Scleroderma-like Impairment in the Network of Telocytes/CD34+ Stromal Cells in the Experimental Mouse Model of Bleomycin-Induced Dermal Fibrosis

Considerable evidence accumulated over the past decade supports that telocytes (TCs)/CD34⁺ stromal cells represent an exclusive type of interstitial cells identifiable by transmission electron microscopy (TEM) or immunohistochemistry in various organs of the human body, including the skin. By means of their characteristic cellular extensions (telopodes), dermal TCs are arranged in networks intermingled with a multitude of neighboring cells and, hence, they are thought to contribute to skin homeostasis through both intercellular contacts and releasing extracellular vesicles. In this context, fibrotic skin lesions from patients with systemic sclerosis (SSc, scleroderma) appear to be characterized by a disruption of the dermal network of TCs, which has been ascribed to either cell degenerative processes or possible transformation into profibrotic myofibroblasts. In the present study, we utilized the well-established mouse model of bleomycin-induced scleroderma to gain further insights into the TC alterations found in cutaneous fibrosis. CD34 immunofluorescence revealed a severe impairment in the dermal network of TCs/CD34⁺ stromal cells in bleomycin-treated mice. CD31/CD34 double immunofluorescence confirmed that CD31⁻/CD34⁺ TC counts were greatly reduced in the skin of bleomycin-treated mice compared with control mice. Ultrastructural signs of TC injury were detected in the skin of bleomycin-treated mice by TEM. The analyses of skin samples from mice treated with bleomycin for different times by either TEM or double immunostaining and immunoblotting for the CD34/α-SMA antigens collectively suggested that, although a few TCs may transition to α-SMA⁺ myofibroblasts in the early disease stage, most of these cells rather undergo degeneration, and then are lost. Taken together, our data demonstrate that TC changes in the skin of bleomycin-treated mice mimic very closely those observed in human SSc skin, which makes this experimental model a suitable tool to (i) unravel the pathological mechanisms underlying TC damage and (ii) clarify the possible contribution of the TC loss to the development/progression of dermal fibrosis. In perspective, these findings may have important implications in the field of skin regenerative medicine.     

High Mobility Group Box 1 Protein in Cerebral Thromboemboli

High-mobility group box 1 protein (HMGB1) is a damage-associated molecular pattern (DAMP) involved in neutrophil extracellular trap (NET) formation and thrombosis. NETs are regularly found in cerebral thromboemboli. We here analyzed associated HMGB1 expression in human thromboemboli retrieved via mechanical thrombectomy from 37 stroke patients with large vessel occlusion. HMGB1 was detected in all thromboemboli, accounting for 1.7% (IQR 0.6–6.2%) of the total thromboemboli area and was found to be colocalized with neutrophils and NETs and in spatial proximity to platelets. Correlation analysis revealed that the detection of HMGB1 was strongly related to the number of neutrophils (r = 0.58, p = 0.0002) and platelets (r = 0.51, p = 0.001). Our results demonstrate that HMGB1 is a substantial constituent of thromboemboli causing large vessel occlusion stroke.     

Characterization of high molecular weight coffee fractions and their fermentation by human intestinal microbiota

To investigate the structure and fermentability of high M(r) components of coffee brews by human gut bacteria Arabica coffee samples of three different degrees of roast (light, medium, and dark) were used for drip brew preparations and fractionation by ultrafiltration with different M(r) cut-offs. Total carbohydrates of the fractions ranged from 28.6 g/100 g to 56.7 g/100 g. Galactomannans and arabinogalactans were the main polysaccharides and made up between one-fourth and one-half of the respective coffee fraction. After 24 h of incubation with a human fecal suspension the polysaccharides of all fractions were extensively degraded. A decrease in the absorbance values at 405 and 280 nm, respectively, indicated that also chemically noncharacterized UV-active components such as Maillard reaction products, had been partially degraded or modified by the human gut bacteria. The remainder after 24 h of fermentation still showed antioxidant activity. Bacterial cells belonging to the Bacteroides-Prevotella group increased 2- to 40-fold during fermentation depending on the M(r) range of the fraction and the degree of roast. The production of high amounts of acetate and propionate is in accordance with a role of these bacteria in the degradation of high M(r) components from coffee.     

Space-time image sequence analysis: object tunnels and occlusion volumes.

We address the issue of image sequence analysis jointly in space and time. While typical approaches to such an analysis consider two image frames at a time, we propose to perform this analysis jointly over multiple frames. We concentrate on spatiotemporal segmentation of image sequences and on analysis of occlusion effects therein. The segmentation process is three-dimensional (3-D); we search for a volume carved out by each moving object in the image sequence domain, or "object tunnel," a new space-time concept. We pose the problem in variational framework by using only motion information (no intensity edges). The resulting formulation can be viewed as volume competition, a 3-D generalization of region competition. We parameterize the unknown surface to be estimated, but rather than using an active-surface approach, we embed it into a higher dimensional function and apply the level-set methodology. We first develop simple models for the detection of moving objects over static background; no motion models are needed. Then, in order to improve segmentation accuracy, we incorporate motion models for objects and background. We further extend the method by including explicit models for occluded and newly exposed areas that lead to "occlusion volumes," another new space-time concept. Since, in this case, multiple volumes are sought, we apply a multiphase variant of the level-set method. We present various experimental results for synthetic and natural image sequences.     

Endwall convective heat transfer for bluff bodies

The endwall heat transfer characteristics of forced flow past bluff bodies have been investigated using liquid crystal thermography (LCT). The bluff body is placed in a rectangular channel with both its ends attached to the endwalls. The Reynolds number varies from 50,000 to 100,000. In this study, a single bluff body and two bluff bodies arranged in tandem are considered. Due to the formation of horseshoe vortices, the heat transfer is enhanced appreciably for both cases. However, for the case of two bluff bodies in tandem, it is found that the presence of the second bluff body decreases the heat transfer as compared to the case of a single bluff body. In addition, the results show that the heat transfer exhibits Reynolds number similarity. For a single bluff body, the Nusselt number profiles collapse well when the data are scaled by Re^0.55; for two bluff bodies arranged in tandem, the heat transfer scaling is changed to Re^0.51, indicating that the power index of Reynolds number is flow dependent.     

Development of benzophenone-based farnesyltransferase inhibitors as novel antimalarials

The development of farnesyltransferase inhibitors directed against Plasmodium falciparum is a strategy towards new drugs against malaria. Previously, we described benzophenone-based farnesyltransferase inhibitors with high in vitro antimalarial activity but no in vivo activity. Through the introduction of a methylpiperazinyl moiety, farnesyltransferase inhibitors with in vivo antimalarial activity were obtained. Subsequently, a structure-based design approach was chosen to further improve the antimalarial activity of this type of inhibitor. As no crystal structure of the farnesyltransferase of the target organism is available, homology modeling was used to reveal differences between the active sites of the rat/human and the P. falciparum farnesyltransferase. Based on flexible docking data, the piperazinyl moiety was replaced by a N,N,N'-trimethylethylenediamine moiety. This resulted in an inhibitor with significantly improved in vitro and in vivo antimalarial activity. Furthermore, this inhibitor displayed a notable increase in selectivity towards malaria parasites relative to human cells.     

An efficient numerical technique for solving population balance equation involving aggregation, breakage, growth and nucleation

A new discretization for simultaneous aggregation, breakage, growth and nucleation is presented. The new discretization is an extension of the cell average technique developed by the authors [J. Kumar, M. Peglow, G. Warnecke, S. Heinrich, and L. Mörl. Improved accuracy and convergence of discretized population balance for aggregation: The cell average technique. Chemical Engineering Science 61 (2006) 3327-3342.]. It is shown that the cell average scheme enjoys the major advantage of simplicity for solving combined problems over other existing schemes. This is done by a special coupling of the different processes that treats all processes in a similar fashion as it handles the individual process. It is demonstrated that the new coupling makes the technique more useful by being not only more accurate but also computationally less expensive. At first, the coupling is performed for combined aggregation and breakage problems. Furthermore, a new idea that considers the growth process as aggregation of existing particle with new small nuclei is presented. In that way the resulting discretization of the growth process becomes very simple and consistent with first two moments. Additionally, it becomes easy to combine the growth discretization with other processes. The new discretization of pure growth is a little diffusive but it predicts the first two moments exactly without any computational difficulties like appearance of negative values or instability etc. The numerical scheme proposed in this work is consistent only with the first two moments but it can easily be extended to the consistency with any two or more than two moments. Finally, the discretization of pure and coupled problems is tasted on several analytically solvable problems.