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The development of mammalian cell perfusion cultures is still laborious and complex to perform due to the limited availability of scale-down models and limited knowledge of time- and cost-effective procedures. The maximum achievable viable cell density (VCDmax ), minimum cell-specific perfusion rate (CSPRmin ), cellular growth characteristics, and resulting bleed rate at steady-state operation are key variables for the effective development of perfusion cultures. In this study, we developed a stepwise procedure to use shake tubes (ST) in combination with benchtop (BR) bioreactors for the design of a mammalian cell perfusion culture at high productivity (23 pg·cell-1 ·day-1 ) and low product loss in the bleed (around 10%) for a given expression system. In a first experiment, we investigated peak VCDs in STs by the daily discontinuous medium exchange of 1 reactor volume (RV) without additional bleeding. Based on this knowledge, we performed steady-state cultures in the ST system using a working volume of 10 ml. The evaluation of the steady-state cultures allowed performing a perfusion bioreactor run at 20 × 106 cells/ml at a perfusion rate of 1 RV/day. Constant cellular environment and metabolism resulted in stable product quality patterns. This study presents a promising strategy for the effective design and development of perfusion cultures for a given expression system and underlines the potential of the ST system as a valuable scale-down tool for perfusion cultures.

Laparoscopic Roux-en-Y gastric bypass (RYGB) and sleeve gastrectomy (SG) are common and effective methods to treat severe obesity, but these procedures can adversely influence bone metabolism and areal bone mineral density (aBMD). This was a prospective 24-month single-center interventional two-arm study in 220 women and similarly aged men (median age 40.7 years) with a body mass index (BMI) >38 kg/m(2) after RYGB and SG procedures. Patients were randomized into: 1) an intervention group receiving: 28,000 IU cholecalciferol/wk for 8 weeks before bariatric surgery, 16,000 IU/wk and 1000 mg calciummonocitrate/d after surgery, daily BMI-adjusted protein supplementation and physical exercise (Nordic walking, strength perseverance, and equipment training); 2) a non-intervention group: no preoperative loading, nutritional supplementation, or obligatory physical exercise. At study endpoint, when comparing the intervention group to the non-intervention group, the relative percentage changes of serum levels of sclerostin (12.1% versus 63.8%), cross-linked C-telopeptide (CTX, 82.6% versus 158.3%), 25-OH vitamin D (13.4% versus 18.2%), phosphate (23.7% versus 32%, p < 0.001 for all), procollagen type 1 amino-terminal propeptide (P1NP, 12% versus 41.2%), intact parathyroid hormone (iPTH, -17.3% versus -7.6%), and Dickkopf-1 (-3.9% versus -8.9%, p < 0.05 for all) differed. The decline in lumbar spine, total hip and total body aBMD, changes in BMI, lean body mass (LBM), as well as changes in trabecular bone score (TBS) values (p < 0.005 for all) were less, but significantly, pronounced in the intervention group. We conclude that vitamin D loading and ongoing vitamin D, calcium, and BMI-adjusted protein supplementation in combination with physical exercise decelerates the loss of aBMD and LBM after bariatric surgery. Moreover, the well-known increases of bone turnover markers are less pronounced.

It doesn't take much to oxidize cycloalkanes directly to the corresponding cyclic ketones: molecular oxygen as the oxidant, water as the solvent, the cofactor NADP(+) (and a little 2-propanol to reduce it), as well as two catalytic enzymes-a hydroxylating P450 monooxygenase and an alcohol dehydrogenase. (Copyright 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background Investigate the usefulness of echocardiography and acoustic cardiography to monitor patients exposed to anthracycline chemotherapy. Hypothesis Serial echocardiographies to monitor systolic function may not be neccessary in all patients undergoing anthracycline chemotherapy. Methods In a prospective study, consecutive patients undergoing anthracycline-containing chemotherapy were evaluated with echocardiography and acoustic cardiography at baseline, after completion of chemotherapy, and after a median follow-up of 3.8 years. Systolic dysfunction was defined as a left ventricular ejection fraction ≤50%. Results A total of 187 patients (83% female) with a mean age of 55 ± 14 years underwent chemotherapy for breast cancer (73%), malignant lymphoma (23%), and sarcoma (4%). None of the patients had systolic dysfunction at baseline. Patients were treated with doxorubicin 276 ± 74 mg/m2 or epirubicin 317 ± 55 mg/m2. After chemotherapy, 170 (91%) had normal systolic function, 8 (4%) developed systolic dysfunction, and 9 (5%) had died. Of those 8 patients with systolic dysfunction, 4 (50%) improved to normal systolic function, 1 (13%) remained unchanged, and 3 (37%) died. Patients with normal systolic function after chemotherapy had a mortality rate of 3.5%, and 1.8% developed late systolic dysfunction. Acoustic cardiography-derived percent electromechanical activation time >12.4% had a sensitivity of 88% and a specificity of 84% to identify patients with systolic dysfunction (area under the receiver operating characteristic curve 0.87). Conclusions Patients with systolic dysfunction early after anthracycline treatment had worse outcome. Acoustic cardiography was able to identify these patients with a high sensitivity and specificity. Based on the findings of this study, we propose a simple algorithm to monitor patients undergoing anthracycline-containing chemotherapy. The authors have no funding, financial relationships, or conflicts of interest to disclose.

Summary: Purpose: Valproic acid (VPA) is an antiepileptic drug (AED) commonly used for generalized and focal epilepsies. We provide an update on hepatotoxic side effects in Germany between 1994 and 2003. Methods: We mailed a questionnaire to all members of the German Section of the International League Against Epilepsy, asking for VPA-induced side effects, especially severe side effects such as hepatopathy. Results: As a result of our questionnaire, we found 31 cases of reversible hepatotoxicity and nine cases of lethal hepatopathies in Germany from 1994 to 2003. Conclusions: The outcome of patients with severe hepatotoxicity is better than that in the past. The risk of a VPA-induced hepatopathy is not limited to patients younger than 2 years, receiving polytherapy, or patients with congenital or acquired metabolic diseases.

Air particulate matter (PM) and bound chemicals are potential mediators for adverse health effects. The cytotoxicity and changes in energy-providing processes caused by chemical compounds bound to PM of different size fractions were investigated in Tetrahymena pyriformis. The PM samplings were carried out using a high volume cascade impactor (6 size fractions between 10 μm and less than 0.49 μm) at three points of La Plata, Argentina: in an industrial area, a traffic-influenced urban area, and a control area. Extracts from respirable particles below 1 μm initiated the highest cytotoxic effects, demonstrating their higher risk. In contrast, an increase on oxygen consumption was observed especially in tests of extracts from particles less than 1 μm from urban and industrial areas. The increase on oxygen consumption could be caused by decoupling processes in the respiratory chain. Otherwise the ATP concentration was increased too, even though to a lower extent. The observed imbalance between oxygen consumption and ATP concentration in exposed T. pyriformis cells may be due to oxidative stress, caused by chemical compounds bound to the particles. Owing to the complexity of effects related to PM and their associated chemical compounds, various physiological parameters necessarily need to be investigated to obtain more information about their possible involvement in human relevant pathogenic processes. As shown here, effects on cell proliferation and on energy-providing processes are suitable indicators for the different impact of PM and adsorbed chemicals from various sampling locations. © 2006 Wiley Periodicals, Inc. Environ Toxicol 21: 457–463, 2006.

Airborne particulates, especially fine particles and bound chemical compounds, are a potential mediator of adverse health effects. In this study an analysis was done of the concentration and size distribution of air particulate matter, the content of bound polycyclic aromatic hydrocarbons (PAHs), and the biological effects of organic extracts from different fractions of dust that had been influenced by urban and industrial emissions from the regions of La Plata, Argentina, and Leipzig, Germany, along with samples from control areas. Air particulate matter was sampled in summer and winter in each region using a high-volume sampler with a six-stage cascade impactor, classifying dust in six size fractions from 10-μm particles to those less than 0.49 μm in size. Organic extracts of dust were tested for mutagenicity (Ames test, Salmonella typhimurium TA98 strain, S9+) and cytotoxicity (Tetrahymena pyriformis test system, growth rate, cell respiration). The content of PAHs was analyzed by high-performance liquid chromatography (HPLC) with diode array and fluorescence detection. Mutagenic and cytotoxic effects were found to be associated with very fine (

Endolymphatic ion composition in the adult inner ear is characterized by high K(+) and low Na(+) concentration. This unique ion composition is essential for proper functioning of sensory processing. Although a lot has been learned in recent years about molecules involved in K(+) transport in inner ear, the molecules involved in Na(+) transport are only beginning to emerge. The epithelial Na(+) channel (ENaC) is a highly selective Na(+) channel that is expressed in many Na(+)-reabsorbing tissues. The aim of our study was to investigate whether ENaC is expressed in inner ear of rats and could account for Na(+) reabsorption from endolymph. We detected mRNA for the three channel-forming subunits (alpha, beta and gamma ENaC) in cochlea, vestibular system and endolymphatic sac. mRNA abundance increased during the first 12 days of life in cochlea and vestibular system, coinciding with decreasing Na(+) concentration in endolymph. Expression was strongest in epithelial cells lining scala media, most notably Claudius' cells. As these cells are characterized by a very negative resting potential they would be ideally suited for reabsorption of Na(+). mRNA abundance in endolymphatic sac decreased during the first 6 days of life, suggesting that ENaC might be implicated in reabsorption of endolymph in the endolymphatic sac of neonatal animals. Together, our results suggest that the epithelial Na+ channel is a good candidate for a molecule involved in Na(+) homeostasis in inner ear.

Chemical characterization and effects assessment of semivolatile organic compounds in organic extracts from air particulate matter from the region of the greater La Plata area was undertaken. Effects covered the study of mutagenicity with the Ames test (Salmonella typhimurium TA100 and TA98 strains with metabolic activation by S9) and cytotoxicity using the Tetrahymena pyriformis test system (growth rate, cell volume, and cell respiration). Chemical analysis of organic extracts was done using gas chromatography. Results demonstrate the presence of polycyclic aromatic hydrocarbons (PAH) in the matrix, high mutagenicity, and cytotoxicity. A higher mutagenic activity detected with TA98 and TA100 strains is associated with an increment of total PAH and total five or six ring PAH content, respectively. Linked with it, a PAH dependent toxicity on Tetrahymena pyriformis has been observed. This cell system proved to be very sensitive. From the results obtained with the cell respiration assay with T. pyriformis it appears that uncoupling agents are present in the samples. The results of this study indicate that air particulate matter from the Rio de La Plata area contains significant genotoxic and cytotoxic activity probably due to the presence of PAHs.

Networks of 'flexible' and 'rigid' chains were synthesized. As 'flexible' component we have used different partially hydrogenated cis-1,4-polybutadiene. The 'rigid' component was synthesized from bis(1,2,4-triazoline-3,5-dione)s and biscyclohexadienes via repetitive Diels-Alder reaction. A slight excess of bis(1,2, 4-triazoline-3,5-dione)s leads to polymers with 1,2,4-triazoline-3,5-dione end groups, which can easily react with the partially hydrogenated cis-1,4-polybutadiene. The influence of the extent of hydrogenation and the amount of crosslinker on the mechanical and thermal behaviour is described.

“Rigid-rod” telechelics were synthesized via repetitive Diels-Alder reaction of biscyclohexadienes and bis(1,2,4-triazoline-3,5-diones). The bistriazolinediones were used in slight excess. The rigidity of the building blocks and the molecular weight were varied. Various amounts of these reactive telechelics are used to crosslink cis-1,4-polybutadiene. The mechanical and the thermal properties of these two-component networks were investigated by stress-strain, differential scanning calorimetry and dynamic-mechanical measurements. The occurrence of microphase separation between the “rigid” and the “flexible” components is discussed to explain the properties of these materials.

The purpose of this paper is the empirical investigation of the status quo of green supply chain management in the German automotive industry from a practitioner's point of view. Several aspects of green supply chain management, such as the point of time of implementation, the driving forces, the relevance of intended goals and their particular realization and the adoption of eco-programs with suppliers and customers as well as internal and external barriers are analyzed. Furthermore, the link to different performance criteria is tested by inductive statistics in order to show the potential of green supply chain management for competitiveness. For this study, managers from the automotive supply industry in Germany were asked about their estimations in this regard. The main conclusion is that managers express the need for green supply chain management on the one hand but also see corresponding problems in terms of required resources on the other hand. Furthermore, the analyses show that green supply chain management leads to higher performance in terms of several performance criteria. Copyright © 2009 John Wiley & Sons, Ltd and ERP Environment.