INITIAL EVALUATION OF CYCLIC ADENOSINE MONOPHOSPHATE ENZYME IMMUNOASSAY FOR USE WITH CRANE SEMEN SAMPLES

The management of ex situ and in situ populations of many wildlife species requires detailed knowledge of a species’ reproductive biology. For species such as cranes, where artificial insemination is a critical component of ex situ management strategies, understanding normal sperm function is especially important. Previous research has shown that captive cranes exhibit highly variable production and quality of semen samples produced by individual males and high levels of variation of cell concentration and motility across different species. Cyclic adenosine monophosphate (cyclic AMP) has been implicated in regulating sperm function, such as cell motility, and may affect an individual’s ability to successfully fertilize. Here we demonstrate the feasibility of an enzyme-linked immunosorbent assay (ELISA) for measuring cyclic AMP produced by crane sperm to facilitate future research into its role in sperm function and fertilization

A Search for Distant Solar System Bodies in the Region of Sedna

We present the results of a wide-field survey for distant Sedna-like bodies in the outer solar system using the 1.2-m Samuel Oschin Telescope at Palomar Observatory. We searched ~12,000 square degrees down to a mean limiting magnitude of 21.3 in R. A total number of 53 Kuiper belt objects and Centaurs have been detected; 25 of which were discovered in this survey. No additional Sedna-like bodies with perihelia beyond 70 AU were found despite a sensitivity to motions out to ~1000 AU. We place constraints on the size and distribution of objects on Sedna orbits.Comment: 10 pages, 3 figures, accepted by ApJ Letter

How to… support others in developing a career in clinical education research

The Incubator for Clinical Education Research (ClinEdR) is a UK-wide network, established with support from the National Institute for Health Research, to lead initiatives to build capacity in the field. A key barrier identified by this group is that many experienced educators, clinicians, and researchers, who may be committed to supporting others, have little guidance on how best to do this. In this “How to …” paper, we draw on relevant literature and our individual and collective experiences as members of the National Institute for Health Research ClinEdR incubator to offer suggestions on how educators can support others to develop successful careers involving ClinEdR. This article offers guidance and inspiration for all professionals whose role involves research and scholarship and who encounter aspiring or developing clinical education researchers in the course of their work. It will also be of interest to researchers who are starting out and progressing in the field

Repeated high flows drive morphological change in rivers in recently deglaciated catchments

Climate change is decreasing glacier cover and increasing the frequency and magnitude of precipitation-driven high flows and floods in many regions of the world. Precipitation may become the dominant water source for river systems in recently deglaciated catchments, with major rainfall events driving significant changes in river channel morphology. Few studies, however, have examined river channel response to repeated precipitation-driven high flows. In this study, we measured the geomorphological condition of four low-order rivers in recently deglaciated catchments (70–210 years ice free) before and after a series of repeated precipitation-driven high flows during summer 2014. High flows drove substantial initial morphological change, with up to 75% change in baseflow channel planform position and active channel form change from pre- to post-high flow. Post-high flow years were associated with increased instream wood and geomorphological complexity at all but the youngest river. Channel changes were part of an active relaxation stage at all rivers, where channels continued to migrate, and complexity varied through time. Overall, these measurements permit us to propose a conceptual model of the role of geomorphologically effective high flows in the context of paraglacial adjustment theory. Specifically, we suggest that older rivers in recently deglaciated catchments can undergo a short-term (&lt;10 years) increase in the rate of geomorphological development as a result of the recruitment of instream wood and channel migration during and following repeated precipitation-driven high flows. Enhancing our knowledge of these geomorphological and paraglacial processes in response to high flows is important for the effective management of riverine water and ecosystem resources in rapidly changing environments.</p

Mitochondrial Dysfunction is Evident in Lewis Lung Carcinoma-Induced Muscle Wasting

Cancer cachexia is a paraneoplastic syndrome associated with adverse prognosis and shortened survival. The defining feature of cachexia is extensive muscle atrophy leading to progressive functional impairments. The molecular mechanisms responsible for the rapid muscle wasting are not fully elucidated. Based on emerging evidence, we developed the hypothesis cachectic muscle wasting is caused by mitochondrial dysfunction increasing reactive oxygen species production leading to global oxidative stress. To test this hypothesis we utilized the well-established Lewis-Lung Carcinoma (LLC) model of cancer cachexia. The time-course study consisted of one, two, three and four week LLC tumor bearing mice and age-matched four week saline (PBS) control (Ctrl) mice. Tumors were implanted into the hind flank at 1X106 cells in 100 µL PBS. The plantaris was weighed for wet mass then teased into small fiber bundles and permeabilized for the quantification of mitochondrial function. Mitochondrial dysfunction was classified by a decrease in the respiratory control ratio (RCR), which is the ratio of state 3 (maximal ADP stimulated respiration) to state 4 (oligomycin-induced leak respiration). Muscle mass progressively declined over the time-course, reaching significance at 4 weeks (Ctrl vs 4-week, p\u3c0.05). Mitochondrial function was not different among groups, however individual a priori comparison between groups revealed that 4wk cancer animals exhibited marked mitochondrial dysfunction compared to all other groups (p\u3c0.05). These data demonstrate that late stage cancer-induced muscle wasting is associated with significant mitochondrial dysfunction

Properties of the Distant Kuiper Belt: Results from the Palomar Distant Solar System Survey

We present the results of a wide-field survey using the 1.2-m Samuel Oschin Telescope at Palomar Observatory. This survey was designed to find the most distant members of the Kuiper belt and beyond. We searched ~12,000 deg2 down to a mean limiting magnitude of 21.3 in R. A total number of 52 KBOs and Centaurs have been detected, 25 of which were discovered in this survey. Except for the re-detection of Sedna, no additional Sedna-like bodies with perihelia greater than 45 AU were detected despite sensitivity out to distances of 1000 AU. We discuss the implications for a distant Sedna- like population beyond the Kuiper belt, focusing on the constraints we can place on the embedded stellar cluster environment the early Sun may be have been born in, where the location and distribution of Sedna-like orbits sculpted by multiple stellar encounters is indicative of the birth cluster size. We also report our observed latitude distribution and implications for the size of the plutino population.Comment: 40 pages, 12 figures, 3 tables Accepted by Ap

Mitochondrial Dysfunction in Diaphragm Muscle Precedes the Cachectic Phenotype in LLC Tumor-Bearing Mice.

The defining feature of cancer cachexia is extensive weight loss and skeletal muscle atrophy. It is clinically important because cachexia reduces patient survival, results in functional impairment, and is estimated to be directly responsible for 20-40% of cancer deaths. Unfortunately, no clinical therapy exists and therefore, it is important to understand the molecular mechanisms responsible for rapid muscle wasting. Compared to limb muscles, the diaphragm is relatively understudied in cancer cachexia, but is likely to be adversely affected because cachexia is a systemic disease. Wasting of the primary inspiratory muscle may result in difficulty breathing and inability to adjust minute ventilation in response to a respiratory challenge. Based on emerging evidence, it is clear that oxidative stress is present in cachexia-induced wasting of the diaphragm; PURPOSE: we developed the hypothesis that mitochondrial dysfunction in the diaphragm precedes cachexia. METHODS: 1X106 Lewis Lung Carcinoma cells (LLC) or Phosphate-Buffered Saline (PBS, control) were implanted to the hind-flank of C57BL6/J mice at 8 wks of age. Tumors were allowed to develop for 1, 2, 3, or 4 wks. At designated time points diaphragms were collected and mitochondrial function was assessed by respiration and ROS production. RESULTS: Cancer cachexia was evident only at the 4 wk time point demonstrated by decrease in body mass and muscle atrophy in several limb muscles. Mitochondrial respiration, assessed by respiratory control ratio (state3/state 4 respiration), was significantly lower at 1 wk (pCONCLUSIONS:The molecular events that lead to muscle atrophy in cancer cachexia are unknown. We demonstrate that two hallmarks of mitochondrial dysfunction, altered respiration and ROS production, occur in the diaphragm well before the cancer cachexia phenotype is evident in the LLC model. These data suggest that the mitochondria are likely a suitable target to treat or prevent cancer cachexia-induced muscle wasting in the diaphragm

Disuse Atrophy Occurs Without a Change in Mitochondrial Respiratory Control Ratio During Hindlimb Unloading in Mice

Skeletal muscle atrophy commonly occurs during prolonged periods of inactivity, however, the precise mechanisms that cause muscle atrophy have yet to be determined. Specifically, a controversy exists on whether mitochondrial dysfunction is a cause or consequence of disuse muscle atrophy. PURPOSE: The purpose of this study was to determine if a change in the respiratory control ratio, which is a ratio of maximal O2 respiration to leak respiration, could be detected prior to muscle atrophy in a time-course study in mice. METHODS: Disuse atrophy was induced using hindlimb unloading (HU) in adult, C57BL/6J male mice for 0 (control), 1, 2, 3, or 7 days (n=~6-8/group). Following completion, gastrocnemius and soleus muscles were weighed and assessed for mitochondrial function in permeabilized muscle fibers. Here, we define mitochondrial function as the respiratory control ratio (RCR) determined by maximal ADP stimulated respiration (State 3) divided by leak or ATP synthase inhibited (state 4) respiration. A one-way ANOVA was used to determine differences between means. When significant F ratios were found, a Tukey post-hoc was used to compare differences between means. Values presented are mean ± standard error RESULTS: In both the gastrocnemius and soleus, muscle mass was not significantly different from control at day 1, but was significantly lower at 2, 3, and 7-day timepoints. In contrast, there was no significant difference in RCR in gastrocnemius (control 3.11±0.20, 1 day 2.96±0.60, 2 day 3.07±0.31, 3 day 3.08±0.25, 7 day 3.41± .29) or soleus (control 2.33±0.33, 1 day 2.77±0.33, 2 day 3.03±0.51, 3 day 2.93±0.30, 7 day 2.78±0.48). CONCLUSION: It is well established that HU causes rapid muscle atrophy. These data support mitochondrial RCR does decrease before muscle atrophy in either gastrocnemius or soleus muscle, and therefore may not be a primary cause of HU-induced muscle atrophy in mice

Fractional Synthetic Rate and Markers of Protein Turnover are Altered in the Diaphragms of Cachectic Mice

Cancer cachexia, a wasting syndrome characterized by rapid skeletal muscle wasting and fat loss, directly accounts for up to 20-40% of cancer-related deaths. All muscles, including respiratory muscles, are susceptible to atrophy because cancer cachexia is a systemic disease. Atrophy of the primary breathing muscle, the diaphragm, can lead to respiratory distress, which is commonly associated with a cachectic phenotype. Indeed, the diaphragm is more susceptible to atrophy in certain conditions, but little is known about the effects of cancer-cachexia on protein turnover in the diaphragm. Therefore, investigations into the alterations in protein turnover could provide insight to the molecular events and provide valuable information in the search for therapeutic targets. PURPOSE: The purpose of this study was to describe changes in diaphragmatic protein synthesis and molecular markers of synthesis and degradation during the progression of cancer cachexia. METHODS: C57BL6/J mice (8 wks old) were implanted with 1X106 Lewis Lung Carcinoma cells (LLC) or Phosphate-Buffered Saline (PBS, control). Tumors developed over a 1-4 wk time course and diaphragms were harvested at each time point (1, 2, 3, or 4 wks). Fractional synthetic rates (FSR) were determined using deuterium incorporation into muscle. Selected markers of protein synthesis and degradation pathways were analyzed by immunoblot analysis. One-Way ANOVA was used for statistical analyses, with significance set at pRESULTS: FSR trended downward over time, but did not reach significance. Similar to FSR, anabolic signaling markers (4EBP-1, ERK1/2, Deptor) did not demonstrate significant differences. p62, an autophagic degradation marker, was significantly less than PBS in 3 wk diaphragms (