiology of skin inflammation in the IMQ model, adding to previous studies on cytokine levels using conventional immunochemical methods. Specifically, the changes in the MMP profiles observed in the IMQ-treated skin resemble the MMP patterns found in skin lesions of individuals with psoriasis. Ultimately, the differences in MMP abundance under IMQ-induced inflammation as compared to non-inflamed control skin can be exploited as a model to investigate drug efficacy or performance of drug delivery systems.

Protease-Responsive Hydrogel Microparticles for Intradermal Drug Delivery.

Protease-responsive multi-arm polyethylene glycol-based microparticles with biscysteine peptide crosslinkers (CGPGG↓LAGGC) were obtained for intradermal drug delivery through inverse suspension photopolymerization. The average size of the spherical hydrated microparticles was ∼40 µm after crosslinking, making them attractive as a skin depot and suitable for intradermal injections, as they are readily dispensable through 27G needles. The effects of exposure to matrix metalloproteinase 9 (MMP-9) on the microparticles were evaluated by scanning electron microscopy and atomic force microscopy, demonstrating partial network destruction and decrease in elastic moduli. Given the recurring course of many skin diseases, the microparticles were exposed to MMP-9 in a flare-up mimicking fashion (multiple-time exposure), showing a significant increase in release of tofacitinib citrate (TC) from the MMP-responsive microparticles, which was not seen for the non-responsive microparticles (polyethylene glycol dithiol crosslinker). It was found that the degree of multi-arm complexity of the polyethylene glycol building blocks can be utilized to tune not only the release profile of TC but also the elastic moduli of the hydrogel microparticles, with Young's moduli ranging from 14 to 140 kPa going from 4-arm to 8-arm MMP-responsive microparticles. Finally, cytotoxicity studies conducted with skin fibroblasts showed no reduction in metabolic activity after 24 h exposure to the microparticles. Overall, these findings demonstrate that protease-responsive microparticles exhibit the properties of interest for intradermal drug delivery.

Design and characterization of matrix metalloproteinase-responsive hydrogels for the treatment of inflammatory skin diseases.

Enzyme-responsive hydrogels, formed by step growth photopolymerization of biscysteine peptide linkers with alkene functionalized polyethylene glycol, provide interesting opportunities as biomaterials and drug delivery systems. In this study, we developed stimuli-responsive, specific, and cytocompatible hydrogels for delivery of anti-inflammatory drugs for the treatment of inflammatory skin diseases. We designed peptide linkers with optimized sensitivity towards matrix metalloproteinases, a family of proteolytic enzymes overexpressed in the extracellular matrix of the skin during inflammation. The peptide linkers were crosslinked with branched 4-arm and 8-arm polyethylene glycols by thiol-norbornene photopolymerization, leading to the formation of a hydrogel network, in which the anti-inflammatory Janus kinase inhibitor tofacitinib citrate was incorporated. The hydrogels were extensively characterized by physical properties, in vitro release studies, cytocompatibility with fibroblasts, and anti-inflammatory efficacy testing in both an atopic dermatitis-like keratinocyte assay and an activated T-cell assay. The drug release was studied after single and multiple-time exposure to matrix metalloproteinase 9 to mimic inflammatory flare-ups. Drug release was found to be triggered by matrix metalloproteinase 9 and to depend on type of crosslinker and of the polyethylene glycol polymer, due to differences in architecture and swelling behavior. Moreover, swollen hydrogels showed elastic properties similar to those of extracellular matrix proteins in the dermis. Cell studies revealed limited cytotoxicity when fibroblasts and keratinocytes were exposed to the hydrogels or their enzymatic cleavage products. Taken together, our results suggest multi-arm polyethylene glycol hydrogels as promising matrix metalloproteinase-responsive drug delivery systems, with potential in the treatment of inflammatory skin disease. STATEMENT OF SIGNIFICANCE: Smart responsive drug delivery systems such as matrix metalloproteinase-responsive hydrogels are excellent candidates for the treatment of inflammatory skin diseases including psoriasis. Their release profile can be optimized to correspond to the patient's individual disease state by tuning formulation parameters and disease-related stimuli, providing personalized treatment solutions. However, insufficient cross-linking efficiency, low matrix metalloproteinase sensitivity, and undesirable drug release kinetics remain major challenges in the development of such drug delivery systems. In this study, we address shortcomings of previous work by designing peptide linkers with optimized sensitivity towards matrix metalloproteinases and high cross-linking efficiencies. We further provide a proof-of-concept for the usability of the hydrogels in inflammatory skin conditions by employing a drug release set-up simulating inflammatory flare-ups.

The role of lipid components in lipid nanoparticles for vaccines and gene therapy.

Lipid nanoparticles (LNPs) play an important role in mRNA vaccines against COVID-19. In addition, many preclinical and clinical studies, including the siRNA-LNP product, Onpattro®, highlight that LNPs unlock the potential of nucleic acid-based therapies and vaccines. To understand what is key to the success of LNPs, we need to understand the role of the building blocks that constitute them. In this Review, we discuss what each lipid component adds to the LNP delivery platform in terms of size, structure, stability, apparent pKa, nucleic acid encapsulation efficiency, cellular uptake, and endosomal escape. To explore this, we present findings from the liposome field as well as from landmark and recent articles in the LNP literature. We also discuss challenges and strategies related to in vitro/in vivo studies of LNPs based on fluorescence readouts, immunogenicity/reactogenicity, and LNP delivery beyond the liver. How these fundamental challenges are pursued, including what lipid components are added and combined, will likely determine the scope of LNP-based gene therapies and vaccines for treating various diseases.

Future Perspectives of Oral Delivery of Next Generation Therapies for Treatment of Skin Diseases.

Gene therapies have conspicuously bloomed in recent years as evidenced by the increasing number of cell-, gene-, and oligo-based approved therapies. These therapies hold great promise for dermatological disorders with high unmet need, for example, epidermolysis bullosa or pachyonychia congenita. Furthermore, the recent clinical success of clustered regularly interspaced short palindromic repeats (CRISPR) for genome editing in humans will undoubtedly contribute to defining a new wave of therapies. Like biologics, naked nucleic acids are denatured inside the gastrointestinal tract and need to be administered via injections. For a treatment to be effective, a sufficient amount of a given regimen needs to reach systemic circulation. Multiple companies are racing to develop novel oral drug delivery approaches to circumvent the proteolytic and acidic milieu of the gastrointestinal tract. In this review, we provide an overview of the evolution of the gene therapy landscape, with a deep focus on gene and oligonucleotide therapies in clinical trials aimed at treating skin diseases. We then examine the progress made in drug delivery, with particular attention on the peptide field and drug-device combinations that deliver macromolecules into the gastrointestinal tract. Such novel devices could potentially be applied to administer other therapeutics including genes and CRISPR-based systems.

Imageability ratings across languages.

Imageability is a psycholinguistic variable that indicates how well a word gives rise to a mental image or sensory experience. Imageability ratings are used extensively in psycholinguistic, neuropsychological, and aphasiological studies. However, little formal knowledge exists about whether and how these ratings are associated between and within languages. Fifteen imageability databases were cross-correlated using nonparametric statistics. Some of these corresponded to unpublished data collected within a European research network-the Collaboration of Aphasia Trialists (COST IS1208). All but four of the correlations were significant. The average strength of the correlations (rho = .68) and the variance explained (R 2 = 46%) were moderate. This implies that factors other than imageability may explain 54% of the results. Imageability ratings often correlate across languages. Different possibly interacting factors may explain the moderate strength and variance explained in the correlations: (1) linguistic and cultural factors; (2) intrinsic differences between the databases; (3) range effects; (4) small numbers of words in each database, equivalent words, and participants; and (5) mean age of the participants. The results suggest that imageability ratings may be used cross-linguistically. However, further understanding of the factors explaining the variance in the correlations will be needed before research and practical recommendations can be made.

Lexical access in a bilingual speaker with dementia: Changes over time.

In this article, we explore the naming skills of a bilingual English-Norwegian speaker diagnosed with Primary Progressive Aphasia, in each of his languages across three different speech contexts: confrontation naming, semi-spontaneous narrative (picture description), and conversation, and at two points in time: 12 and 30 months post diagnosis, respectively. The results are discussed in light of two main theories of lexical retrieval in healthy, elderly speakers: the Transmission Deficit Hypothesis and the Inhibitory Deficit Theory. Our data show that, consistent with the participant's premorbid use of and proficiency in the two languages, his performance in his L2 is lower than in his L1, but this difference diminishes as the disease progresses. This is the case across the three speech contexts; however, the difference is smaller in the narrative task, where his performance is very low in both languages already at the first measurement point. Despite his word finding problems, he is able to take active part in conversation, particularly in his L1 and more so at the first measurement point. In addition to the task effect, we find effects of word class, frequency, and cognateness on his naming skills. His performance seems to support the Transmission Deficit Hypothesis. By combining different tools and methods of analysis, we get a more comprehensive picture of the impact of the dementia on the speaker's languages from an intra-individual as well as an inter-individual perspective, which may be useful in research as well as in clinical practice.

Corticosterone Response as an Age-Specific Mediator of Nestling Body Mass in a Wild Passerine.

The environment can affect individual performance directly via resource availability or indirectly through resource allocation among competing fitness components, such as body growth and maintenance activities related to short-term survival. Corticosterone (CORT), the main glucocorticoid in birds, may be an important mediator of energy allocation to different organismal functions, but its effect on the plasticity of fitness-related traits has rarely been investigated at different ontogenetic stages. Here, we evaluated the role of baseline and stress-induced CORT on nestling development of wild great tits (Parus major) under different growth conditions and at different developmental stages. We found that nestling mass and condition were positively related to stress-induced levels of total CORT and free CORT in the middle of nestling stage but negatively related at the prefledging stage. Our results also showed that stress-induced levels of total CORT can be used as a proxy for bioavailable free CORT. No significant relationship of body mass or condition in either stage was shown with levels of baseline CORT or stress-induced CORT-binding globulin capacity. None of these age-specific relationships depend on brood size manipulation. We conclude that the effects of glucocorticoids on nestling performance are stage specific. The ability to secrete CORT may have beneficial effects on nestling development in the middle of the nestling stage, while it can be adaptive to keep CORT levels at optimal ranges before fledging to give priority to growth-related processes that are important for long-term fitness.

Cross-linguistic adaptations of The Comprehensive Aphasia Test: Challenges and solutions.

Comparative research on aphasia and aphasia rehabilitation is challenged by the lack of comparable assessment tools across different languages. In English, a large array of tools is available, while in most other languages, the selection is more limited. Importantly, assessment tools are often simple translations and do not take into consideration specific linguistic and psycholinguistic parameters of the target languages. As a first step in meeting the needs for comparable assessment tools, the Comprehensive Aphasia Test is currently being adapted into a number of languages spoken in Europe. In this article, some key challenges encountered in the adaptation process and the solutions to ensure that the resulting assessment tools are linguistically and culturally equivalent, are proposed. Specifically, we focus on challenges and solutions related to the use of imageability, frequency, word length, spelling-to-sound regularity and sentence length and complexity as underlying properties in the selection of the testing material.

Supersaturation of Calcipotriene and Betamethasone Dipropionate in a Novel Aerosol Foam Formulation for Topical Treatment of Psoriasis Provides Enhanced Bioavailability of the Active Ingredients.

INTRODUCTION: Previous studies have demonstrated the superior efficacy of a novel aerosol foam formulation of fixed combination calcipotriene 0.005% (Cal) and betamethasone dipropionate 0.064% (BD), compared with the ointment formulation. The aim of this study is to ascertain whether enhanced bioavailability of the active ingredients due to supersaturation and/or occlusive properties can explain the observed greater clinical efficacy. METHODS: Solubility and evaporation experiments were conducted to examine the abilities of Cal/BD aerosol foam ingredients to create a supersaturated environment. Optical microscopy, Raman imaging and X-ray powder diffraction were used to examine the physical state of Cal and BD in the formulations after application, and determine whether a supersaturated state remained stable for clinically relevant time periods. In vitro skin penetration and ex vivo biomarker assays were conducted to compare the skin penetration and bioavailability of Cal and BD from the aerosol foam and ointment formulations, respectively. Occlusive properties were examined via transepidermal water loss. RESULTS: Solubility studies showed that Cal and BD solubility increased with increasing dimethyl ether (DME) content. Both active ingredients are completely dissolved in the final aerosol foam formulation. DME rapidly evaporates after spraying, and the amount was reduced to 0.5% of the initial amount after 2 min. This led to the formation of a supersaturated environment, where Cal and BD crystals were absent for at least 26 h after application. Cal/BD aerosol foam had significantly greater in vitro skin penetration and had increased bioavailability compared with Cal/BD ointment. Both formulations effectively occluded the skin. CONCLUSION: A stable supersaturated solution of Cal/BD in the aerosol foam leads to increased bioavailability and explains the improved clinical effect when compared to the Cal/BD ointment. FUNDING: The studies included in the paper are all conducted by LEO Pharma A/S or CROs on behalf of LEO Pharma A/S.

Acute embryonic exposure to corticosterone alters physiology, behaviour and growth in nestlings of a wild passerine.

Maternally-derived glucocorticoids can modify the normal development of young animals. To date, little is known about maternal effects that are mediated by acute embryonic exposure to glucocorticoids. In birds, elevated maternal transmission of corticosterone (CORT) to egg albumen is mainly dependent on acute stress. In this study, we increased CORT levels in the egg albumen of a wild passerine, the great tit (Parus major), breeding in favourable deciduous and less suitable coniferous habitat. Subsequently we measured the somatic growth, baseline and acute glucocorticoid responses, immunity and behaviour of prenatally manipulated offspring with respect to control siblings. We found that prenatally CORT-exposed nestlings had lower baseline CORT levels, a more rapid decline in CORT during recovery from a standardized stressor, and a reduced heterophil/lymphocyte ratio compared with controls. Although stress-induced total CORT levels remained unchanged, free CORT levels were significantly lower and the levels of corticosteroid binding globulins (CBG) significantly higher in experimental offspring. Prenatally CORT-exposed offspring begged longer after hatching than controls. Stress-induced behavioural activity of fledglings did not differ between treatments, while its association with baseline CORT levels was significant in the control group only. The body mass and tarsus length of fledglings was positively affected by manipulation in unfavourable coniferous habitat only. We conclude that maternal effects related to elevated levels of albumen CORT modify diverse aspects of offspring phenotype and potentially increase offspring performance in resource poor environments. Moreover, our results indicate that maternal glucocorticoids may suppress the effect of hormones on behavioural responses.

Towards a phronetic space for responsible research (and innovation).

The term Responsible Research and Innovation has recently gained currency, as it has been designated to be a key-term in the European research framework Horizon 2020. At the level of European research policy, Responsible Research and Innovation can be viewed as an attempt to reach a broader vision of research and innovation as a public good. The current academic debate may be fairly enriched by considering the role that phronesis may have for RRI. Specifically, in this paper we argue that the current debate might be fruitfully enriched by making a categorial shift. Such a categorial shift involves moving away from the temptation to interpret responsible research and innovation in a technocratic way towards a more pluralistic vision that is rooted in the idea of phronesis. In the present context phronesis points the attention to the cultivation and nurturement of the researcher's formation as a type of engagement with the actual practice of researching, a practice in which researchers (and other parties concerned) are called to apply judgment and exercise discretion in specific and often unique situations without the re-assuring viewpoint of the technician.

Effect of Intraoperative Three-Dimensional Imaging During the Reduction and Fixation of Displaced Calcaneal Fractures on Articular Congruence and Implant Fixation.

BACKGROUND: Operative treatment of displaced calcaneal fractures should restore joint congruence, but conventional fluoroscopy is unable to fully visualize the subtalar joint. We questioned whether intraoperative 3-dimensional (3D) imaging would aid in the reduction of calcaneal fractures, resulting in improved articular congruence and implant positioning. METHOD: Sixty-two displaced calcaneal fractures were operated on using standard fluoroscopic views. When the surgeon had achieved a satisfactory reduction, an intraoperative 3D scan was conducted, malreductions or implant imperfections were revised, the calcaneus was rescanned, and this sequence was repeated until the optimal operative result was achieved. RESULTS: Five fractures underwent 1 intraoperative scan, 39 fractures underwent 2 scans, 13 fractures underwent 3 scans, and 5 fractures underwent 4 scans. The average number of scans was 2.3. Intraoperative scanning led to re-reduction and improvement of reduction in 13 fractures, change of plate position in 1 patient, optimizing of the screw directions in 5 fractures, and shortening of screws that were intra-articular or protruding medially in 6 fractures. The postoperative articular displacement was 0 mm in 69% of the Sanders type 2 fractures and 57% of the Sanders type 3 fractures. Operation duration averaged 118 minutes, and there were no reoperations due to misplaced screws or plates. The average absorbed radiation dose per patient was 288 mGy·cm. CONCLUSION: Intraoperative 3D imaging improved the articular reduction of the posterior facet and secured optimal implant position in displaced calcaneal fractures. Radiation dose to the patient was less than that of a normal foot computed tomography scan. LEVEL OF EVIDENCE: Level IV, case series.

Norwegian words: A lexical database for clinicians and researchers.

All words have properties linked to form, meaning and usage patterns which influence how easily they are accessed from the mental lexicon in language production, perception and comprehension. Examples of such properties are imageability, phonological and morphological complexity, word class, argument structure, frequency of use and age of acquisition. Due to linguistic and cultural variation the properties and the values associated with them differ across languages. Hence, for research as well as clinical purposes, language specific information on lexical properties is needed. To meet this need, an electronically searchable lexical database with more than 1600 Norwegian words coded for more than 12 different properties has been established. This article presents the content and structure of the database as well as the search options available in the interface. Finally, it briefly describes some of the ways in which the database can be used in research, clinical practice and teaching.

[High complication rate after surgical treatment of ankle fractures]. / Høj komplikationsrate efter kirurgisk behandling af malleolfraktur.

The purpose of this study was to determine the quality and re-operation rate of the surgical treatment of ankle fractures at a large university hospital. X-rays and patient records of 137 patients surgically treated for ankle fractures were analyzed for: 1) correct classification according to Lauge-Hansen, 2) if congruity of the ankle joint was achieved, 3) selection and placement of the hardware, and 4) the surgeon's level of education. Totally 32 of 137 did not receive an optimal treatment, 11 were re-operated. There was no clear correlation between incorrect operation and the surgeon's level of education.

Spina bifida and sexuality.

OBJECTIVE: To evaluate sexual function amongst adult individuals with spina bifida and to register their subjective satisfaction with their sexual life and relationships. SETTING: Department for Spinal Cord Injuries, East Denmark. STUDY DESIGN AND METHODS: Cohort study. Medical record information, neurological examination, personal interview, Functional Independence Measure (FIMTM), Medical Outcome Study Short Form 36 (SF-36) on quality of life, and questions on sexual function and related topics. Study cohort: Fifty-three participants (27 women, 26 men) with spina bifida (mean age 27.1, range 18-35) years. Response rate 74%. RESULTS: Fifty-one percent of subjects regarded their sexual life as a failure or dysfunctional. However, 45% reported being satisfied with their sexual life. Participants with partners were more satisfied with their sexual life than those without partners. Faecal, but not urinary, incontinence was associated with poorer sexual function and less satisfaction. Forty-nine percent of subjects indicated that the sexual education they received at puberty was useful; however, 32% lacked knowledge about their sexual functioning with regard to their disability. CONCLUSION: There is a need for further sexual education and counselling for adults with spina bifida in order to improve their sexuality and quality of life.

Imageability of Norwegian nouns, verbs and adjectives in a cross-linguistic perspective.

In this article, we present a study of imageability ratings for a set of 1599 Norwegian words (896 nouns, 483 verbs and 220 adjectives) from a web-based survey. To a large extent, the results are in accordance with previous studies of other languages: high imageability scores in general, higher imageability scores for nouns than for verbs, and an inverse relation between frequency and imageability. A more surprising finding is the low imageability of low-frequency verbs. Also, imageability ratings increase systematically and significantly with informant age, reminding us that conceptual learning continues and changes throughout life. This has consequences for our expectations of different linguistic skills in a life span perspective. These findings have an obvious clinical relevance both for choice of items in test construction, for evaluation of performance in clinical groups and for development of therapy material.

[Atrial myxom is a rare cause of peripheral embolus]. / Atrialt myksom er en sjælden årsag til perifer embolus.

Two case stories of seemingly healthy individuals, a 45 year-old male and a 54 year-old woman, with acute lower extremity ischemia caused by cardiac myxomas. Both patients had a lower extremity amputated and a removal of myxomas by surgery. In patients presenting with acute ischemia of the lower extremity surgeons need to consider this rare, but important diagnosis; especially in middle-aged, otherwise healthy patients. Performing an ultrasound sonography of the heart is the best way to make a diagnosis.

Compound nouns in spoken language production by speakers with aphasia compared to neurologically healthy speakers: an exploratory study.

Compounds are words that are made up of at least two other words (lexemes), featuring lexical and syntactic characteristics and thus particularly interesting for the study of language processing. Most studies of compounds and language processing have been based on data from experimental single word production and comprehension tasks. To enhance the ecological validity of morphological processing research, data from other contexts, such as discourse production, need to be considered. This study investigates the production of nominal compounds in semi-spontaneous spoken texts by a group of speakers with fluent types of aphasia compared to a group of neurologically healthy speakers. The speakers with aphasia produce significantly fewer nominal compound types in their texts than the non-aphasic speakers, and the compounds they produce exhibit fewer different types of semantic relations than the compounds produced by the non-aphasic speakers. The results are discussed in relation to theories of language processing.

Semi-spontaneous oral text production: measurements in clinical practice.

Functionally relevant assessment of the language production of speakers with aphasia should include assessment of connected speech production. Despite the ecological validity of everyday conversations, more controlled and monological types of texts may be easier to obtain and analyse in clinical practice. This article discusses some simple measurements for the analysis of semi-spontaneous oral text production by speakers with aphasia. Specifically, the measurements are related to the production of verbs and nouns, and the realization of different sentence types. The proposed measurements should be clinically relevant, easily applicable, and linguistically meaningful. The measurements have been applied to oral descriptions of the 'Cookie Theft' picture by eight monolingual Norwegian speakers, four with an anomic type of aphasia and four without any type of language impairment. Despite individual differences in both the clinical and the non-clinical group, most of the measurements seem to distinguish between speakers with and without aphasia.