Development of health diagnostics based on personalized medical models.

Due to rapid evolution of new technologies the concept of personalized medicine has evolved. Components include molecular biology, proteomics, metabolomic analysis, genetic testing, and molecular medicine for diagnostics. In addition to diagnostics these methods can be used to determine individual susceptibility to diseases and conditions. In conjunction with new diagnostic methods, new therapies can be tailored to the individual. These new technologies present a challenge in terms of the expansion of the medical record as well as the development of new methods for creating disease profiles. This article focuses on a computer-aided support for personalized medicine. Specific approaches are explored that permit automated data analysis for prognosis and treatment based on analysis methods for numeric and pictorial data. Although personalized medicine based on the genome of the patient are occasionally performed, because of the large amount of data new methods are needed to form general disease models as well as specific profiles of the individual patient.

Perceptions of underweight images: are women with anorexia nervosa perceived as attractive and healthy?

The current study examined the impact of receiving information about a woman's eating disorder status on perceptions of the woman's health and attractiveness. A total of 99 females and 84 males viewed a photo of a model who had disclosed her diagnosis of anorexia nervosa. Participants were randomly divided into three groups: model (M) group (those who were informed that the photo showed a model), eating disorders (ED) group (those who were informed that the photo showed a woman with an eating disorder), and no description control (C) group. Male and female participants in the ED group rated the woman in the photo as less healthy than did participants in the M and C groups. However, there were no differences between groups for ratings of attractiveness or the participants' desire to achieve a similar look (for females). Additionally, male participants rated the photo as less attractive than female participants had predicted. Finally, internalization of the thin ideal was a significant predictor of ratings of health and attractiveness of the woman in the photo.

TEAD1 and c-Cbl are novel prostate basal cell markers that correlate with poor clinical outcome in prostate cancer.

Prostate cancer is the most frequently diagnosed male cancer, and its clinical outcome is difficult to predict. The disease may involve the inappropriate expression of genes that normally control the proliferation of epithelial cells in the basal layer and their differentiation into luminal cells. Our aim was to identify novel basal cell markers and assess their prognostic and functional significance in prostate cancer. RNA from basal and luminal cells isolated from benign tissue by immunoguided laser-capture microdissection was subjected to expression profiling. We identified 112 and 267 genes defining basal and luminal populations, respectively. The transcription factor TEAD1 and the ubiquitin ligase c-Cbl were identified as novel basal cell markers. Knockdown of either marker using siRNA in prostate cell lines led to decreased cell growth in PC3 and disrupted acinar formation in a 3D culture system of RWPE1. Analyses of prostate cancer tissue microarray staining established that increased protein levels of either marker were associated with decreased patient survival independent of other clinicopathological metrics. These data are consistent with basal features impacting on the development and clinical course of prostate cancers.

Intervening with coaches to promote awareness and prevention of weight pressures in cheerleaders.

Research has found that athletes, particularly those involved in "aesthetically oriented" sports, experience significant pressures for thinness and are at increased risk for developing eating disorders. This study targeted cheerleading coaches as potential change agents by training them to recognize the symptoms of eating disorders and reduce the pressures for thinness among their squads. Cheerleading coaches at national or regional conferences attended an intervention workshop or a control workshop. Coaches who attended the intervention workshop received information regarding negative coaching behaviors, the symptoms of eating disorders, and ways to manage athletes with eating disorders. In addition, intervention coaches were encouraged to participate in six intervention strategies (e.g., reading materials, video, parent handouts, etc.) after attending the workshop. Eight months following the workshop, the coaches completed an assessment battery designed to test the effectiveness of the entire intervention. The results indicated that the intervention was successful in producing behavior changes in coaches. However, the intervention was less successful in producing long-term change in knowledge about eating disorders. These findings imply that interventions can be implemented by important adult figures (e.g., coaches, teachers) but the overall effectiveness of these interventions must be enhanced in order to have a significant and long-term impact.

The expression of xenobiotic-metabolizing enzymes in human prostate and in prostate epithelial cells (PECs) derived from primary cultures.

BACKGROUND: Dietary heterocyclic amines (HCAs) are carcinogenic in rodent prostate requiring activation by enzymes such as cytochrome P450 (CYP) and N-acetyltransferase (NAT). METHODS: We investigated by Western blotting and immunohistochemistry the expression of CYP1A1, CYP1A2, and NAT1 in human prostate and in prostate epithelial cells (PECs) derived from primary cultures and tested their ability to activate the dietary carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and its N-hydroxy metabolite (N-OH-IQ) to DNA-damaging moieties. RESULTS: Western blotting identified CYP1A1, CYP1A2, and NAT1. Immunohistochemistry localized NAT1 to the cytoplasm of PECs. Inter-individual variation was observed in the expression levels of CYP1A1, 1A2, and NAT1 (11, 75, and 35-fold, respectively). PECs expressed CYP1A1 and NAT1 but not CYP1A2. When incubated with IQ or N-OH-IQ, PECs formed DNA adducts indicating their ability to metabolically activate these compounds. CONCLUSIONS: Prostate cells possess the capacity to activate dietary carcinogens. PECs may provide a useful model system to study their role in prostate carcinogenesis.

Intelligent agent model for remote support of rural healthcare for the elderly.

With the aging population, the number of individuals requiring long-term care is expected to dramatically increase in the next twenty years, placing an increasing burden on healthcare. Many patients are admitted to assisted living facilities at a fairly early stage due to their inability to perform normal daily living activities. The purpose of this study is to determine if the use of technology for both monitoring and intervention can permit elderly patients to remain in their homes for longer periods of time with the benefit of the comfort of familiar surroundings while at the same time reducing the burden on caregivers. In addition, remote access to healthcare can improve monitoring of the patient's physical and mental condition and involve the patient in his or her own care. The home monitoring and intervention system is based on intelligent agent methodology developed by the authors.

Prostate epithelial stem cells.

The prostate gland is the site of the most commonly diagnosed cancer in men in USA and UK, accounting for one in five of new cases of male cancer. Common with many other cancer types, prostate cancer is believed to arise from a stem cell that shares characteristics with the normal stem cell. Normal prostate epithelial stem cells were recently identified and found to have a basal cell phenotype together with expression of CD133. Preliminary data have now emerged for a prostate cancer stem cell that also expresses cell surface CD133 but lacks expression of the androgen receptor. Here we examine the evidence supporting the existence of prostate cancer stem cells and discuss possible mechanisms of stem cell maintenance.

Epithelial stem cells in human prostate growth and disease.

Benign prostatic hyperplasia and prostate cancer arise as a consequence of changes in the balance between cell division and differentiation. Little, however, is known about the control of this process. Stem cells are a small population of cells that divide occasionally to produce transit-amplifying cells that in turn produce the differentiated cell types of the tissue. It is believed that cancer cell proliferation is also driven by stem cells. We have shown that around one in 200 prostate epithelial cells have characteristics of stem cells and that these cells are contained within a population with a distinct keratin expression pattern. Work is now ongoing to identify markers for these cells that will allow us to study the role they play in prostatic disease.

Non-linear analysis using continuous chaotic modeling.

Recent research in nanotechnology is opening exciting new avenues not only for understanding the human body but also for creating devices that can effectively interact with it to alleviate the effects of disease. These new developments present both challenges and opportunities for adaptation of existing methodologies to create new approaches for analysis and modeling of nanotechnology-based systems. The concept of continuous chaotic modeling presents an avenue for a paradigm shift away from traditional digital computing to take advantage of analog models that are more compatible with biological systems. The theoretical basis of continuous chaotic modeling is summarized, followed by illustrations of applications of this methodology.

The role of networks and artificial intelligence in nanotechnology design and analysis.

Techniques with their origins in artificial intelligence have had a great impact on many areas of biomedicine. Expert-based systems have been used to develop computer-assisted decision aids. Neural networks have been used extensively in disease classification and more recently in many bioinformatics applications including genomics and drug design. Network theory in general has proved useful in modeling all aspects of biomedicine from healthcare organizational structure to biochemical pathways. These methods show promise in applications involving nanotechnology both in the design phase and in interpretation of system functioning.

Inclusion of signal analysis in a hybrid medical decision support system.

OBJECTIVES: Signal analysis has played an important role in cardiac diagnosis, both as a separate entity and in conjunction with clinical parameters. Hybrid systems are an effective method for developing higher-order decision models in which biomedical signal data can be incorporated. METHODS: The hybrid system components include a knowledge-based system that utilizes approximate reasoning techniques, a neural network model based on a potential function approach to supervised learning that uses the general class of Cohen orthogonal functions as potential functions, and a signal analysis component that relies on continuous chaotic modeling to produce a degree of variability in the time series. The hybrid system is illustrated in an application for differentiation among different types of dementia. RESULTS: Application of this method to cardiac diagnosis shows that chaotic parameters alone contribute significantly to correct classification while the addition of clinical parameters increases the sensitivity, specificity, and accuracy. Applications to electroencephalogram analysis indicate that the second-order difference plots display significant differences for the different types of EEG waves identifiable by frequency, both in shape and degree of dispersion. Hence the identification of these waves, and the duration of their occurrence, may provide suitable variables for chaotic analysis. CONCLUSIONS: Results from studies in cardiology demonstrate that using chaotic measures for ECG analysis provide useful information for classification. Sensitivity, specificity, and accuracy are increased if these methods are combined with other clinical parameters in a hybrid system. This approach has been extended to new applications based on EEG analysis combined with other relevant information.

The use of consequential reasoning in cancer chemotherapy.

Knowledge-based decision support systems traditionally rely on condition-action rule structures, an adequate representation for simple decisions. In complex domains an important part of decision-making includes analysis of the consequences of a decision. Consequential reasoning is particularly important in medicine as potential risk and/or benefit can be included. In this paper, a knowledge structure and inference engine is described that permits the representation and analysis of consequential reasoning in a computer-assisted decision support system. The use of consequential reasoning is then illustrated in an application designed to assist in cancer chemotherapy decisions. The result is a method that is sensitive to individual patient reactions to chemotherapy agents, permitting an individualized approach to therapy. Individualized drug therapy is becoming increasingly feasible due to advances made in the field of genomics. The system is structured so that new information can be incorporated easily. Although the application shown here is to chemotherapy, the general methodology can be used in any area in which the consequences should significantly influence the decision.

A novel mechanism of matrix metalloproteinase-9 gene expression implies a role for keratinization.

To investigate the pathophysiological role of matrix metalloproteinase (MMP)-9 in the skin, we analyzed MMP-9 expression from human keratinocytes in culture. MMP-9 and the terminal differentiation marker involucrin were co-localized in the same keratinocytes with a high concentration of Ca(2+), a potent stimulator of differentiation. We identified the novel KRE-M9 element, further downstream to the previously reported TPA responsive element in the MMP-9 promoter, and both of these two elements were shown to be important for MMP-9 transcription and Ca(2+) induction. The concomitant upregulation of MMP-9 and involucrin transcripts was probably due to the very similar gene regulatory elements, KRE-M9 and KRE-4, in their respective promoters. These results indicate a novel mechanism of transcriptional regulation for MMP-9 in the process of keratinization, implying the probable association of apoptosis and differentiation of keratinocytes in epidermal skin tissue.

Epithelial cell differentiation pathways in the human prostate: identification of intermediate phenotypes by keratin expression.

The prostate grows slowly throughout adult life, leading to benign prostatic hyperplasia (BPH), which often results in urethral obstruction in later years. The symptoms of BPH are the second most common reason for surgery in men over 65. The aim of this study was to determine the relationship between cell proliferation and cell differentiation in BPH tissue. Using multiple antibodies, simultaneously detected with different fluorophore-conjugated secondary antibodies, several subpopulations of epithelial cells were detected. In addition to K14, basal cells also expressed keratins 15, 17, and 19 in various combinations, and some of the luminal cells also expressed K19 together with K8 and K18. Co-staining for cytokeratins and Ki-67 indicated that 44% of proliferative cells expressed K14 and 36% K19, although the difference was not statistically significant. This report provides a detailed description of the relationship between keratin expression and cell proliferation in the prostate and indicates that K19-positive cells form the link between the basal and luminal layers of the epithelium. (J Histochem Cytochem 49:271-278, 2001)

Proliferative heterogeneity in the human prostate: evidence for epithelial stem cells.

Clonal analysis of human prostate epithelial cells was undertaken in order to identify stem cells. Two types of colony were distinguished, termed type I and type II. Type I colonies were relatively small and irregular and contained a loose mixture of differentiated and undifferentiated cells. In contrast, type II colonies were large, round, and homogeneous, consisting almost exclusively of small undifferentiated and dividing cells. The colony-forming efficiency was 5.8% +/- 1.8 for freshly isolated epithelial cells. There were approximately 10 times as many type I as type II colonies and about 1 in 200 of the plated cells was capable of forming a type II colony. In three-dimensional culture on Matrigel, the type II colonies produced structures reminiscent of prostate epithelium, with luminal cells expressing markers of prostate epithelial differentiation, including the androgen receptor. On the basis of their proliferative characteristics and pluripotency, the type II colonies may be the progeny of stem cells and the type I colonies of a more differentiated transit-amplifying population.

Bone morphogenetic protein 1 is an extracellular processing enzyme of the laminin 5 gamma 2 chain.

Epithelial cells maintained in culture medium containing low calcium proteolytically process laminin 5 (alpha3beta3gamma2) within the alpha3 and gamma2 chains (). Experiments were designed to identify the enzyme(s) responsible for the laminin 5 processing and the sites of proteolytic cleavage. To characterize the nature of laminin 5 processing, we determined the N-terminal amino acid sequences of the proteolytic fragments produced by the processing events. The results indicate that the first alpha3 chain cleavage (200-l65 kDa alpha3) occurs within subdomain G4 of the G domain. The second cleavage (l65-l45 kDa alpha3) occurs within the lIla domain, 11 residues N-terminal to the start of domain II. The gamma chain is cleaved within the second epidermal growth factor-like repeat of domain Ill. The sequence cleaved within the gamma2 chain matches the consensus sequence for the cleavage of type I, II, and III procollagens by bone morphogenetic protein-1 (BMP-1), also known as type I procollagen C-proteinase (). Recombinant BMP-1 cleaves gamma2 in vitro, both within intact laminin 5 and at the predicted site of a recombinant gamma2 short arm. alpha3 is also cleaved by BMP-1 in vitro, but the cleavage site is yet to be determined. These results show the laminin alpha3 and gamma2 chains to be substrates for BMP-1 in vitro. We speculate that gamma2 cleavage is required for formation of the laminin 5-6 complex and that this complex is directly involved in assembly of the interhemidesmosomal basement membrane. This further suggests that BMP-1 activity facilitates basement membrane assembly, but not hemidesmosome assembly, in the laminin 5-rich dermal-epidermal junction basement membrane in vivo.

Comparison of marker protein expression in benign prostatic hyperplasia in vivo and in vitro.

OBJECTIVE: To use multiple immunofluorescence to compare the in vivo and in vitro expression of tissue-specific proteins in BPH. Materials and methods Pure populations of prostate epithelial and stromal cells were produced using standard methods. Serum-free media for epithelial cells were compared. Co-localization of proteins was compared in frozen-tissue sections and cultured cells by simultaneous multiple immunofluorescence, and recorded using a high-resolution charge-coupled device camera. RESULTS: In contrast to the other serum-free media tested, epithelial cells grew without squamous differentiation or vacuolation in prostate epithelial growth medium (PrEGM, Clonetics, BioWhittaker UK Ltd., Berks, UK). These cells were predominantly of a basal phenotype, with some cells showing a luminal phenotype. Most of the stromal cells had features of myofibroblasts, but smooth muscle cells and fibroblasts also were present. CONCLUSION: PrEGM is a commercially available serum-free medium in which primary cultures of prostate epithelial cells can be propagated reproducibly. This study provides a comprehensive description of tissue-specific protein expression in BPH in vivo and in vitro. The use of simultaneous multiple immunofluorescence to study co-localization has resulted in a more precise definition of phenotype than has previously been possible, thereby establishing the relevance of the in vitro model system BPH.

Complete primary structure of two splice variants of collagen XII, and assignment of alpha 1(XII) collagen (COL12A1), alpha 1(IX) collagen (COL9A1), and alpha 1(XIX) collagen (COL19A1) to human chromosome 6q12-q13.

Overlapping cDNA clones that encode the full-length human alpha 1(XII) collagen polypeptides were isolated. The long variant molecule cDNA of 9750 nucleotides (nt) contains a 9189-nt open reading frame encoding 3063 amino acid residues. The short variant molecule cDNA of 6258 nt contains a 5697-nt open reading frame encoding 1899 amino acid residues. At the amino terminus of each variant is a 24-residue signal peptide that is followed by the mature polypeptides of 3039 amino acid residues with a calculated molecular mass of 330,759 Da for the long variant and 1875 amino acid residues with a calculated molecular mass of 203,163 Da for the short variant polypeptide. The human collagen XII chains are predicted to have all the structural domains described for the molecules in chicken and mouse, including, fibronectin type III repeats, von Willebrand factor A domains, and two triple-helical domains similar to those of all the other collagen family members. The amino acid residue sequence of human alpha 1(XII) collagen showed 92% identity to the mouse chain and 78% identity to the chicken chain. The sequence of three peptide fragments of collagen XII isolated from human placenta was identical to the sequence predicted from the deduced cDNA sequence and confirms that the cDNA encodes human alpha 1(XII) collagen. An isolated genomic clone was used to map the locus of the COL12A1 gene to chromosome 6q12-q13, very close to the locus of the FACIT collagen genes COL9A1 and COL19A1. RT-PCR on a variety of cDNAs demonstrates that both variant transcripts appear in human amnion, chorion, skeletal muscle, small intestine, and in cell cultures of human dermal fibroblasts, keratinocytes, and endothelial cells. Only the small variant transcript is apparent in human lung, placenta, kidney, and a squamous cell carcinoma cell line. These results confirm the previous observations showing that collagen XII is found in collagen I-containing tissues.