The phageome in normal and inflamed human skin.

Dysbiosis of skin microbiota drives the progression of atopic dermatitis (AD). The contribution of bacteriophages to bacterial community compositions in normal and inflamed skin is unknown. Using shotgun metagenomics from skin swabs of healthy individuals and patients with AD, we found 13,586 potential viral contiguous DNA sequences, which could be combined into 164 putative viral genomes including 133 putative phages. The Shannon diversity index for the viral metagenome-assembled genomes (vMAGs) did not correlate with AD. In total, we identified 28 vMAGs that differed significantly between normal and AD skin. Quantitative polymerase chain reaction validation of three complete vMAGs revealed their independence from host bacterium abundance. Our data indicate that normal and inflamed skin harbor distinct phageomes and suggest a causative relationship between changing viral and bacterial communities as a driver of skin pathology.

Optimization of in vitro cultivation strategies for human adipocyte derived stem cells.

With adipose-derived stem cells being in the focus of research in regenerative medicine, the need arises for fast reliable cultivation protocols. We have tested the cultivation of human adipose-derived stem cells in endothelial cell growth medium prior to induction and differentiation, against the long-established use of DMEM/F12 medium-based cultivation protocols. We found that cultivation in endothelial cell growth medium not only accelerates growth before induction and differentiation, but also allows shorter induction and differentiation times than those following precultivation with DMEM/F12 medium with regard to the formation of mature adipocytes and to the viability undifferentiated cells. These results were first observed morphologically but could be confirmed by performing adiponectin ELISA and cell proliferation assays.

Total reconstruction of the auricle: our experiences on indications and recent techniques.

INTRODUCTION: Auricular reconstruction is a great challenge in facial plastic surgery. With the advances in surgical techniques and biotechnology, different options are available for consideration. The aim of this paper is to review the knowledge about the various techniques for total auricular reconstruction based on the literature and our experience. METHODS: Approximately 179 articles published from 1980 to 2013 were identified, and 59 articles were included. We have focused on the current status of total auricular reconstruction based on our personal experience and on papers of particular interest, published within the period of review. We have also included a prospective view on the tissue engineering of cartilage. RESULTS: Most surgeons still practice total auricular reconstruction by employing techniques developed by Brent, Nagata, and Firmin with autologous rib cartilage. Within the last years, alloplastic frameworks for reconstruction have become well established. Choosing the reconstruction techniques depends mainly on the surgeon's preference and experience. Prosthetic reconstruction is still reserved for special conditions, even though the material is constantly improving. Tissue engineering has a growing potential for clinical applicability. CONCLUSION: Auricular reconstruction still receives attention of plastic/maxillofacial surgeons and otolaryngologists. Even though clinical applicability lags behind initial expectations, the development of tissue-engineered constructs continues its potential development.

SEE++: a biomechanical model of the oculomotor plant.

The consequences of changes in the oculomotor system on the three-dimensional eye movements are difficult to grasp. Although changes to the rectus muscles can still be approximately understood with simplified geometric models, this approach no longer works with the oblique muscles. It is shown how SEE++, a biomechanical model of the oculomotor plant that was built on the ideas of Miller and Robinson (1984) can improve the understanding of the effects of changes to the oblique eye muscles. By displaying only selected muscles, and by illustrating the relative contribution of these muscles through color-coding the bulb surface, the functional properties of the oblique muscles can be presented in a much clearer way. Investigating the effects of a hyperactive inferior oblique muscle shows that this type of model can help to clarify the functional cause of a pathology, which can otherwise be unclear, even for common pathologies.

Functional maturation of dendritic cells by exposure to CD40L transgenic tumor cells, fibroblasts or keratinocytes.

Tumor antigen pulsed dendritic cells (DCs) can induce anti-tumor immunity. We studied strategies for the reliable generation of such a tumor vaccine by functional maturation of DCs via interaction of CD40 with its ligand (CD40L, CD154). Exposure of immature DCs to CD40L transgenic cells, soluble recombinant human CD40L molecules or lipopolysaccharide induced expression of the co-stimulatory molecules, CD80 and CD86, and supported an allogeneic mixed leukocyte reaction. In contrast, the release of IL-12, an important mediator of anti-tumor immunity, and antigen-specific expansion and IFNgamma secretion of lymphocytes, was strongly triggered only by DCs exposed to CD40L transgenic cells.

Xenogenization by tetanus toxoid loading into lymphoblastoid cell lines and primary human tumor cells mediated by polycations and liposomes.

We explored the potential of the xenogenization concept as an adjuvant procedure in anti-tumor immunity. To mediate effective loading we used polyarginine (pArg) molecules of various degrees of polymerization, cationic liposomes, or chimeric molecules of transferrin (Tf) and the polycation polyethyleneimine (PEI). Tetanus toxoid (TT) was loaded onto primary human leukemia cells, culture adapted primary human neuroblastoma cells, and human lymphoblastoid cell lines (LCLs) with high efficiency by all procedures. Trypsin treatment of loaded cells provided evidence that only liposomes and Tf-PEI mediated internalization of TT. Lymphocytes primed with xenogenized LCLs and challenged with unmodified LCLs showed increased IFNgamma secretion compared with lymphocytes primed with non-xenogenized LCLs.

Coupling of cell-binding ligands to polyethylenimine for targeted gene delivery.

Recently the high transfection potential of the cationic polymer polyethylenimine (PEI) was described (Boussif O et al. Proc Natl Acad Sci USA 1995; 92: 7297-7301). To combine the promising DNA delivering activity of PEI with the concept of receptor-mediated gene delivery, cell-binding ligands (transferrin or antiCD3 antibody) were incorporated by covalent linkage to PEI. DNA complexes of PEI or ligand-PEI conjugates were tested for transfection of cultured neuroblastoma Neuro 2A cells, melanoma B16 or H225 cells, erythroid leukemic K562 cells and T cell leukemia Jurkat E6.1 cells. Depending on the cell line, incorporation of the cell-binding ligand resulted in an up to 1000-fold increased transfection efficiency. This activity depends on ligand-receptor interaction and was observed also at low PEI cation:DNA anion ratios where ligand-free PEI lacks efficiency. Depending on the cell-binding ligand, specific targeting (CD3 antibody, Jurkat cells) can be achieved. Gene transfer can be augmented by the addition of an endosome-destabilizing influenza peptide, but is not dependent on the presence of additional endosomolytic agents. Application of transferrin-PEI for the production of murine interleukin-2 in B16 cells resulted in exceptionally high secretion rates of 19 micrograms IL-2 protein per 10(6) cells per 24 h.

Comparison of epidural analgesia and cryoanalgesia in thoracic surgery.

A prospective study was carried out in 120 patients undergoing elective thoracotomy for parenchymal disease. Patients were randomized into three groups: A (control group), B (epidural analgesia), C (freezing of intercostal nerves). Subjective pain relief was assessed on a linear visual analog scale. Analgesic requirements were evaluated during the 12 days following surgery, or until discharge if earlier. The vital capacity (VC) and forced expiratory volume in 1 s (FEV1) were measured on the day before operation and on the 1st, 2nd, 3rd and 7th postoperative days (POD). Subjective pain relief was significantly better in Group B in comparison with Group A (P < 0.05) or C (P < 0.05). Group C had the lowest score on the 11th and 12th POD but differences were not statistically significant. Requirements for intravenous analgesics were lower in Group B than in the control group (P < 0.05) during the first 3 POD, and in group C than in the control group the day of operation (P < 0.05). Oral analgesic requirements, when compared with controls, were lower in group B during the first 5 POD, and lower in group C on the 3rd and the 4th POD (P < 0.05). Cryoanalgesia led to a slight but not significant increase in VC and FEV1. Epidural analgesia led to a significant increase when compared with controls in FEV1 during the first 3 POD, and in FVC on the 7th POD (P < 0.05). It is concluded that epidural analgesia led to the best pain relief and restoration of pulmonary function after thoracotomy.(ABSTRACT TRUNCATED AT 250 WORDS)