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Legume proteins are increasingly used in structuring various foods under the influence of heating and stirring energy. Based on available studies, this structuring potential is not yet fully understood. This raises the question of the suitability of legume isolates and concentrates for structuring in emulsion gels and the effect of heat and oil on the gel properties. In this study, soy- and pea-based suspensions and emulsions were prepared with the least gelling concentration using different oil concentrations (0%, 7.5%, 15%, 22.5%, and 30%). The viscoelastic properties were measured before and after heating cycles (65 °C and 95 °C). Scanning electron microscopy images complemented the results. All gels measured showed viscoelastic solid behavior. Thermal treatment showed a positive effect on the gel properties for most samples, especially for concentrates (reduction in the loss factor and networking factor > 1). The concentrates showed much higher networking factors and tighter cross-linking than the isolates. The rheological and microstructural properties of the emulsion gels are influenced by a number of factors, such as carbohydrate content, protein chemistry, the protein purification method, and initial viscosity. Moreover, the influence of oil on the rheological properties depends on the material used and whether oil droplets act as an active or inactive filler., Competing Interests: Author Andreas Schubert is employed by Anton Paar Germany GmbH. He participated in 2024 in the study. The role of the company was to support the realization of the method. Author Peter Bauermann was employed by the company EVONIK Operations GmbH. He participated in 2024 in the study. The role of the company was to support the method realization. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Ultrasonic vibration superimposed face milling enables the generation of predefined surface microstructures by an appropriate setting of the process parameters. The geometrical reproducibility of the surface characteristics depends strongly on the plastic material deformation. Thus, the precise prediction of the emerging surface microstructures using kinematic simulation models is limited, because they ignore the influence of material flow. Consequently, the effects of plastic as well as elastic deformation are investigated in depth by finite element analysis. Microstructured surfaces resulting from these numerical models are characterized quantitatively by areal surface parameters and compared to those from a kinematical simulation and a real machined surface. A high degree of conformity between the values of the simulated surfaces and the measured values is achieved, particularly with regard to material distribution. Deficits in predictability exist primarily due to deviations in plastic deformation. Future research can address this, either by implementing a temperature consideration or adapting specific modeling aspects like an adjusted depth of cut or experimental validated material parameters.

Periprosthetic joint infections (PJI) are a common reason for orthopedic revision surgeries. It has been shown that the silver surface modification of a titanium alloy (Ti-6Al-4V) by PMEDM (powder mixed electrical discharge machining) exhibits an antibacterial effect on Staphylococcus spp. adhesion. Whether the thickness of the silver-modified surface influences the adhesion and proliferation of bacteria as well as the ossification processes and in-vivo antibacterial capacity has not been investigated before. Therefore, the aim of this work is to investigate the antibacterial effect as well as the in vitro ossification process depending on the thickness of PMEDM silver modified surfaces. The attachment of S. aureus on the PMEDM modified surfaces was significantly lower than on comparative control samples, independently of the tested surface properties. Bacterial proliferation, however, was not affected by the silver content in the surface layer. We observed a long-term effect of antibacterial capacity in vitro , as well as in vivo . An induction of ROS, as indicator for oxidative stress, was observed in the bacteria, but not in osteoblast-like cells. No influence on the in vitro osteoblast function was observed, whereas osteoclast formation was drastically reduced on the silver surface. No changes in cell death, the metabolic activity and oxidative stress was measured in osteoblasts. We show that already small amounts of silver exhibit a significant antibacterial capacity while not influencing the osteoblast function. Therefore, PMEDM using silver nano-powder admixed to the dielectric represents a promising technology to shape and concurrently modify implant surfaces to reduce infections while at the same time optimizing bone ingrowth of endoprosthesis., Competing Interests: The authors have no financial or personal interest or belief that could affect their objectivity with regard to the present manuscript., (© 2023 The Authors.)

Local anodization with a free electrolyte jet is a suitable solution for locally confined surface functionalization without additionally required preparation of the parts. However, the geometrical formation of the anodic oxide layer in jet-based anodization is not yet sufficiently understood. In this study, numerical calculations based on physical descriptions are used to describe the lateral and vertical oxide formation on aluminum alloy EN AW-7075. The required electrical resistance and capacitance were determined by immersion-based anodization and implemented into the numerical simulation model to evaluate the electrical conductivity of the porous layer. The simulation results showed an electrical conductivity of 2.6 × 10 -6 S/m for the porous layer. Subsequently, a model for jet-based anodization was developed and the previous results were implemented to calculate the oxide formation. The simulation results showed decreasing oxide layer thickness at increasing radial distance from the center of the jet, which corresponds to experimental results. The simulation model was validated by varying the current efficiency from 5% to 90%, where similar developments of the anodic oxide layer thickness compared with experimental results were determined at 5%.

AlSi7Mg/SiC p aluminium matrix composites (AMCs) with a high ceramic content (35 vol.%) that were produced by using the field-assisted sintering technique (FAST) were subjected to tribological preconditioning and evaluated as a potential lightweight material to substitute grey cast iron brake discs. However, since an uncontrolled running-in process of the AMC surface can lead to severe wear and thus to failure of the friction system, AMC surfaces cannot be used directly after finishing and have to be preconditioned. A defined generation of a tribologically conditioned surface (tribosurface) is necessary, as was the aim in this study. To simulate tribological conditions in automotive brake systems, the prepared AMC samples were tested in a pin-on-disc configuration against conventional brake lining material under dry sliding conditions. The influence of the surface topography generated by face turning using different indexable inserts and feeds or an additional plasma electrolytic treatment was investigated at varied test pressures and sliding distances. The results showed that the coefficient of friction remained nearly constant when the set pressure was reached, whereas the initial topography of the samples studied by SEM varied substantially. A novel approach based on analysing the material ratio determined by 3D surface measurement was developed in order to obtain quantitative findings for industrial application.

Laser structuring is by far the most investigated metal surface-pretreatment method for creating adhesion in polymer-metal hybrids. Especially, cone-like protrusions show excellent wetting behaviour as well as high compound strength. However, the processing time is extremely high. Therefore, this paper assesses a process strategy for creating pin structures with scalable height by single pulse drilling with an Nd/YVO 4 nanosecond laser system on EN AW-6082 aluminium alloy. The strength testing is carried out by butt-bonded hollow cylinder torsion. The samples are manufactured by heat-conduction thermal joining with polyamide 6. Ten different surface structures with two different ablation diameters are investigated and compared to cone-like protrusions in terms of processing time, wetting behaviour, shear strength and fracture behaviour. The experimental results show that pulse drilling pins structures with high aspect ratio reach-strength values close to cone-like protrusions but with 31 times higher processing rate.

Introduction: Many people living with HIV/AIDS (PLHIV) in Uganda widely use herbal medicines. However, their toxicity and safety have not been investigated. The use of these plants can potentially cause harmful effects to the health of patients. The purpose of this study was to determine the cytotoxicity of some commonly used medicinal plant species used by PLHIV. Methods: The cytotoxicity of the plant extracts was determined with the AlamarBlue cell viability assay using the human glioblastoma cell line U87.CD4.CXCR4. The cells were treated with varying concentrations of extracts of Warburgia ugandensis , Erythrina abyssinica , Cryptolepis sanguinolenta , Albizia coriaria , Psorospermum febrifugium , Gymnosporia senegalensis , Zanthoxylum chalybeum , Securidaca longipendunculata , Vachellia hockii , Gardenia ternifolia , and Bridelia micrantha reconstituted with ethanol and dimethyl sulfoxide (DMSO). Using regression analysis, the half maximal cytotoxic concentration (CC 50 ) of the plant extracts were calculated from exponential curve fits, since they provided the highest coefficient of determination, R 2 . Results: The ethanol extracts of W. ugandensis (CC 50 = 7.6 μg/ml) and A. coriaria (CC 50 = 1.5 μg/ml) as well as the DMSO-reconstituted extracts of W. ugandensis (CC 50 = 6.4 μg/ml) and A. coriria (CC 50 = < 4 μg/ml) were highly cytotoxic. The cytotoxicity of W. ugandensis and A. coriaria compared well with the indigenous traditional knowledge of the toxic effects experienced when the plants were not used correctly. However, the cytotoxicity of most of the plant extracts (15/22) was low to moderate (CC 50 = 21-200 μg/ml). Conclusion: Most of the plant species tested in this study had low to moderate cytotoxicity against U87.CD4.CXCR4 cells, except W. ugandensis and A. coriria which were highly cytotoxic., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Anywar, Kakudidi, Oryem-Origa, Schubert and Jassoy.)

In organic viticulture, copper-based fungicides are commonly used to suppress Downy Mildew infection, caused by the oomycete Plasmopara viticola . However, the frequent and intensive use of such fungicides leads to accumulation of the heavy metal in soil and nearby waters with adverse effects on the ecosystem. Therefore, alternative, organic fungicides against Downy Mildew are urgently needed to reduce the copper load in vineyards. In this study, the use of Warburgia ugandensis Sprague (Family Canellacea) leaf and bark extracts as potential fungicides against Downy Mildew were evaluated. In vitro (microtiter) and in vivo (leaf discs, seedlings) tests were conducted, as well as field trials to determine the efficacy of the extracts against Downy Mildew. The results revealed an MIC 100 of 500 µg/mL for the leaf extract and 5 µg/mL for the bark extract. Furthermore, experiments with leaf discs and seedlings demonstrated a strong protective effect of the extracts for up to 48 h under (semi-) controlled conditions. However, in field trials the efficacy of the extracts distinctly declined, regardless of the extracts' origin and concentration.

The leaves of Piliostigma thonningii are used in traditional medicine in Benin to treat inflammatory skin diseases and infections. So far, pharmacological studies of the anti-inflammatory and anti-infective effects of phytochemically characterized extracts of P. thonningii have been very limited. Therefore, we investigated the in vitro anti-inflammatory and antimicrobial effect of P. thonningii leaf extracts and analyzed the phytochemical composition of extracts of different polarities (water, 50% ethanol, and n -hexane). Quercetin-3- O -rhamnoside was confirmed as the main flavonoid in the polar extracts. GC-MS analysis identified 20 constituents of the aqueous extract and 28 lipophilic compounds of the n -hexane extract by comparison with authentic standards and spectral library data. The aqueous P. thonningii leaf extract inhibited the IL-8 and IL-6 secretion in TNF- α -stimulated HaCaT cells in a concentration-dependent manner with IC 50 values of 74 µg/mL for IL-8 and 89 µg/mL for IL-6. However, an inhibitory effect of the identified quercetin-3- O -rhamnoside and its aglycone, quercetin, on the release of IL-8 and IL-6 could not be demonstrated. In the antimicrobial screening, inhibition zones for a 50% EtOH leaf extract of P. thonningii were found for Staphylococcus epidermidis, Staphylococcus aureus , methicillin-resistant Staphylococcus aureus , and Escherichia coli . For none of the microbial strains, however, the MIC was below 500 µg/mL, so that the antibacterial activity must be classified as low. As a result, our investigations primarily support the ethnomedical use of P. thonningii leaf extracts in topical inflammatory conditions. Further studies are required to identify the compounds responsible for the in vitro anti-inflammatory effects., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)

Competing Interests: Disclosure The authors report no conflicts of interest in this work.

The fungus growing termite species Macrotermes bellicosus ( M. bellicosus ) is used in nutrition and traditional medicine in the Republic of Benin for the treatment of infectious and inflammatory diseases. Previous findings demonstrated evidence of anti-inflammatory and spasmolytic properties of M. bellicosus . The aim of the present study was to evaluate the antimicrobial potential of different extracts of M. bellicosus samples and determine the chemical profile of an ethanolic M. bellicosus extract. Chemical profiling was conducted using centrifugal partition chromatography and 13 C-NMR, followed by MALDI-TOF MS. Major identified compounds include hydroquinone (HQ), methylhydroquinone (MHQ), 3,4-dihydroxyphenethyl glycol (DHPG), N -acetyldopamine (NADA) and niacinamide. The fatty acid mixture of the extract was mainly composed of linoleic and oleic acid and highlights the nutritional purpose of M. bellicosus . Using the Kirby-Bauer disc diffusion and broth microdilution assay, an antibacterial activity of M. bellicosus samples was observed against various clinical strains with a highest growth inhibition of S. aureus . In addition, HQ and MHQ as well as fractions containing DHPG, niacinamide and NADA inhibited S. aureus growth. The reported antimicrobial activity of M. bellicosus and identified active substances provide a rationale for the traditional medicinal use of M. bellicosus .

UV-C exposure is an effective disinfectant for a range of bacteria and viruses. As such, UV-C treatment, in combination with a chemical wipe, is a common cleaning protocol in medical facilities. Given the increase in severe bacterial and viral agents in society, having access to environmentally friendly disinfectant methods is of increasing interest. In response, we designed, constructed, and validated a UV-C disinfection system from readily accessible components. To improve the UV-C intensity, the enclosure interior was coated with chrome paint. The system is validated using Bacillus cereus , a gram-positive endospore-forming bacteria., Competing Interests: The authors declare no conflicts of interest., (© 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement.)

The interlaminar strength of mechanically interlocked polymer-metal interfaces is strongly dependent on the surface structure of the metal component. Therefore, this contribution assesses the suitability of the fractal dimension for quantification of the surface structure, as well as interlaminar strength prediction of aluminum/polyamide 6 polymer-metal hybrids. Seven different surface structures, manufactured by mechanical blasting, combined mechanical blasting and etching, thermal spraying, and laser ablation, are investigated. The experiments are carried out on a butt-bonded hollow cylinder testing method that allows shear and tensile strength determination with one specific specimen geometry. The fractal dimension of the metal surfaces is derived from cross-sectional images. For comparison, the surface roughness slope is determined and related to the interlaminar strength. Finally, a fracture analysis is conducted. For the investigated material combination, the experimental results indicate that the fractal dimension is an appropriate measure for predicting the interlaminar strength.

Leaves from Combretum collinum Fresen (Combretaceae) are commonly used for the treatment of inflammatory conditions, wound healing and bacterial infections in traditional West African medicine. This research focuses on the characterization of the phenolic profile and lipophilic compounds of leaves extracts of C. collinum . Studies of the in vitro anti-inflammatory activity were performed in TNFα stimulated HaCaT cells and antibacterial activity was evaluated with agar well diffusion and microdilution assays. Antioxidant activity was determined by DPPH and ABTS assays and compared to standards. The phytochemical studies confirmed myricetin-3- O -rhamnoside and myricetin-3- O -glucoside as major components of the leaves extracts, each contributing significantly to the antioxidant activity of the hydrophilic extracts. GC-MS analysis identified 19 substances that were confirmed by comparison with spectral library data and authentic standards. Combretum collinum aqueous leaves extract decreased pro-inflammatory mediators in TNFα stimulated HaCaT cells. Further investigations showed that myricetin-3- O -rhamnoside has an anti-inflammatory effect on IL-8 secretion. In the antimicrobial screening, the largest inhibition zones were found against S. epidermidis , MRSA and S. aureus . MIC values resulted in 275.0 µg/mL for S. epidermidis and 385.5 µg/mL for MRSA . The in vitro anti-inflammatory, antibacterial and antioxidant activity supports topical use of C. collinum leaves extracts in traditional West African medicine., Competing Interests: The authors declare no conflict of interest.

This Data in Brief article provides supplementary information to or earlier Ethnobotanical survey on medicinal plants used by traditional medicine practitioners to boost the immune system in people living with HIV/AIDS in Uganda [1]. We identified 71 medicinal plant species from 37 families and 64 genera. The data were analysed using descriptive statistics such as frequencies and percentages. Most of the plant species used were trees (27) and herbs (25) from the Fabaceae (15.7%) Asteraceae Phyllanthaceae (8.6%), Rubiaceae (5.7%) and Rubiaceae (5.7%) families. Additionally, we conducted a detailed literature review of the documented species to justify their use as immunostimulants. This data is derived from a larger survey to document the use of medicinal plant species in treating opportunistic infections in Uganda by Anywar et al. [2]., (© 2020 The Author(s).)

Mechanical interlocking has been proven to be an effective bonding mechanism for dissimilar material groups like polymers and metals. Therefore, this contribution assesses several surface pretreatments for the metallic adherent. Blasting, etching, combined blasting and etching, thermal spraying, and laser structuring processes are investigated with regard to the achievable interlaminar strength and the corresponding surface roughness parameters. The experiments are carried out on EN AW-6082/polyamide 6 polymer-metal-hybrids, utilizing a novel butt-bonded hollow cylinder specimen geometry for determining the shear and tensile strength. The experimental results indicate that the surface roughness slope has a major impact on the interlaminar strength. A laser-generated pin structure is found to provide the best mechanical performance as well as the highest surface slope of all investigated structuring methods.

Jet electrochemical machining (Jet-ECM) is a flexible method for machining complex microstructures in high-strength and hard-to-machine materials. Contrary to mechanical machining, in Jet-ECM there is no mechanical contact between tool and workpiece. This enables Jet-ECM, like other electrochemical machining processes, to realize surface layers free of mechanical residual stresses, cracks, and thermal distortions. Besides, it causes no burrs and offers long tool life. This paper presents selected features of Jet-ECM, with special focus on the analysis of the current density during the machining of single grooves in stainless steel EN 1.4301. Especially, the development of the current density resulting from machining grooves intersecting previous machining steps was monitored in order to derive systematic influences. The resulting removal geometry is analyzed by measuring the depth and the roughness of the machined grooves. The correlation between the measured product features and the monitored current density is investigated. This correlation shows that grooves with the desired depth and surface roughness can be machined by controlling current density through the adjustment of process parameters. On the other hand, current density is sensitive to the changes of working gap. As a consequence of the changes of workpiece form and size for the grooves intersecting premachined grooves as well as the grooves with a lateral gap, working gap, and current density change. By analyzing monitoring data and removal geometry results, the suitability of current density inline monitoring to enable process control is shown, especially with regards to manufacture products that should comply with tight predefined specifications.

Currently, the demand for surface treatment methods like plasma electrolytic polishing (PeP)-a special case of electrochemical machining-is increasing. This paper provides a literature review on the fundamental mechanisms of the plasma electrolytic polishing process and discusses simulated and experimental results. The simulation shows and describes a modelling approach of the polishing effect during the PeP process. Based on the simulation results, it can be assumed that PeP can be simulated as an electrochemical machining process and that the simulation can be used for roughness and processing time predictions. The simulation results exhibit correlations with the experimentally-achieved approximation for roughness decrease. The experimental part demonstrates the results of the PeP processing for different times. The results for different types of roughness show that roughness decreases exponentially. Additionally, a current efficiency calculation was made. Based on the experimental results, it can be assumed that PeP is a special electrochemical machining process with low passivation., Competing Interests: The authors declare no conflict of interest.

Oral administration of preformed specific antibodies is an attractive approach against infections of the digestive system in humans and animals in times of increasing antibiotic resistances. Previous studies showed a positive effect of egg yolk IgY antibodies on bacterial intoxications in animals and humans. Immunization of chickens with specific antigens offers the possibility to create various forms of antibodies. Research shows that orally applied IgY's isolated from egg yolks can passively cure or prevent diseases of the digestive system. The use of these alternative therapeutic drugs provides further advantages: (1) The production of IgY's is a non-invasive alternative to current methods; (2) The keeping of chickens is inexpensive; (3) The animals are easy to handle; (4) It avoids repetitive bleeding of laboratory animals; (5) It is also very cost effective regarding the high IgY concentration within the egg yolk. Novel targets of these antigen specific antibodies are Helicobacter pylori and also molecules involved in signaling pathways in gastric cancer. Furthermore, also dental caries causing bacteria like Streptococcus mutans or opportunistic Pseudomonas aeruginosa in cystic fibrosis patients are possible targets. Therefore, IgY's included in food for human consumption may be able to prevent or cure human diseases.

Tyrosine kinase inhibitors (TKI), erlotinib and gefitinib are small molecule inhibitors which are used for the treatment of lung cancer. But, the development of drug resistance has been reported as one of the major setbacks in oncology. This study focused on the mechanisms leading to secondary resistance by assessing the gene expression of BCL2 family proteins which are associated with the intrinsic apoptotic signaling pathway. 8 genes were investigated in erlotinib and gefitinib treated cells by real time PCR and protein analysis by western blotting. The cells were exposed to the test drugs 48h prior to RNA or protein isolation. It was observed that BIM-EL, a pro-apoptotic protein was up-regulated in cells sensitive to the drugs but not in the resistant cells. On the other hand BCL2-α, an anti-apoptotic protein was up-regulated in the resistant cells and not in the sensitive cells. BCL2-α revealed a counter-regulation effect on BIM-EL and this effect is probably one of the causes of secondary resistance to erlotinib and gefitinib., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The electrical current exclusion (ECE) principle provides an alternative to common methods of cell diameter measurement and especially in atrophy and cancer associated cachexia research. C2C12 myoblasts were differentiated into myotubes and treated with 100 μM dexamethasone to induce atrophy in vitro. Subsequently, they were incubated for 24 h with media containing different concentrations of curcumin and/or branched-chain amino acids (BCAAs) in order to counteract atrophy. After treatment with curcumin, an increase in cell diameter was detectable; the highest increase with 13.9 ± 0.4% was seen with 10 μM curcumin. The combination of curcumin and BCAAs showed an increase of 13.4 ± 1.2 %. Cell diameter measurement via the ECE showed that curcumin, and curcumin in combination with BCAAs, were able to restore atrophic C2C12 myotubes. Therefore, the application of ECE in muscle atrophy and also cancer-associated cachexia research allows rapid screening of novel compounds in order to test their efficacy in vitro., (Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Although cancer belongs to one of the leading causes of death around the world, fortunately studies have shown that tumor cells have various targets that are susceptible to attack. Interestingly, tumor cells are comprised of cellular membranes, which are altered in chemical composition relative to non-neoplastic cells, giving them an increased net negative charge. These altered membranes are ideal targets for antimicrobial peptides (AMPs) shown to have additional tumoricidal properties and, hence, named anticancer peptides (ACPs). Several hundred ACPs have been explored in vitro and in vivo on various types of cancer. Novel anticancer agents are supposed not to cause serious side effects and the formation of multidrug-resistant tumor cells. During the quest for potent ACPs, promising candidates were isolated from skin secretions of amphibians, such as the granular glands of the Chinese brown frog, Rana chensinensis. ACPs have to be selective to cancer cells and should not induce strong immune responses or be cleared from the body rapidly. Several modifications can improve ACPs either by optimizing the primary structure rationally or randomly or even by introducing other chemical modifications., (Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Curcumin, the extract of the rhizome of Curcuma longa, is known for its health-promoting properties in traditional medicine. It has anti-inflammatory, antitumor and antioxidant properties and stimulates appetite. In the present study, we investigated the stability of curcumin and its effect on cytotoxicity, apoptosis and melanin content in melanoma cells and the effect on atrophic C2C12 muscle cells. Cytotoxicity of curcumin was dose-dependent and the EC50 for 24-h incubation was 69 μM. Saturation was reached at 30 μM for a 48-h incubation. The EC50 for 24-h incubation with degraded curcumin solution was 116 μM and that for 48-h was 94 μM. Curcumin induced a strong increase in caspase-3/7 activity at 30-40 μM. Electrical impedance measurements showed that sub-toxic doses of curcumin counteracted atrophy in an in vitro model system. These findings indicate not only the positive effects of curcumin on melanoma cells in vitro, but also that curcumin was able to considerably trigger anti-cachectic effects in vitro. However, the importance of the stability of curcumin and its tumoricidal and anti-cachectic potential might play a pivotal role in its use in the nutrition and health industrie since it degrades rapidly in aqueous solutions., (Copyright© 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Background: Alveolar echinococcosis (AE) is a life-threatening disease caused by larvae of the fox-tapeworm Echinococcus multilocularis. Crucial to AE pathology is continuous infiltrative growth of the parasite's metacestode stage, which is driven by a population of somatic stem cells, called germinative cells. Current anti-AE chemotherapy using benzimidazoles is ineffective in eliminating the germinative cell population, thus leading to remission of parasite growth upon therapy discontinuation., Methodology/principal Findings: We herein describe the characterization of EmPlk1, encoded by the gene emplk1, which displays significant homologies to members of the Plk1 sub-family of Polo-like kinases that regulate mitosis in eukaryotic cells. We demonstrate germinative cell-specific expression of emplk1 by RT-PCR, transcriptomics, and in situ hybridization. We also show that EmPlk1 can induce germinal vesicle breakdown when heterologously expressed in Xenopus oocytes, indicating that it is an active kinase. This activity was significantly suppressed in presence of BI 2536, a Plk1 inhibitor that has been tested in clinical trials against cancer. Addition of BI 2536 at concentrations as low as 20 nM significantly blocked the formation of metacestode vesicles from cultivated Echinococcus germinative cells. Furthermore, low concentrations of BI 2536 eliminated the germinative cell population from mature metacestode vesicles in vitro, yielding parasite tissue that was no longer capable of proliferation., Conclusions/significance: We conclude that BI 2536 effectively inactivates E. multilocularis germinative cells in parasite larvae in vitro by direct inhibition of EmPlk1, thus inducing mitotic arrest and germinative cell killing. Since germinative cells are decisive for parasite proliferation and metastasis formation within the host, BI 2536 and related compounds are very promising compounds to complement benzimidazoles in AE chemotherapy.

Background: Tyrosine kinase inhibitors (TKI) have emerged as important therapeutic agents for the treatment of several types of cancer including lung cancer. Recent research attempts show that only a small population of cancer patients responds to TKI and furthermore, these patients eventually develop resistance. Studies support the classification of resistance in primary and secondary resistance., Materials and Methods: In the present study the differentiation between primary and secondary resistance to TKI in lung cancer cell lines was investigated. Lung cancer cell lines were tested for viability, apoptosis and cell cycle after exposure to the TKI erlotinib and gefitinib., Results: Cells with primary resistance showed similar cell-cycle patterns to those with secondary resistance but differences were observed between the two groups in the viability and apoptosis assays., Conclusion: Understanding the effects of TKI on cell signaling pathways would shed light on the mechanisms of acquired resistance and the differences between primary and secondary resistance., (Copyright© 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Copeptin, in addition to troponin, has recently been suggested for non-invasive differentiation between Tako-Tsubo cardiomyopathy (TTC) and acute myocardial infarction (AMI). In order to test this hypothesis, we investigated release patterns of pituitary copeptin and cardiac troponin in 49 patients with TTC and 49 age-, gender-, and ECG-matched control patients with AMI. Elevated copeptin levels (i.e. >12 pmol/l) at cardiac catheterization were found in 23/49 (47%) TTC patients and 25/49 (51%) AMI patients. Of these, median copeptin levels were almost identical in both cohorts (34.1 vs. 35.4 pmol/l). Subgroup analysis according to ECG changes revealed that AMI patients with ST-segment elevation had 3.6-fold higher copeptin levels than AMI patients without ST-segment elevation (p<0.05). Furthermore, in patients with TTC and atypical (midventricular) ballooning on left ventricular angiography, copeptin levels were 5.7-fold higher than in TTC patients with a typical (apical) type of ballooning (p<0.01). Elevated troponin T levels (i.e. >0.01 μg/l) at catheterization were detectable in 47/49 (96%) TTC patients and 45/49 (92%) AMI patients; however, peak levels did not differ significantly between both cohorts (median 0.35 vs. 0.27 μg/l). Subgroup analysis according to ECG changes revealed 2-fold higher peak troponin T levels in TTC patients presenting with ST-segment elevation than non-ST-segment elevation (p<0.05). In conclusion, copeptin does not seem to significantly increase non-invasive differentiation between TTC and AMI. At present, coronary angiography, specifically in patients with ST-segment elevation at presentation remains absolutely mandatory., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The removal of biofilms from microstructured titanium used for dental implants is a still unresolved challenge. This experimental study investigated disinfection and removal of in situ formed biofilms from microstructured titanium using cold atmospheric plasma in combination with air/water spray. Titanium discs (roughness (Ra): 1.96 µm) were exposed to human oral cavities for 24 and 72 hours (n = 149 each) to produce biofilms. Biofilm thickness was determined using confocal laser scanning microscopy (n = 5 each). Plasma treatment of biofilms was carried out ex vivo using a microwave-driven pulsed plasma source working at temperatures from 39 to 43°C. Following plasma treatment, one group was air/water spray treated before re-treatment by second plasma pulses. Vital microorganisms on the titanium surfaces were identified by contact culture (Rodac agar plates). Biofilm presence and bacterial viability were quantified by fluorescence microscopy. Morphology of titanium surfaces and attached biofilms was visualized by scanning electron microscopy (SEM). Total protein amounts of biofilms were colorimetrically quantified. Untreated and air/water treated biofilms served as controls. Cold plasma treatment of native biofilms with a mean thickness of 19 µm (24 h) to 91 µm (72 h) covering the microstructure of the titanium surface caused inactivation of biofilm bacteria and significant reduction of protein amounts. Total removal of biofilms, however, required additional application of air/water spray, and a second series of plasma treatment. Importantly, the microstructure of the titanium discs was not altered by plasma treatment. The combination of atmospheric plasma and non-abrasive air/water spray is applicable for complete elimination of oral biofilms from microstructured titanium used for dental implants and may enable new routes for the therapy of periimplant disease.

Atmospheric plasma jets are being intensively studied with respect to potential applications in medicine. The aim of this in vitro study was to test a microwave-powered non-thermal atmospheric plasma jet for its antimicrobial efficacy against adherent oral micro-organisms. Agar plates and dentin slices were inoculated with 6 log(10) c.f.u. cm(-2) of Lactobacillus casei, Streptococcus mutans and Candida albicans, with Escherichia coli as a control. Areas of 1 cm(2) on the agar plates or the complete dentin slices were irradiated with a helium plasma jet for 0.3, 0.6 or 0.9 s mm(-2), respectively. The agar plates were incubated at 37 degrees C, and dentin slices were vortexed in liquid media and suspensions were placed on agar plates. The killing efficacy of the plasma jet was assessed by counting the number of c.f.u. on the irradiated areas of the agar plates, as well as by determination of the number of c.f.u. recovered from dentin slices. A microbe-killing effect was found on the irradiated parts of the agar plates for L. casei, S. mutans, C. albicans and E. coli. The plasma-jet treatment reduced the c.f.u. by 3-4 log(10) intervals on the dentin slices in comparison to recovery rates from untreated controls. The microbe-killing effect was correlated with increasing irradiation times. Thus, non-thermal atmospheric plasma jets could be used for the disinfection of dental surfaces.

Hyperplasia and hypertrophy of fat cells can be found in obesity and increased adiposity is associated with endothelial dysfunction as an early event of atherosclerosis. However, it is unclear whether human adipocytes directly influence endothelial protein secretion. To study the crosstalk between fat and endothelial cells, human umbilical venous endothelial cells (HUVECs) were cultured in infranatants (Adipo) of primary differentiated human adipocytes. Interestingly, significantly increased secretion of 23 cytokines and chemokines from HUVECs was detected in four independent experiments after Adipo stimulation by protein array analysis detecting a total of 174 different proteins. Among those, time-dependent Adipo-induced upregulation of cytokine secretion in HUVECs was confirmed by ELISA for interleukin (IL)-8, monokine induced by gamma interferon, macrophage inflammatory protein (MIP)-1beta, MIP-3alpha, monocyte chemoattractant protein-1, and IL-6. Factors besides adiponectin, leptin, resistin, and tumor necrosis factor alpha appear to mediate these stimulatory effects. Our findings suggest that endothelial cell secretion is significantly influenced towards a proinflammatory pattern by adipocyte-secreted factors.

A common problem in comparative bibliometric studies at the meso and micro level is the differentiation and specialization of research profiles of the objects of analysis at lower levels of aggregation. In this study, institutional profile clusters are used to examine which level of the hierarchical subject classification should preferably be used to build subject-normalized citation indicators. It is shown that a set of properly normalized indicators can serve as a basis of comparative assessment within and even among different clusters, provided that their profiles still overlap and such comparison is thus meaningful. Using the example of 24 European universities, a new version of relational charts is presented for the comparative assessment of citation impact.

Hyperplasia and hypertrophy of fat cells can be found in obesity, and increased adiposity is associated with endothelial dysfunction as an early event of atherosclerosis. However, it is unclear whether human adipocytes directly influence endothelial function. To study the crosstalk between fat and endothelial cells, human umbilical venous endothelial cells (HUVECs), and human coronary artery endothelial cells (HCAECs) were cultured in infranatants (Adipo) of primary differentiated human adipocytes. Interestingly, incubation of HUVECs and HCAECs with Adipo significantly increased monocyte adhesion 7.3 and 2.2-fold, respectively. VCAM-1, ICAM-1, and E-selectin in HUVECs were upregulated 3.9, 3.0, and 9.5-fold, respectively, under these conditions. Furthermore, Adipo significantly stimulated NFkappaB activity 1.9-fold. The NFkappaB inhibitor MG-132 and heat inactivation significantly reversed Adipo-stimulated monocyte adhesion. TNFalpha-neutralizing antibodies partly reversed Adipo-induced monocyte adhesion. In contrast, thiazolidinedione-pretreatment of human adipocytes did not alter the effects of Adipo. Adipo did not show cytotoxic effects. Taken together, we demonstrate that endothelial dysfunction is induced by adipocyte-secreted factors via NFkappaB partly dependent on TNFalpha.

Background: Tobacco abuse is still among the most important cardiovascular risk factors in modern society. We investigated whether sub-chronic nicotine exposure can induce endothelial dysfunction and communication failure., Methods and Results: Primary human umbilical vein endothelial cells (HUVEC) were cultured with or without 1 microM nicotine given for the entire cell culture passage until confluence (5+/-0.5 days). Cells were cultured on special Petri dishes consisting of two compartments which communicated only via a small cellular bridge. We determined the propagation of the NO signal after stimulation of compartment A with ATP by simultaneous spectrophotometric measurement of ATP and methemoglobin formation indicating NO release in compartment B. In HUVECs grown under nicotine we found significantly reduced NO formation in compartment B 5 min after ATP stimulation of compartment A. At that time, there was no ATP detectable in compartment B. The difference in NO-signal-propagation could be abolished with the gap junction blocker Na-propionate. Western blot and immunohistochemistry indicated significantly reduced levels of endothelial gap junction proteins Cx37 and Cx43, but not Cx40. Dye transfer experiments revealed reduced number of communicating cells in nicotine exposed cells indicating the functional relevance., Conclusions: These results - for the first time - show that nicotine induces functional intercellular communication failure in endothelial cells probably resulting from down-regulated Cx37 and Cx43 expression.

Molecular diagnostic tests based on the PCR or alternative nucleic acid amplification technologies are commonly used for pathogen screening at blood drawing centers. Contrived process surveillance using test-specific external and internal controls is critical for the efficient leverage of PCR power. We describe here novel control constructs for use in nucleic acid amplification assays for pathogens with a single-stranded DNA genome, e.g., parvovirus B19. These controls are derived from a deletion mutant of the filamentous phage fd-tet, fKN16, and consist of single-stranded DNA packaged in a protein coat. They are essentially noninfectious to Escherichia coli and highly resistant to nuclease degradation. fKN16 based controls can be readily manufactured and highly purified. Despite their confirmed filamentous morphology, they can be precisely and accurately diluted over a wide range. Stability studies reveal that the novel control constructs are highly resistant to temperature stress, regardless of whether they are tested as concentrated stocks in storage buffer or diluted in buffer or human plasma. Real-time amplification curves derived from recombinant control constructs containing a parvovirus B19 specific sequence fragment match those derived from native virus. In summary, our data demonstrate the feasibility of novel nuclease-resistant single-stranded DNA controls as surrogates for parvovirus B19 and their applicability in routine molecular diagnostics.

Background and Aim of the Study: Reversal of myocardial collagen gene expression was examined in parallel to left ventricular reverse remodeling after surgical correction of experimental aortic stenosis., Methods: A standard growing sheep model (age at baseline 6-8 months) was used. Measurements were performed at baseline (point A) when inducing left ventricular hypertrophy (LVH) through supracoronary banding, at 8.3 +/- 1.0 months later at surgical correction (point B), and after another 10.1 +/- 2.0 months during final examinations (point C). Gene expression for collagen I and III was also studied., Results: Left ventricular function was stable throughout the study. The left ventricular mass index (LVMI) was 82 +/- 21 g/m2 at point A, 150 +/- 33 g/m2 at B, and 78 +/- 18 g/m2 at C (p <0.01). Myocardial fiber diameter was 11.3 +/- 0.8, 15.9 +/- 1.2 and 11.4 +/- 1 microm at points A, B and C, respectively (p <0.01). Protein expression for collagen I was 0.71 +/- 0.2 (at A), 1.13 +/- 0.3 (at B) and 0.85 +/- 0.4 (at C) (p <0.01), while that for collagen III was 0.72 +/- 0.4 (at A), 1.26 +/- 0.8 (at B) and 0.83 +/- 0.3 (at C) (p <0.01). There was a significant correlation between changes in LVMI and myocardial collagen expression., Conclusion: Complete reverse remodeling with regression of LVH and myocardial collagen protein expression can be anticipated after surgical correction of experimental aortic stenosis.

Background: Statins and angiotensin type 1 (AT1) receptor blockers reduce cardiovascular mortality and morbidity. In the Endothelial Protection, AT1 blockade and Cholesterol-Dependent Oxidative Stress (EPAS) trial, impact of independent or combined statin and AT1 receptor blocker therapy on endothelial expression of anti-atherosclerotic and proatherosclerotic genes and endothelial function in arteries of patients with coronary artery disease were tested., Methods and Results: Sixty patients with stable coronary artery disease undergoing elective coronary artery bypass grafting (CABG) surgery were randomized 4 weeks before surgery to: (A) control without inhibition of renin-angiotensin system or statin; (B) statin (pravastatin 40 mg/d); (C) AT1 blockade (irbesartan 150 mg/d); or (D) combination of statin and AT1 blocker in same dosages. Primary end point was a priori therapy-dependent regulation of an anti-atherosclerotic endothelial expression quotient Q including mRNA expression (in arbitrary units measured by real-time polymerase chain reaction) of endothelial nitric oxide synthase and C-type natriuretic peptide, divided by expression of oxidized low-density lipoprotein receptor LOX-1 and NAD(P)H oxidase subunit gp91phox in left internal mammary arteries biopsies obtained by CABG surgery; 49 patients completed the study. Statin therapy increased lnQ from 3.2+/-0.4 to 4.4+/-0.4 significantly versus control. AT(1) blockade showed a trend to increase lnQ to 4.2+/-0.5. Combination of statin and AT1 blocker further increased lnQ to 5.1+/-0.6, but a putative interaction of both therapies in lnQ was not significant. Furthermore, preoperative therapy with statin, AT1 blocker and their combination improved endothelial function in internal mammary artery rings., Conclusions: Statin and AT1 blocker therapy independently and in combination improve an anti-atherosclerotic endothelial expression quotient and endothelial function.

Objective: Recent studies have demonstrated that transplanted bone marrow-derived stem cells (BMCs) possess a broad differentiation potential and are able to form new cardiomyocytes. However, the identity of BMCs as true cardiomyocytes is still ambiguous. Therefore, we investigated the fate of transplanted fluorescence labeled BMCs and cardiomyocytes in co-culture., Methods: For cell tracking we used two different fluorescent probes, Vybrant/DiO and Vybrant/DiI. BMCs were taken from human sternal marrow, purified using a Ficoll-gradient-centrifugation, treated with 5-azacytidine and stained with Vybrant/DiO. Furthermore, isolated spontaneous beating cardiomyocytes of neonatal rats (CM) were labeled with Vybrant/DiI. Thereafter, the BMCs were transplanted into CM-cultures and investigated on day 1, 4, 7, 14 and 28 using two-color fluorescence phenotyping by laser-scanning-cytometry (LSC). Two-color positive cells were harvested by patch-clamp technique and beta-MHC mRNA expression was analyzed by single-cell PCR., Results: Two different morphological phenotypes were observed by LSC. First, isolated DiO labeled BMCs without contact or with direct cell contact to DiI labeled CMs. Second, some BMCs and CMs were double positive for DiO/DiI spontaneously forming hybrids. This population increased by 18% from day 1 to 4 and decreased only slightly until day 28. Additionally, few two-color positive cell formations expressed both human and rat specific beta-MHC mRNA as well as only human beta-MHC mRNA indicating that cell-fusion and transdifferentiation has occurred., Conclusion: These observations provide in vitro evidence for spontaneous cell fusion and transdifferentiation of BMCs in co-culture, raising the possibility that the observed phenomenons may contribute to development or maintenance of these cell types.

Background: Chronic heart failure (CHF) is accompanied by an inflammatory activation which occurs both systemically and in the skeletal muscle. Exercise training has been shown to reduce the local expression of cytokines and inducible nitric oxide synthase (iNOS) in muscle biopsies of CHF patients. INOS-derived NO can inhibit oxidative phosphorylation and contribute to skeletal muscle dysfunction in CHF., Design: To investigate the correlation between changes in local iNOS expression associated with regular exercise and changes in aerobic enzyme activities in the skeletal muscle of patients with CHF. Twenty male CHF patients [ejection fraction 25% (SE 2), age 54 (SE 2) years] were randomized to a training (n=10) or a control group (C, n=10)., Methods: At baseline and after 6 months skeletal muscle iNOS expression was measured by real-time polymerase chain reaction. INOS protein and protein nitrosylation were assessed by immunohistochemistry. Cytochrome c oxidase (COX) activity was quantified electrochemically using the Clark oxygen electrode., Results: Exercise training led to a 27% increase in cytochrome c oxidase activity [from 21.8 (SE 3.2) to 27.7 (SE 3.5) nmol O2/mg per min, P=0.02 versus baseline]. Changes in iNOS expression and iNOS protein content were inversely correlated with changes in COX-activity (r=-0.60, P=0.01; r=-0.71, P<0.001)., Conclusions: The inverse correlation between iNOS expression/iNOS protein content and COX-activity indicates that local anti-inflammatory effects may contribute to improved muscular oxidative metabolism.

Background: Chronic heart failure (CHF) is associated with progressive muscle atrophy and reduced local expression of insulin-like growth factor I (IGF-I)., Design: The present study was designed to test the hypothesis that the local deficiency of IGF-I in the skeletal muscle of patients with CHF would respond to a 6-months aerobic training intervention. Therefore, 18 patients [mean age 52.4 (SD 4.8) years, left ventricular ejection function (LVEF) 27 (SD 6)%] were prospectively randomized to either 6 months of training or sedentary lifestyle., Methods: Serum levels of growth hormone (GH) were measured by immunofluorometric assay, IGF-I by competitive solid phase immunoassay. IGF-I expression was assessed in vastus lateralis biopsies by real-time PCR., Results: Exercise training led to a significant increase in peak oxygen uptake by 26% [from 20.3 (SD 3.3) ml/kg per min to 25.5 (SD 5.7) ml/kg per min, P=0.003 versus control]. Local expression of IGF-I increased significantly after exercise training by 81% [from 6.3 (SE 0.8) to 11.4 (SE 1.4) relative units, P=0.007 versus control] while IGF-I receptor expression was reduced by 33% [from 20.0 (SE 2.1) to 13.8 (SE 1.7) relative units, P=0.008 versus control]. Serum growth hormone (GH) rose modestly from 0.12 (SE 0.07) to 0.65 (SE 0.37) ng/ml in the training group (P=0.043 versus baseline), however, this change was not significant compared to the control group (P=0.848). IGF-I serum levels remained virtually unchanged., Conclusions: Exercise training improves local IGF-I expression without significant changes of systemic parameters of the GH/IGF-I axis. These findings indicate that exercise training has the therapeutic potential to attenuate peripheral skeletal muscle alterations in particular with respect to local IGF-I expression in patients with moderate CHF.

Background: In chronic heart failure (CHF), cross-talk between inflammatory activation and oxidative stress has been anticipated in skeletal muscle (SM). The role of the radical scavenger enzymes superoxide dismutase (SOD), catalase (Cat), and glutathione peroxidase (GPX), which remove oxygen radicals, has never been assessed in the SM in this context. Moreover, it remains unknown whether exercise training augments the activity of these enzymes in CHF., Methods and Results: Twenty-three patients with CHF were randomized to either 6 months of exercise training (T) or a sedentary lifestyle (C); 12 age-matched healthy subjects (HS) were studied in parallel. Activity of Cat, SOD, and GPX was assessed in SM biopsies before and after 6 months (6 months). Oxidative stress was determined by measuring nitrotyrosine formation. SOD, Cat, and GPX activity was reduced by 31%, 57%, and 51%, respectively, whereas nitrotyrosine formation was increased by 107% in SM in CHF (P<0.05 versus HS). In CHF, exercise training augmented GPX and Cat activity in SM by 41% (P<0.05 versus before and group C) and 42% (P<0.05 versus before and group C), respectively, and decreased nitrotyrosine production by 35% (from 3.8+/-0.4% tissue area before to 2.5+/-0.3% after 6 months; P<0.05 versus before)., Conclusions: The reduced activity of major antioxidative enzymes in the SM of CHF patients is associated with increased local oxidative stress. Exercise training exerts antioxidative effects in the SM in CHF, in particular, due to an augmentation in activity of radical scavenger enzymes.

Objective: Transplantation of bone marrow-derived mononuclear cells (BMC) may be suitable to prevent myocardial remodeling and improve left ventricular function after myocardial infarction. However, it is unknown whether or not cardiomyocytes and BMCs can form functioning cell-to-cell coupling and develop adequate electrophysiological properties., Methods: BMCs were isolated from minipig leg bones, treated with 5-azacytidine (10 microM) for 24 h, cultured for 7 days and labeled with a fluoroscopic dye (DIL). BMCs were cocultured with spontaneously beating cardiomyocytes of neonatal rats. On days 4, 7 and 14 cocultured cells were analyzed. Immunhistochemistry (Connexin 43, alpha-actinin) was used to assess cardiomyogenic differentiation. Action potential characteristics were recorded in whole cell patch clamp mode and to investigate intercellular communication a second gap junction permeable fluoroscopic dye was brought into BMCs by microinjection (Lucifer yellow, LY)., Results: From day 7 in coculture BMCs beated synchronously with neonatal rat cardiomyocytes. On day 14, 55.9% of BMCs expressed alpha-actinin and 98.3% were positive for gap junction protein connexin 43. BMC action potential duration (APD90) was mean 11.1 ms with dV/dt(max) of 26.8 V/s and similar to atrial cardiac type. However, microinjection of LY revealed only little dye transfer into adjacent rat cardiomyocytes., Conclusions: Cocultured BMCs have the potential for early expression of muscle specific proteins in about 60% after 14 days and for cardiac gap junction proteins. Synchronous beating indicates an effective electromechanical coupling. In this heterologous setting we could prove only weak metabolic coupling.

Objectives: The aim of this study was to assess the effects of regular physical exercise on local inflammatory parameters in the skeletal muscle of patients with chronic heart failure (CHF)., Background: Inflammatory activation with increased serum cytokine levels and expression of inducible nitric oxide synthase (iNOS) in the myocardium and peripheral skeletal muscles has been described in CHF., Methods: Twenty male patients with stable CHF (left ventricular ejection fraction 25 +/- 2%; age 54 +/- 2 years) were randomized to a training group (n = 10) or a control group (n = 10). At baseline and after six months, serum samples and vastus lateralis muscle biopsies were obtained. Serum tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-1-beta levels were measured by enzyme-linked immunosorbent assay, local cytokine, and iNOS expression by real-time polymerase chain reaction., Results: Exercise training improved peak oxygen uptake by 29% in the training group (from 20.3 +/- 1.0 to 26.1 +/- 1.5 ml/kg. min; p < 0.001 vs. control group). While serum levels of TNF-alpha, IL-6, and IL-1-beta remained unaffected by training, local skeletal muscle TNF-alpha decreased from 1.9 +/- 0.4 to 1.2 +/- 0.3 relative U (p < 0.05 for change vs. control group), IL-6 from 71.3 +/- 16.5 to 41.3 +/- 8.8 relative U (p < 0.05 vs. begin), and IL-1-beta from 2.7 +/- 1.1 to 1.4 +/- 0.6 relative U (p = 0.02 vs. control group). Exercise training also reduced local iNOS expression by 52% (from 6.3 +/- 1.2 to 3.0 +/- 1.0 relative U; p = 0.007 vs. control group)., Conclusions: Exercise training significantly reduced the local expression of TNF-alpha, IL-1-beta, IL-6, and iNOS in the skeletal muscle of CHF patients. These local anti-inflammatory effects of exercise may attenuate the catabolic wasting process associated with the progression of CHF.

Background: Surgical therapy for aortic stenosis leads to reverse remodeling, with normalization of left ventricular hypertrophy (LVH). The aim of this study was to examine Renin-Angiotensin system (RAS) gene expression in this setting., Methods: Growing sheep (n=44) underwent supracoronary aortic banding for controlled induction of LVH at the age of 6 to 8 months (A=baseline). Surgical revision to completely release the pressure gradient was performed 8.3+/-1 months later (B). The animals were sacrificed after another 10.1+/-2 months (C). Along with hemodynamic measurements, subtractive hybridization and competitive polymerase chain reaction were applied to quantify mRNA expression for angiotensin-converting enzyme (ACE) and angiotensin receptors 1 and 2 (AT1-R and AT2-R)., Results: Left ventricular mass index was 82+/-21 g (A), 150+/-33 g (B), and 78+/-18 g (C), P<0.01. Left ventricular function and cardiac index remained stable. Myocardial fiber diameter was 11.3+/-0.8 (A), 15.9+/-1.2 (B), and 11.4+/-1 (C) microm, P<0.01. Gene expression was as follows: ACE 0.8+/-0.05 (A), 1.3+/-0.08 (B), and 0.9+/-0.06 (C), P<0.01; AT1-R 0.7+/-0.06 (A), 0.9+/-0.07 (B), and 0.3+/-0.04 (C), P<0.01; AT2-R 0.5+/-0.05 (A), 0.2+/-0.04 (B), and 0.5+/-0.05 (C), P<0.01., Conclusion: LVH in aortic stenosis coincides with significant alterations of the RAS. Surgical therapy leads to reverse remodeling, which is paralleled by regression of RAS gene expression.

Objectives: We sought to assess the role of insulin-like growth factor-I (IGF-I) in muscle wasting in chronic heart failure (CHF), serum concentrations and local muscular IGF-I expression were determined in patients with severe CHF., Background: Chronic heart failure is associated with progressive muscle atrophy, leading to cardiac cachexia. Skeletal muscle disuse and inflammatory activation with elevated cytokine levels have been implicated; however, the pathomechanism involved remains largely unknown., Methods: Serum levels of IGF-I were measured by competitive solid phase immunoassay in 47 patients with severe CHF (left ventricular ejection fraction < or =30%) and 15 age-matched healthy subjects (HS). Insulin-like growth factor-I and IGF-I receptor expression were assessed in vastus lateralis biopsies by real-time PCR and Western blot analysis., Results: Although serum IGF-I was not significantly different (175 +/- 10 ng/ml in CHF vs. 170 +/- 12 ng/ml in HS, p = NS), local muscle IGF-I mRNA expression was reduced by 52% in CHF (6.7 +/- 0.4 vs. 14.0 +/- 0.9 arbitrary units in HS, p < 0.001). This was accompanied by an increase in IGF-I receptor mRNA expression (86.8 +/- 5.4 in CHF vs. 23.1 +/- 1.8 arbitrary units in HS, p < 0.001). Local IGF-I expression was significantly correlated with muscle cross-sectional area (R = 0.75, p = 0.01). Chronic heart failure patients with a body mass index of < 25 kg/m(2) showed signs of peripheral growth hormone (GH) resistance, as indicated by elevated serum GH, and reduced IGF-I levels., Conclusions: In CHF patients, muscle IGF-I expression is considerably reduced in the presence of normal serum IGF-I levels, possibly contributing to early loss of muscle mass. These findings are consistent with a potential role of IGF-I for skeletal muscle atrophy in CHF.