Pathological Angiogenesis Requires Syndecan-4 for Efficient VEGFA-Induced VE-Cadherin Internalization.

[Figure: see text].

Exposed CendR Domain in Homing Peptide Yields Skin-Targeted Therapeutic in Epidermolysis Bullosa.

Systemic skin-selective therapeutics would be a major advancement in the treatment of diseases affecting the entire skin, such as recessive dystrophic epidermolysis bullosa (RDEB), which is caused by mutations in the COL7A1 gene and manifests in transforming growth factor-ß (TGF-ß)-driven fibrosis and malignant transformation. Homing peptides containing a C-terminal R/KXXR/K motif (C-end rule [CendR] sequence) activate an extravasation and tissue penetration pathway for tumor-specific drug delivery. We have previously described a homing peptide CRKDKC (CRK) that contains a cryptic CendR motif and homes to angiogenic blood vessels in wounds and tumors, but it cannot penetrate cells or tissues. In this study, we demonstrate that removal of the cysteine from CRK to expose the CendR sequence confers the peptide novel ability to home to normal skin. Fusion of the truncated CRK (tCRK) peptide to the C terminus of an extracellular matrix protein decorin (DCN), a natural TGF-ß inhibitor, resulted in a skin-homing therapeutic molecule (DCN-tCRK). Systemic DCN-tCRK administration in RDEB mice led to inhibition of TGF-ß signaling in the skin and significant improvement in the survival of RDEB mice. These results suggest that DCN-tCRK has the potential to be utilized as a novel therapeutic compound for the treatment of dermatological diseases such as RDEB.

R-Ras regulates vascular permeability, but not overall healing in skin wounds.

Wounds close by keratinocytes migrating from the edge of the wound and re-epithelializing the epidermis. It has been proposed that the major stimuli for wound closure are blood-derived growth factors, chemokines and cytokines. The small GTPase R-Ras, a known integrin activator, also regulates vascular permeability during angiogenesis, and blood vessels lacking R-Ras leak plasma proteins constantly. We explored whether the access to blood-derived proteins influences skin wound healing in R-Ras knockout (KO) mice. In skin wounds, R-Ras expression was mostly restricted to the vasculature in the granulation tissue. Angiogenic blood vessels in the R-Ras KO mice were significantly more permeable than in wild-type (WT) controls. Although the distances between epidermal tongues, and the panniculus carnosus muscles, were significantly longer in R-Ras KO than WT controls before the granulation tissue formation took place, there were no differences in the wound closure or re-epithelialization rates or granulation tissue formation. These findings were also corroborated in a special splint excision wound model. Our study shows that although R-Ras does not influence the skin wound healing itself, the blood vessels lacking R-Ras are leaky and thus could facilitate the access of blood-derived proteins to the wound.

[From anamnesis to the art of interpretation, or: What is a "genuine" psychoanalyst? Wolfgang Warda, Ludwig Binswanger, Wilhelm Strohmayer and the origins of psychoanalysis in Thuringia]. / Vom Wachexamen zur Deutungskunst, oder: Was ist ein "richtiger" Psychoanalytiker. Wolfgang Warda, Ludwig Binswanger, Wilhelm Strohmayer und die Anfänge der Psychoanalyse in Thüringen.

Warda and Strohmayer from Thuringia were among the first German physicians who developed an interest in Freuds theory and therapeutic method around 1900. Their contributions reflect the influence of Otto Binswanger, professor of psychiatry in Jena, a representative of the "psychological direction" in psychiatry which in the beginning was relatively receptive to Freud. The paper discusses their rapprochement to, and detachment from, the Freudian school, including also the work of a third young physician: Ludwig Binswanger, Otto's nephew, who was active in Jena at the same time. It points to certain factors contributing to the increasing rejection Freud met in academic circles which have been underrated to date: (1) the transformation of psychoanalysis into an art of interpretation; (2) the introduction of transference. Both factors which were elaborated by Freud as essentials of his theoretical and practical approach around 1900 and published in 1904/05, undermined the claim of academic medicine to objectivity. The paper describes how psychoanalysis officially abandoned the scientific standards of contemporary medicine at the Weimar congress in 1911, at the same time as Warda and Strohmayer left the Freudian group.

Trans-signaling is a dominant mechanism for the pathogenic actions of interleukin-6 in the brain.

IL-6 is implicated in the pathogenesis of various neuroinflammatory and neurodegenerative disorders of the CNS. IL-6 signals via binding to either the membrane bound IL-6Rα (classic signaling) or soluble (s)IL-6Ra (trans-signaling) that then form a complex with gp130 to activate the JAK/STAT signaling pathway. The importance of classic versus trans-signaling in mediating IL-6 actions in the living CNS is relatively unknown and was the focus of this investigation. Bigenic mice (termed GFAP-IL6/sgp130 mice) were generated with CNS-restricted, astrocyte-targeted production of IL-6 and coproduction of the specific inhibitor of IL-6 trans-signaling, human sgp130-Fc. Transgene-encoded IL-6 mRNA levels were similar in the brain of GFAP-IL6 and GFAP-IL6/sgp130 mice. However, GFAP-IL6/sgp130 mice had decreased pY(705)-STAT3 in the brain due to a reduction in the total number of pY(705)-STAT3-positive cells and a marked loss of pY(705)-STAT3 in specific cell types. Blockade of trans-signaling in the brain of the GFAP-IL6 mice significantly attenuated Serpina3n but not SOCS3 gene expression, whereas vascular changes including angiogenesis and blood-brain barrier leakage as well as gliosis were also reduced significantly. Hippocampal neurogenesis which was impaired in GFAP-IL6 mice was rescued in young GFAP-IL6 mice with cerebral sgp130 production. Finally, degenerative changes in the cerebellum characteristic of GFAP-IL6 mice were absent in GFAP-IL6/sgp130 mice. The findings indicate that in the CNS: (1) sgp130 is able to block IL-6 trans-signaling, (2) trans-signaling is important for IL-6 cellular communication with selective cellular and molecular targets, and (3) blocking of trans-signaling alleviates many of the detrimental effects of IL-6.

Systemically Administered, Target Organ-Specific Therapies for Regenerative Medicine.

Growth factors and other agents that could potentially enhance tissue regeneration have been identified, but their therapeutic value in clinical medicine has been limited for reasons such as difficulty to maintain bioactivity of locally applied therapeutics in the protease-rich environment of regenerating tissues. Although human diseases are treated with systemically administered drugs in general, all current efforts aimed at enhancing tissue repair with biological drugs have been based on their local application. The systemic administration of growth factors has been ruled out due to concerns about their safety. These concerns are warranted. In addition, only a small proportion of systemically administered drugs reach their intended target. Selective delivery of the drug to the target tissue and use of functional protein domains capable of penetrating cells and tissues could alleviate these problems in certain circumstances. We will present in this review a novel approach utilizing unique molecular fingerprints ("Zip/postal codes") in the vasculature of regenerating tissues that allows target organ-specific delivery of systemically administered therapeutic molecules by affinity-based physical targeting (using peptides or antibodies as an "address tag") to injured tissues undergoing repair. The desired outcome of targeted therapies is increased local accumulation and lower systemic concentration of the therapeutic payload. We believe that the physical targeting of systemically administered therapeutic molecules could be rapidly adapted in the field of regenerative medicine.

Systemically administered wound-homing peptide accelerates wound healing by modulating syndecan-4 function.

CAR (CARSKNKDC) is a wound-homing peptide that recognises angiogenic neovessels. Here we discover that systemically administered CAR peptide has inherent ability to promote wound healing: wounds close and re-epithelialise faster in CAR-treated male mice. CAR promotes keratinocyte migration in vitro. The heparan sulfate proteoglycan syndecan-4 regulates cell migration and is crucial for wound healing. We report that syndecan-4 expression is restricted to epidermis and blood vessels in mice skin wounds. Syndecan-4 regulates binding and internalisation of CAR peptide and CAR-mediated cytoskeletal remodelling. CAR induces syndecan-4-dependent activation of the small GTPase ARF6, via the guanine nucleotide exchange factor cytohesin-2, and promotes syndecan-4-, ARF6- and Cytohesin-2-mediated keratinocyte migration. Finally, we show that genetic ablation of syndecan-4 in male mice eliminates CAR-induced wound re-epithelialisation following systemic administration. We propose that CAR peptide activates syndecan-4 functions to selectively promote re-epithelialisation. Thus, CAR peptide provides a therapeutic approach to enhance wound healing in mice; systemic, yet target organ- and cell-specific.

Karl Abraham's revolution of 1916: from sensual sucking to the oral-aggressive wish of destruction.

The author argues that "The First Pregenital Stage of the Libido" (Abraham 1916-1917) expounds a new conception of orality, i.e., of purposeful oral aggression directed against an object during the first stage of psychic development. This conception is shown to be contrary to Freud's view of orality as elaborated in Three Essays on the Theory of Sexuality (1905), as well as in other writings of late 1914 and 1915. Abraham's conception ignores fundamental dimensions of Freud's thinking during these years, namely, the difference between autoerotism/narcissism and object love, on the one hand, and also between the leading role of sexuality and the secondary role of aggression, on the other. Thus, Abraham's thinking represents a basic theoretical change that had far-reaching consequences for psychoanalytic practice.

We are looking deeper than Freud On the departure from the primacy of the sexual in Berlin and London between 1920 and 1925.

Between 1920 and 1925 a shift occurred in psychoanalytic theory that related primarily to the status of aggression. The author shows which authors in Berlin, Amsterdam and London participated in this change and to what extent they prepared the ground for the reception of Melanie Klein, who developed in her analyses of children a technique that applied pre-eminently to the perception and interpretation of aggression. The way in which Freud distanced himself from this departure from the primacy of the sexual is described, as well as the waning importance of his perspective following the establishment of the Institutes in Berlin and London.

Dual drug delivery collagen vehicles for modulation of skin fibrosisin vitro.

Single molecule drug delivery systems have failed to yield functional therapeutic outcomes, triggering investigations into multi-molecular drug delivery vehicles. In the context of skin fibrosis, although multi-drug systems have been assessed, no system has assessed molecular combinations that directly and specifically reduce cell proliferation, collagen synthesis and transforming growth factorß1 (TGFß1) expression. Herein, a core-shell collagen type I hydrogel system was developed for the dual delivery of a TGFßtrap, a soluble recombinant protein that inhibits TGFßsignalling, and Trichostatin A (TSA), a small molecule inhibitor of histone deacetylases. The antifibrotic potential of the dual delivery system was assessed in anin vitroskin fibrosis model induced by macromolecular crowding (MMC) and TGFß1. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and high performance liquid chromatography analyses revealed that ∼50% of the TGFßtrap and ∼30% of the TSA were released from the core and shell compartments, respectively, of the hydrogel system after 10 d (longest time point assessed) in culture. As a direct consequence of this slow release, the core (TGFßtrap)/shell (TSA) hydrogel system induced significantly (p< 0.05) lower than the control group (MMC and TGFß1) collagen type I deposition (assessed via SDS-PAGE and immunocytochemistry),αsmooth muscle actin (αSMA) expression (assessed via immunocytochemistry) and cellular proliferation (assessed via DNA quantification) and viability (assessed via calcein AM and ethidium homodimer-I staining) after 10 d in culture. On the other hand, direct TSA-TGFßsupplementation induced the lowest (p< 0.05) collagen type I deposition,αSMA expression and cellular proliferation and viability after 10 d in culture. Our results illustrate the potential of core-shell collagen hydrogel systems for sustained delivery of antifibrotic molecules.

[Freud's abstract of "A contribution to the option of neurosis" (1913). Publication and commentary]. / Freuds Autoreferat von "Ein Beitrag zum Problem der Neurosenwahl" (1913). Erstpublikation des Textes und Kommentar.

The text, which is published here for the first time, summarizes the paper Freud presented to the IPA congress in Munich. A short commentary highlights the circumstances under which the paper was written and the reasons why the abstract remained unpublished. It also sketches the historical significance of the paper in terms of the problem of the option of neurosis, in particular obsessional neurosis, of the development of Freud's sexual theory and of the relation of his views to those of Ernest Jones.

Systemically Administered Homing Peptide Targets Dystrophic Lesions and Delivers Transforming Growth Factor-ß (TGFß) Inhibitor to Attenuate Murine Muscular Dystrophy Pathology.

Muscular dystrophy is a progressively worsening and lethal disease, where accumulation of functionality-impairing fibrosis plays a key pathogenic role. Transforming growth factor-ß1 (TGFß1) is a central signaling molecule in the development of fibrosis in muscular dystrophic humans and mice. Inhibition of TGFß1 has proven beneficial in mouse models of muscular dystrophy, but the global strategies of TGFß1 inhibition produce significant detrimental side effects. Here, we investigated whether murine muscular dystrophy lesion-specific inhibition of TGFß1 signaling by the targeted delivery of therapeutic decorin (a natural TGFß inhibitor) by a vascular homing peptide CAR (CARSKNKDC) would reduce skeletal muscle fibrosis and pathology and increase functional characteristics of skeletal muscle. We demonstrate that CAR peptide homes to dystrophic lesions with specificity in two muscular dystrophy models. Recombinant fusion protein consisting of CAR peptide and decorin homes selectively to sites of skeletal muscle damage in mdxDBA2/J and gamma-sarcoglycan deficient DBA2/J mice. This targeted delivery reduced TGFß1 signaling as demonstrated by reduced nuclear pSMAD staining. Three weeks of targeted decorin treatment decreased both membrane permeability and fibrosis and improved skeletal muscle function in comparison to control treatments in the mdxD2 mice. These results show that selective delivery of decorin to the sites of skeletal muscle damage attenuates the progression of murine muscular dystrophy.

Adapting the Scar-in-a-Jar to Skin Fibrosis and Screening Traditional and Contemporary Anti-Fibrotic Therapies.

Skin fibrosis still constitutes an unmet clinical need. Although pharmacological strategies are at the forefront of scientific and technological research and innovation, their clinical translation is hindered by the poor predictive capacity of the currently available in vitro fibrosis models. Indeed, customarily utilised in vitro scarring models are conducted in a low extracellular matrix milieu, which constitutes an oxymoron for the in-hand pathophysiology. Herein, we coupled macromolecular crowding (enhances and accelerates extracellular matrix deposition) with transforming growth factor ß1 (TGFß1; induces trans-differentiation of fibroblasts to myofibroblasts) in human dermal fibroblast cultures to develop a skin fibrosis in vitro model and to screen a range of anti-fibrotic families (corticosteroids, inhibitors of histone deacetylases, inhibitors of collagen crosslinking, inhibitors of TGFß1 and pleiotropic inhibitors of fibrotic activation). Data obtained demonstrated that macromolecular crowding combined with TGFß1 significantly enhanced collagen deposition and myofibroblast transformation. Among the anti-fibrotic compounds assessed, trichostatin A (inhibitors of histone deacetylases); serelaxin and pirfenidone (pleiotropic inhibitors of fibrotic activation); and soluble TGFß receptor trap (inhibitor of TGFß signalling) resulted in the highest decrease of collagen type I deposition (even higher than triamcinolone acetonide, the gold standard in clinical practice). This study further advocates the potential of macromolecular crowding in the development of in vitro pathophysiology models.

[Karl Abraham's revolution. From sensual sucking to oral-aggressive destruction wishes]. / Karl Abrahams Revolution: Vom Wonnesaugen zum oral-aggressiven Vernichtungswunsch.

The author argues that Abraham's paper on "The first pregenital stage of the libido" (1916-17) expounds a new conception of orality, i. e. the idea of a purposeful oral aggression directed against an object. This conception is shown to be contrary to Freud's view of orality as elaborated in his Three Essays as well as in other writings of late 1914 and early 1915. It ignores fundamental dimensions of Freud's thinking, namely the difference between autoerotism/narcissism and object love on the one hand and between libido and aggression on the other, thus representing a basic theoretical change that also had consequences for analytical practice.

[1400 hours of analysis with Freud: Viktor von Dirsztay. A biographical sketch]. / Vierzehnhundert Stunden Analyse bei Freud: Viktor von Dirsztay. Eine biographische Skizze.

On the basis of mostly unpublished sources, the author reconstructs the life of the Hungarian writer Viktor von Dirsztay (1884?-1935) who was personally acquainted with many expressionist artists and writers, e. g. with Karl Kraus, Oskar Kokoschka, Herwarth Walden, Walter Hasenclever, Hermann Broch and Arthur Schnitzler. This association puts Freud into closer proximity with the cultural avantgarde of his times than previously realized. Between 1910 and 1920 Dirsztay underwent several phases of analysis with Freud; then he was treated by Theodor Reik. The overall length of his analysis with Freud is almost unparalleled. The article discusses whether and in which way Dirsztay's writings might have been influenced by his analyses and how Freud and Reik might have drawn upon their experiences with this patient. It is argued that likely references can be discovered in both authors' theories of masochism. There is an intriguing late remark of Dirsztay's that he was "ruined by analysis".

In conversation: Freud, Abraham and Ferenczi on "Mourning and Melancholia" (1915-1918).

This article concentrates on Freud's draft of "Mourning and Melancholia," written in 1915 and published in 1996. After presenting a summary of the main theses of Freud's draft, Abraham's and Ferenczi's reactions to the text are discussed as well as Freud's response to their comments. In addition to reviewing Freud's partial adoption of Ferenczi's introjection and his reluctance towards Abraham's "mouth eroticism and sadism," the article considers the question of whether and to what extent his disciples' interjections-particularly Abraham's approach-made their way into the final version of "Mourning and Melancholia." The article closes by integrating the notion of narcissistic identification, which forms the core of Freud's understanding of depression, and his study of the "preliminary stages of love," written the same year, into a conceptualization of the narcissistic relationship between subject and object. Special attention is paid to the clinical relevance of the difference between narcissistic and libidinal object cathexis, which Freud had introduced.

Interleukin 27 induces differentiation of neural C6-precursor cells into astrocytes.

Interleukin 6 (IL6)-type cytokines are major regulators of inflammation and thereby contribute to the neuropathology and pathophysiology associated with inflammation of the central nervous system (CNS). Furthermore, astrocyte development which is a key process in the development of the CNS is also controlled by cytokines of the IL6-family. Interleukin 27 (IL27) is a recently identified member of this family and has been implicated in the inhibition of TH17 T-cell-responses. Here we show that IL27 and the HHV8 encoded viral IL6 (vIL6) induce C6 glioma cells to differentiate into an astrocyte-like state. Cytokine stimulation led to STAT-factor phosphorylation and consequently to protein expression of the astrocyte marker glial fibrillary acidic protein (GFAP). These data could be confirmed by GFAP-immunostaining of stimulated cells. Taken together, IL27 and vIL6 can be considered as new astrocyte-inducing cytokines of the brain.

[Remarks on the Introduction to Karl Abraham's collected works (1969-71)]. / Ober Johannes Cremerius' Einleitung zu den Gesammelten Werken von Karl Abraham (1969-1971).

The introduction of Johannes Cremerius to his two-volumed edition of Abraham's writings had a great impact on how Abraham's work was received in Germany. The present article shows that this introduction is essentially a plagiarism of Ernest Jones's classical appraisal (1926) of Abraham's work. This fact is connected with the historical situation of German psychoanalysis after the Second World War.

Systemically Administered, Target-Specific Therapeutic Recombinant Proteins and Nanoparticles for Regenerative Medicine.

Growth factors, chemokines, and cytokines responsible for tissue regeneration have been identified. Their therapeutic usage in humans is almost nonexistent because of the difficulty in maintaining their bioactivity in the protease-rich milieu of injured tissues. Safety concerns have ruled out the systemic administration of growth factors. Angiogenic vasculature forming in the regenerating tissues has unique molecular structures, so-called "zip/postal codes". These unique vascular zip codes provide an opportunity for target-specific delivery of systemically administered therapeutics to tissue injuries by ligands (using peptides or antibodies as a delivery vehicle) binding to these specific structures. Molecules with therapeutic potential can also be packaged into nanocarriers which then can be targeted to the desired location by placing large number of peptides on the nanoparticle. The targeted delivery of systemically administered recombinant proteins to the injured tissue is hopefully rapidly advanced to provide new therapeutics to regenerative medicine.