Modeling demyelination and endogenous remyelination in spinal cord ex vivo rat organotypic slice cultures

IntroductionDemyelination of the spinal cord is a prominent feature of multiple sclerosis (MS) and spinal cord injuries (SCI), where impaired neuronal communication between the brain and periphery has devastating consequences on neurological function. Demyelination precedes remyelination, an endogenous process in which oligodendrocyte precursor cells (OPCs) differentiate into mature, myelinating oligodendrocytes with the ability to restore the myelin sheath and reinstate functional nerve signaling. However, in MS or SCI, demyelination is more severe, persistent, and inhibitory to OPC-mediated remyelination, leading to a permanent loss of neuronal function. Currently, there are no effective treatments for demyelination, and existing pre-clinical models typically focus on brain tissue with little characterization of demyelination within the spinal cord. Organotypic slice cultures are a useful tool to study neurological disease, providing a more complex 3-dimensional system than standard 2-dimensional in vitro cell cultures.MethodsBuilding on our previously developed rat brain slice culture protocol, we have extended our findings to develop a rat longitudinal spinal cord ex vivo model of demyelination.ResultsWe generated rat longitudinal spinal cord slice cultures that remain viable for up to 6 weeks in culture and retain key anatomical features of the spinal cord’s cytoarchitecture. We show that treating longitudinal spinal cord slices with lysolecithin (LPC) induced robust demyelination with some endogenous remyelination, which was not seen following exposure to lipopolysaccharide (LPS).DiscussionOur ex vivo organotypic spinal cord slice culture system provides a platform to model demyelination and endogenous remyelination long-term, mimicking that observed in LPC-induced rodent models of demyelination. This platform is suitable for the development and testing of novel therapeutic strategies with ease of manipulation prior to in vivo experimentation

Effects of OmniGen® PRO on ruminal fermentation, stress, and inflammation of Holstein cattle induced with a subacute ruminal acidosis

The aim of our study was to examine the effects of an immunomodulatory feed additive (OmniGen® PRO, Phibro Animal Health, Teaneck, NJ) on ruminal fermentation homeostasis and biomarkers of stress and inflammation in dairy heifers following subacute ruminal acidosis (SARA). Holstein heifers (n = 32, 8.4 ± 0.3 months old) were allocated to receive two treatments based on body weight (BW). Treatments were (1) control (CON; n = 16, no additive) and (2) OmniGen® PRO (OGPRO, n = 16, 10 g/100 kg BW, top-dressed). From d 1 to 69, and d 72 to 77, heifers were fed ad libitum a basal TMR formulated for early lactation. On d 70, the TMR offerings were reduced by 50% based on the intake of the previous 3 days. On d 71, heifers were fed ad libitum a starch challenge diet, which was a 50/50 mix steam-rolled barley and the basal TMR on a DM basis, to induce SARA. Rumen fluid and blood samples were collected on d 66, 71, and 73. Prior to the challenge, DMI, ADG, and feed efficiency were not affected by treatment. Following the challenge, all heifers experienced a 43% decline in DMI. Rumen pH was lower on the challenge day than pre and post challenge but was unaffected by treatment. Ruminal lactate was negligible pre and post challenge but increased on the challenge day; OGPRO reduced ruminal lactate compared to CON. At all sampling points, rumen total VFA were higher in OGPRO than in CON. The challenge caused fluctuations in the acetate to propionate ratio in CON, while OGPRO heifers had less variation. Two days post challenge, OGPRO heifers tended to have lower plasma cortisol, haptoglobin, and non-esterified fatty acids (NEFA) than CON heifers. The OGPRO heifers maintained the mean corpuscular hemoglobin concentration (MCHC) and platelet concentration after the challenge, while their levels declined in CON. In this study, supplementing OGPRO to heifers fed an early lactation diet improved rumen fermentation measures prior to the starch challenge and reduced the negative effects of the challenge on rumen fermentation. In addition, following the challenge, indicators of systemic inflammation tended to be lower in heifers supplemented with OGPRO

Engaging Patients with Late-Stage Non-Small Cell Lung Cancer in Shared Decision Making about Treatment

Few treatment decision support interventions (DSIs) are available to engage patients diagnosed with late-stage non-small cell lung cancer (NSCLC) in treatment shared decision making (SDM). We designed a novel DSI that includes care plan cards and a companion patient preference clarification tool to assist in shared decision making. The cards answer common patient questions about treatment options (chemotherapy, chemotherapy plus immunotherapy, targeted therapy, immunotherapy, clinical trial participation, and supportive care). The form elicits patient treatment preference. We then conducted interviews with clinicians and patients to obtain feedback on the DSI. We also trained oncology nurse educators to implement the prototype. Finally, we pilot tested the DSI among five patients with NSCLC in treatment SDM at the beginning of an office visit scheduled to discuss Treatment with an oncologist. Analyses of pilot study baseline and exit survey data showed that DSI use was associated with increased patient awareness of the alternatives’ treatment options and benefits/risks. In contrast, patient concern about treatment costs and uncertainty in treatment decision making decreased. All patients expressed a treatment preference. Future randomized controlled trials are needed to assess DSI implementation feasibility and efficacy in clinical care

The Spitzer Gould Belt Survey of Large Nearby Interstellar Clouds: Discovery of A Dense Embedded Cluster in the Serpens-Aquila Rift

We report the discovery of a nearby, embedded cluster of young stellar objects, associated filamentary infrared dark cloud, and 4.5 mu m shock emission knots from outflows detected in Spitzer IRAC mid-infrared imaging of the Serpens-Aquila Rift obtained as part of the Spitzer Gould Belt Legacy Survey. We also present radial velocity measurements of the region from molecular line observations obtained with the Submillimeter Array (SMA) that suggest the cluster is comoving with the Serpens Main embedded cluster to the north. We therefore assign it 3 degrees the same distance, 260 pc. The core of the new cluster, which we call Serpens South, is composed of an unusually large fraction of protostars (77%) at high mean surface density (> 430 pc(-2)) and short median nearest neighbor spacing (3700 AU). We perform basic cluster structure characterization using nearest neighbor surface density mapping of the YSOs and compare our findings to other known clusters with equivalent analyses available in the literature.Astronom

The Spitzer c2d Survey of Large, Nearby, Interstellar Clouds. VIII. Serpens Observed with MIPS

We present maps of 1.5 deg^2 of the Serpens dark cloud at 24, 70, and 160 μm observed with the Spitzer Space Telescope MIPS camera. We describe the observations and briefly discuss the data processing carried out by the c2d team on these data. More than 2400 compact sources have been extracted at 24 μm, nearly 100 at 70 μm, and four at 160 μm. We estimate completeness limits for our 24 μm survey from Monte Carlo tests with artificial sources inserted into the Spitzer maps. We compare source counts, colors, and magnitudes in the Serpens cloud to two reference data sets: a 0.50 deg^2 set on a low-extinction region near the dark cloud, and a 5.3 deg^2 subset of the SWIRE ELAIS N1 data that was processed through our pipeline. These results show that there is an easily identifiable population of young stellar object candidates in the Serpens cloud that is not present in either of the reference data sets. We also show a comparison of visual extinction and cool dust emission illustrating a close correlation between the two and find that the most embedded YSO candidates are located in the areas of highest visual extinction

The Spitzer c2d Survey of Large, Nearby, Interstellar Clouds. I. Chamaeleon II Observed with MIPS

We present maps of over 1.5 square degrees in Chamaeleon (Cha) II at 24, 70, and 160 micron observed with the Spitzer Space Telescope Multiband Imaging Photometer for Spitzer (MIPS) and a 1.2 square degree millimeter map from SIMBA on the Swedish-ESO Submillimetre Telescope (SEST). The c2d Spitzer Legacy Team's data reduction pipeline is described in detail. Over 1500 24 micron sources and 41 70 micron sources were detected by MIPS with fluxes greater than 10-sigma. More than 40 potential YSOs are identified with a MIPS and 2MASS color-color diagram and by their spectral indices, including two previously unknown sources with 24 micron excesses. Our new SIMBA millimeter map of Cha II shows that only a small fraction of the gas is in compact structures with high column densities. The extended emission seen by MIPS is compared with previous CO observations. Some selected interesting sources, including two detected at 1 mm, associated with Cha II are discussed in detail and their SEDs presented. The classification of these sources using MIPS data is found to be consistent with previous studies.Comment: 44 pages, 12 figures (1 color), to be published in Ap

The Spitzer c2d Survey of Large, Nearby, Interstellar Clouds. VII. Ophiuchus Observed with MIPS

We present maps of 14.4 deg^2 of the Ophiuchus dark clouds observed by the Spitzer Space Telescope Multiband Imaging Photometer for Spitzer (MIPS). These high-quality maps depict both numerous point sources and extended dust emission within the star-forming and non–star-forming portions of these clouds. Using PSF-fitting photometry, we detect 5779 sources at 24 μm and 81 sources at 70 μm at the 10 σ level of significance. Three hundred twenty-three candidate young stellar objects (YSOs) were identified according to their positions on the MIPS/2MASS K versus color-magnitude diagrams, as compared to 24 μm detections in the SWIRE extragalactic survey. We find that more than half of the YSO candidates, and almost all those with protostellar Class I spectral energy distributions, are confined to the known cluster and aggregates

The Spitzer c2d Survey of Weak-line T Tauri Stars II: New Constraints on the Timescale for Planet Building

One of the central goals of the Spitzer Legacy Project ``From Molecular Cores to Planet-forming Disks'' (c2d) is to determine the frequency of remnant circumstellar disks around weak-line T Tauri stars (wTTs) and to study the properties and evolutionary status of these disks. Here we present a census of disks for a sample of over 230 spectroscopically identified wTTs located in the c2d IRAC (3.6, 4.5, 4.8, and 8.0 um) and MIPS (24 um) maps of the Ophiuchus, Lupus, and Perseus Molecular Clouds. We find that ~20% of the wTTs in a magnitude limited subsample have noticeable IR-excesses at IRAC wavelengths indicating the presence of a circumstellar disk. The disk frequencies we find in these 3 regions are ~3-6 times larger than that recently found for a sample of 83 relatively isolated wTTs located, for the most part, outside the highest extinction regions covered by the c2d IRAC and MIPS maps. The disk fractions we find are more consistent with those obtained in recent Spitzer studies of wTTs in young clusters such as IC 348 and Tr 37. From their location in the H-R diagram, we find that, in our sample, the wTTs with excesses are among the younger part of the age distribution. Still, up to ~50% of the apparently youngest stars in the sample show no evidence of IR excess, suggesting that the circumstellar disks of a sizable fraction of pre-main-sequence stars dissipate in a timescale of ~1 Myr. We also find that none of the stars in our sample apparently older than ~10 Myrs have detectable circumstellar disks at wavelengths < 24 um. Also, we find that the wTTs disks in our sample exhibit a wide range of properties (SED morphology, inner radius, L_DISK/L*, etc) which bridge the gaps observed between the cTTs and the debris disk regimes.Comment: 54 pages, 13 figures, Accepted by Ap

The Spitzer c2d Survey of Weak-Line T Tauri Stars. III. The Transition from Primordial Disks to Debris Disks

We present 3.6 to 70 {\mu}m Spitzer photometry of 154 weak-line T Tauri stars (WTTS) in the Chamaeleon, Lupus, Ophiuchus and Taurus star formation regions, all of which are within 200 pc of the Sun. For a comparative study, we also include 33 classical T Tauri stars (CTTS) which are located in the same star forming regions. Spitzer sensitivities allow us to robustly detect the photosphere in the IRAC bands (3.6 to 8 {\mu}m) and the 24 {\mu}m MIPS band. In the 70 {\mu}m MIPS band, we are able to detect dust emission brighter than roughly 40 times the photosphere. These observations represent the most sensitive WTTS survey in the mid to far infrared to date, and reveal the frequency of outer disks (r = 3-50 AU) around WTTS. The 70 {\mu}m photometry for half the c2d WTTS sample (the on-cloud objects), which were not included in the earlier papers in this series, Padgett et al. (2006) and Cieza et al. (2007), are presented here for the first time. We find a disk frequency of 19% for on-cloud WTTS, but just 5% for off- cloud WTTS, similar to the value reported in the earlier works. WTTS exhibit spectral energy distributions (SEDs) that are quite diverse, spanning the range from optically thick to optically thin disks. Most disks become more tenuous than Ldisk/L* = 2 x 10^-3 in 2 Myr, and more tenuous than Ldisk/L* = 5 x 10^-4 in 4 Myr.Comment: 40 pages, 13 figures, 4 tables. Accepted for publication in ApJ on September 20, 201