Micropositioning and microscopic observation of individual picoliter-sized containers within SU-8 microchannels

We describe the fabrication and application of a bioanalytical chip, made of SU-8 photoresist, comprising integrated, high aspect-ratio microfluidic channels, suitable to manipulate and investigate vesicles, cell fragments and biological cells. A central micrometer-sized aperture allows electrical particle counting and planar membrane experiments, microfluids allow (sub)micrometer-sized objects to be transported and addressed with different chemicals. Here we show how lipid vesicles are positioned with micrometer precision within the micro-channels by means of pressure and electrophoretic movement. Our approach is suited for controlling and investigating (bio)chemical synthesis and cellular signalling processes in ultrasmall individual vesicles by electro-optical techniques.     

Disambiguating Visual Motion by Form-Motion Interaction—a Computational Model

The neural mechanisms underlying motion segregation and integration still remain unclear to a large extent. Local motion estimates often are ambiguous in the lack of form features, such as corners or junctions. Furthermore, even in the presence of such features, local motion estimates may be wrong if they were generated near occlusions or from transparent objects. Here, a neural model of visual motion processing is presented that involves early stages of the cortical dorsal and ventral pathways. We investigate the computational mechanisms of V1-MT feedforward and feedback processing in the perception of coherent shape motion. In particular, we demonstrate how modulatory MT-V1 feedback helps to stabilize localized feature signals at, e.g. corners, and to disambiguate initial flow estimates that signal ambiguous movement due to the aperture problem for single shapes. In cluttered environments with multiple moving objects partial occlusions may occur which, in turn, generate erroneous motion signals at points of overlapping form. Intrinsic-extrinsic region boundaries are indicated by local T-junctions of possibly any orientation and spatial configuration. Such junctions generate strong localized feature tracking signals that inject erroneous motion directions into the integration process. We describe a simple local mechanism of excitatory form-motion interaction that modifies spurious motion cues at T-junctions. In concert with local competitive-cooperative mechanisms of the motion pathway the motion signals are subsequently segregated into coherent representations of moving shapes. Computer simulations demonstrate the competency of the proposed neural model.     

Scintillation light produced by low-energy beams of highly-charged ions

Measurements have been performed of scintillation light intensities emitted from various inorganic scintillators irradiated with low-energy beams of highly-charged ions from an electron beam ion source (EBIS) and an electron cyclotron resonance ion source (ECRIS). Beams of xenon ions Xe^q+ with various charge states between q = 2 and q = 18 have been used at energies between 5 and 17.5 keV per charge generated by the ECRIS. The intensity of the beam was typically varied between 1 and 100 nA. Beams of highly charged residual gas ions have been produced by the EBIS at 4.5 keV per charge and with low intensities down to 100 pA. The scintillator materials used are flat screens of P46 YAG and P43 phosphor. In all cases, scintillation light emitted from the screen surface was detected by a CCD camera. The scintillation light intensity has been found to depend linearly on the kinetic ion energy per time deposited into the scintillator, while up to q = 18 no significant contribution from the ions’ potential energy was found. We discuss the results on the background of a possible use as beam diagnostics, e.g. for the new HITRAP facility at GSI, Germany.     

Male gender role conflict and willingness to seek counseling: Testing a mediation model on college-aged men.

Men who experience negative consequences of their socialized gender roles--that is, have greater gender role conflict--report less positive attitudes and willingness to seeking counseling. Using structural equation modeling with data from 575 undergraduate men, the authors examined 3 mediators (self-stigma associated with seeking counseling, tendency to disclose distressing information, and attitudes toward seeking counseling) regarding the link between gender role conflict and willingness to seek counseling for psychological and interpersonal concerns. Results indicated that this link was partially mediated by these 3 factors. Men experiencing greater gender role conflict were more likely to self-stigmatize and less likely to self-disclose. High self-stigma and less disclosure then led to less positive attitudes and subsequently to less willingness to seek counseling. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Evaluation of high‐resolution and mobile display systems for digital radiology in dark and bright environments using human and computational observers

Abstract— As digital display systems replace film traditionally used for reading radiographic images, resource‐intensive acceptance testing must be performed to ensure that quality meets and maintains desired specifications. If machine observers can replace human readers, whose performances are highly variable, the results will be more consistent and less costly. To be effective, however, the automated observers must track human performance. An approach for a model observer, validated with human readers, for the evaluation of the visibility of low‐contrast small targets in high‐resolution and mobile displays under different ambient illumination, will be described. The displays were tested using CDMAM‐like digital phantoms containing disks of varying diameters and contrasts on a flat background. For this task, we find the best indicator of display performance to be the display's ability to represent small luminance contrast, not resolution or pixel size. The results confirm that high‐resolution systems perform better under low illumination while illuminance has a minor impact on the mobile‐display performance. Finally, the results show that the machine observer tracks the performance of human readers. Machine observers with proper validation can replace humans in the acceptance testing procedures, saving the testers both time and money.     

ER Stress in ERp57 Knockout Knee Joint Chondrocytes Induces Osteoarthritic Cartilage Degradation and Osteophyte Formation

Ageing or obesity are risk factors for protein aggregation in the endoplasmic reticulum (ER) of chondrocytes. This condition is called ER stress and leads to induction of the unfolded protein response (UPR), which, depending on the stress level, restores normal cell function or initiates apoptotic cell death. Here the role of ER stress in knee osteoarthritis (OA) was evaluated. It was first tested in vitro and in vivo whether a knockout (KO) of the protein disulfide isomerase ERp57 in chondrocytes induces sufficient ER stress for such analyses. ER stress in ERp57 KO chondrocytes was confirmed by immunofluorescence, immunohistochemistry, and transmission electron microscopy. Knee joints of wildtype (WT) and cartilage-specific ERp57 KO mice (ERp57 cKO) were analyzed by indentation-type atomic force microscopy (IT-AFM), toluidine blue, and immunofluorescence/-histochemical staining. Apoptotic cell death was investigated by a TUNEL assay. Additionally, OA was induced via forced exercise on a treadmill. ER stress in chondrocytes resulted in a reduced compressive stiffness of knee cartilage. With ER stress, 18-month-old mice developed osteoarthritic cartilage degeneration with osteophyte formation in knee joints. These degenerative changes were preceded by apoptotic death in articular chondrocytes. Young mice were not susceptible to OA, even when subjected to forced exercise. This study demonstrates that ER stress induces the development of age-related knee osteoarthritis owing to a decreased protective function of the UPR in chondrocytes with increasing age, while apoptosis increases. Therefore, inhibition of ER stress appears to be an attractive therapeutic target for OA.     

Kinetic masks: a new approach and device for dispersing biologically relevant fluids

The controlled dispersion of fluids, particularly biologically relevant solutions in micro-volumes, is of high practical interest in biotechnology and medicine. Pharmaceutical test assays, for example, need a method for the fast and defined deposition of fluid samples. Most current micro-dispensing methods, i.e. contact-based pin printing, have problems such as time delays, limited dosing velocity, minimum volume or high interference that limit biological applications. Spraying techniques suffer from a lack of reproducibility; a defined deposition of samples on targets is not possible. Here, we introduce a new method for the parallel and spatially defined dispersion of many micro-volumes that overcomes disadvantages of common micro-dispensers. The overall approach is that a fluid drop, produced by a droplet generator, falls on a free trajectory with a defined kinetic energy, and is split by a masking unit placed perpendicular to the flight direction into at least two smaller droplets (Zimmermann et al. in Method and device for dosing fluid media, WO/2002/102515, Germany, 2002). On the target, the resulting droplets form reproducible patterns, which are enlarged and scalable images of the grid pattern. Possible applications for this method are non-contact cell patterning, cell encapsulation, cryopreservation and fast mixing processes in micro-volumes. Here, we use this method for the direct and defined parallel positioning of cell suspensions on specific substrates, which can be useful for test assays, tissue engineering and cryopreservation.     

Mitochondrial Mechanisms in Septic Cardiomyopathy

Sepsis is the manifestation of the immune and inflammatory response to infection that may ultimately result in multi organ failure. Despite the therapeutic strategies that have been used up to now, sepsis and septic shock remain a leading cause of death in critically ill patients. Myocardial dysfunction is a well-described complication of severe sepsis, also referred to as septic cardiomyopathy, which may progress to right and left ventricular pump failure. Many substances and mechanisms seem to be involved in myocardial dysfunction in sepsis, including toxins, cytokines, nitric oxide, complement activation, apoptosis and energy metabolic derangements. Nevertheless, the precise underlying molecular mechanisms as well as their significance in the pathogenesis of septic cardiomyopathy remain incompletely understood. A well-investigated abnormality in septic cardiomyopathy is mitochondrial dysfunction, which likely contributes to cardiac dysfunction by causing myocardial energy depletion. A number of mechanisms have been proposed to cause mitochondrial dysfunction in septic cardiomyopathy, although it remains controversially discussed whether some mechanisms impair mitochondrial function or serve to restore mitochondrial function. The purpose of this review is to discuss mitochondrial mechanisms that may causally contribute to mitochondrial dysfunction and/or may represent adaptive responses to mitochondrial dysfunction in septic cardiomyopathy.