The differentiated state of mature cells of adult organisms is achieved and maintained through the epigenetic regulation of gene expression, which consists of several mechanisms including DNA methylation. The advent of induced pluripotent stem cell technology enabled the conversion of adult cells into any other cell type passing through a stable pluripotency state. However, indefinite pluripotency is unphysiological, inherently labile, and makes cells prone to culture-induced alterations. The direct conversion of one cell type to another without an intermediate pluripotent stage is also possible but, at present, requires the viral transfection of appropriate transcription factors, limiting its therapeutic potential. The aim of this study was to investigate whether it is possible to achieve the direct conversion of an adult cell by exposing it to a demethylating agent immediately followed by differentiating culture conditions. Adult human skin fibroblasts were exposed for 18 h to the DNA methyltransferase inhibitor 5-azacytidine, followed by a three-step protocol for the induction of endocrine pancreatic differentiation that lasted 36 d. At the end of this treatment, 35 ± 8.9% fibroblasts became pancreatic converted cells that acquired an epithelial morphology, produced insulin, and then released the hormone in response to a physiological glucose challenge in vitro. Furthermore, pancreatic converted cells were able to protect recipient mice against streptozotocin-induced diabetes, restoring a physiological response to glucose tolerance tests. This work shows that it is possible to convert adult fibroblasts into insulin-secreting cells, avoiding both a stable pluripotent stage and any transgenic modification.

Clustered, regularly interspaced, short palindromic repeat (CRISPR) loci, found in prokaryotes, are transcribed to produce CRISPR RNAs (crRNAs). The Cmr proteins (Cmr1-6) and crRNA form a ribonucleoprotein complex that degrades target RNAs derived from invading genetic elements. Cmr2dHD, a Cmr2 variant lacking the N-terminal putative HD nuclease domain, and Cmr3 were co-expressed in Escherichia coli cells and co-purified as a complex. The Cmr2dHD-Cmr3 complex was co-crystallized with 3'-AMP by the vapour-diffusion method. The crystals diffracted to 2.6 Å resolution using synchrotron radiation at the Photon Factory. The crystals belonged to the orthorhombic space group I222, with unit-cell parameters a = 103.9, b = 136.7, c = 192.0 Å. The asymmetric unit of the crystals is expected to contain one Cmr2dHD-Cmr3 complex with a Matthews coefficient of 3.0 Å(3) Da(-1) and a solvent content of 59%.

OBJECTIVES:

The purpose of this study was to investigate the use of Gd-DTPA shortly before magnetic resonance guided high-intensity focused ultrasound MR-HIFU thermal ablation therapy with respect to dissociation, trapping, and long-term deposition of gadolinium (Gd) in the body.

MATERIALS AND METHODS:

Magnetic resonance-HIFU ablation treatment was conducted in vivo on both rat muscle and subcutaneous tumor (9L glioma) using a clinical 3T MR-HIFU system equipped with a small-animal coil setup. A human equivalent dose of gadopentetate dimeglumine (Gd-DTPA) (0.6 mmol/kg of body weight) was injected via a tail vein catheter just before ablation (≤5 minutes). Potential trapping of the contrast agent in the ablated area was visualized through the acquisition of R1 maps of the target location before and after therapy. The animals were sacrificed 2 hours or 14 days after the injection (n = 4 per group, a total of 40 animals). Subsequently, the Gd content in the tissue and carcass was determined using inductively coupled plasma techniques to investigate the biodistribution.

RESULTS:

Temporal trapping of Gd-DTPA in the coagulated tissue was observed on the R1 maps acquired within 2 hours after the ablation, an effect confirmed by the inductively coupled plasma analysis (3 times more Gd was found in the treated muscle volume than in the control muscle tissue). Two weeks after the therapy, the absolute amount of Gd present in the coagulated tissue was low compared with the amount present in the kidneys 14 days after the injection (ablated muscle, 0.009% ± 0.002% ID/g; kidney, 0.144% ± 0.165% ID/g). There was no significant increase in Gd content in the principal target organs for translocated Gdions (liver, spleen, and bone) or in the entire carcasses between the HIFU- and sham-treated animals. Finally, an in vivo relaxivity of 4.6 mmols was found in the HIFU-ablated volume, indicating intact Gd-DTPA.

CONCLUSIONS:

Magnetic resonance-HIFU treatment does not induce the dissociation of Gd-DTPA. In small-tissue volumes, no significant effect on the long-term in vivo Gd retention was found. However, care must be taken with the use of proton resonance frequency shift-based MR thermometry for HIFU guidance in combination with Gd because the susceptibility artifact induced by Gd can severely influence treatment outcome.

Chitinase A1 (ChiA1) from Bacillus circulans WL-12 comprises an N-terminal catalytic domain, two fibronectin type III domains (FnIIIDs), and a C-terminal chitin-binding domain (ChBD). The chitin-binding domain of ChiA1 (ChBDChiA1) belongs to carbohydrate-binding module (CBM) family 12 and specifically binds to insoluble or crystalline chitin. It has been suggested that tryptophan-687 (Trp687) is involved in the chitin-binding activity of this ChBD. Site-directed mutagenesis was used to identify additional amino acid residues required for chitin-binding activity of this domain. Furthermore, a total of 14 amino acid residues in ChBDChiA1 were carefully selected, and it was found that mutation of Gln679, which is not well-conserved in CBM family 12, significantly decreased the binding activity to colloidal chitin. A nuclear magnetic resonance (NMR) study demonstrated that neither the Q679A nor the W687A mutation altered the overall structure of ChBDChiA1. Therefore, Gln679 was identified as a new residue that is involved in the chitin-binding activity of ChBDChiA1 in addition to Trp687. However, the mechanism of chitin binding by ChBD is still unknown.

Prenatal cocaine exposure (PCE) is associated with arousal dysregulation, and alterations of amygdala activity in response to emotional arousal have previously been reported. However, voluntary regulation of emotional affect, enabling appropriate neural response to different streams of stimuli, must also engage prefrontal regions, yet the impact of PCE on these prefrontal mechanisms has not been investigated. Recent neuroimaging studies have shown the involvement of ventral prefrontal cortex (vPFC) in the modulation of amygdala reactivity and the mediation of effective emotional regulation. Based on these findings, using functional magnetic resonance imaging (fMRI) and diffusion tensor imaging (DTI), the present study compared functional activations of the vPFC as well as its structural connectivity with the amygdala between groups of PCE and control adolescents. In a working memory task with emotional distracters, the PCE adolescents exhibited less capability of increasing their vPFC activation in response to increased memory load, which corresponded with their less suppressed amygdala activation. Reduced structural connectivity between the vPFC and the amygdala was also observed from DTI measurement in the PCE group. In addition, correlations between amygdala activation and (i) vPFC activation, as well as (ii) amygdala-vPFC structural connectivity, were observed in the control but not in the PCE group. These data complement previous findings of the impact of PCE on the activity of the amygdala and extend our understanding of the neurobiological mechanisms underlying the effect of PCE on arousal dysregulation reported in human and animal studies.

The aim of this study was to examine the association between different types of exercise loading and femoral neck cross-sectional geometry. Our data comprised proximal femur magnetic resonance (MR) images obtained from 91 female athletes and their 20 age-matched controls. The athletes were categorized according to typical training activity - high impact (high and triple jumping), odd impact (racket and soccer playing), high magnitude (power lifting), repetitive low impact (endurance running) and repetitive non-impact (swimming). Segmented MR images at two locations, narrowest cross-section of the femoral neck (narrowFN) and the cross-section at insertion of articular capsule (distalFN), were investigated to detect between group differences in shape, curvature and buckling ratio derived using image and signal analysis tools. The narrowFN results indicated that the high-impact group had weaker antero-superior (33% larger buckling ratio than controls) but stronger inferior weight-bearing region (32% smaller than controls), while the odd-impact group had stronger superior, posterior and anterior region (21% smaller buckling ratio than controls). The distalFN results indicated that the high-impact group had stronger inferior region (37% smaller buckling ratio), but the odd-impact group had stronger superior region (22% smaller buckling ratio) than the controls. Overall, the results point towards odd-impact exercise loading, with inherently varying directions of impact, associated with more robust cross-sectional geometry along the femoral neck. In conclusion, our one-dimensional polar treatment for geometrical traits and intuitive presentation of differences in trends between exercise groups and controls provides a basis for analysis with high angular accuracy.

Segmentation of nuclei from images of tissue sections is important for many biological and biomedical studies. Many existing image segmentation algorithms may lead to oversegmentation or undersegmentation for clustered nuclei images. In this paper, we proposed a new image segmentation algorithm based on concave curve expansion to correctly and accurately extract markers from the original images. Marker-controlled watershed is then used to segment the clustered nuclei. The algorithm was tested on both synthetic and real images and better results are achieved compared with some other state-of-the-art methods.

The biomedical model is the foundation upon which current evaluations in clinical practice are based. In the quest for objective evidence to support clinical interventions, the patient is reduced to a number of technologically generated variables that serve as a surrogate for the patient herself. The biomedical model, as a reflection of Gestell or the essence of technology, carries with it the danger that it may overwhelm the practitioner's perspective so that other perspectives that may include the lived experience of the patient are actively suppressed. An ontology of the patient based on a Heideggerian exegesis is developed as a response to this concern. Morris has suggested that the most fundamental disturbance in the lived experience of the patient is an alteration in the patient's relationship to ecstatic temporality. In ecstatic temporality, the past, present and future are experienced as a unity in which the patient sees herself as her own possibility. Access to this experience is disturbed in illness; the future is no longer experienced as the patient's own possibility but rather as a series of predetermined external events that dictate the patient's affairs. By developing a biologically plausible model of ecstatic temporality, the lived experience of the patient does not have to be considered separate from the physical mechanisms involved in the disease state. As a consequence, the biomedical model cannot suppress the practitioner's humane perspective since the latter is explicitly brought under its purview.

Due to the relatively recent emergence of the human T-lymphotropic and the human immunodeficiency viruses, enthusiasm for the identification of novel viruses, especially retroviruses, with pathogenic potential in humans, remains high. Novel technologies are now available with the ability to search for unknown viruses, such as gene arrays and new generation sequencing of tissue and other samples. In 2006, chip technology identified a novel retrovirus in human prostate cancer (PCa) tissue samples. Due to close homology to a mouse retrovirus, the virus was named xenotropic murine leukaemia virus-related virus (XMRV). Ever since the initial disease association with PCa, XMRV has stirred a lot of attention and concern worldwide for the medical community, public health officials and in particular global transfusion services. Public response, in this new era of electronic communication and advocacy was rapid, wide and unprecedented. In this review, we outline the course of biomedical research efforts that were put forward internationally in the process of determining the risk to the human population, the response of the blood banking community and review the current state of knowledge of xenotropic murine retroviruses. Although XMRV is no longer regarded as an infection of humans, a lesson was learnt in modern virology that holds deeper implications for biomedical research, particularly stem cell generation and transplantation practices.

The goal of this study was to investigate the influence of the brain-to-skull conductivity ratio (BSCR) on EEG source localization accuracy. In this study, we evaluated four BSCRs: 15, 20, 25, and 80, which were mainly discussed according to the literature. The scalp EEG signals were generated by BSCR-related forward computation for each cortical dipole source. Then, for each scalp EEG measurement, the source reconstruction was performed to identify the estimated dipole sources by the actual BSCR and the misspecified BSCRs. The estimated dipole sources were compared with the simulated dipole sources to evaluate EEG source localization accuracy. In the case of considering noise-free EEG measurements, the mean localization errors were approximately equal to zero when using actual BSCR. The misspecified BSCRs resulted in substantial localization errors which ranged from 2 to 16 mm. When considering noise-contaminated EEG measurements, the mean localization errors ranged from 8 to 18 mm despite the BSCRs used in the inverse calculation. The present results suggest that the localization accuracy is sensitive to the BSCR in EEG source reconstruction, and the source activity can be accurately localized when the actual BSCR and the EEG scalp signals with high signal-to-noise ratio (SNR) are used.