Mesenchymal stem cells support human vascular endothelial cells to form vascular sprouts in human platelet lysate-based matrices.

During tissue regeneration, mesenchymal stem cells can support endothelial cells in the process of new vessel formation. For a functional interaction of endothelial cells with mesenchymal stem cells a vascular inductive microenvironment is required. Using a cellular model for neo-vessel formation, we could show that newly formed vascular structures emanated from the embedded aggregates, consisting of mesenchymal stem cells co-cultured with autologous human umbilical vein endothelial cells, into avascular human platelet lysate-based matrices, bridging distances up to 5 mm to join with adjacent aggregates with the same morphology forming an interconnected network.

Dependence of mitochondrial function on the filamentous actin cytoskeleton in cultured mesenchymal stem cells treated with cytochalasin B.

Owing to their self-renewal and multi-lineage differentiation capability, mesenchymal stem cells (MSCs) hold enormous potential in regenerative medicine. A prerequisite for a successful MSC therapy is the rigorous investigation of their function after in vitro cultivation. Damages introduced to mitochondria during cultivation adversely affect MSCs function and can determine their fate.

Signs of sympathetic and endothelial cell activation in the skin of patients with restless legs syndrome.

Objectives: To evaluate skin biopsies of patients with early- and late onset restless legs syndrome (RLS) for concomitant small fiber neuropathy (SFN) and to determine cutaneous sympathetic innervation and microvascularization in comparison to healthy individuals.

Methods: Density of intraepidermal nerve fibers (IENFD), adrenergic nerve fibers and dermal capillaries was analyzed by immunofluorescence for PGP9.5, tyrosine hydroxylase and endothelial markers CD31 and CD105 in skin biopsies of 11 individuals with RLS and 8 age- and sex-matched controls.

Influence of Platelet Lysate on 2D and 3D Amniotic Mesenchymal Stem Cell Cultures.

The mechanobiological behavior of mesenchymal stem cells (MSCs) in two- (2D) or three-dimensional (3D) cultures relies on the formation of actin filaments which occur as stress fibers and depends on mitochondrial dynamics involving vimentin intermediate filaments. Here we investigate whether human platelet lysate (HPL), that can potentially replace fetal bovine serum for clinical-scale expansion of functional cells, can modulate the stress fiber formation, alter mitochondrial morphology, change membrane elasticity and modulate immune regulatory molecules IDO and GARP in amnion derived MSCs. We can provide evidence that culture supplementation with HPL led to a reduction of stress fiber formation in 2D cultured MSCs compared to a conventional growth medium (MSCGM).

Bai Hu Tang, Si Ni Tang, and Xue Bi Tang amplify pro-inflammatory activities and reduce apoptosis in endothelial cells in a cell culture model of sepsis.

Ethnopharmacological Relevance: Sepsis is a systemic inflammatory response of the body to a severe infection or massive tissue injury. Despite intensive research, sepsis continues to have a high mortality rate and successful treatment options are strongly needed. Bai Hu Tang (BHT), Si Ni Tang (SNT), and Xue Bi Tang (XBT) are ancient traditional Chinese formulas derived from Chinese herbs that are used to treat Sepsis, but their mechanisms of activity are largely unknown.

miR-146a, miR-146b, and miR-155 increase expression of IL-6 and IL-8 and support HSP10 in an In vitro sepsis model.

microRNAs (miRNAs) play an essential role in inflammation processes including sepsis. This study aimed to identify miRNAs as candidates for therapies that are involved in the innate immune response and to assess their potential functions in the activation of the endothelium. We stimulated THP-1 monocytes with 10 ng/ml LPS for 4 h and used the supernatant for the stimulation of human umbilical vein endothelial cells (HUVEC) or human pulmonary microvascular endothelial cells (HPMEC) for 16 h.

Marker profile for the evaluation of human umbilical artery smooth muscle cell quality obtained by different isolation and culture methods.

Even though umbilical cord arteries are a common source of vascular smooth muscle cells, the lack of reliable marker profiles have not facilitated the isolation of human umbilical artery smooth muscle cells (HUASMC). For accurate characterization of HUASMC and cells in their environment, the expression of smooth muscle and mesenchymal markers was analyzed in umbilical cord tissue sections. The resulting marker profile was then used to evaluate the quality of HUASMC isolation and culture methods.

Monocytes, peripheral blood mononuclear cells, and THP-1 cells exhibit different cytokine expression patterns following stimulation with lipopolysaccharide.

THP-1 cells are widely applied to mimic monocytes in cell culture models. In this study, we compared the cytokine release from THP-1, peripheral blood mononuclear cells (PBMC), monocytes, or whole blood after stimulation with lipopolysaccharide (LPS) and investigated the consequences of different cytokine profiles on human umbilical vein endothelial cell (HUVEC) activation. While Pseudomonas aeruginosa-stimulated (10 ng/mL) THP-1 secreted similar amounts of tumor necrosis factor alpha (TNF- α ) as monocytes and PBMC, they produced lower amounts of interleukin(IL)-8 and no IL-6 and IL-10.

Characterization of SVEP1, KIAA, and SRPX2 in an in vitro cell culture model of endotoxemia.

To assess the influence of unknown factors in endotoxemia, a conditioned medium, achieved by the stimulation of THP1 monocytes with lipopolysaccharide (LPS) [4h], was used for the stimulation of human umbilical vein endothelial cells (HUVECs) [16h]. SVEP1, KIAA0247, and SRPX2 were selected after microarray analysis. To study their possible functions, siRNAs of SVEP1, KIAA0247, or SRPX2 were used for the transfection of HUVECs and cells were stimulated with conditioned medium [16h].

Monitoring of endothelial cell activation in experimental sepsis with a two-step cell culture model.

The aim of this work was to establish and characterize a cell culture model for lipopolysaccharide (LPS)-induced activation of human endothelial cells. Monocytic THP-1 cells were stimulated for 4 h with 10 ng/ml LPS from Pseudomonas aeruginosa in media containing 10% human plasma. Culture supernatants containing LPS and factors secreted by THP-1 in response to stimulation were applied to human umbilical vein endothelial cells (HUVECs).

Kinematical multiple diffraction: the width of psi-scan intensity profiles.

It was shown in Rossmanith [Acta Cryst. (1992), A48, 596-610] that the peak width of intensity profiles of multiple diffraction events can be calculated in a simple manner from the divergence delta and the wavelength spread Deltalambda/lambda of the incident beam and from the mosaic spread eta and the magnitude r of the ideally perfect crystallites in the sample. In the present paper, an improvement of the concept is given.

Comparison of specific adsorbents for tumor necrosis factor-alpha and therapeutic anti-tumor necrosis factor-alpha antibodies: an in vitro sepsis model.

Background: Tumor necrosis factor alpha (TNF-alpha), as a key mediator, represents a major point of attack in sepsis. Since it has been shown that systemic anti-TNF-alpha antibodies do not improve the situation of septic patients, the use of specific adsorption technology in the treatment of sepsis could have beneficial effects.

Methods: Magnetic beads coated with polyclonal or with monoclonal anti-TNF-alpha antibodies were investigated in vitro in order to analyze their ability to prevent TNF-alpha induced adhesion of peripheral blood mononuclear cells (PBMCs) at human umbilical vein endothelial cells (HUVECs).

Behavioural flexibility in the mating system buffers population extinction: lessons from the lesser spotted woodpecker Picoides minor.

1. In most stochastic models addressing the persistence of small populations, environmental noise is included by imposing a synchronized effect of the environment on all individuals. However, buffer mechanisms are likely to exist that may counteract this synchronization to some degree.

Multiple diffraction in the kinematical approach.

The expressions derived for the calculation of multiple diffraction patterns for a perfect crystal in the framework of the kinematical theory are compared with the relevant result of the kinematical power-transfer equation deduced for a mosaic crystal. It is shown that the results of the two concepts differ appreciably.

Pyrogen transfer across high- and low-flux hemodialysis membranes.

The extent to which bacterial products from contaminated dialysate enter a patient's blood depends upon the type and permeability of the hemodialysis membrane in use. This study was performed to assess the transfer of pyrogenic substances across both high- and low-flux membranes (DIAPES, Fresenius Polysulfone, Helixone, Polyamide S). All experiments were carried out in the saline-saline model.

Concerning intensity profiles observed in the 'real experiment'.

The modifications of the kinematical and extinction-corrected intensity profiles caused by the divergence of the incident synchrotron-radiation beam and the diffraction by the monochromator system are considered. Expressions for the integral width as well as the full width at half-maximum are presented and discussed. The theoretical results are compared with observations.

On the structure of aragonite -- Lawrence Bragg revisited.

The structure of aragonite was first determined by Lawrence Bragg in 1924 in what is the now standard space-group setting Pnma (No. 62). Subsequent studies have all taken his structure as their starting points, despite Bragg's own stated doubts and some earlier etching studies which indicated that the underlying symmetry may really be polar.

Concerning intensity profiles.

The kinematical derivation of intensity profiles is considered. It is shown that the shapes of the profiles depend not only on the shape of the crystal but also on the crystal system and axial ratios of the unit cell and on the angles between the direction of the diffracted beam and the cell axes. For all crystal shapes, the widths of the profiles are determined by the Lorentz factor and the mean thickness of the crystal in the direction of the diffracted beam.

Improved antigen and nucleic acid detection in a bovine virus diarrhoea eradication program.

A bovine viral diarrhoea/mucosal disease (BVD/MD) control and eradication program was introduced in Lower Austria in 1996, according to the Swedish model. An important risk factor for BVD transmission under local conditions is communal grazing where susceptible pregnant cattle from several herds may be mixed with unrecognised persistently infected (PI) animals. A reliable system for identification of PI animals is therefore essential for BVD eradication and steps were taken to improve a commercially available antigen-capture ELISA (Ag-ELISA) by modifying the method for leukocyte preparation and adjusting the negative cut-off value.

Rotary cell culture system (RCCS): a new method for cultivating hepatocytes on microcarriers.

The Rotary Cell Culture System (RCCS) is a new technology for growing anchorage dependent or suspension cells in the laboratory. The RCCS is a horizontally rotated, bubble free disposable culture vessel with diffusion gas exchange. The system provides a reproducible, complex 3D in vitro culture system with large cell masses.

In vitro cell culture systems as the basis for an extracorporeal blood purification strategy in multiorgan failure treatment.

Multiorgan failure (MOF) based on septic processes is very common but prognostically an extremely severe disease that has to be treated exclusively under intensive care conditions. Extracorporeal blood purification (ECBP) using specific and efficient systems such as the microspheres based detoxification system (MDS) (Artif Organs 1996;20:420) could improve significantly the situation of MOF in terms of the efficient removal of endotoxins as well as key mediators such as tumor necrosis factor alpha (TNF alpha). The purpose of the study was to test the effectiveness of endotoxin and cytokine removal to blunt cellular response.

Equine lentivirus, comparative studies on four serological tests for the diagnosis of equine infectious anaemia.

Serological diagnosis of equine infectious anemia is of necessity group-reactive, i.e. based on viral core protein p26, because viral envelope components as well as the host's immune response to them undergo rapid antigenic change.

Attempts to immunoprotect adult horses, specifically pregnant mares, with commercial vaccines against clinical disease induced by equine herpesvirus-1.

In a project lasting 4 years more than 300 Lipizzans, around 180 of them adults, were vaccinated systematically against Equine Herpesvirus-1 (EHV-1) and representative groups thereof were serologically controlled for their antibody responses. In part, vaccination intervals recommended on packing slips were followed, in part other intervals, implicated by intermediary results, were used. A live virus vaccine proved ineffective if humoral antibodies were present.

Comparative evaluation of the agar gel immunodiffusion test and two commercial ELISA kits for the serodiagnosis of equine infectious anemia.

Selected sets of serum samples of horses were tested blindly in a comparative investigation for antibodies against Equine Infectious Anemia (EIA) virus. Three commercial kits were used, a well-established agar-gel immuno-diffusion kit which our laboratory has been using routinely for 14 years on one hand, a competitive ELISA kit (CELISA) and a non-competitive ELISA kit on the other hand. The American EIA Reference Laboratory in Ames cotested 56 serum samples with the same 3 products, with highest-level correlation, thereby ascertaining full dependability of our own results.