The role of neoadjuvant chemotherapy in the management of primary breast cancer.

Neoadjuvant chemotherapy for primary breast cancer is the gold standard in the treatment of locally advanced, inoperable breast cancer, but based on a large body of evidence has become a standard treatment option for patients with operable disease, who are clear candidates for adjuvant chemotherapy. Neoadjuvant chemotherapy aims at reducing mortality and improving surgical options and offers an in vivo chemosensitivity testing at the same time. It is the ideal setting for clinical and translational research. Administering chemotherapy before surgery raises some important issues concerning the choice of specific treatment regimens as well as the management of the axilla and postoperative radiotherapy. A reliable high quality diagnostic and pathological work-up is mandatory for an ideal tailoring of neoadjuvant chemotherapy. This review gives an outline of the state-of-the-art management of primary breast cancer in the setting of neoadjuvant chemotherapy.

Effect of different gamma-irradiation doses on cytotoxicity and material properties of porous polyether-urethane polymer.

Biomaterials respond to sterilization methods differently. Steam sterilization might decrease the performance of thermoplastic polyether-urethane (TPU); however, the effect of different gamma-radiation doses on this polymer is contradictory in present literature. The purpose of this study was to investigate the differences between irradiative doses in comparison with steam sterilization on a porous TPU scaffold produced by a new processing method. No significant differences in the surface chemical structure were found with attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) analysis when comparing with the sterilization methods. The molecular weight (M(w)) had a net increase from 11.5 +/- 0.039 to 13.2 +/- 0.072 kDa by gamma-sterilization from 10 to 60 kGy. The samples that were irradiated (>60 kGy) had also an increase in polydispersity index (PDI; 1.45 +/- 0.007) in comparison with the nonsterile ones (1.31 +/- 0.017), which indicate branching. Liquid chromatography/mass spectroscopy (LC/MS) analysis showed that there was a correlation between the concentration of the breakdown product, methyl dianiline, and cytotoxicity. The concentration of this compound was found to be four times higher in steam-sterilized sample (1.3 +/- 0.01 ppb) compared with that of the polymer sample gamma-sterilized at 10 kGy (0.3 +/- 0.01 ppb). The cytotoxicity of TPU was found to decrease with higher radiation doses, and was significantly higher for the steam-sterilized samples. It is recommended that TPU produced with the described foaming method should be sterilized by gamma-irradiation at 25 kGy or higher doses.

Brown Marmorated Stink Bug (Hemiptera: Pentatomidae) Infestations in Tree Borders and Subsequent Patterns of Abundance in Soybean Fields.

The invasive brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), is an important pest of soybean (Glycine max L. Merr.) in the Mid-Atlantic United States. In order to assess the influence of nonmanaged wooded borders on H. halys infestation patterns in soybean, 12 soybean fields in Orange and Madison Counties, VA, were sampled each week from July to October in 2013 or 2014 for H. halys. At each location, five 2-min visual counts of H. halys life stages were made on tree of heaven (Ailanthus altissima Mill.) and other favorable host trees along a wooded border, on the adjacent soybean edge, 15 m into the soybean field, and 30 m into the field. Seasonal data showed a clear trend at all locations of H. halys densities building up on A. altissima-dominated wooded borders in July, then, gradually moving into adjacent soybean field edges later in the summer. Halyomorpha halys did not move far from the invading field edge, with approximately half as many bugs being present at 15 m into the field and very few being detected 30 m into the field. These results have implications for continued monitoring and management using field border sprays, particularly on edges adjacent to woods.

A novel processing method for injection-molded polyether-urethane scaffolds. Part 2: cellular interactions.

A large-scale scaffold processing method with injection molding has been successfully developed. Water was used as a foaming agent for the new technique. NaCl was used as a porogen to achieve an open-cell structure. Organic solvents, which are common foaming agents for polyurethane, where not used. Toxic remains in the polymer were therefore prevented. Biocompatibility tested gave a mean optical density of 81% from WST-1 proliferation assay. In comparison to the previously study processing method, hot pressing (Haugen H, Ried V, Brunner M, Will J, Wintermantel E. J Mater Sci: Mater Med2004;15:343-346), the current scaffolds had an increase of 20% of the mean optical density. Cell seeding showed that human fibroblasts adhered to the surface and proliferated. The spread of the adhered fibroblasts was uniform on the surface. A quantitative MTT analysis proved that there was a significant (p < 0.01) increase in the OD level after 7 and 14 days of incubation. This cell layer thickened with increased incubation time from 7 to 14 days (p < 0.05) and had typical fibroblast morphology.

Endotoxin-inducible granulocyte-mediated hepatocytotoxicity requires adhesion and serine protease release.

In primary cultures of Kupffer cells and hepatocytes, human granulocytes potentiated toxicity of endotoxin about 1000-fold. Granulocyte elastase activity was found to correlate with toxicity. The serine protease inhibitors alpha1-antitrypsin, eglin C, and aprotinin protected against toxicity. Tumor necrosis factor-alpha (TNF-alpha) induced cytotoxicity and elastase release, whereas neutralization of TNF-alpha blocked both events. We conclude that TNF-alpha formed by Kupffer cells activates granulocytes. Experiments in cultures where cells were separated by membranes permeable to mediators indicated that cell contact is needed for toxicity. Scanning electron microscopy showed granulocytes adhering to and interdigitating with hepatocytes. Using liver cells from ICAM-1-deficient mice had no effect on toxicity. However, neutralizing CD31 inhibited toxicity and elastase release but not granulocyte adhesion. Our findings demonstrate that adhesion of granulocytes is a necessary but not sufficient condition for the synergistic interaction of endotoxin-stimulated liver macrophages and granulocytes in the proteolytic killing of hepatocytes.

[Techniques for functional tissue and organ replacement using postnatal stem cells]. / Funktioneller Gewebe- und Organersatz mit postnatalen Stammzellen. Technologische Grundlagen.

Postnatal stem cells play a decisive role in cell-based therapies due to their high proliferation activity and functional plasticity. On the one hand, basic research in cell biological processes of adult stem cells is crucial in order to establish them as therapeutic tools. On the other hand, development and enhancements of appropriate techniques are required: we need to establish defined technologies for extraction and differentiation of stem cells and to develop adequate cell carrier devices, scaffolds, and bioreactors for in vitro purposes. Furthermore, it is an interdisciplinary challenge to consider logistical aspects concerning isolation, transport, and storage of stem cells in order to use them in a wide range of activities in regenerative medicine. In this review we present the current methods of work and research on adult stem cells. We explain their therapeutic use and define requirements for future technological developments for work with postnatal stem cells.

Approach to an organo-typical environment for cultured cells and tissues.

If cells or tissues are taken out of an organ and put in culture, normally they lose morphological, physiological and biochemical features. This dedifferentiation process starts during the isolation procedure and continues during the whole culture period. It is caused by the stagnant liquid condition and the inadequate anchorage of cells at the bottom of tissue culture plasticware. The use of filters as basement membrane substitutes and the coating of cultureware with extracellular matrix proteins improve the environmental factors for cultured cells but do not consider the paracrine influence of cytokines or the nutritional needs of individual cell types. To limit cellular dedifferentiation in culture, we constructed a new system, which adapts, as far as possible, cell and tissue cultures to an organo-typical environment. The system is based on a compatible cell carrier arrangement, which allows individual selection of supports for optimal cell anchorage and differentiation. The cell carriers are placed in a newly constructed container, which is permanently perfused with fresh culture medium. The system runs outside an incubator with simple laboratory tools; only a peristaltic pump, a warming table and pH-stabilized media are necessary. Without any subculturing, acute and chronic influences of drugs or the quality of medical implantation grafts can be studied over months.

Tissue-engineered cartilage using serially passaged articular chondrocytes. Chondrocytes in alginate, combined in vivo with a synthetic (E210) or biologic biodegradable carrier (DBM).

In vitro multiplication of isolated autologous chondrocytes is required to obtain an adequate number of cells to generate neo-cartilage, but is known to induce cell-dedifferentiation. The aim of this study was to investigate whether multiplied chondrocytes can be used to generate neo-cartilage in vivo. Adult bovine articular chondrocytes, of various differentiation stages, were suspended in alginate at densities of 10 or 50 million/ml, either directly after isolation (P0) or after multiplication in monolayer for one (P1) or three passages (P3). Alginate with cells was seeded in demineralized bovine bone matrix (DBM) or a fleece of polylactic/polyglycolic acid (E210) and implanted in nude mice for 8 weeks. The newly formed tissue was evaluated by Alcian Blue and immunohistochemical staining for collagen type-II and type-I. Structural homogeneity of the tissue, composed of freshly isolated as well as serially passaged cells, was found to be enhanced by high-density seeding (50 million/ml) and the use of E210 as a carrier. The percentage of collagen type-II positive staining P3-cells was generally higher when E210 was used as a carrier. Furthermore, seeding P3-chondrocytes at the highest density (50 million/ml) enhanced collagen type-II expression. This study shows promising possibilities to generate structurally regular neo-cartilage using multiplied chondrocytes in alginate in combination with a fleece of polylactic/polyglycolic acid.

Sleep deprivation in depression stabilizing antidepressant effects by repetitive transcranial magnetic stimulation.

Partial sleep deprivation (PSD) has a profound and rapid effect on depressed mood. However, the transient antidepressant effect of PSD - most patients relapse after one night of recovery sleep - is limiting the clinical use of this method. Using a controlled, balanced parallel design we studied, whether repetitive transcranial magnetic stimulation (rTMS) applied in the morning after PSD is able to prevent this relapse. 20 PSD responders were randomly assigned to receive either active or sham stimulation during the following 4 days after PSD. Active stimulation prolonged significantly (p < 0.001) the antidepressant effect of PSD up to 4 days. This finding indicates that rTMS is an efficacious method to prevent relapse after PSD.

Detection of polymorphic triplet repeats in the genomes of patients suffering from bipolar affective disorder.

Evidence for the operation of expanded trinucleotide repeats in the pathogenesis of bipolar affected disorder has recently been found at the molecular genetic level. For the screening of these repeat motifs in genomes of patients with bipolar affective disorder, we established a modified PCR-based fingerprinting technique, called triplet repeat enhanced arbitrarily primed PCR (TREAP-PCR). Using this approach, 40 patients suffering from bipolar affective disorder (ICD10: F31) and 15 healthy controls were investigated. Interindividual polymorphisms generated by TREAP-PCR seemed to depend on the type of triplet. Using CCG triplet primers, polymorphisms could be observed more often in the genomes of patients compared with controls, whereas no significant differences could be found using primers of the CAG or AAT type. These data might indicate the existence of subgroups of manic-depressive patients based on molecular genetic differences.

Relapsing polychondritis: a course over 20 years with cerebral involvement.

Relapsing polychondritis is a chronic inflammatory disease associated with an autoimmune disorder in cartilaginous tissue, eyes, labyrinth, blood vessels, and central nervous system. We describe a 75-year-old woman who presented with a 20-year history of dyspnea, inspiratory stridor, and polyarthritis. She developed dysmorphism of both ears and a saddle nose approximately 10 years earlier. Subsequently, she suffered from hearing loss and a tremor. A T2-weighted magnetic resonance imaging scan of the brain revealed multiple, spotted signal intensities. Immunohistochemical analysis of a serum sample showed antibodies to cartilaginous tissue, which were further identified on immunoblotting as antibodies to type II collagen. The extremely prolonged course of disease (>20 years) until a correct diagnosis was made is remarkable. Also, cerebral involvement, which was most likely caused by cerebral angiitis, and which, to our knowledge, has never previously been reported in this form, was detected. Arch Otolaryngol Head Neck Surg. 2000;126:1495-1498

[Microtia: technique for external ear reconstruction with autologous rib cartilage]. / Mikrotie: Technik zur Ohrmuschelrekonstruktion mit autologem Rippenknorpel.

Various methods for treatment of classic microtia are known. Beside a prosthesis, the most common way of auricle reconstruction is the use of autogenous rib cartilage; a process that requires two to three operations. In the first operation, rib cartilage is harvested from the 6th to the 9th rib. The base of the framework is the 6th and 7th rib cartilage which is taken under preservation of the synchondrosis. To mimic a 3-dimensional structure, the triangular fossa and scapha are carved into the groundplate and the 8th rib is fixed as a helical rim. After optimising the framework, it is placed in a subcutaneous pocket on the mastoid plane. In a second operation, approximately three months later, the auriculocephalic angle is reconstructed with a cartilage wedge, which is covered by a temporalis fascia flap and split skin-graft from the hairbearing skull. Commonly, a third operation is needed for minor refinements. Currently, autogenous rib cartilage is the ideal material available for ear reconstruction resulting in an excellent cosmetical outcome, although harvesting of the cartilage causes a specific donor-site morbidity. Operations improving the hearing ability by building up the external hearing channel and middle ear are mainly done in cases of bilateral microtia. Ear reconstruction with autogenous rib cartilage produces a replicable aesthetic result. The patients should be at least eight years old.

Vascular tissue engineering with magnetic nanoparticles: seeing deeper.

The endothelium bares a paramount therapeutic and diagnostic significance in vascular disease. The current work presents a novel strategy based on the use of superparamagnetic nanoparticles to obtain an endothelial cell lining on the luminal surface of vascular conduits, which can be detected non-invasively in a clinical Magnetic Resonance Imaging (MRI) scanner. Human umbilical vein endothelial cells (HUVECs) were prelabeled with clinically approved superparamagnetic nanoparticles. Cell viability and eNOS expression were not affected by the labelling procedure. Magnetically labelled cells were delivered onto the lumen of a PTFE tubular graft by a customised electromagnet. The endothelium was detected in a 1,5T MRI scanner. Magnetic cell delivery provides an efficient technique to seed tubular scaffolds enabling the non-invasive depiction of the cells from the substrate, thus providing a reliable tool to assess the quality of cell delivery procedures.

[The STEMMAT-project as part of health initiative BayernAktiv: adult stem cells from umbilical cord and cord blood as alternative to embryonic stem cell research]. / Das STEMMAT-Projekt als Teil der Gesundheitsinitiative BayernAktiv: Adulte Stammzellen aus Nabelschnur und -blut als Alternative zur embryonalen Stammzellforschung.

Adult stem cells from umbilical cord and cord blood are an interesting alternative to embryonic stem cells because such research is commonly recognized as ethical undisputed and many aspects are still insufficiently investigated. In the context of the STEMMAT research project (STEM = Stem Cell and MAT = Material) different aspects of stem cells from umbilical cord and cord blood are investigated, to improve basic science understanding and potentially leading someday to a clinical application.

[Realistic imaging of cell systems using confocal laser scanning microscopy exemplified by 3-dimensional chondrocyte culture]. / Die realitätsgetreue Darstellung von Zellsystemen mit Hilfe der Konfokalen-Laser-Scanning-Mikroskopie am Beispiel dreidimensional kultivierter Chondrozyten.

BACKGROUND: We have developed a three-dimensional model for tissue engineering of cartilage. Chondrocytes were isolated and first multiplied in conventional monolayer cultures. Then the cells are seeded with or without agarose on special absorbable scaffolds that provided stability and enabled three-dimensional cell distribution of the tissue-engineered cartilage. The aim of the study was to investigate the possibility of avoiding agarose in tissue engineering because of the potential risk of causing an inflammatory process in later human implantation. METHOD: For the first time we investigated cell distribution combined with vitality directly in the cell carrier under the conditions described by using confocal laser-scanning microscopy. Working with unfixed cells, this method enables the reconstruction of three-dimensional cell cultures with suitable cell markers that closely simulates the physiologic situation, thereby exceeding each other method. RESULTS: It was evident that agarose had no positive effect on cell distribution and vitality. CONCLUSION: Further experiments concerning the effect of agarose on synthesis of cartilage-specific matrix are in progress.

Tissue engineering of autologous cartilage transplants for rhinology.

In reconstructive surgery there is increasing demand for cartilage transplants to fill defects, especially nose and/or outer ear defects. Tissue engineering is one of the most modern pathways to generate autologous cartilage transplants. Isolated chondrocytes obtained from a tiny patient's biopsy were seeded on bioresorbable preshaped cell carriers to provide a 3-dimensional cell arrangement as in vivo. The combined use of these cell carriers in form of a non-woven mesh and a constant medium perfusion was performed to generate a cartilage-like cell-polymer-construct, which was finally subcutanously implanted in nude mice for full maturation. After explantation of 6 months, expression of cartilage specific extracellular matrix molecules was obvious by using histochemical and immunohistochemical methods. These data show that tissue engineering with isolated multiplied human chondrocytes from a tiny biopsy seeded on bioresorbable polymer is a promising system to generate autologous cartilage transplants for replacements in reconstructive surgery.

Interrelationship of renal vascular development and nephrogenesis.

Within the cortex region of the neonatal rabbit kidney the developing microvasculature was investigated by means of two endothelium-detecting antibodies (EnPo 1 and EC1). Rows of antibody-labelled cells were found within tissue regions that had previously been described as avascular. We conclude that these vessel-like structures detected by EnPo 1 and EC1 are capillary precursors without lumina. Furthermore, beneath the fibrous capsule within the morphologically homogeneous mesenchyme two cell populations can be discriminated by use of differential antigen expression. The EnPo 1 antigen, which is abundant on endothelial cells and podocytes at different developmental stages, was detected on a subpopulation of mesenchymal cells. These cells were exclusively detected surrounding the tip of the collecting duct ampulla. Due to the unique specificity of EC1 and EnPo 1 the process of microvascular development can be readily followed on serial optical sections gained by laser scan microscopy. (1) Adjacent to EnPo 1-positive mesenchymal cell islets vessel-like structures are found that are in contact with the differentiated vasculature. (2) The renal vesicle is enclosed by a network of vessel-like structures establishing contact with differentiated vessels. (3) No guidance of invading capillary sprouts toward the developing glomerulus and nephron is required, since vascular elements already accompany the earliest detectable nephron stage.