Present state and future perspectives of using pluripotent stem cells in toxicology research

The use of novel drugs and chemicals requires reliable data on their potential toxic effects on humans. Current test systems are mainly based on animals or in vitro–cultured animal-derived cells and do not or not sufficiently mirror the situation in humans. Therefore, in vitro models based on human pluripotent stem cells (hPSCs) have become an attractive alternative. The article summarizes the characteristics of pluripotent stem cells, including embryonic carcinoma and embryonic germ cells, and discusses the potential of pluripotent stem cells for safety pharmacology and toxicology. Special attention is directed to the potential application of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) for the assessment of developmental toxicology as well as cardio- and hepatotoxicology. With respect to embryotoxicology, recent achievements of the embryonic stem cell test (EST) are described and current limitations as well as prospects of embryotoxicity studies using pluripotent stem cells are discussed. Furthermore, recent efforts to establish hPSC-based cell models for testing cardio- and hepatotoxicity are presented. In this context, methods for differentiation and selection of cardiac and hepatic cells from hPSCs are summarized, requirements and implications with respect to the use of these cells in safety pharmacology and toxicology are presented, and future challenges and perspectives of using hPSCs are discussed

Spectroscopic Studies of the Iron and Manganese Reconstituted Tyrosyl Radical in Bacillus Cereus Ribonucleotide Reductase R2 Protein

Ribonucleotide reductase (RNR) catalyzes the rate limiting step in DNA synthesis where ribonucleotides are reduced to the corresponding deoxyribonucleotides. Class Ib RNRs consist of two homodimeric subunits: R1E, which houses the active site; and R2F, which contains a metallo cofactor and a tyrosyl radical that initiates the ribonucleotide reduction reaction. We studied the R2F subunit of B. cereus reconstituted with iron or alternatively with manganese ions, then subsequently reacted with molecular oxygen to generate two tyrosyl-radicals. The two similar X-band EPR spectra did not change significantly over 4 to 50 K. From the 285 GHz EPR spectrum of the iron form, a g1-value of 2.0090 for the tyrosyl radical was extracted. This g1-value is similar to that observed in class Ia E. coli R2 and class Ib R2Fs with iron-oxygen cluster, suggesting the absence of hydrogen bond to the phenoxyl group. This was confirmed by resonance Raman spectroscopy, where the stretching vibration associated to the radical (C-O, ν7a = 1500 cm−1) was found to be insensitive to deuterium-oxide exchange. Additionally, the 18O-sensitive Fe-O-Fe symmetric stretching (483 cm−1) of the metallo-cofactor was also insensitive to deuterium-oxide exchange indicating no hydrogen bonding to the di-iron-oxygen cluster, and thus, different from mouse R2 with a hydrogen bonded cluster. The HF-EPR spectrum of the manganese reconstituted RNR R2F gave a g1-value of ∼2.0094. The tyrosyl radical microwave power saturation behavior of the iron-oxygen cluster form was as observed in class Ia R2, with diamagnetic di-ferric cluster ground state, while the properties of the manganese reconstituted form indicated a magnetic ground state of the manganese-cluster. The recent activity measurements (Crona et al., (2011) J Biol Chem 286: 33053–33060) indicates that both the manganese and iron reconstituted RNR R2F could be functional. The manganese form might be very important, as it has 8 times higher activity

Strength Training for Arthritis Trial (START): design and rationale

Background Muscle loss and fat gain contribute to the disability, pain, and morbidity associated with knee osteoarthritis (OA), and thigh muscle weakness is an independent and modifiable risk factor for it. However, while all published treatment guidelines recommend muscle strengthening exercise to combat loss of muscle mass and strength in knee OA patients, previous strength training studies either used intensities or loads below recommended levels for healthy adults or were generally short, lasting only 6 to 24 weeks. The efficacy of high-intensity strength training in improving OA symptoms, slowing progression, and affecting the underlying mechanisms has not been examined due to the unsubstantiated belief that it might exacerbate symptoms. We hypothesize that in addition to short-term clinical benefits, combining greater duration with high-intensity strength training will alter thigh composition sufficiently to attain long-term reductions in knee-joint forces, lower pain levels, decrease inflammatory cytokines, and slow OA progression. Methods/Design This is an assessor-blind, randomized controlled trial. The study population consists of 372 older (age ≥ 55 yrs) ambulatory, community-dwelling persons with: (1) mild-to-moderate medial tibiofemoral OA (Kellgren-Lawrence (KL) = 2 or 3); (2) knee neutral or varus aligned knee ( -2° valgus ≤ angle ≤ 10° varus); (3) 20 kg.m-2 ≥ BMI ≤ 45 kg.m-2; and (3) no participation in a formal strength-training program for more than 30 minutes per week within the past 6 months. Participants are randomized to one of 3 groups: high-intensity strength training (75-90% 1Repetition Maximum (1RM)); low-intensity strength training (30-40%1RM); or healthy living education. The primary clinical aim is to compare the interventions’ effects on knee pain, and the primary mechanistic aim is to compare their effects on knee-joint compressive forces during walking, a mechanism that affects the OA disease pathway. Secondary aims will compare the interventions’ effects on additional clinical measures of disease severity (e.g., function, mobility); disease progression measured by x-ray; thigh muscle and fat volume, measured by computed tomography (CT); components of thigh muscle function, including hip abductor strength and quadriceps strength, and power; additional measures of knee-joint loading; inflammatory and OA biomarkers; and health-related quality of life. Discussion Test-retest reliability for the thigh CT scan was: total thigh volume, intra-class correlation coefficients (ICC) = 0.99; total fat volume, ICC = 0.99, and total muscle volume, ICC = 0.99. ICC for both isokinetic concentric knee flexion and extension strength was 0.93, and for hip-abductor concentric strength was 0.99. The reliability of our 1RM testing was: leg press, ICC = 0.95; leg curl, ICC = 0.99; and leg extension, ICC = 0.98. Results of this trial will provide critically needed guidance for clinicians in a variety of health professions who prescribe and oversee treatment and prevention of OA-related complications. Given the prevalence and impact of OA and the widespread availability of this intervention, assessing the efficacy of optimal strength training has the potential for immediate and vital clinical impact

The Timing of Daily Demand for Goods and Services – Multivariate Probit Estimates and Microsimulation Results for an Aged Population with German Time Use Diary Data

Though consumption research provides a broad spectrum of theoretical and empirical founded results, studies based on a daily focus are missing. Knowledge about the individual timing of daily demand for goods and services, opens – beyond a genuine contribution to consumption research – interesting societal and macro economic as well as individual personal and firm perspectives: it is important for an efficient timely coordination of supply and demand in the timing perspective as well as for a targeted economic, social and societal policy for a better support of the every day coordination of life. Last not least, the individual daily public and private living situations will be visible, which are of particular importance for the social togetherness in family and society. Our study contributes to the timing of daily consumption for goods and services with an empirical founded microanalysis on the basis of more than 37.000 individual time use diaries of the nationwide Time Budget Survey of the German Federal Statistical Office 2001/02. We describe the individual timing of daily demand for goods and services for important socio-demographic groups like for women and men, the economic situation with income poverty and daily working hour arrangements. The multivariate microeconometric explanation of the daily demand for goods and services is based on a latent utility maximizing approach over a day. We estimate an eight equation Multivariate/Simultaneous Probit Model, which allows the decision for multiple consumption activities in more than one time period a day. The estimates quantify effects on the timing of daily demand by individual socio-economic variables, which encompasses, personal, household, regional characteristics as well as daily working hour arrangements within a flexible labour market. The question about individual effects of an aged society on the timing of daily demand for goods and services is analyzed with our microsimulation model ServSim and a population forecast for 2020 by the German Federal Statistical Office. Main result: There are significant differences in explaining the timing of daily demand for goods compared to services on the one hand and in particular for different daily time periods. The conclusion: without the timing aspects an important and significant dimension for understanding individual consumption behaviour and their impacts on other individual living conditions would be missing

Timing, Fragmentation of Work and Income Inequality - An Earnings Treatment Effects Approach

Traditional welfare analyses based on money income needs to be broadened by its time dimension. In the course of time the traditional full-time work is diminishing and new labour arrangements are discussed (keyword: flexible labour markets). Our study is contributing to economic well-being by adding insights into particular work effort characteristics - the daily timing of work and its fragmentation - and its resulting income distribution. With our focus on ‘who is working when within a day with which earnings consequences’ we go beyond traditional labour market analyses with its working time division into aggregated full and part time work, working hours spread across a week and weekend, life time working etc. Whereas the first part of our study is describing the distribution of timing and fragmentation of daily work time and its resulting income based on more than 35.000 diaries of the recent German Time Budget Survey 2001/2002, the second part of our study quantifies determinants of arrangement specific earnings functions detecting significant explanatory pattern of what is behind. The economic theory behind is a human capital approach in a market and non-market context, extended by non-market time use, the partner’s working condition, social networking as well as household and regional characteristics. The econometrics use a treatment effects type interdependent estimation of endogenous participation (selection) in a daily working hour pattern (self-selection)and pattern specific earnings function explanation. The overall result: Individual earnings in Germany are dependent on and significant different with regard to the daily working hour arrangement capturing timing and fragmentation of work time. Market and non-market factors are important and significant in explaining earnings