Quantification of dynamic morphological drug responses in 3D organotypic cell cultures by automated image analysis.

Glandular epithelial cells differentiate into complex multicellular or acinar structures, when embedded in three-dimensional (3D) extracellular matrix. The spectrum of different multicellular morphologies formed in 3D is a sensitive indicator for the differentiation potential of normal, non-transformed cells compared to different stages of malignant progression. In addition, single cells or cell aggregates may actively invade the matrix, utilizing epithelial, mesenchymal or mixed modes of motility. Dynamic phenotypic changes involved in 3D tumor cell invasion are sensitive to specific small-molecule inhibitors that target the actin cytoskeleton. We have used a panel of inhibitors to demonstrate the power of automated image analysis as a phenotypic or morphometric readout in cell-based assays. We introduce a streamlined stand-alone software solution that supports large-scale high-content screens, based on complex and organotypic cultures. AMIDA (Automated Morphometric Image Data Analysis) allows quantitative measurements of large numbers of images and structures, with a multitude of different spheroid shapes, sizes, and textures. AMIDA supports an automated workflow, and can be combined with quality control and statistical tools for data interpretation and visualization. We have used a representative panel of 12 prostate and breast cancer lines that display a broad spectrum of different spheroid morphologies and modes of invasion, challenged by a library of 19 direct or indirect modulators of the actin cytoskeleton which induce systematic changes in spheroid morphology and differentiation versus invasion. These results were independently validated by 2D proliferation, apoptosis and cell motility assays. We identified three drugs that primarily attenuated the invasion and formation of invasive processes in 3D, without affecting proliferation or apoptosis. Two of these compounds block Rac signalling, one affects cellular cAMP/cGMP accumulation. Our approach supports the growing needs for user-friendly, straightforward solutions that facilitate large-scale, cell-based 3D assays in basic research, drug discovery, and target validation.

Feasibility of a personal health technology-based psychological intervention for men with stress and mood problems: randomized controlled pilot trial.

BACKGROUND: Work-related stress is a significant problem for both people and organizations. It may lead to mental illnesses such as anxiety and depression, resulting in increased work absences and disabilities. Scalable interventions to prevent and manage harmful stress can be delivered with the help of technology tools to support self-observations and skills training. OBJECTIVE: The aim of this study was to assess the feasibility of the P4Well intervention in treatment of stress-related psychological problems. P4Well is a novel intervention which combines modern psychotherapy (the cognitive behavioral therapy and the acceptance and commitment therapy) with personal health technologies to deliver the intervention via multiple channels, includinggroup meetings, Internet/Web portal, mobile phone applications, and personal monitoring devices. METHODS: This pilot study design was a small-scale randomized controlled trial that compared the P4Well intervention with a waiting list control group. In addition to personal health technologies for self-assessment, the intervention consisted of 3 psychologist-assisted group meetings. Self-assessed psychological measures through questionnaires were collected offline pre- and post-intervention, and 6 months after the intervention for the intervention group. Acceptance and usage of technology tools were measured with user experience questionnaires and usage logs. RESULTS: A total of 24 subjects were randomized: 11 participants were followed up in the intervention group (1 was lost to follow-up) and 12 participants did not receive any intervention (control group). Depressive and psychological symptoms decreased and self-rated health and working ability increased. All participants reported they had benefited from the intervention. All technology tools had active users and 10/11 participants used at least 1 tool actively. Physiological measurements with personal feedback were considered the most useful intervention component. CONCLUSIONS: Our results confirm the feasibility of the intervention and suggest that it had positive effects on psychological symptoms, self-rated health, and self-rated working ability. The intervention seemed to have a positive impact on certain aspects of burnout and job strain, such as cynicism and over-commitment. Future studies need to investigate the effectiveness, benefits, and possible problems of psychological interventions which incorporate new technologies. TRIAL REGISTRATION: The Finnish Funding Agency for Technology and Innovation (TEKES), Project number 40011/08.

A comprehensive panel of three-dimensional models for studies of prostate cancer growth, invasion and drug responses.

Prostate epithelial cells from both normal and cancer tissues, grown in three-dimensional (3D) culture as spheroids, represent promising in vitro models for the study of normal and cancer-relevant patterns of epithelial differentiation. We have developed the most comprehensive panel of miniaturized prostate cell culture models in 3D to date (n = 29), including many non-transformed and most currently available classic prostate cancer (PrCa) cell lines. The purpose of this study was to analyze morphogenetic properties of PrCa models in 3D, to compare phenotypes, gene expression and metabolism between 2D and 3D cultures, and to evaluate their relevance for pre-clinical drug discovery, disease modeling and basic research. Primary and non-transformed prostate epithelial cells, but also several PrCa lines, formed well-differentiated round spheroids. These showed strong cell-cell contacts, epithelial polarization, a hollow lumen and were covered by a complete basal lamina (BL). Most PrCa lines, however, formed large, poorly differentiated spheroids, or aggressively invading structures. In PC-3 and PC-3M cells, well-differentiated spheroids formed, which were then spontaneously transformed into highly invasive cells. These cell lines may have previously undergone an epithelial-to-mesenchymal transition (EMT), which is temporarily suppressed in favor of epithelial maturation by signals from the extracellular matrix (ECM). The induction of lipid and steroid metabolism, epigenetic reprogramming, and ECM remodeling represents a general adaptation to 3D culture, regardless of transformation and phenotype. In contrast, PI3-Kinase, AKT, STAT/interferon and integrin signaling pathways were particularly activated in invasive cells. Specific small molecule inhibitors targeted against PI3-Kinase blocked invasive cell growth more effectively in 3D than in 2D monolayer culture, or the growth of normal cells. Our panel of cell models, spanning a wide spectrum of phenotypic plasticity, supports the investigation of different modes of cell migration and tumor morphologies, and will be useful for predictive testing of anti-cancer and anti-metastatic compounds.

A concept to empower self-management of psychophysiological wellbeing: preliminary user study experiences.

In prevention of chronic diseases, health promotion and early interventions based on self-management should be emphasized. Mental health problems and stress cause a significant portion of healthcare costs, and also complicate the management of other chronic conditions. In addition to physical health, psychophysiological and social wellbeing should be equally promoted. Thus, we have previously designed and reported the P4Well or Pervasive Personal and PsychoPhysiological management of WELLness concept for working-age citizens. The concept supports the stress and recovery management on a daily basis through improved health management strategies, and combines psychological methods with personal health technologies. In this paper, we discuss the preliminary user study experiences of ongoing evaluations with two different user groups consisting of: 1) middle-aged men who are using the concept for managing their mental wellbeing or mild depression; and 2) entrepreneurs who are using the concept for coping with stress. Our results provide a preliminary assessment of the role and importance of experts, technologies, and peer-support in the concept.

Assessment of automated scintigram alignment in the stackgram domain for cardiac stress-rest SPECT studies.

Single photon emission computed tomography (SPECT) provides a method to obtain three-dimensional images of blood perfusion. In cardiac stress-rest SPECT studies, coronary artery disease can be examined by comparing intensity differences of the acquired images of the heart at stress and rest. The stress-rest image pair needs to be aligned into the same spatial position and orientation in order to compare the images in an automated and reproducible manner. In this work, we study a novel stackgram-based alignment algorithm for cardiac stress-rest SPECT data. The stackgram approach allows the alignment of the acquired raw data prior to image reconstruction, unlike conventional alignment techniques.

Experimental investigation of angular stackgram filtering for noise reduction of SPECT projection data: study with linear and nonlinear filters.

We discuss data filtering prior to image reconstruction. For this kind of filtering, the radial direction of the sinogram is routinely employed. Recently, we have introduced an alternative approach to sinogram data processing, exploiting the angular information in a novel way. This new stackgram representation can be regarded as an intermediate form of the sinogram and image domains. In this experimental study, we compare the radial sinogram and angular stackgram filtering methods using physical SPECT phantoms. Our study is carried out by employing simple linear and nonlinear filters with ten different Gaussian kernels, in order to provide a comparable investigation. According to our results, angular stackgram filtering with the nonlinear filters provides the best resolution-noise tradeoff of the compared methods. Besides, stackgram filtering with these filters seems to preserve the resolution in an exceptional way. Visually, noise in the reconstructed images after stackgram filtering appears more "powdery" in comparison with radial sinogram filtering.