Pharmacodynamic differences between species exemplified by the novel anticancer agent CHS 828

When a candidate drug enters clinical trials, decisions regarding dosing are mainly based on animal data. Occasionally, toxicity problems are faced in the clinic because of unexpected species differences in pharmacokinetics or pharmacodynamics between humans and preclinical species. Fludarabine and topotecan are examples of such drugs. In the first clinical trials of the new agent CHS 828, the maximum tolerated dose was reached earlier than expected from animal data. This paper discusses the issue of species differences in the development of anticancer drugs, and preclinical models for detection and quantification of such differences. Pharmacokinetic and hematological toxicity data of CHS 828 from studies in rats and humans are presented. In vitro sensitivity to CHS 828 and some established cytotoxic agents was measured in lymphocytes from humans and rats and in a panel of human and rodent cell‐lines. 10–100 times higher CHS 828 exposure was tolerated by rats than by patients. In both in vitro cell systems, CHS 828 showed higher potency in human cells compared to rodent cells. A species difference was evident also for fludarabine, but not for doxorubicin and cisplatin. CHS 828 pharmacokinetics were similar across species. In conclusion, the lower tolerance of CHS 828 in humans than in rats could be detected in vitro in cultures of peripheral lymphocytes. Preclinical studies of species differences could help the interpretation of in vivo effect studies as well as the choice of starting dose for clinical trials. We suggest peripheral lymphocytes from different species as a potential model system for such studies. Drug Dev. Res. 61:218–226, 2004. © 2004 Wiley‐Liss, Inc.     

Promoting and sustaining health through increased vegetable and fruit consumption among European schoolchildren: The Pro Children Project

Aim The Pro Children consortium consists of the following partners: Knut-Inge Klepp (Coordinator), Department of Nutrition, University of Oslo, Norway; Carmen Perez Rodrigo, Unidad de Nutricion Comunitaria, Bilbao, Spain; Inga Thorsdottir, Unit for Nutrition Research, Landspitali University Hospital, Reykjavik, Iceland; Pernille Due, Department of Social Medicine, University of Copenhagen, Denmark; Maria Daniel Vaz de Almeida, Faculdade de Ciências da Nutrição e Alimentação da Universidade do Porto, Portugal; Ibrahim Elmadfa and Alexandra Wolf, Institute of Nutrition, University of Vienna, Austria; Jóhanna Haraldsdóttir, Research Department of Human Nutrition, Royal Veterinary and Agricultural University, Copenhagen, Denmark; Johannes Brug, Erasmus Medical Center Rotterdam, Department of Public Health, The Netherlands; Michael Sjöström and Agneta Yngve, Unit for Preventive Nutrition, Karolinska Institutet, Stockholm, Sweden; Ilse De Bourdeaudhuij, Department of Movement and Sport Sciences, Ghent University, Belgium.The Pro Children study is designed to assess vegetable and fruit consumption and determinants of the consumption patterns among European school children and their parents. A second objective is to develop and test strategies for promoting increased consumption of vegetables and fruits among school children and their parents.Subjects and methods Surveys of national, representative samples of 11-year-old school children and their parents were conducted in nine countries during October–November 2003, i.e. in Austria, Belgium, Denmark, Iceland, The Netherlands, Norway, Portugal, Spain and Sweden. Comprehensive school-based educational programmes were developed and tested in three settings, i.e. in the Bilbao region, Spain, in Rotterdam, The Netherlands, and in Buskerud county of Norway. A 24-h recall format and frequency items assessing regular intake were used to assess vegetable and fruit consumption. Determinants were assessed employing the theoretical framework of the ASE model (Attitudes, Social Influences and Self-Efficacy), including cognitive factors, normative influences, skills and environmental barriers related to vegetable and fruit consumption. The intervention programmes were tested employing a group-randomized trial design where schools were randomly allocated to an intervention arm and a delayed intervention arm. Surveys among all participating children and their parents were conducted prior to the initiation of the intervention (September 2003; month 0), immediately after the end of the intervention (at month 8) and at the end of the subsequent school year (month 20).Results Preliminary data from the project indicate that girls eat vegetables and fruit significantly more often than do boys across all participating countries. There are no sex differences, however, with respect to perceived availability of vegetables and fruit at home and outside the home setting. In all countries, perceived availability appears to be significantly associated with reported frequency of both vegetable and fruit consumption.Conclusion Experience so far indicates that the Pro Children Project will succeed in producing valid and reliable research instruments for assessing vegetable and fruit consumption among school children and their parents and that comparable, comprehensive intervention programmes can be implemented across geographic and cultural settings within Europe.     

Bromobenzene-induced hepatotoxicity at the transcriptome level.

Rats were exposed to three levels of bromobenzene, sampled at 6, 24, and 48 h, and liver gene expression profiles were determined to identify dose and time-related changes. Expression of many genes changed transiently, and dependent on the dose. Few changes were identified after 6 h, but many genes were differentially expressed after 24 h, while after 48 h, only the high dose elicited large effects. Differentially expressed genes were involved in drug metabolism (upregulated GSTs, mEH, NQO1, Mrps, downregulated CYPs, sulfotransferases), oxidative stress (induced HO-1, peroxiredoxin, ferritin), GSH depletion (induced GCS-l, GSTA, GSTM) the acute phase response, and in processes like cholesterol, fatty acid and protein metabolism, and intracellular signaling. Trancriptional regulation via the electrophile and sterol response elements seemed to mediate part of the response to bromobenzene. Recovery of the liver was suggested in response to BB by the altered expression of genes involved in protein synthesis and cytoskeleton rearrangement. Furthermore, after 48 h, rats in the mid dose group showed no toxicity, and gene expression patterns resembled the normal situation. For certain genes (e.g., CYP4A, metallothioneins), intraday variation in expression levels was found, regardless of the treatment. Selected cDNA microarray measurements were confirmed using the specific and sensitive branched DNA signal amplification assay.     

Social-cognitive and school factors in lifetime smoking among adolescents

BACKGROUND: Smoking is a serious health threat and identifying risk factors for smoking is thus of great importance. The aim of the study was to examine the effects of social-cognitive factors and school factors on lifetime smoking status among adolescents. METHODS: The study was based on cross-sectional data on 2,913 Danish adolescents in grade 7 attending 118 randomly selected public schools. Social-cognitive factors were examined with five measures: self-efficacy to resist pressure to smoke, social influence (norms), social influence (behavior), social influence (pressure), and attitude. We used multilevel analyses to estimate the associations between social-cognitive factors and lifetime smoking status as well as the group-level effects of school, school class, and gender group in the school class. RESULTS: Each social-cognitive factor was significantly associated with lifetime smoking status, even when several potential confounders and the effects of school, school class, and gender group were taken into account. Of the three group-level school factors, gender group in the school class had the strongest effect on smoking status. CONCLUSION: We conclude that self-efficacy to resist pressure to smoke, attitude, and the three types of social influence are significantly associated with lifetime smoking status, even when the effects of group-level school factors are taken into account. The strong effect of gender group in school class on lifetime smoking status indicates that prevention actions should address the social context of adolescents.     

Mechanical coupling of supracellular stress amplification and tissue fluidization during exit from quiescence

Cellular quiescence is a state of reversible cell cycle arrest that is associated with tissue dormancy. Timely regulated entry into and exit from quiescence is important for processes such as tissue homeostasis, tissue repair, stem cell maintenance, developmental processes, and immunity. However, little is known about processes that control the mechanical adaption to cell behavior changes during the transition from quiescence to proliferation. Here, we show that quiescent human keratinocyte monolayers sustain an actinomyosin-based system that facilitates global cell sheet displacements upon serum-stimulated exit from quiescence. Mechanistically, exposure of quiescent cells to serum-borne mitogens leads to rapid amplification of preexisting contractile sites, leading to a burst in monolayer tension that subsequently drives large-scale displacements of otherwise motility-restricted monolayers. The stress level after quiescence exit correlates with the level of quiescence depth at the time of activation, and a critical stress magnitude must be reached to overcome the cell sheet displacement barrier. The study shows that static quiescent cell monolayers are mechanically poised for motility, and it identifies global stress amplification as a mechanism for overcoming motility restrictions in confined confluent cell monolayers.

Quiescence refers to a state of cell cycle arrest in which cells are retained in a standby mode, ready to re-enter the cell cycle upon activation by a given physiological stimuli. The pool of quiescent cells in the human body is typically represented by tissue-specific stem and progenitor cells, naive immune cells, fibroblasts, and epithelial cells (1, 2). In addition, certain cancer cells have the ability to evade cancer therapy by entering a dormant quiescence-like state (1, 2). Accordingly, careful regulation of entry into and exit out of quiescence is important for several physiological processes such as tissue homeostasis and repair, stem cell maintenance, immunity, reproduction, and development (1, 2).During homeostasis, the balance between quiescent and proliferating cells is controlled by constituents of the microenvironment such as soluble factors, extracellular matrix components, blood vessels, and neighboring cells. On the other hand, during episodes that require extensive tissue renewal and remodeling, for example after injury, coordinated stimulation of quiescent cells into proliferation is facilitated by increased exposure to blood-borne and cell-secreted mitogens through local inflammatory responses such as increased blood flow, increased vascular permeability (vasodilation), and immune cell recruitment (3, 4). Accordingly, a commonly used methodology for studies of quiescence in cultured mammalian cells involves consecutive treatments with serum-free and serum-containing growth medium (1).Quiescent cells are required to maintain a high level of preparedness in order to facilitate rapid activation of specialized cell functions once cell division is stimulated. In agreement with this, quiescent stem cells and naive immune cells have been shown to possess multiple epigenetic and posttranslation mechanisms that facilitate the rapid expression of linage-specific genes following stimulation of quiescence exit (2, 5–14). However, little is known about mechanical forces that facilitate adaptation to cell cycle–activated behaviors.Quiescence exit is frequently associated with activation of cell motility. For example, quiescent stem and naive immune cells migrate out of their niches in response to cell cycle activation in order to support tissue homeostasis, repopulate injured tissue, or to perform immune surveillance at distal locations (15–18). In addition, reawakening of dormant quiescent cancer cells can cause tumor relapse and formation of metastases years after remission (19). In multilayered epithelial tissue, like the skin, exit from quiescence during homeostasis is associated with lateral migration to suprabasal regions, while skin injury evokes massive reawakening of basally localized keratinocytes concomitant with activation of cell sheet displacement by collective migration to restore damaged epidermal surfaces (20–23). The strong correlation between quiescence exit and cell migration in multiple physiological settings suggests the existence of mechanisms that link quiescence exit to activation of cell motility.The dynamics of epithelial collectives is largely regulated by mechanical forces generated through cell–cell interactions as well as interactions between cells and the extracellular environment (24). Key components involved in controlling these forces are cytoskeletal components such as actinomyosin and adhesion complexes such as adherent junctions and focal adhesion complexes (25). Additional factors that have been reported to influence the dynamic behavior of epithelial monolayers include the presence of epithelial edges (24, 26), mechanical stretching or compression (27, 28), expression of the endosomal Rab5 protein (29), exposure of cells to growth factors (30–32), local changes in cell shape (33), and the ability of cells to undergo neighbor exchange (34, 35). In addition, recent studies have also identified a functional link between cell cycle progression and force fluctuation leading to dynamic behavior of cultured epithelial monolayers (36, 37).In this study, we have investigated a mechanical link between quiescence exit and activation of large-scale cell sheet displacements. Using traction force microscopy (TFM), we found that confluent cell monolayers install an actinomyosin-based system during quiescence that produces a coordinated burst of contractile forces and intercellular tension across the epithelial monolayer immediately following exposure to serum-borne mitogens. By combining experiments and theoretical modeling, we show that the amplified forces are essential for driving coordinated cell sheet displacements within otherwise motility-restricted cell monolayers. Furthermore, the magnitude of mechanical forces created during quiescence exit and the extent of cell sheet displacement correlate with quiescence depth. Our study provides evidence that quiescent keratinocyte monolayers possess mechanical preparedness for motility and establish monolayer stress amplification as a strategy for overcoming the motility barrier in confined cell sheets.