Defining early steps in mRNA transport: mutant mRNA in myotonic dystrophy type I is blocked at entry into SC-35 domains

In myotonic dystrophy type 1 (DM1), triplet repeat expansion in the 3′ untranslated region of dystrophia myotonica protein kinase (DMPK) causes the nuclear retention of mutant messenger RNA (mRNA). Although the DMPK gene locus positions precisely at the outer edge of a factor-rich SC-35 domain, the normal mRNA consistently accumulates within the domain, and this RNA is depleted upon transcriptional inhibition. In DM1, mutant transcripts detach from the gene but accumulate in granules that abut but do not enter SC-35 domains, suggesting that RNA entry into the domain is blocked. Despite their exclusion from these compartments, mutant transcripts are spliced. MBNL1 (muscleblind-like protein 1) is an alternative splicing factor that becomes highly concentrated with mutant RNA foci. Small interfering RNA–mediated knockdown of MBNL1 promotes the accumulation or entry of newly synthesized mutant transcripts in the SC-35 domain. Collectively, these data suggest that an initial step in the intranuclear path of some mRNAs is passage from the gene into an SC-35 domain and implicate these structures in postsplicing steps before export

Clustering of multiple specific genes and gene-rich R-bands around SC-35 domains: evidence for local euchromatic neighborhoods

Typically, eukaryotic nuclei contain 10–30 prominent domains (referred to here as SC-35 domains) that are concentrated in mRNA metabolic factors. Here, we show that multiple specific genes cluster around a common SC-35 domain, which contains multiple mRNAs. Nonsyntenic genes are capable of associating with a common domain, but domain “choice” appears random, even for two coordinately expressed genes. Active genes widely separated on different chromosome arms associate with the same domain frequently, assorting randomly into the 3–4 subregions of the chromosome periphery that contact a domain. Most importantly, visualization of six individual chromosome bands showed that large genomic segments (∼5 Mb) have striking differences in organization relative to domains. Certain bands showed extensive contact, often aligning with or encircling an SC-35 domain, whereas others did not. All three gene-rich reverse bands showed this more than the gene-poor Giemsa dark bands, and morphometric analyses demonstrated statistically significant differences. Similarly, late-replicating DNA generally avoids SC-35 domains. These findings suggest a functional rationale for gene clustering in chromosomal bands, which relates to nuclear clustering of genes with SC-35 domains. Rather than random reservoirs of splicing factors, or factors accumulated on an individual highly active gene, we propose a model of SC-35 domains as functional centers for a multitude of clustered genes, forming local euchromatic “neighborhoods.

SeaWiFS Postlaunch Technical Report Series

The SeaWiFS Transfer Radiometer (SXR) was built for the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) Project as part of an Interagency Agreement with the National Aeronautics and Space Administration (NASA). The SXR is a multichannel radiometer designed to verify and compare measurements of spectral radiance at six discrete wavelengths in the visible and near infrared for various calibration sources in the SeaWiFS Project. In addition, the SXR is used to compare these sources to standards of spectral radiance maintained at the National Institute of Standards and Technology (NIST). The SXR was designed, built, and thoroughly characterized in the Optical Technology Division at NIST. A unique optical design provides six independent optical paths, each equipped with a temperature stabilized interference filter and silicon photodiode. A separate beam path through the input lens is used to visually align the SXR. The entrance windows for each channel overlap at the source, with each channel sampling a unique solid angle within the field of view of the SXR; this allows for simultaneous sampling of all channels. The combined standard relative uncertainty of spectral radiance measurements with the SXR is estimated to be between 0.6% and 1.3%. This report describes the design and construction of the SXR in detail, and gives the results of the optical characterization and calibrations done at NIST. The SXR has been used for several intercomparisons which include several SeaWiFS Intercalibration Round-Robin Experiments (SIRREXs); those done at the Marine Optical Buoy (MOBY) laboratories in Honolulu, Hawaii; at the NEC Corporation in Yokohama, Japan; and Orbital Sciences Corporation (OSC) in Germantown, Maryland. Thorough optical characterization and calibration of the SXR was essential to the successful application of the radiometer for these measurements

No Significant Differences in Muscle Growth and Strength Development When Consuming Soy and Whey Protein Supplements Matched for Leucine Following a 12 Week Resistance Training Program in Men and Women: A Randomized Trial

There are conflicting reports regarding the efficacy of plant versus animal-derived protein to support muscle and strength development with resistance training. The purpose of this study was to determine whether soy and whey protein supplements matched for leucine would comparably support strength increases and muscle growth following 12 weeks of resistance training. Sixty-one untrained young men (n = 19) and women (n = 42) (18–35 year) enrolled in this study, and 48 completed the trial (17 men, 31 women). All participants engaged in supervised resistance training 3×/week and consumed 19 grams of whey protein isolate or 26 grams of soy protein isolate, both containing 2 g (grams) of leucine. Multi-level modeling indicated that total body mass (0.68 kg; 95% CI: 0.08, 1.29 kg; p \u3c 0.001), lean body mass (1.54 kg; 95% CI: 0.94, 2.15 kg; p \u3c 0.001), and peak torque of leg extensors (40.27 Nm; 95% CI: 28.98, 51.57 Nm, p \u3c 0.001) and flexors (20.44 Nm; 95% CI: 12.10, 28.79 Nm; p \u3c 0.001) increased in both groups. Vastus lateralis muscle thickness tended to increase, but this did not reach statistical significance (0.12 cm; 95% CI: −0.01, 0.26 cm; p = 0.08). No differences between groups were observed (p \u3e 0.05). These data indicate that increases in lean mass and strength in untrained participants are comparable when strength training and supplementing with soy or whey matched for leucine

Total versus superficial parotidectomy for stage III melanoma

BackgroundThe primary purpose of this study was to describe the parotid recurrence rates after superficial and total parotidectomy.MethodsA retrospective cohort study was performed on patients with cutaneous melanoma metastatic to the parotid gland who underwent parotidectomy from 1998 through 2014. Primary outcome was parotid bed recurrence. Secondary outcomes were facial nerve function postoperatively and at last follow‐up.ResultsOne hundred twenty‐nine patients were included in the study. Thirty‐four patients (26%) underwent a total parotidectomy and 95 patients underwent superficial parotidectomy. Twelve patients (13%) developed parotid bed recurrence after superficial parotidectomy alone versus zero after total parotidectomy (P = .035). Facial nerve function, clinically detected disease, stage, and adjuvant treatment were not statistically different between the groups (P = .32, .32, .13, and 0.99, respectively).ConclusionParotid bed melanoma recurrence was more common after superficial parotidectomy compared to total parotidectomy, and recurrence resulted in significant facial nerve functional deficit. Our results support total parotidectomy when metastatic melanoma involves the parotid nodal basin.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/137735/1/hed24810_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/137735/2/hed24810.pd

The Epithelial Sodium Channel (ENaC) Establishes a Trafficking Vesicle Pool Responsible for Its Regulation

The epithelial sodium channel (ENaC) is the rate-limiting step for sodium reabsorption across tight epithelia. Cyclic-AMP (cAMP) stimulation promotes ENaC trafficking to the apical surface to increase channel number and transcellular Na+ transport. Removal of corticosteroid supplementation in a cultured cortical collecting duct cell line reduced ENaC expression. Concurrently, the number of vesicles trafficked in response to cAMP stimulation, as measured by a change in membrane capacitance, also decreased. Stimulation with aldosterone restored both the basal and cAMP-stimulated ENaC activity and increased the number of exocytosed vesicles. Knocking down ENaC directly decreased both the cAMP-stimulated short-circuit current and capacitance response in the presence of aldosterone. However, constitutive apical recycling of the Immunoglobulin A receptor was unaffected by alterations in ENaC expression or trafficking. Fischer Rat Thyroid cells, transfected with α,β,γ-mENaC had a significantly greater membrane capacitance response to cAMP stimulation compared to non-ENaC controls. Finally, immunofluorescent labeling and quantitation revealed a smaller number of vesicles in cells where ENaC expression was reduced. These findings indicate that ENaC is not a passive passenger in regulated epithelial vesicle trafficking, but plays a role in establishing and maintaining the pool of vesicles that respond to cAMP stimulation. © 2012 Edinger et al

SeaWiFS Postlaunch Technical Report Series

The Sea-viewing Wide Field-of-view Sensor (SeaWiFS) was originally calibrated by the instrument's manufacturer, Santa Barbara Research Center (SBRC), in November 1993. In preparation for an August 1997 launch, the SeaWiFS Project and the National Institute of Standards and Technology (NIST) undertook a second calibration of SeaWiFS in January and April 1997 at the facility of the spacecraft integrator, Orbital Sciences Corporation (OSC). This calibration occurred in two phases, the first after the final thermal vacuum test, and the second after the final vibration test of the spacecraft. For the calibration, SeaWiFS observed an integrating sphere from the National Aeronautics and Space Administration (NASA) Goddard Space Flight Center (GSFC) at four radiance levels. The spectral radiance of the sphere at these radiance levels was also measured by the SeaWiFS Transfer Radiometer (SXR). In addition, during the calibration, SeaWiFS and the SXR observed the sphere at 16 radiance levels to determine the linearity of the SeaWiFS response. As part of the calibration analysis, the GSFC sphere was also characterized using a GSFC spectroradiometer. The 1997 calibration agrees with the initial 1993 calibration to within +/- 4%. The new calibration coefficients, computed before and after the vibration test, agree to within 0.5%. The response of the SeaWiFS channels in each band is linear to better than 1%. In order to compare to previous and current methods, the SeaWiFS radiometric responses are presented in two ways: using the nominal center wave-lengths for the eight bands; and using band-averaged spectral radiances. The band-averaged values are used in the flight calibration table. An uncertainty analysis for the calibration coefficients is also presented

SeaWiFs Technical Report Series. Volume 34: The Third SeaWiFS Intercalibration Round-Robin Experiment (SIRREX-3), 19-30 September 1994

This report presents results of the third Sea-viewing Wide Field-of-view Sensor (SeaWiFS) Intercalibration Round- Robin Experiment (SIRREX-3), which was held at the San Diego State University (SDSU) Center for Hydro-Optics and Remote Sensing (CHORS) on 19-30 September 1994. Spectral irradiances of FEL lamps belonging to each participant were intercompared by reference to the National Institute of Standards and Technology (NIST) scale of spectral irradiance using secondary standard lamps F268, F269, and F182, with a Type A uncertainty between 1.1-1.5%. This level of uncertainty was achieved despite difficulties with lamp F269. The average spectral irradiances of FEL lamps, compared in both SIRREX-2 and SIRREX-3, differed between the two experiments by 1.5%, which probably indicates that the values assigned to the secondary standard lamp at the time of SIRREX-2 were in error. With two exceptions, spectral radiance values of integrating sphere sources were measured during SIRREX-3 with uncertainties in temporal stability of less than 0.3% and absolute uncertainties of 1.5-2.0%. This is a significant improvement over similar intercomparisons in SIRREX- I and SIRREX-2. Plaque reflectances were intercompared with an uncertainty of about 1-2%, but the absolute uncertainty is undefined. Although this is an improvement over results of previous SIRREXS, the sources and magnitude of uncertainty associated with transfers of spectral radiance using plaques requires further evaluation in future experiments

SeaWiFS Technical Report Series

This document provides brief reports, or case studies, on a number of investigations sponsored by the Calibration and Validation Team (CVT) within the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) Project. Chapter I describes the calibration and characterization of the GSFC sphere, which was used in the recent recalibration of the SeaWiFS instrument. Chapter 2 presents a revision of the diffuse attenuation coefficient, K(490), algorithm based on the SeaWiFS wavelengths. Chapter 3 provides an implementation scheme for an algorithm to remove out-of-band radiance when using a sensor calibration based on a finite width (truncated) spectral response function, e.g., between the 1% transmission points. Chapter 4 describes the implementation schemes for the stray light quality flag (local area coverage [LAC] and global area coverage [GAC]) and the LAC stray light correction

Sea WiFS Technical Report Series: The fourth SeaWIFS Intercalibration Round-Robin Experiment (SIRREX-4), May 1995

This report documents the fourth Sea-viewing Wide Field-of-view Sensor (SeaWiFS) Intercalibration Round-Robin Experiment (SIRREX-4), which was held at the National Institute of Standards and Technology (NIST) on 3-10 May 1995. The agenda for SIRREX-4 was established by a consensus reached at the conclusion of SIRREX-3: there should be an emphasis on training and work to foster and encourage uniform use of accepted protocols for calibrating radiometric instruments in the laboratory. The goal was to host the activity in a setting where proper techniques could be discussed and demonstrated. It seemed appealing to split the day between morning lectures and afternoon laboratory exercises or practicals. The former gave the user community a chance to present what was important to them and discuss it with acknowledged experts in radiometry, while the latter presented a unique opportunity for training and evaluation in the presence of these same experts. The five laboratory sessions were concerned with (1) determining the responsivity of a spectroradiometer and the spectral radiance of an unknown integrating sphere source, (2) demonstrating spectral field calibration procedures for an integrating sphere using three different instruments, (3) measuring spectral radiance using the plaque method, (4) setting up and aligning lamp calibration transfer standards using the NIST specifications for irradiance measurements, and (5) characterizing radiometric instruments. In addition to documenting some supplemental studies performed outside the laboratory sessions, this report includes an evaluation of the hardware that has been used during the SIRREX activities plus a critical evaluation of SIRREX objectives