The Structural Basis for Phospholamban Inhibition of the Calcium Pump in Sarcoplasmic Reticulum

P-type ATPases are a large family of enzymes that actively transport ions across biological membranes by interconverting between high (E1) and low (E2) ion-affinity states; these transmembrane transporters carry out critical processes in nearly all forms of life. In striated muscle, the archetype P-type ATPase, SERCA (sarco(endo)plasmic reticulum Ca2+-ATPase), pumps contractile-dependent Ca2+ ions into the lumen of sarcoplasmic reticulum, which initiates myocyte relaxation and refills the sarcoplasmic reticulum in preparation for the next contraction. In cardiac muscle, SERCA is regulated by phospholamban (PLB), a small inhibitory phosphoprotein that decreases the Ca2+ affinity of SERCA and attenuates contractile strength. cAMP-dependent phosphorylation of PLB reverses Ca2+-ATPase inhibition with powerful contractile effects. Here we present the long sought crystal structure of the PLB-SERCA complex at 2.8-Å resolution. The structure was solved in the absence of Ca2+ in a novel detergent system employing alkyl mannosides. The structure shows PLB bound to a previously undescribed conformation of SERCA in which the Ca2+ binding sites are collapsed and devoid of divalent cations (E2-PLB). This new structure represents one of the key unsolved conformational states of SERCA and provides a structural explanation for how dephosphorylated PLB decreases Ca2+ affinity and depresses cardiac contractility

The Effect of GnRH at Time of Insemination on Initiation of LH Pulses and Subsequent Progesterone

Research has indicated that luteinizing hormone (LH) pulses play a vital role in corpus luteum (CL) formation and subsequent progesterone concentrations. Therefore, our objectives were to determine: 1) when LH pulses begin following onset of estrus, 2) the effect an injection of gonadotropin releasing hormone (GnRH) would have on initiation of LH pulses, and 3) the effect LH pulse initiation had on subsequent plasma progesterone concentrations. Cows were synchronized with the Select Synch + Controlled Internal Drug Releasing device (CIDR) protocol (d -7 100 μg GnRH and CIDR; d 0 25 mg prostaglandin (PG) and removal of CIDR; estrus detected with HeatWatch). Following detection in estrus, a jugular catheter was inserted in each cow (n = 10). Based on initiation of estrus, cows were allotted into two treatments: 1) GnRH given 12 h (12.5 ± 1.2 h) after the initiation of estrus (n = 5; 100 μg) and 2) Control (n = 5). Blood samples were collected at 15-min intervals for 6 h at 12 h (bleed 1), 26 h (bleed 2), 40 h (bleed 3), 54 h (bleed 4), and 68 h (bleed 5) after the onset of estrus. The interval from onset of estrus to bleed 1 and ovulation was similar between treatments. The GnRH cows tended to have a greater area under the LH curve for bleed 1 compared to control cows. No differences were detected in bleeds 2, 3, 4, or 5. Average concentration of LH for GnRH cows in bleed 1 tended to be greater than control. No differences were detected in bleeds 2, 3, 4, or 5. No differences were detected in pulse frequency between treatments in bleeds 1, 3, 4, or 5, but in bleed 2, control tended to have more pulses than GnRH (2.5 ± 0.5 vs 1.4 ± 0.4). The GnRH-treated cows tended to have greater subsequent progesterone concentrations; however, GnRH-treated cows that had no LH pulses during bleed 2 had lower progesterone concentrations than cows with pulses (control or GnRH). In summary, injecting cows with GnRH approximately 12 h after the onset of estrus tended to reduce LH pulses 26-32 h following initiation of estrus, and elimination of LH pulses between 26-32 h resulted in decreased concentrations of progesterone during the subsequent cycle

Doctoral student reflections of blended learning before and during covid-19

Purpose: Our study aimed to address the central research question: how were our experiences as graduate students in a blended learning professional doctoral program changed by the COVID-19 crisis? The study adds to a growing body of literature on blended learning graduate programs. Methods: We employed action research as our central methodology and leveraged narrative inquiry to elevate our (students’) voices. The two participant-researchers responded to a series of questions supported by narrative reflections from their common academic supervisor. Emergent themes were identified in the data using narrative analysis techniques for coding qualitative data into themes. This was followed by a second phase of data collection and analysis after the emergence of the COVID-19 pandemic. Results: The researchers identified four themes within the data: 1. balancing doctoral work with professional and personal responsibilities; 2. cohort provides formal and informal support; 3. individuality of the experience; and 4. supervisory group support. Implications: Our study offers a number of key learnings that may benefit researchers studying blended learning programs. The key learnings suggest benefits to cohort-based, blended learning programs, as well as difficulties that may emerge in the individuality of the experience, when encountering crises, as well as more generally. (DIPF/Orig.

Doctoral student reflections of blended learning before and during covid-19

Purpose: Οur study aimed to address the central research question: how were our experiences as graduate students in a blended learning professional doctoral program changed by the COVID-19 crisis? The study adds to a growing body of literature on blended learning graduate programs. Methods: We employed action research as our central methodology and leveraged narrative inquiry to elevate our (students’) voices. The two participant-researchers responded to a series of questions supported by narrative reflections from their common academic supervisor. Emergent themes were identified in the data using narrative analysis techniques for coding qualitative data into themes. This was followed by a second phase of data collection and analysis after the emergence of the COVID-19 pandemic. Results: Τhe researchers identified four themes within the data: 1. balancing doctoral work with professional and personal responsibilities; 2. cohort provides formal and informal support; 3. individuality of the experience; and 4. supervisory group support. Implications: Οur study offers a number of key learnings that may benefit researchers studying blended learning programs. The key learnings suggest benefits to cohort-based, blended learning programs, as well as difficulties that may emerge in the individuality of the experience, when encountering crises, as well as more generally

The Effects of Suspended Sediment on Japanese Medaka (Oryzias latipes) and Mosquitofish (Gambusia affinis) Metabolism

Sedimentation is recognized as a significant environmental stressor in aquatic ecosystems and high amounts of suspended sediments (SS) in streams are known to negatively affect aquatic organisms. In particular, it has been hypothesized that many fish species mayexhibit increased respiration rates when exposed to elevated SS. To evaluate this hypothesis, we evaluated the acute response (3 hour exposure) of two small, freshwater fish species exposed to high suspended sediment loads using experimental respirometry chambers which measure oxygen consumption as a proxy of metabolism. Our results indicate that Japanese Medaka (Oryzias latipes) did not exhibit a significantly greater oxygen consumption, as compared to control fish, when exposed to chamber sediment of 0.17 g/L (p-value of 0.41), though oxygen consumption was higher for sediment treatments. However, mosquitofish (Gambusia affinis) exposed to 0.17 g/L did exhibit noticeably greater oxygen consumption compared to fish in no-sediment control test, but not at statistically significant levels (p-value of 0.07). Further study is needed with increased sample sizes and across SS levels to determine the threshold of increased metabolic rate among freshwater species exposed to SS

Biogeochemical processes at hydrothermal vents : microbes and minerals, bioenergetics, and carbon fluxes

Author Posting. © The Oceanography Society, 2012. This article is posted here by permission of The Oceanography Society for personal use, not for redistribution. The definitive version was published in Oceanography 25, no. 1 (2012): 196–208, doi:10.5670/oceanog.2012.18.Hydrothermal vents are among the most biologically active regions of the deep ocean. However, our understanding of the limits of life in this extreme environment, the extent of biogeochemical transformation that occurs in the crust and overlying ocean, and the impact of vent life on regional and global ocean chemistry is in its infancy. Recently, scientific studies have expanded our view of how vent microbes gain metabolic energy at vents through their use of dissolved chemicals and minerals contained in ocean basalts, seafloor sulfide deposits, and hydrothermal plumes and, in turn, how they catalyze chemical and mineral transformations. The scale of vent environments and the difficulties inherent in the study of life above, on, and below the deep seafloor have led to the development of geochemical and bioenergetic models. These models predict habitability and biological activity based on the chemical composition of hydrothermal fluids, seawater, and the surrounding rock, balanced by the physiological energy demand of cells. This modeling, coupled with field sampling for ground truth and discovery, has led to a better understanding of how hydrothermal vents affect the ocean and global geochemical cycles, and how they influence our views of life on the early Earth and the search for life beyond our own planet.Research for this paper was supported by the National Science Foundation (NSF) Division of Ocean Sciences grants 0732611 for JFH, 0926805 and 1038055 for JAB, and 1038055 for BMT; and by the University of Missouri Research Board for KLR

A Comparative Study of Cellular Diversity between the Xenopus Pronephric and Mouse Metanephric Nephron

The kidney is an essential organ that ensures bodily fluid homeostasis and removes soluble waste products from the organism. Nephrons, the functional units of the kidney, comprise a blood filter, the glomerulus or glomus, and an epithelial tubule that processes the filtrate from the blood or coelom and selectively reabsorbs solutes, such as sugars, proteins, ions, and water, leaving waste products to be eliminated in the urine. Genes coding for transporters are segmentally expressed, enabling the nephron to sequentially process the filtrate. The Xenopus embryonic kidney, the pronephros, which consists of a single large nephron, has served as a valuable model to identify genes involved in nephron formation and patterning. Therefore, the developmental patterning program that generates these segments is of great interest. Prior work has defined the gene expression profiles of Xenopus nephron segments via in situ hybridization strategies, but a comprehensive understanding of the cellular makeup of the pronephric kidney remains incomplete. Here, we carried out single-cell mRNA sequencing of the functional Xenopus pronephric nephron and evaluated its cellular composition through comparative analyses with previous Xenopus studies and single-cell mRNA sequencing of the adult mouse kidney. This study reconstructs the cellular makeup of the pronephric kidney and identifies conserved cells, segments, and associated gene expression profiles. Thus, our data highlight significant conservation in podocytes, proximal and distal tubule cells, and divergence in cellular composition underlying the capacity of each nephron to remove wastes in the form of urine, while emphasizing the Xenopus pronephros as a model for physiology and disease

Effect of Feeding Different Types of Byproducts and Concentrations Throughout a Beef Growing System on Ground Beef Color and Lipid Oxidation

The objective of this trial was to evaluate the effect of feeding different concentrations of wet distillers grains during winter backgrounding and either modified wet distillers grains or Sweet Bran® during the finishing phase on ground beef color and lipid oxidation. After a 14 day aging period, ground beef patties were made and placed in a simulated retail display for seven days. There were no overall differences in lipid oxidation between treatments but was a treatment by day interaction for discoloration. Ground beef from heifers finished with modified wet distillers grains discolored at a greater extent when compared to ground beef from heifers finished with Sweet Bran

NEBRASKA AGRICULTURAL WATER MANAGEMENT DEMONSTRATION NETWORK (NAWMDN): INTEGRATING RESEARCH AND EXTENSION/OUTREACH

Maximizing the net benefits of irrigated plant production through appropriately designed agricultural water management programs is of growing importance in Nebraska, and other western and Midwestern states, because many areas are involved in management and policy changes to conserve irrigation water. In Nebraska, farmers are being challenged to practice conservation methods and use water resources more efficiently while meeting plant water requirements and maintaining high yields. Another challenge Nebraska experiences in it\u27s approximately 3.5‐million‐ha irrigated lands is limited adoption of newer technologies/tools to help farmers better manage irrigation, conserve water and energy, and increase plant water use efficiency. In 2005, the Nebraska Agricultural Water Management Demonstration Network (NAWMDN or Network) was formed from an interdisciplinary team of partners including the Natural Resources Districts (NRD); USDA‐NRCS; farmers from south central, northeast, west central, and western Nebraska; crop consultants; and University of Nebraska‐Lincoln faculty. The main goal of the Network is to enable the transfer of high quality research‐based information to Nebraskans through a series of demonstration projects established in farmers\u27 fields and implement newer tools and technologies to address and enhance plant water use efficiency, water conservation, and reduce energy consumption for irrigation. The demonstration projects are supported by the scientifically‐based field research and evaluation projects conducted at the University of Nebraska‐Lincoln, South Central Agricultural Laboratory located near Clay Center, Nebraska. The Network was formed with only 15 farmers as collaborators in only one of the 23 NRDs in 2005. As of late 2009, the number of active collaborators has increased to over 300 in 12 NRDs and 35 of 93 counties. The Network is impacting both water and energy conservation due to farmers adopting information and newer technologies for irrigation management. The NAWMDN is helping participants to improve irrigation management and efficiency by monitoring plant growth stages and development, soil moisture, and crop evapotranspiration. As a result, they are reducing irrigation water application amounts and associated energy savings is leading to greater profitability to participating farmers. This article describes the goals and objectives of the Network, technical and educational components, operational functions, and procedures used in the NAWMDN. The quantitative impacts in terms of water and energy conservation are reported