Multi-parametric evaluation of sit-to-stand and stand-to-sit transitions in elderly people.

The aim of this study was to extract multi-parametric measures characterizing different features of sit-to-stand (Si-St) and stand-to-sit (St-Si) transitions in older persons, using a single inertial sensor attached to the chest. Investigated parameters were transition's duration, range of trunk tilt, smoothness of transition pattern assessed by its fractal dimension, and trunk movement's dynamic described by local wavelet energy. A measurement protocol with a Si-St followed by a St-Si postural transition was performed by two groups of participants: the first group (N=79) included Frail Elderly subjects admitted to a post-acute rehabilitation facility and the second group (N=27) were healthy community-dwelling elderly persons. Subjects were also evaluated with Tinetti's POMA scale. Compared to Healthy Elderly persons, frail group at baseline had significantly longer Si-St (3.85��1.04 vs. 2.60��0.32, p=0.001) and St-Si (4.08��1.21 vs. 2.81��0.36, p=0.001) transition's duration. Frail older persons also had significantly decreased smoothness of Si-St transition pattern (1.36��0.07 vs. 1.21��0.05, p=0.001) and dynamic of trunk movement. Measurements after three weeks of rehabilitation in frail older persons showed that smoothness of transition pattern had the highest improvement effect size (0.4) and discriminative performance. These results demonstrate the potential interest of such parameters to distinguish older subjects with different functional and health conditions.

Control of pancreatic β-cell mass and function by gluco-incretin hormones : identification of novel regulatory mechanisms for the treatment of diabetes

Summary : Control of pancreatic ��-cell mass and function by gluco-incretin hormones: Identification of novel regulatory mechanisms for the treatment of diabetes The ��-cells of islets of Langerhans secrete insulin to reduce hyperglycemia. The number of pancreatic islet ��-cells and their capacity to secrete insulin is modulated in normal physiological conditions to respond to the metabolic demand of the organism. A failure of the endocrine pancreas to maintain an adequate insulin secretory capacity due to a reduced ��-cell number and function underlies the pathogenesis of both type 1 and type 2 diabetes. The molecular mechanisms controlling the glucose competence of mature ��-cells, i.e., the magnitude of their insulin secretion response to glucose, ��-cell replication, their differentiation from precursor cells and protection against apoptosis are poorly understood. To investigate these mechanisms, we studied the effects on ��-cells of the gluco-incretin hormones, glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) which are secreted by intestinal endocrine cells after food intake. Besides acutely potentiating glucose-stimulated insulin secretion, these hormones induce ��-cell differentiation from precursor cells, stimulate mature ��-cell replication, and protect them against apoptosis. Therefore, understanding the molecular basis for gluco-incretin action may lead to the uncovering of novel ��-cell regulatory events with potential application for the treatment or prevention of diabetes. Islets from mice with inactivation of both GIP and GLP-1 receptor genes (dK0) present a defect in glucose-induced insulin secretion and are more sensitive than control islets to cytokine-induced apoptosis. To search for regulatory genes, that may control both glucose competence and protection against apoptosis, we performed comparative transcriptomic analysis of islets from control and dK0 mice. We found a strong down-regulation of the IGF1 Rexpression in dK0 islets. We demonstrated in both a mouse insulin-secreting cell line and primary islets, that GLP-1 stimulated IGF-1R expression and signaling. Importantly, GLP-1induced IGF-1R-dependent Akt phosphorylation required active secretion, indicating the presence of an autocrine activation mechanism. We further showed that activation of IGF-1R signaling was dependent on the secretion of IGF-2 and IGF-2 expression was regulated by nutrients. Finally, we demonstrated that the IGF-Z/IGF-1R autocrine loop was required for GLP-1 i) to protect ��-cells against cytokine-induced apoptosis, ii) to enhance their glucose competence and iii) to increase ��-cell proliferation. R��sum�� : Contr��le de la masse des cellules �� pancr��atiques et de leur fonction par les hormones glucoincr��tines: Identification de nouveaux m��canismes r��gulateurs pour le traitement du diab��te Les cellules �� des ��lots de Langerhans s��cr��tent l'insuline pour diminuer l'hyperglyc��mie. Le nombre de cellules �� et leur capacit�� �� s��cr��ter l'insuline sont modul��s dans les conditions physiologiques normales pour r��pondre �� la demande m��tabolique de l'organisme. Un ��chec du pancr��as endocrine �� maintenir sa capacit�� s��cr��toire d'insuline d�� �� une diminution du nombre et de la fonction des cellules �� conduit au diab��te de type 1 et de type 2. Les m��canismes mol��culaires contr��lant la comp��tence au glucose des cellules �� matures, tels que, l'augmentation de la s��cr��tion d'insuline en r��ponse au glucose, la r��plication des cellules ��, leur diff��rentiation �� partir de cellules pr��curseurs et la protection contre l'apoptose sont encore peu connus. Afin d'examiner ces m��canismes, nous avons ��tudi�� les effets sur les cellules �� des hormones gluco-incr��tines, glucose-d��pendent insulinotropic polypeptide (G1P) et glucagon-like peptide-1 (GLP-1) qui sont s��cr��t��es par les cellules endocrines de l'intestin apr��s la prise alimentaire. En plus de potentialiser la s��cr��tion d'insuline induite par le glucose, ces hormones induisent la diff��rentiation de cellules �� �� partir de cellules pr��curseurs, stimulent leur prolif��ration et les prot��gent contre l'apoptose. Par cons��quent, comprendre les m��canismes d'action des gluco-incr��tines permettrait de d��couvrir de nouveaux processus r��gulant les cellules �� avec d'��ventuelles applications dans le traitement ou la pr��vention du diab��te. Les ��lots de souris ayant une double inactivation des g��nes pour les r��cepteurs du GIP et du GLP-1 (dK0) pr��sentent un d��faut de s��cr��tion d'insuline stimul��e par le glucose et une sensibilit�� accrue �� l'apoptose induite par les cytokines. Afin de d��terminer les g��nes r��gul��s, qui pourraient contr��ler �� la fois la comp��tence au glucose et la protection contre l'apoptose, nous avons effectu�� une analyse comparative transcriptomique sur des ��lots de souris contr��les et dKO. Nous avons constat�� une forte diminution de l'expression d'IGF-1R dans les ��lots dKO. Nous avons d��montr��, �� la fois dans une lign��e cellulaire murine s��cr��tant l'insuline et dans ��lots primaires, que le GLP-1 stimulait l'expression d'IGF-1R et sa voie de signalisation. Par ailleurs, la phosphorylation d'Akt d��pendante d'IGF1-R induite parle GLP-1 n��cessite une s��cr��tion active, indiquant la pr��sence d'un m��canisme d'activation autocrine. Nous avons ensuite montr�� que l'activation de la voie de signalisation d'IGF-1R ��tait d��pendante de la s��cr��tion d'IGF-2, dont l'expression est r��gul��e par les nutriments. Finalement, nous avons d��montr�� que la boucle autocrine IGF-2/IGF-1R est n��cessaire pour le GLP-1 i) pour prot��ger les cellules �� contre l'apoptose induite par les cytokines, ii) pour am��liorer la comp��tence au glucose et iii) pour augmenter la prolif��ration des cellules ��. R��sum�� tout public : Contr��le de la masse des cellules �� pancr��atiques et de leur fonction par les hormones gluco-incr��tines: Identification de nouveaux m��canismes r��gulateurs pour le traitement du diab��te Chez les mammif��res, la concentration de glucose sanguine (glyc��mie) est r��gul��e et maintenue �� une valeur relativement constante d'environ 5 mM. Cette r��gulation est principalement contr��l��e par 2 hormones produites par les ��lots pancr��atiques de Langerhans: l'insuline s��cr��t��e par les cellules �� et le glucagon s��cr��t�� par les cellules a. A la suite d'un repas, l'augmentation de la glyc��mie entra��ne la s��cr��tion d'insuline ce qui permet le stockage du glucose dans le foie, les muscles et le tissu adipeux afin de diminuer le taux de glucose circulant. Lors d'un je��ne, la diminution de la glyc��mie permet la s��cr��tion de glucagon favorisant alors la production de glucose par le foie, normalisant ainsi la glyc��mie. Le nombre de cellules �� et leur capacit�� s��cr��toire s'adaptent aux variations de la demande m��tabolique pour assurer une normoglyc��mie. Une destruction compl��te ou partielle des cellules �� conduit respectivement au diab��te de type 1 et de type 2. Bien que l'augmentation de la glyc��mie soit le facteur stimulant de la s��cr��tion d'insuline, des hormones gluco-incr��tines, principalement le GLP-1 (glucagon-like peptide-1) et le GIP (glucose-dependent insulinotropic polypeptide) sont lib��r��es par l'intestin en r��ponse aux nutriments (glucose, acides gras) et agissent au niveau des cellules ��, potentialisant la s��cr��tion d'insuline induite par le glucose, stimulant leur prolif��ration, induisant la diff��rentiation de cellules pr��curseurs en cellules �� matures et les prot��gent contre la mort cellulaire (apoptose). Afin d'��tudier plus en d��tail ces m��canismes, nous avons g��n��r�� des souris d��ficientes pour les r��cepteurs du GIP et du GLP-l. Les ��lots pancr��atiques de ces souris pr��sentent un d��faut de s��cr��tion d'insuline stimul��e par le glucose et une sensibilit�� accrue �� l'apoptose par rapport aux ��lots de souris contr��les. Nous avons donc cherch�� les g��nes r��gul��s pas ces hormones contr��lant la s��cr��tion d'insuline et la protection contre l'apoptose. Nous avons constat�� une forte diminution de l'expression du r��cepteur �� l'IGF-1 (IGF-1R) dans les ��lots de souris d��ficientes pour les r��cepteurs des gluco-incr��tines. Nous avons d��montr�� dans un model de cellules �� en culture et d'��lots que le GLP-1 augmentait l'expression d'IGF-1R et la s��cr��tion de son ligand (IGF-2) permettant l'activation de la voie de signalisation. Finalement, nous avons montr�� que l'activation de la boucle IGF-2/IGF-1R induite par le GLP-1 ��tait n��cessaire pour la protection contre l'apoptose, l'augmentation de la s��cr��tion et la prolif��ration des cellules ��.

Molecular markers allow sibling species identification in red wood ants (Formica rufa group)

Red wood ants (Formica rufa group) constitute a group of species that are considered to be among the most promising bioindicators in forest ecosystems. However, because of their morphological similarity and intraspecific variability, morphological species identification can be difficult. Considerable expertise is necessary to discriminate between the sibling species F. lugubris and F. paralugubris, two species that often live in sympatry in the same Alpine forests. New taxonomic tools providing rapid and reliable species identification are needed. We present a simple and reliable molecular technique based on mtDNA (COI gene) and a restriction enzyme for discriminating between F. lugubris and F. paralugubris. We confirm the validity of this method with a Bayesian analysis based on microsatellites. This new molecular tool represents a clear breakthrough for discriminating between F. lugubris and F. paralugubris and is likely to be helpful in large-scale biomonitoring.

Reverse immunology approach for the identification of CD8 T-cell-defined antigens: advantages and hurdles.

One of the challenges of tumour immunology remains the identification of strongly immunogenic tumour antigens for vaccination. Reverse immunology, that is, the procedure to predict and identify immunogenic peptides from the sequence of a gene product of interest, has been postulated to be a particularly efficient, high-throughput approach for tumour antigen discovery. Over one decade after this concept was born, we discuss the reverse immunology approach in terms of costs and efficacy: data mining with bioinformatic algorithms, molecular methods to identify tumour-specific transcripts, prediction and determination of proteasomal cleavage sites, peptide-binding prediction to HLA molecules and experimental validation, assessment of the in vitro and in vivo immunogenic potential of selected peptide antigens, isolation of specific cytolytic T lymphocyte clones and final validation in functional assays of tumour cell recognition. We conclude that the overall low sensitivity and yield of every prediction step often requires a compensatory up-scaling of the initial number of candidate sequences to be screened, rendering reverse immunology an unexpectedly complex approach.

Identification and antifungal susceptibility of a large collection of yeast strains isolated in Tunisian hospitals.

Abstract In this study, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used as a rapid method to identify yeasts isolated from patients in Tunisian hospitals. When identification could not be exstablished with this procedure, sequencing of the internal transcribed spacer with 5.8S ribosomal DNA (rDNA) (ITS1-5.8S-ITS2) and D1/D2 domain of large-subunit (LSU rDNA) were employed as a molecular approach for species differentiation. Candida albicans was the dominant species (43.37% of all cases), followed by C. glabrata (16.55%), C. parapsilosis (13.23%), C. tropicalis (11.34%), C. dubliniensis (4.96%), and other species more rarely encountered in human diseases such as C. krusei, C. metapsilosis, C. lusitaniae, C. kefyr, C. palmioleophila, C. guilliermondii, C. intermedia, C. orthopsilosis, and C. utilis. In addition, other yeast species were obtained including Saccharomyces cerevisiae, Debaryomyces hansenii (anamorph known as C. famata), Hanseniaspora opuntiae, Kodamaea ohmeri, Pichia caribbica (anamorph known as C. fermentati), Trichosporon spp. and finally a novel yeast species, C. tunisiensis. The in vitro antifungal activities of fluconazole and voriconazole were determined by the agar disk diffusion test and Etest, while the susceptibility to additional antifungal agents was determined with the Sensititre YeastOne system. Our results showed low incidence of azole resistance in C. albicans (0.54%), C. tropicalis (2.08%) and C. glabrata (4.28%). In addition, caspofungin was active against most isolates of the collection with the exception of two K. ohmeri isolates. This is the first report to describe caspofungin resistant isolates of this yeast.

A clinical prognostic model for the identification of low-risk patients with acute symptomatic pulmonary embolism and active cancer.

BACKGROUND: Physicians need a specific risk-stratification tool to facilitate safe and cost-effective approaches to the management of patients with cancer and acute pulmonary embolism (PE). The objective of this study was to develop a simple risk score for predicting 30-day mortality in patients with PE and cancer by using measures readily obtained at the time of PE diagnosis. METHODS: Investigators randomly allocated 1,556 consecutive patients with cancer and acute PE from the international multicenter Registro Informatizado de la Enfermedad TromboEmb��lica to derivation (67%) and internal validation (33%) samples. The external validation cohort for this study consisted of 261 patients with cancer and acute PE. Investigators compared 30-day all-cause mortality and nonfatal adverse medical outcomes across the derivation and two validation samples. RESULTS: In the derivation sample, multivariable analyses produced the risk score, which contained six variables: age > 80 years, heart rate &#8805; 110/min, systolic BP < 100 mm Hg, body weight < 60 kg, recent immobility, and presence of metastases. In the internal validation cohort (n = 508), the 22.2% of patients (113 of 508) classified as low risk by the prognostic model had a 30-day mortality of 4.4% (95% CI, 0.6%-8.2%) compared with 29.9% (95% CI, 25.4%-34.4%) in the high-risk group. In the external validation cohort, the 18% of patients (47 of 261) classified as low risk by the prognostic model had a 30-day mortality of 0%, compared with 19.6% (95% CI, 14.3%-25.0%) in the high-risk group. CONCLUSIONS: The developed clinical prediction rule accurately identifies low-risk patients with cancer and acute PE.

Identification concept and the use of probabilities in forensic odontology--an approach by philosophical discussion.

This paper questions the practitioners' deterministic approach(es) in forensic identification and notes the limits of their conclusions in order to encourage a discussion to question current practices. With this end in view, a hypothetical discussion between an expert in dentistry and an enthusiastic member of a jury, eager to understand the scientific principles of evidence interpretation, is presented. This discussion will lead us to regard any argument aiming at identification as probabilistic.

Identification of potential rockfall source areas at a regional scale using a DEM-based geomorphometric analysis

The availability of high resolution Digital Elevation Models (DEM) at a regional scale enables the analysis of topography with high levels of detail. Hence, a DEM-based geomorphometric approach becomes more accurate for detecting potential rockfall sources. Potential rockfall source areas are identified according to the slope angle distribution deduced from high resolution DEM crossed with other information extracted from geological and topographic maps in GIS format. The slope angle distribution can be decomposed in several Gaussian distributions that can be considered as characteristic of morphological units: rock cliffs, steep slopes, footslopes and plains. A terrain is considered as potential rockfall sources when their slope angles lie over an angle threshold, which is defined where the Gaussian distribution of the morphological unit "Rock cliffs" become dominant over the one of "Steep slopes". In addition to this analysis, the cliff outcrops indicated by the topographic maps were added. They contain however "flat areas", so that only the slope angles values above the mode of the Gaussian distribution of the morphological unit "Steep slopes" were considered. An application of this method is presented over the entire Canton of Vaud (3200 km2), Switzerland. The results were compared with rockfall sources observed on the field and orthophotos analysis in order to validate the method. Finally, the influence of the cell size of the DEM is inspected by applying the methodology over six different DEM resolutions.

Expression of neuroectodermal antigens common to melanomas, gliomas, and neuroblastomas. I. Identification by monoclonal anti-melanoma and anti-glioma antibodies.

The reactivity spectrum of five different monoclonal anti-melanoma antibodies cross-reacting with gliomas and neuroblastomas and one monoclonal anti-glioma antibody cross-reacting with melanomas and neuroblastomas was investigated. Comparison of the binding activity of these monoclonal antibodies for 11 melanoma, seven glioma, and three neuroblastoma cell lines showed that each of these clones had a different pattern of cross-reactivity. The results indicated that the antigenic determinants detected by these antibodies were not associated with the same antigen and thus suggested the existence of at least six different antigens common to melanomas, gliomas, and neuroblastomas. Since all these tumors are known to derive from cells originating embryologically from the neural crest, it can be assumed that the antigens recognized by our monoclonal antibodies are neuroectodermal differentiation antigens. However, absorption with fetal brain homogenates abolished only the binding of monoclonal anti-glioma antibody, but did not modify the binding of monoclonal anti-melanoma antibodies.

Direct identification of recombinant vaccinia virus plaques by PCR.

A fast method for the identification of recombinant vaccinia viruses directly from individual plaques is described. Plaques are picked, resuspended in PBS-A and processed for PCR using two 'universal' primers. The amplified sequences are analyzed by agarose gel electrophoresis. This procedure allows discrimination between spontaneously arising TK-negative mutants, which do not carry the inserted gene, and the desired TK-negative recombinants resulting from insertional inactivation of the TK gene.

Identification et caract��risation de g��nes suppresseurs de tumeurs dans le glioblastome

Introduction: Glioblastoma (WHO Grade IV glioma) is the most frequent and most��malignant primary tumor of the brain. With a mean survival of 15 months despite��multidisciplinary management combining surgery, chemo- and radiotherapy, the prognosis��is poor. Different studies measured a down-regulation of Wnt Inhibitory Factor 1 (WIF1)��expression in a majority of gliobastoma due to genetic and epigenetic regulation. Recently,��a focus on chromosome 12 identified WIF1 as a potential tumor suppressor gene. In��previous results, transfected glioblastoma cells with ectopic expression of WIF1 had a��decreased growth rate and adopted a senescence-like phenotype. In this report, we first��investigated the effect of WIF1 re-expression in glioblastoma cell lines to see if Wnt��inhibition by WIF1 can lead to senescence. To look further, we assessed p21 and c-Myc��expression. p21 has a key role in senescence onset and is directly inhibited by c-Myc,��itself a target of Wnt-pathway. We thus looked if a variation of expression of these genes is��triggered by WIF1 activity. Finally, as autophagy is thought to play a role in senescence��onset, we analyzed the expression of different autophagy genes. We therefore looked for��an association between autophagy activity and senescent phenotype in WIF1-��overexpressing cell lines.��Methods: WIF1-overexpressing clones were selected after transfection of stable��glioblastoma cell lines. Analysis were made through quantitative Polymerase Chain��Reaction (qPCR), Fluorescence-activated Cell Sorting (FACS) and histochemistry.��IGFBP7 and ALDH1A3 have been selected to reflect senescence. ATG5, ATG7 and ULK3��have been selected to reflect autophagy activity.��Results: Using FACS analysis, we found a higher percentage of large cells with increased��granularity amongst WIF1-overexpressing cell lines, which are characteristics of��senescence. In addition, histochemistry showed a higher percentage of multi-nucleated,��beta-galactosidase positive cells in the same cell lines. An increased expression of genes��associated with senescence was found as well. All characteristics were correlated with��levels of WIF1 expression. We did not find any association between p21 and WIF1��expression. No correlation between WIF1 and c-Myc expression was noticed either. In one��of the two cell lines analyzed, the expression of autophagy genes showed some��correlation with expression of WIF1 and expression of genes associated with senescence.��Discussion: After investigations and characterizations on multiple levels, we have��evidence for a senescence phenotype upon WIF1-overexpressing cell lines. This gives a��role to Wnt pathway in the tumorigenicity of glioblastoma. Further experiments are��required to investigate how Wnt inhibition leads to senescence. The role of autophagy in��our senescent cells is here still unclear. Some correlations can be found, letting us think��that there is indeed some involvement of autophagy. However, it is yet to soon to explain��this relationship. Further experiments are required again to confirm the preliminary results��and analyze the variations of autophagy activity within time.

Cultural self-identification(s) and personal social networks among first and second-generation immigrants

L���estudi examina les relacions entre (1) les xarxes socials personals de la poblaci�� immigrant resident a Barcelona i (2) les seves identitats culturals m��ltiples. L���objectiu principal de l���estudi ��s entendre com el contingut i l���estructura de les relacions socials dels immigrants facilita o dificulta (1) tenir un sentiment de pertinen��a a les noves cultures d���acollida, la catalana i la espanyola, i (2) la integraci�� d���aquestes noves identitats socioculturals amb la seva identitat d���origen en una nova identitat bicultural cohesiva. El nostre plantejament inicial era que els immigrants amb xarxes socials m��s diverses des del punt de vista de la seva composici�� cultural tindrien m��s recursos socials i experi��ncies cognitives m��s diverses , factors que afavoreixen les identificacions m��ltiples i la participaci�� c��vica. Els resultats de l���estudi mostren que el grau d���identificaci�� dels participants amb la seva cultura ��tnica o d���origen ��s for��a alt i, en certa mesura, m��s alt en comparaci�� amb les cultures d���acollida ( catalana, c��vica i espanyola). Tanmateix, el vincle dels participants amb les cultures d���acollida (p. ex., la cultura catalana) ��s prou rellevant per a indicar una orientaci�� bicultural (catalana i ��tnica). Les an��lisis de correlacions revelen que sentir-se catal�� no impedeix sentir-se part de la comunitat etnocultural d���origen. A m��s, existeix una interrelaci�� entre l'orientaci�� cultural catalana i la identificaci�� amb les comunitats c��viques locals. De la mateixa manera, tenir compet��ncies en llengua catalana no va en detriment de les compet��ncies en llengua castellana. Les an��lisis tamb�� mostren que factors com l���orientaci�� cultural catalana, l�����s del catal�� i la identificaci�� amb la cultura catalana tenen una correlaci�� positiva amb el grau de chohesio de la indentitat bicultural, afavoreixen el benestar psicol��gic i disminueixen l���estr��s aculturatiu. L���an��lisi de les xarxes socials mostra que la identificaci�� amb la cultura catalana, l���orientaci�� cultural catalana i la integraci�� de la identitat s��n factors clau per tenir xarxes socials m��s diverses des del punt de vista ��tnic i ling����stic, amb menys membres del col���lectiu d���origen, i amb subgrups o ���cliques��� culturalment m��s heterogenis. La identificaci�� espanyola tamb�� prediu, en mesura m��s redu��da, la diversitat de les xarxes. Els nostres resultats contribueixen a la recerca actual i les teories sobre interculturalitat i identitat cultural.

Hours worked - Productivity puzzle: identification in fractional integration settings

A recent finding of the structural VAR literature is that the response of hours worked to a technology shock depends on the assumption on the order of integration of the hours. In this work we relax this assumption, allowing for fractional integration and long memory in the process for hours and productivity. We find that the sign and magnitude of the estimated impulse responses of hours to a positive technology shock depend crucially on the assumptions applied to identify them. Responses estimated with short-run identification are positive and statistically significant in all datasets analyzed. Long-run identification results in negative often not statistically significant responses. We check validity of these assumptions with the Sims (1989) procedure, concluding that both types of assumptions are appropriate to recover the impulse responses of hours in a fractionally integrated VAR. However, the application of longrun identification results in a substantial increase of the sampling uncertainty. JEL Classification numbers: C22, E32. Keywords: technology shock, fractional integration, hours worked, structural VAR, identification

Identification of potential small molecule binding pockets on Rho family GTPases

Rho GTPases are conformational switches that control a wide variety of signaling pathways critical for eukaryotic cell development and proliferation. They represent attractive targets for drug design as their aberrant function and deregulated activity is associated with many human diseases including cancer. Extensive high-resolution structures (.100) and recent mutagenesis studies have laid the foundation for the design of new structure-based chemotherapeutic strategies. Although the inhibition of Rho signaling with drug-like compounds is an active area of current research, very little attention has been devoted to directly inhibiting Rho by targeting potential allosteric non-nucleotide binding sites. By avoiding the nucleotide binding site, compounds may minimize the potential for undesirable off-target interactions with other ubiquitous GTP and ATP binding proteins. Here we describe the application of molecular dynamics simulations, principal component analysis, sequence conservation analysis, and ensemble small-molecule fragment mapping to provide an extensive mapping of potential small-molecule binding pockets on Rho family members. Characterized sites include novel pockets in the vicinity of the conformationaly responsive switch regions as well as distal sites that appear to be related to the conformations of the nucleotide binding region. Furthermore the use of accelerated molecular dynamics simulation, an advanced sampling method that extends the accessible time-scale of conventional simulations, is found to enhance the characterization of novel binding sites when conformational changes are important for the protein mechanism.