Operation of rain gauge and ground-water monitoring networks for the Imperial Valley Water Authority, year six: September 1997 - August 1998

The Illinois State Water Survey (ISWS), under contract to the Imperial Valley Water Authority (IVWA), has operated a network of rain gauges in Mason and Tazewell Counties since August 1992. The ISWS also established a network of ground-water observation wells in the Mason-Tazewell area. The networks are located in the most heavily irrigated region of the state. Ground water in various aquifers are the major source of the irrigation and domestic water supplies in the region. Recent extreme weather events (e.g., the drought of 1988 and the great flood of 1993) resulted in large fluctuations of the ground-water levels in the Imperial Valley area. The purpose of the rain gauge network and the ground-water observation well network is to collect a long-term series of data to determine the rate of ground-water drawdown in dry periods and during the growing season, and the rate at which the aquifers recharge. Precipitation is recorded for each storm that traverses the Imperial Valley, and ground-water levels at the 13 observation wells are measured the first of each month. The database from these networks consists of 6 years of precipitation data and 4 years of ground-water observations. At the beginning of the ground-water observations in 1995, the water levels were at the highest in the four years of observation. These high ground-water levels were the result of the very wet 1992-1995 period when annual precipitation was above the 30-year normals at both Havana and Mason City. From September 1995-August 1997 precipitation in the region was below the 30-year normal. The 1997-1998 observation year had rainfall above the 30-year normal. Ground-water levels in the observation wells mirrored these rainfall patterns, showing a general downward trend during the dry years and a recovery in the wet 1997-1998 year. Seasonal increases in the ground-water levels were observed at most wells during the late spring and early summer, followed by decreases in August-November ground water levels. Analysis indicates that the ground-water levels are affected by both the precipitation in the Imperial Valley area and the Illinois River stages. The observation wells closest to the Illinois River show an increase in water levels whenever the river stage is high. Generally, the water levels in the wells correlate best with precipitation and Illinois River stages one to two months before the water levels are measured, i.e., the June ground-water levels are most highly correlated with the Illinois River stage or precipitation that occurs in either April or May. The analyses conducted indicate the need for continued operation of both networks due to inconsistencies associated with ground-water levels, precipitation, and the Illinois River stage. For instance, the Mason-Tazwell observation well number 2 (MTOW-2) is located near the center of Mason County well away from the Illinois River, but it has an equal correlation with the Illinois River stage and the precipitation in the area. Additional analysis needs to be undertaken to explain this unusual finding

Visualizing Interactions along the Escherichia coli Twin-Arginine Translocation Pathway Using Protein Fragment Complementation

The twin-arginine translocation (Tat) pathway is well known for its ability to export fully folded substrate proteins out of the cytoplasm of Gram-negative and Gram-positive bacteria. Studies of this mechanism in Escherichia coli have identified numerous transient protein-protein interactions that guide export-competent proteins through the Tat pathway. To visualize these interactions, we have adapted bimolecular fluorescence complementation (BiFC) to detect protein-protein interactions along the Tat pathway of living cells. Fragments of the yellow fluorescent protein (YFP) were fused to soluble and transmembrane factors that participate in the translocation process including Tat substrates, Tat-specific proofreading chaperones and the integral membrane proteins TatABC that form the translocase. Fluorescence analysis of these YFP chimeras revealed a wide range of interactions such as the one between the Tat substrate dimethyl sulfoxide reductase (DmsA) and its dedicated proofreading chaperone DmsD. In addition, BiFC analysis illuminated homo- and hetero-oligomeric complexes of the TatA, TatB and TatC integral membrane proteins that were consistent with the current model of translocase assembly. In the case of TatBC assemblies, we provide the first evidence that these complexes are co-localized at the cell poles. Finally, we used this BiFC approach to capture interactions between the putative Tat receptor complex formed by TatBC and the DmsA substrate or its dedicated chaperone DmsD. Our results demonstrate that BiFC is a powerful approach for studying cytoplasmic and inner membrane interactions underlying bacterial secretory pathways

Intranasal “painless” Human Nerve Growth Factors Slows Amyloid Neurodegeneration and Prevents Memory Deficits in App X PS1 Mice

Nerve Growth Factor (NGF) is being considered as a therapeutic candidate for Alzheimer's disease (AD) treatment but the clinical application is hindered by its potent pro-nociceptive activity. Thus, to reduce systemic exposure that would induce pain, in recent clinical studies NGF was administered through an invasive intracerebral gene-therapy approach. Our group demonstrated the feasibility of a non-invasive intranasal delivery of NGF in a mouse model of neurodegeneration. NGF therapeutic window could be further increased if its nociceptive effects could be avoided altogether. In this study we exploit forms of NGF, mutated at residue R100, inspired by the human genetic disease HSAN V (Hereditary Sensory Autonomic Neuropathy Type V), which would allow increasing the dose of NGF without triggering pain. We show that “painless” hNGF displays full neurotrophic and anti-amyloidogenic activities in neuronal cultures, and a reduced nociceptive activity in vivo. When administered intranasally to APPxPS1 mice ( n = 8), hNGFP61S/R100E prevents the progress of neurodegeneration and of behavioral deficits. These results demonstrate the in vivo neuroprotective and anti-amyloidogenic properties of hNGFR100 mutants and provide a rational basis for the development of “painless” hNGF variants as a new generation of therapeutics for neurodegenerative diseases

Application guide for omics approaches to cell signaling

Research in signal transduction aims to identify the functions of different signaling pathways in physiological and pathological states. Traditional techniques using biochemical, genetic or cell biological approaches have made important contributions to our understanding of cellular signaling. However, the single-gene approach does not take into account the full complexity of cell signaling. With the availability of omics techniques, great progress has been made in understanding signaling networks. Omics approaches can be classified into two categories: 'molecular profiling', including genomic, proteomic, post-translational modification and interactome profiling; and 'molecular perturbation', including genetic and functional perturbations

Increasing Exogenous Progesterone during Synchronization of Estrus Decreases Endogenous 17β-Estradiol and Increases Conception in Cows

The objectives of this study were to determine the effects of dose of exogenous progesterone (P4) prior to artificial insemination on concentrations of 17β-estradiol (E2) and on conception rates in bovine females. Heifers (n = 100) and cows (n = 100) received P4-releasing intravaginal devices (PRIDs) to produce two different circulating concentrations of P4. All animals received a single PRID 10 days before (Day -10) the start of the breeding season (Day 0). In animals that received the low dose of P4 (1 PRID, target concentration of 2-3 ng/ml of plasma), the original PRID remained in place for 10 days. In animals that received the larger dose of P4 (2 PRIDs, target concentration of 5-8 ng/ml of plasma), an additional PRID was inserted on Day -9. To maintain concentrations of P4 in the 2-PRID group, the PRIDs inserted on Days -10 and -9 were replaced with new PRIDs on Days -5 and -4, respectively. Prostaglandin F2α (25 mg) was administered to all animals on Days -9 and -3 to remove the endogenous source of P4. Following PRID removal, animals were artificially inseminated 12 h after signs of behavioral estrus were observed. A treatment-by-day interaction (p \u3c 0.0001) was observed for concentrations of P4 in circulation of both heifers and cows. Animals that received 2 PRIDs had greater (p \u3c 0.001) concentrations of P4 by Day -8 of treatment than animals that received 1 PRID. In cows that received 1 PRID, concentrations of E2 increased 2.4-fold from Day -10 (6.8 ± 1.0 pg/ml) to Day -2 (16.7 ± 1.4 pg/ml). Cows treated with 2 PRIDs maintained concentrations of E2 similar to those at the initiation of treatment (treatment-by-day, p \u3c 0.0001). In contrast to cows, concentrations of E2 did not increase in heifers treated with 1 PRID. After PRID withdrawal, both heifers and cows receiving 1 PRID exhibited behavioral estrus earlier (p \u3c 0.006) than animals receiving 2 PRIDs. Cows receiving 1 PRID had a lower (p \u3c 0.02) conception rate than did cows receiving 2 PRIDs (53.3% and 76.6%, respectively). Conception rate in cows was negatively correlated (r = -0.39; p \u3c 0.0001) to mean concentration of E2 prior to P4 withdrawal. There were no differences in conception rates between heifers that received 1 or 2 PRIDs. In summary, cows with low circulating concentrations of P4 (2-3 ng/ml) have elevated concentrations of E2. Prolonged exposure to elevated concentrations of E2 may alter the cascade of events required to establish pregnancy and thereby reduce conception rates in cattle

Identification of Beef Heifers with Superior Uterine Capacity for Pregnancy1

Infertility and subfertility represent major problems in domestic animals and humans, and the majority of embryonic loss occurs during the first month of gestation that involves pregnancy recognition and conceptus implantation. The critical genes and physiological pathways in the endometrium that mediate pregnancy establishment and success are not well understood. In study one, predominantly Angus heifers were classified based on fertility using serial embryo transfer to select animals with intrinsic differences in pregnancy loss. In each of the four rounds, a single in vitro-produced, high-quality embryo was transferred into heifers on Day 7 postestrus and pregnancy was determined on Days 28 and 42 by ultrasound and then terminated. Heifers were classified based on pregnancy success as high fertile (HF), subfertile (SF), or infertile (IF). In study two, fertility-classified heifers were resynchronized and bred with semen from a single high-fertility bull. Blood samples were collected every other day from Days 0 to 36 postmating. Pregnancy rate was determined on Day 28 by ultrasound and was higher in HF (70.4%) than in heifers with low fertility (36.8%; SF and IF). Progesterone concentrations in serum during the first 20 days postestrus were not different in nonpregnant heifers and also not different in pregnant heifers among fertility groups. In study three, a single in vivo-produced embryo was transferred into fertility-classified heifers on Day 7 postestrus. The uteri were flushed on Day 14 to recover embryos, and endometrial biopsies were obtained from the ipsilateral uterine horn. Embryo recovery rate and conceptus length and area were not different among the heifer groups. RNA was sequenced from the Day 14 endometrial biopsies of pregnant HF, SF, and IF heifers (n = 5 per group) and analyzed by edgeR-robust analysis. There were 26 differentially expressed genes (DEGs) in the HF compared to SF endometrium, 12 DEGs for SF compared to IF endometrium, and three DEGs between the HF and IF endometrium. Several of the DEG-encoded proteins are involved in immune responses and are expressed in B cells. Results indicate that preimplantation conceptus survival and growth to Day 14 is not compromised in SF and IF heifers. Thus, the observed difference in capacity for pregnancy success in these fertility-classified heifers is manifest between Days 14 and 28 when pregnancy recognition signaling and conceptus elongation and implantation must occur for the establishment of pregnancy

Discovery, synthesis, and molecular pharmacology of selective positive allosteric modulators of the ?-opioid receptor

© 2015 American Chemical Society. Allosteric modulators of G protein-coupled receptors (GPCRs) have a number of potential advantages compared to agonists or antagonists that bind to the orthosteric site of the receptor. These include the potential for receptor selectivity, maintenance of the temporal and spatial fidelity of signaling in vivo, the ceiling effect of the allosteric cooperativity which may prevent overdose issues, and engendering bias by differentially modulating distinct signaling pathways. Here we describe the discovery, synthesis, and molecular pharmacology of δ-opioid receptor-selective positive allosteric modulators (δ PAMs). These δ PAMs increase the affinity and/or efficacy of the orthosteric agonists leu-enkephalin, SNC80 and TAN67, as measured by receptor binding, G protein activation, β-arrestin recruitment, adenylyl cyclase inhibition, and extracellular signal-regulated kinases (ERK) activation. As such, these compounds are useful pharmacological tools to probe the molecular pharmacology of the δ receptor and to explore the therapeutic potential of δ PAMs in diseases such as chronic pain and depression