Cell-type-specific adhesion onto polymer surfaces from mixed cell populations

The targeted adhesion of a specific cell type from a mixed cell suspension via the surface presentation of a cell-specific ligand is demonstrated. This generic strategy is illustrated by the covalent attachment of a galactose derivative to a polylysine backbone via the amine functionality. Following adsorption of the resultant material to a polymer surface, hepatocyte adhesion is increased via the interaction between galactose and asialoglycoprotein receptors in a concentration-dependent manner. The selective nature of the material is demonstrated by the approximate doubling in the adhesion of hepatocytes relative to a nontargeted cell type (hepatic stellate cells), and an inability of the modified polymer surface to attract additional numbers of the nontargeted cells. This strategy provides a mechanism for controlling the ratios of cell types adhering to scaffold supports, thus enabling the rapid creation of defined coculture systems from heterogeneous cell suspensions.     

General growth mixture modeling for randomized preventive interventions

This paper proposes growth mixture modeling to assess intervention effects in longitudinal randomized trials. Growth mixture modeling represents unobserved heterogeneity among the subjects using a finite-mixture random effects model. The methodology allows one to examine the impact of an intervention on subgroups characterized by different types of growth trajectories. Such modeling is informative when examining effects on populations that contain individuals who have normative growth as well as non-normative growth. The analysis identifies subgroup membership and allows theory-based modeling of intervention effects in the different subgroups. An example is presented concerning a randomized intervention in Baltimore public schools aimed at reducing aggressive classroom behavior, where only students who were initially more aggressive showed benefits from the intervention.     

Host-pathogen studies in the post-genomic era

Several studies are starting to show the power of DNA microarrays to identify interactions between animal hosts and their pathogens, and have revealed interesting correlations between host responses to different infectious agents.     

Comparison of different delivery systems of DNA vaccination for the induction of protection against tuberculosis in mice and guinea pigs

The great challenges for researchers working in the field of vaccinology are optimizing DNA vaccines for use in humans or large animals and creating effective single-dose vaccines using appropriated controlled delivery systems. Plasmid DNA encoding the heat-shock protein 65 (hsp65) (DNAhsp65) has been shown to induce protective and therapeutic immune responses in a murine model of tuberculosis (TB). Despite the success of naked DNAhsp65-based vaccine to protect mice against TB, it requires multiple doses of high amounts of DNA for effective immunization. In order to optimize this DNA vaccine and simplify the vaccination schedule, we coencapsulated DNAhsp65 and the adjuvant trehalose dimycolate (TDM) into biodegradable poly (DL-lactide-co-glycolide) (PLGA) microspheres for a single dose administration. Moreover, a single-shot prime-boost vaccine formulation based on a mixture of two different PLGA microspheres, presenting faster and slower release of, respectively, DNAhsp65 and the recombinant hsp65 protein was also developed. These formulations were tested in mice as well as in guinea pigs by comparison with the efficacy and toxicity induced by the naked DNA preparation or BCG. The single-shot prime-boost formulation clearly presented good efficacy and diminished lung pathology in both mice and guinea pigs.     

Growth hormone and gene expression of in vitro‐matured rhesus macaque oocytes

Growth hormone (GH) in rhesus macaque in vitro oocyte maturation (IVM) has been shown to increase cumulus expansion and development of embryos to the 9–16 cell stage in response to 100 ng/ml recombinant human GH (r‐hGH) supplementation during IVM. Although developmental endpoints for metaphase II (MII) oocytes and embryos are limited in the macaque, gene expression analysis can provide a mechanism to explore GH action on IVM. In addition, gene expression analysis may allow molecular events associated with improved cytoplasmic maturation to be detected. In this study, gene expression of specific mRNAs in MII oocytes and cumulus cells that have or have not been exposed to r‐hGH during IVM was compared. In addition, mRNA expression was compared between in vitro and in vivo‐matured metaphase II (MII) oocytes and germinal vesicle (GV)‐stage oocytes. Only 2 of 17 genes, insulin‐like growth factor 2 (IGF2) and steroidogenic acute regulator (STAR), showed increased mRNA expression in MII oocytes from the 100 ng/ml r‐hGH treatment group compared with other IVM treatment groups, implicating insulin‐like growth factor (IGF) and steroidogenesis pathways in the oocyte response to GH. The importance of IGF2 is notable, as expression of IGF1 was not detected in macaque GV‐stage or MII oocytes or cumulus cells. Mol. Reprod. Dev. 77: 353–362, 2010. © 2009 Wiley‐Liss, Inc.     

Composition and preservation of the Chengjiang fauna –a Lower Cambrian soft-bodied biota

The Lower Cambrian Chengjiang fauna is reviewed and shown to be closely comparable with the younger Burgess Shale fauna. but with various differences in detail. A diverse group of more or less annulated lobopod animals including 'armoured lobopods' are regarded as representatives of the phylum Onychophora. 'Trilobitomorphs' include several new types. Probable protaspides of the trilobitomorph Naraoia are described. No molluses or deuterostomes have been identified. The preservational orientations of the various taxa are reviewed and compared with orientations of the Burgess Shale taxa. Orientation in the sediment is found to be closely correlated to the original shape of individuals. Several new genera and species are described: the segmented. worm-shaped Yunnanozoon lividum gen. et sp.n., the 'armoured lobopods' Onychodictyon ferox gen. et sp.n. and Cardiodictyon catenulum gen. et sp.n. and the arthropods Saperion glumaceum gen. et sp.n., Sinoburius Iunaris gen. et sp.n., and Xandarella spectaculum gen. et sp.n.     

Assessment of a 44 gene classifier for the evaluation of chronic fatigue syndrome from peripheral blood mononuclear cell gene expression

Chronic fatigue syndrome (CFS) is a clinically defined illness estimated to affect millions of people worldwide causing significant morbidity and an annual cost of billions of dollars. Currently there are no laboratory-based diagnostic methods for CFS. However, differences in gene expression profiles between CFS patients and healthy persons have been reported in the literature. Using mRNA relative quantities for 44 previously identified reporter genes taken from a large dataset comprising both CFS patients and healthy volunteers, we derived a gene profile scoring metric to accurately classify CFS and healthy samples. This metric out-performed any of the reporter genes used individually as a classifier of CFS.To determine whether the reporter genes were robust across populations, we applied this metric to classify a separate blind dataset of mRNA relative quantities from a new population of CFS patients and healthy persons with limited success. Although the metric was able to successfully classify roughly two-thirds of both CFS and healthy samples correctly, the level of misclassification was high. We conclude many of the previously identified reporter genes are study-specific and thus cannot be used as a broad CFS diagnostic.     

A Membrane Topology Model for Human Interferon Inducible Transmembrane Protein 1

InterFeron Inducible TransMembrane proteins 1–3 (IFITM1, IFITM2 and IFITM3) are a family of proteins capable of inhibiting the cellular entry of numerous human and animal viruses. IFITM1-3 are unique amongst the currently described viral restriction factors in their apparent ability to block viral entry. This restrictive property is dependant on the localisation of the proteins to plasma and endosomal membranes, which constitute the main portals of viral entry into cells. The topology of the IFITM proteins within cell membranes is an unresolved aspect of their biology. Here we present data from immunofluorescence microscopy, protease cleavage, biotin-labelling and immuno-electron microscopy assays, showing that human IFITM1 has a membrane topology in which the N-terminal domain resides in the cytoplasm, and the C-terminal domain is extracellular. Furthermore, we provide evidence that this topology is conserved for all of the human interferon-induced IFITM proteins. This model is consistent with that recently proposed for murine IFITM3, but differs from that proposed for murine IFITM1.