Barium alginate capsules for 3D immobilisation of living cells: morphology, membrane properties and permeability

Encapsulation in a barium alginate membrane is a promising strategy to obtain a three dimensional culture of living cells: membrane properties are crucial for a realistic clinical application. A one-step encapsulation technique, recently developed for controlled release of boar semen, was employed to prepare barium alginate and protamine-alginate membranes: permeability to two model molecules (haemoglobin and glucose) was evaluated. Capsules were evaluated for technological properties and scanning electron microscopy was used to examine the external morphology of the capsules and the 3D distribution of the cells within the core. The results indicate that 3D arrangement and cell shape are maintained, capsule dimensions and mechanical properties can be modulated, as well as their permeability to model molecules such as haemoglobin and glucose

Source extraction and photometry for the far-infrared and sub-millimeter continuum in the presence of complex backgrounds

(Abridged) We present a new method for detecting and measuring compact sources in conditions of intense, and highly variable, fore/background. While all most commonly used packages carry out the source detection over the signal image, our proposed method builds from the measured image a "curvature" image by double-differentiation in four different directions. In this way point-like as well as resolved, yet relatively compact, objects are easily revealed while the slower varying fore/background is greatly diminished. Candidate sources are then identified by looking for pixels where the curvature exceeds, in absolute terms, a given threshold; the methodology easily allows us to pinpoint breakpoints in the source brightness profile and then derive reliable guesses for the sources extent. Identified peaks are fit with 2D elliptical Gaussians plus an underlying planar inclined plateau, with mild constraints on size and orientation. Mutually contaminating sources are fit with multiple Gaussians simultaneously using flexible constraints. We ran our method on simulated large-scale fields with 1000 sources of different peak flux overlaid on a realistic realization of diffuse background. We find detection rates in excess of 90% for sources with peak fluxes above the 3-sigma signal noise limit; for about 80% of the sources the recovered peak fluxes are within 30% of their input values.Comment: Accepted on A&

Fabrication of Ankle-Foot Orthoses using Selective Laser Sintering Technology

Passive dynamic ankle-foot orthoses (AFOs) are often prescribed to improve gait performance for those with various neuromuscular disorders. Designs and materials used for AFOs range from simple polypropylene braces to advanced custom carbon fiber dynamic AFOs that passively store and release mechanical energy during gait. AFO designs vary in the shape and length of the foot component as well as the stiffness and length of the tibial component, depending on the desired functional outcomes. However, the current fabrication technology is not ideally suited for refined customization of AFO characteristics to optimize performance, or for rapid lowcost, high volume manufacturing and global distribution. A promising engineering solution for producing customized dynamic AFOs is the application of Selective Laser Sintering (SLS), which is a versatile manufacturing technology that provides advantages over traditional methods and has already been successfully used to fabricate prosthetic sockets for lower limb amputees (e.g., Faustini et al., 2006). Thus, the primary objective of this study was to explore the feasibility of using an SLSbased design, analysis and manufacturing framework to produce subject-specific passive dynamic AFOs in a cost-effective manner

Variations of the spectral index of dust emissivity from Hi-GAL observations of the Galactic plane

Original article can be found at: http://www.aanda.org/ Copyright The European Southern ObservatoryContext. Variations in the dust emissivity are critical for gas mass determinations derived from far-infrared observations, but also for separating dust foreground emission from the Cosmic Microwave Background (CMB). Hi-GAL observations allow us for the first time to study the dust emissivity variations in the inner regions of the Galactic plane at resolution below 1°. Aims. We present maps of the emissivity spectral index derived from the combined Herschel PACS 160 μm, SPIRE 250 μm, 350 μm, and 500 μm data, and the IRIS 100 μm data, and we analyze the spatial variations of the spectral index as a function of dust temperature and wavelength in the two science demonstration phase Hi-GAL fields, centered at l = 30° and l = 59°. Methods. Applying two different methods, we determine both dust temperature and emissivity spectral index between 100 and 500 μm, at an angular resolution (θ) of 4'. Results. Combining both fields, the results show variations of the emissivity spectral index in the range 1.8–2.6 for temperatures between 14 and 23 K. The median values of the spectral index are similar in both fields, i.e. 2.3 in the range 100–500 μm, while the median dust temperatures are equal to 19.1 K and 16.0 K in the l = 30° and l = 59° field, respectively. Statistically, we do not see any significant deviations in the spectra from a power law emissivity between 100 and 500 μm. We confirm the existence of an inverse correlation between the emissivity spectral index and dust temperature, found in previous analyses.Peer reviewe

Morphological and acrosomal changes of canine spermatozoa during epididymal transit

Background: During epididymal transit, functional and structural modifications leading to full maturation enable male gametes to reach, recognize and fertilize the oocytes. In dogs, little is known on the modifications of spermatozoa during the passage in the epididymis. The aim of this study was to describe the motility, morphology and acrosomal patterns of canine spermatozoa retrieved from the epididymis caput, corpus and cauda.Results: After the dilution required for the collection of epididymal content, sperm motility was significantly higher(P<0.0001) in the cauda compared to corpus and caput.Proportions of spermatozoa with normal morphology were significantly higher in corpus (P =0.02) and cauda(P<0.0001) compared to caput. Overall morphological abnormalities of the head and neck/midpiece were similar in the three different epididymal regions. A significantly increased prevalence of tail defects, mainly represented by single bent tails, was observed in the corpus compared to caput (P<0.0001) and cauda (P=0.006).Numbers of immature sperm with cytoplasmic droplets decreased from the proximal to the distal region of the epididymis. Particularly, proximal cytoplasmic droplets were more frequently found in spermatozoa collected from the caput epididymis than in the corpus (P<0.0001) and in the cauda (P<0.0001), whereas the occurrence of distal cytoplasmic droplets was higher in the corpus than in the caput (P=0.0003) and in the cauda (P<0.05).Significantly higher proportions of spermatozoa with intact acrosomes were retrieved from the cauda epididymis than from the caput (P=0.03) and the corpus (P =0.008). This difference was mainly due to a lower proportion of spermatozoa with abnormal acrosomes (mainly swollen acrosomes) rather than with absent acrosomes. Conclusions: Canine spermatozoa undergo several modifications in the epididymis. The acquisition of progressive motility, migration of the cytoplasmic droplet and acrosomal reshaping lead to mature spermatozoa which are then stored in the cauda epididymis. From this site, spermatozoa can be retrieved and used in assisted reproductive techniques as a valuable tool for propagating genetic traits of high value individuals that dies accidentally or undergoes orchiectomy for medical purposes. Further investigations should be also focused on the potential use of spermatozoa recovered from other epididymal regions

The pre-ZAMS nature of Mol160/IRAS23385+6053 confirmed by Spitzer

Determining the timeline for the formation of massive YSOs requires the identification and characterisation of all the phases that a massive forming YSO undergoes. It is of particular interest to verify the observability of the phase in which the object is rapidly accreting while not yet igniting the fusion of hydrogen that marks the arrival on the ZAMS. One of the candidate prototypical objects for this phase is Mol160/IRAS23385+6053, which previous studies suggest it could be in a pre-Hot Core stage. We further investigate this issue by means of Spitzer imaging and spectroscopy in the 5-70 micron range. The dense core of Mol160/IRAS23385+6053, which up to now had only been detected at submm and mm wavelenghts has been revealed for the first time at 24 and 70 micron by Spitzer. The complete 24 micron -3.4 mm continuum cannot be fitted with a standard model of a Zero-Age Main-Sequence (ZAMS) star embedded in an envelope. A simple greybody fit yields a mass of 220 solar masses. The luminosity is slightly in excess of 3000 solar luminosities, which is a factor of 5 less than previous estimates when only IRAS fluxes were available between 20 and 100 micron. The source is under-luminous by the same factor with respect to UCHII regions or Hot-Cores of similar circumstellar mass, and simple models show that this is compatible with an earlier evolutionary stage. Spectroscopy between 5 and 40 microns revelas typical PDR/PIR conditions, where the required UV illumination may be provided by other sources revealed at 24 microns in the same region, and which can be plausibly modeled as moderately embedded intermediate-mass ZAMS stars. Our results strengthen the suggestion that the central core in Mol160/IRAS23385+6053 is a massive YSO actively accreting from its circumstellar envelope and which did not yet begin hydrogen fusion.Comment: Accepted by A&

Embracing additive manufacture: implications for foot and ankle orthosis design

<p>Abstract</p> <p>Background</p> <p>The design of foot and ankle orthoses is currently limited by the methods used to fabricate the devices, particularly in terms of geometric freedom and potential to include innovative new features. Additive manufacturing (AM) technologies, where objects are constructed via a series of sub-millimetre layers of a substrate material, may present the opportunity to overcome these limitations and allow novel devices to be produced that are highly personalised for the individual, both in terms of fit and functionality.</p> <p>Two novel devices, a foot orthosis (FO) designed to include adjustable elements to relieve pressure at the metatarsal heads, and an ankle foot orthosis (AFO) designed to have adjustable stiffness levels in the sagittal plane, were developed and fabricated using AM. The devices were then tested on a healthy participant to determine if the intended biomechanical modes of action were achieved.</p> <p>Results</p> <p>The adjustable, pressure relieving FO was found to be able to significantly reduce pressure under the targeted metatarsal heads. The AFO was shown to have distinct effects on ankle kinematics which could be varied by adjusting the stiffness level of the device.</p> <p>Conclusions</p> <p>The results presented here demonstrate the potential design freedom made available by AM, and suggest that it may allow novel personalised orthotic devices to be produced which are beyond the current state of the art.</p

Neonatal mortality in dogs : prognostic value of Doppler ductus venosus waveform evaluation - Preliminary results

Aim: To define the prognostic value of Doppler ultrasonographic morphology of ductus venosus (DV) waveform on canine neonatal mortality. Materials and Methods: Fifty-four healthy pregnant bitches underwent fetal ultrasonographic assessment. The DV waveforms were classified as diphasic (dDVw) or triphasic (tDVw) and compared with neonatal mortality. Results: Ninety-three fetuses were evaluated. Twenty fetuses belonged to litters with neonatal mortality, in which tDVw was observed. Seven fetuses belonged to litters without neonatal mortality, in which tDVw was observed. Fifty-eight fetuses belonged to litters without neonatal mortality, in which only dDVw was observed. Eight fetuses belonged to litters with neonatal mortality, in which only dDVw was observed. The correlation between tDVw and neonatal mortality was statistically significant (odds ratio [OR], 20.7; p<0.0001). Considering only pregnancies with one or two fetuses with the same DV waveform: Two fetuses with tDVw belonged to litters with neonatal mortality; 1 foetus with tDVw belonged to litter without neonatal mortality and 26 fetuses showed dDVw without neonatal mortality. The correlation between tDVw and neonatal mortality even in litters up to two pups was statistically significant (OR, 88.3; p=0.01). Conclusion: Echo-Doppler assessment of DV is feasible in canine fetuses, and the presence tDVw seems to be related to neonatal mortality

Transient transfection of porcine granulosa cells after 3D culture in barium alginate capsules

Three-dimensional culture systems in barium alginate capsules can be employed to maintain primary granulosa cells in an undifferentiated state for almost 6 days. This is due to a self-organization of cells in a pseudofollicular structure. The transfection of primary granulosa cells is a necessary condition when employing these culture systems for several purposes, for example as an in vitro toxicity test or the development of oocytes or zygotes. In this work, the feasibility of two transient transfection techniques (liposome-mediated and electroporation) was assessed in primary porcine granulosa cells after a 6-day culture in an artificial extracellular matrix (barium alginate membrane). Human recombinant green fluorescent protein was chosen as a molecular readout, and protein expression was assessed after 48 hours from transfection. Liposome-mediated transfection gave low transfection levels, with increasing yields from 2 to 12 microgDNA/ml of medium; the maximum percentage (85.7%) was reached at 12 microgDNA/ml of medium. Electroporation-mediated transfection yields were higher: the best results (81.7% of transfected cells) were achieved with two 50V pulses and 12 microg/ml DNA. The application of a single or double pulse (50V) at 4 mgDNA/ml gave negligible results. These results indicate that primary granulosa cell cultured in barium alginate capsules can be transfected by electroporation with high transfection yields

Direct Estimate of Cirrus Noise in Herschel Hi-GAL Images

In Herschel images of the Galactic plane and many star forming regions, a major factor limiting our ability to extract faint compact sources is cirrus confusion noise, operationally defined as the "statistical error to be expected in photometric measurements due to confusion in a background of fluctuating surface brightness". The histogram of the flux densities of extracted sources shows a distinctive faint-end cutoff below which the catalog suffers from incompleteness and the flux densities become unreliable. This empirical cutoff should be closely related to the estimated cirrus noise and we show that this is the case. We compute the cirrus noise directly, both on Herschel images from which the bright sources have been removed and on simulated images of cirrus with statistically similar fluctuations. We connect these direct estimates with those from power spectrum analysis, which has been used extensively to predict the cirrus noise and provides insight into how it depends on various statistical properties and photometric operational parameters. We report multi-wavelength power spectra of diffuse Galactic dust emission from Hi-GAL observations at 70 to 500 microns within Galactic plane fields at l= 30 degrees and l= 59 degrees. We find that the exponent of the power spectrum is about -3. At 250 microns, the amplitude of the power spectrum increases roughly as the square of the median brightness of the map and so the expected cirrus noise scales linearly with the median brightness. Generally, the confusion noise will be a worse problem at longer wavelengths, because of the combination of lower angular resolution and the rising power spectrum of cirrus toward lower spatial frequencies, but the photometric signal to noise will also depend on the relative spectral energy distribution of the source compared to the cirrus.Comment: 4 pages (in journal), 3 figures, Astronomy and Astrophysics, accepted for publication 13 May 201