Regulation of sodium transport in the cortical collecting duct : physiological role of GILZ in vitro and in vivo : characterisation of Na+ transport in the mCCDcl1 cell line

SUMMARY Regulation of sodium excretion by the kidney is a key mechanism in the long term regulation of blood pressure, and when altered it constitutes a risk factor for the appearance of arterial hypertension. Aldosterone, which secretion depends upon salt intake in the diet, is a steroid hormone that regulates sodium reabsorption in the distal part of the nephron (functional unit of the kidney) by modulating gene transcription. It has been shown that it can act synergistically with the peptidic hormone insulin through the interaction of their signalisation pathways. Our work consisted of two distinct parts: 1) the in vitro and in vivo characterisation of Glucocorticoid-Induced Leucine Zipper (GILZ) (an aldosterone-induced gene) mechanism of action; 2) the in vitro characterisation of insulin mechanism of action and its interaction with aldosterone. GILZ mRNA, coded by the TSC22D3 gene, is strongly induced by aldosterone in the cell line of principal cells of the cortical collecting duct (CCD) mpkCCDc14, suggesting that GILZ is a mediator of aldosterone response. Co-expression of GILZ and the amiloride-sensitive epithelial sodium channel ENaC in vitro in the Xenopus oocyte expression system showed that GILZ has no direct effect on the ENaC-mediated Na+ current in basal conditions. To define the role of GILZ in the kidney and in other organs (colon, heart, skin, etc.), a conditional knock-out mouse is being produced and will allow the in vivo study of its role. Previous data showed that insulin induced a transepithelial sodium transport at supraphysiological concentrations. Insulin and the insulin-like growth factor 1 (IGF-1) are able to bind to each other receptor with an affinity 50 to 100 times lower than to their cognate receptor. Our starting hypothesis was that the insulin effect observed at these supraphysiological concentrations is actually mediated by the IGF receptor type 1 (IGF-1R). In a new cell line that presents all the characteristics of the principal cells of the CCD (mCCDc11) we have shown that both insulin and IGF-1 induce a physiologically significant increase of Na+ transport through the activation of IGF-1R. Aldosterone and insulin/IGF-1 have an additive effect on Na+ transport, through the activation of the PI3-kinase (PI3-K) pathway and the phosphorylation of the serum- and glucocorticoid-induced kinase 1 (Sgk1) by the IGF-1R, and the induction of Sgk1 expression by aldosterone. Thus, Sgk1 integrates IGF-1/insulin and aldosterone effects. We suggest that IGF-1 is physiologically relevant in the modulation of sodium balance, while insulin can only regulate Na+ transport at supraphysiological conditions. Both hormones would bind to the IGF-1R and induce Na+ transport by activating the PI3-K PDK1/2 - Sgk1 pathway. We have shown for the first time that Sgk1 is expressed and phosphorylated in principal cells of the CCD in basal conditions, although the mechanism that maintains Sgk1 phosphorylation is not known. This new role for IGF-1 suggests that it could be a salt susceptibility gene. In effect, IGF-1 stimulates Na+ and water transport in the kidney in vivo. Moreover, 35 % of the acromegalic patients (overproduction of growth hormone and IGF-1) are hypertensives (higher proportion than in normal population), and genetic analysis suggest a link between the IGF-1 gene locus and blood pressure. RÉSUMÉ La régulation de l'excrétion rénale de sodium (Na+) joue un rôle principal dans le contrôle à long terme de la pression sanguine, et ses altérations constituent un facteur de risque de l'apparition d'une hypertension artérielle. L'aldosterone, dont la sécrétion dépend de l'apport en sel dans la diète, est une hormone stéroïdienne qui régule la réabsorption de Na+ dans la partie distale du nephron (unité fonctionnelle du rein) en contrôlant la transcription de gènes. Elle peut agir de façon synergistique avec l'hormone peptidique insuline, probablement via l'interaction de leurs voies de signalisation cellulaire. Le but de notre travail comportait deux volets: 1) caractériser in vitro et in vivo le mécanisme d'action du Glucocorticoid Induced Leucine Zipper (GILZ) (un gène induit par l'aldosterone); 2) caractériser in vitro le mécanisme d'action de l'insuline et son interaction avec l'aldosterone. L'ARNm de GILZ, codé par le gène TSC22D3, est induit par l'aldosterone dans la lignée cellulaire de cellules principales du tubule collecteur cortical (CCD) mpkCCDc14, suggérant que GILZ est un médiateur potentiel de la réponse à l'aldosterone. La co-expression in vitro de GILZ et du canal à Na+ sensible à l'amiloride ENaC dans le système d'expression de l'oocyte de Xénope a montré que GILZ n'a pas d'effet sur les courants sodiques véhiculées par ENaC en conditions basales. Une souris knock-out conditionnelle de GILZ est en train d'être produite et permettra l'étude in vivo de son rôle dans le rein et d'autres organes. Des expériences préliminaires ont montré que l'insuline induit un transport transépithelial de Na+ à des concentrations supraphysiologiques. L'insuline et l'insulin-like growth factor 1 (IGF-1) peuvent se lier à leurs récepteurs réciproques avec une affinité 50 à 100 fois moindre qu'à leur propre récepteur. Nous avons donc proposé que l'effet de l'insuline soit médié par le récepteur à l'IGF type 1 (IGF-1R). Dans une nouvelle lignée cellulaire qui présente toutes les caractéristiques des cellules principales du CCD (mCCDc11) nous avons montré que les deux hormones induisent une augmentation physiologiquement significative du transport du Na+ par l'activation des IGF-1 R. Aldosterone et insuline/IGF-1 ont un effet additif sur le transport de Na+, via l'activation de la voie de la PI3-kinase et la phosphorylation de la serum- and glucocorticoid-induced kinase 1 (Sgk1) par l'IGF-1R, dont l'expression est induite par l'aldosterone. Sgk1 intègre les effets de l'insuline et l'aldosterone. Nous proposons que l'IGF-1 joue un rôle dans la modulation physiologique de la balance sodique, tandis que l'insuline régule le transport de Na+ à des concentrations supraphysiologiques. Les deux hormones agissent en se liant à l'IGF-1R et induisent le transport de Na+ en activant la cascade de signalisation PI3-K - PDK1/2 - Sgk1. Nous avons montré pour la première fois que Sgk1 est exprimée et phosphorylée dans des conditions basales dans les cellules principales du CCD, mais le mécanisme qui maintient sa phosphorylation n'est pas connu. Ce nouveau rôle pour l'IGF-1 suggère qu'il pourrait être un gène impliqué de susceptibilité au sel. Aussi, l'IGF-1 stimule le transport rénal de Na+ in vivo. De plus, 35 % des patients atteints d'acromégalie (surproduction d'hormone de croissance et d'IGF-1) sont hypertensifs (prévalence plus élevée que la population normale), et des analyses génétiques suggèrent un lien entre le locus du gène de l'IGF-1 et la pression sanguine. RÉSUMÉ GRAND PUBLIC Nos ancêtres se sont génétiquement adaptés pendant des centaines de millénaires à un environnement pauvre en sel (chlorure de sodium) dans la savane équatoriale, où ils consommaient moins de 0,1 gramme de sel par jour. On a commencé à ajouter du sel aux aliments avec l'apparition de l'agriculture (il y a 5000 à 10000 années), et aujourd'hui une diète omnivore, qui inclut des plats préparés, contient plusieurs fois la quantité de sodium nécessaire pour notre fonction physiologique normale (environ 10 grammes par jour). Le corps garde sa concentration constante dans le sang en s'adaptant à une consommation très variable de sel. Pour ceci, il module son excrétion soit directement, soit en sécrétant des hormones régulatrices. Le rein joue un rôle principal dans cette régulation puisque l'excrétion urinaire de sel change selon la diète et peut aller d'une quantité dérisoire à plus de 36 grammes par jour. L'attention qu'on prête au sel est liée à sa relation avec l'hypertension essentielle. Ainsi, le contrôle rénal de l'excrétion de sodium et d'eau est le principal mécanisme dans la régulation de la pression sanguine, et une ingestion excessive de sel pourrait être l'un des facteurs-clé déclenchant l'apparition d'un phénotype hypertensif. L'hormone aldosterone diminue l'excrétion de sodium par le rein en modulant l'expression de gènes qui pourraient être impliqués dans la sensibilité au sel. Dans une lignée cellulaire de rein l'expression du gène TSC22D3, qui se traduit en la protéine Glucocorticoid Induced Leucine Zipper (GILZ), est fortement induite par l'aldosterone. Ceci suggère que GILZ est un médiateur potentiel de l'effet de l'aldosterone, et pourrait être impliqué dans la sensibilité au sel. Pour analyser la fonction de GILZ dans le rein plusieurs approches ont été utilisées. Par exemple, une souris dans laquelle GILZ est spécifiquement inactivé dans le rein est en train d'être produite et permettra l'étude du rôle de GILZ dans l'organisme. De plus, on a montré que GILZ, en conditions basales, n'a pas d'effet direct sur la protéine transportant le sodium à travers la membrane des cellules, le canal sodique épithélial ENaC. On a aussi essayé de trouver des protéines qui interagissent directement avec GILZ utilisant une technique appelée du « double-hybride dans la levure », mais aucun candidat n'a émergé. Des études ont montré que, à de hautes concentrations, l'insuline peut aussi diminuer l'excrétion de sodium. A ces concentrations, elle peut activer son récepteur spécifique, mais aussi le récepteur d'une autre hormone, l'Insulin-Like Growth Factor 1 (IGF-1). En plus, l'infusion d'IGF-1 augmente la rétention rénale de sodium et d'eau, et des mutations du gène codant pour l'IGF-1 sont liées aux différents niveaux de pression sanguine. On a utilisé une nouvelle lignée cellulaire de rein développée dans notre laboratoire, appelée mCCDc11, pour analyser l'importance relative des deux hormones dans l'induction du transport de sodium. On a montré que les deux hormones induisent une augmentation significative du transport de sodium par l'activation de récepteurs à l'IGF-1 et non du récepteur à l'insuline. On a montré qu'à l'intérieur de la cellule leur activation induit une augmentation du transport sodique par le biais du canal ENaC en modifiant la quantité de phosphates fixés sur la protéine Serumand Glucocorticoid-induced Kinase 1 (Sgk1). On a finalement montré que l'IGF-1 et l'aldosterone ont un effet additif sur le transport de sodium en agissant toutes les deux sur Sgk1, qui intègre leurs effets dans le contrôle du transport de sodium dans le rein.

Cell death and autophagy after severe hypoxic-ischemic encephalopathy in term newborns

Introduction: Various studies from hypoxic-ischemic animals haveinvestigated neuroprotection by targeting necrosis and apoptosis with inconclusive results. Three types of cell death have been described: apoptosis, necrosis and more recently, autophagic cell death. While autophagy is a physiological process of degradation of cellular components, excessive autophagy may be involved in cell death. Recent studies showed that inhibition of autophagy is neuroprotective in rodent neonatal models of cerebral ischemia. Furthermore, neonatal hypoxia-ischemia strongly increased neuronal autophagic flux which is linked to cell death in a rat model of perinatal asphyxia. Following our observations in animals, the aim of the present study was to characterize the different neuronal death phenotypes and to clarify whether autophagic cell death could be also involved in neuronal death in the human newborns after perinatal asphyxia. Methods: we selected retrospectively and anonymously all newborns who died in our unit of neonatology between 2004 and 2009, with the following criteria: gestational age >36 weeks, diagnosis of perinatal asphyxia (Apgar <5 at 5 minutes, arterial pH <7.0 at 1 hour of life and encephalopathy Sarnat III) and performed autopsy. The brain of 6 cases in asphyxia group and 6 control cases matching gestational age who died of pulmonary or other malformations were selected. On histological sections of thalamus, frontal cortex and hippocampus, different markers of apoptosis (caspase 3, TUNEL), autophagosomes (LC3-II) and lysosomes (LAMP1, Cathepsin D) were tested by immunohistochemistry. Results: Preliminary studies on markers of apoptosis (TUNEL, caspase 3) and of autophagy (Cathepsin D, LC3II, LAMP1) showed an expected increase of apoptosis, but also an increase of neuronal autophagic flux in the selected areas. The distribution seems to be region specific. Conclusion: This is the first time that autophagic flux linked with cell death is shown in brain of human babies, in association with hypoxicischemic encephalopathy. This work leads to a better understanding of the mechanisms associated with neuronal death following perinatal asphyxia and determines whether autophagy could be a promising therapeutic target.

Cardiologie. Tests de réactivité plaquettaire: mise à jour pour le praticien [Cardiology. Platelet function testing for clinicians].

Platelet P2YI2 receptor inhibition with clopidogrel, prasugrel or ticagrelor plays a key role to prevent recurrent ischaemic events after percutaneous coronary intervention in acute coronary syndromes or elective settings. The degree of platelet inhibition depends on the antiplatelet medication used and is influenced by clinical and genetic factors. A concept of therapeutic window exists. On one side, efficient anti-aggregation is required in order to reduce cardio-vascular events. On the other side, an excessive platelet inhibition represents a risk of bleeding complications. This article describes the current knowledge about some platelet function tests and genetic tests and summarises their role in the clinical practice.

Road Safety Audit for US 6 from the ECL of Iowa City to the WCL of West Liberty in Johnson and Muscatine Counties, Iowa, 2008

Approximately 13.2 miles of US 6 in eastern Iowa extends from the east corporate limits of Iowa City, Iowa, to the west corporate limits of West Liberty, Iowa. This segment of US 6 is a service level B primary highway, with an annual daily traffic volume varying from 3,480 vehicles per day (vpd) to 5,700 vpd. According to 2001–2007 crash density data from the Iowa Department of Transportation (Iowa DOT), the corridor is currently listed among the top 5% of non-freeway Iowa DOT roads in several crash categories, including crashes involving excessive speed, impaired drivers, single-vehicle run-off-road, and multiple-vehicle crossed centerline. A road safety audit of this corridor was deemed appropriate by the Iowa Department of Transportation’s Office of Traffic and Safety. Staff and officials from the Iowa DOT, Iowa State Patrol, Governor’s Traffic Safety Bureau, Federal Highway Administration, Center for Transportation Research and Education, and several local law enforcement and transportation agencies met to review crash data and discuss potential safety improvements to this segment of US 6. This report outlines the findings and recommendations of the road safety audit team to address the safety concerns on this US 6 corridor and explains several selected mitigation strategies.

Road Safety Audit for the US 61/Harrison Street and West Locust Street Intersection in Davenport-Scott County, Iowa, 2008

On the October 7 and 8, 2008, a road safety audit was conducted for the intersection of US 61/Harrison Street and West Locust Street in Davenport, Iowa. US 61/Harrison Street is a one-way street and a principal arterial route through Davenport, with three southbound lanes. Locust Street is a four-lane, two-way minor arterial running across the city from west to east. The last major improvement at this intersection was implemented approximately 20 years ago. The Iowa Department of Transportation requested a safety audit of this intersection in response to a high incidence of crashes at the location over the past several years, in view of the fact that no major improvements are anticipated for this intersection in the immediate future. The road safety audit team discussed current conditions at the intersection and reviewed the last seven years of crash data. The team also made daytime and nighttime field visits to the intersection to examine field conditions and observe traffic flow and crossing guard operations with younger pedestrians. After discussing key issues, the road safety audit team drew conclusions and suggested possible enforcement, engineering, public information, and educational strategies for mitigation.

Field Evaluation of Elliptical Steel Dowel Performance, 2006

Joints are always a concern in the construction and long-term performance of concrete pavements. Research has shown that we need some type of positive load transfer across transverse joints. The same research has directed pavement designers to use round dowels spaced at regular intervals across the transverse joint to distribute the vehicle loads both longitudinally and transversely across the joint. The goal is to reduce bearing stresses on the dowels and the two pavement slab edges and erosion of the underlying surface, hence improved long-term joint and pavement structure performance. Road salts cause metal corrosion in doweled joints, excessive bearing stresses hollow dowel ends, and construction processes are associated with cracking pavement at the end of dowels. Dowels are also a cost factor in the pavement costs when joint spacing is reduced to control curling and warping distress in pavements. Designers desire to place adequate numbers of dowels spaced at the proper locations to handle the anticipated loads and bearing stresses for the design life of the pavement. This interim report is the second of three reports on the evaluation of elliptical steel dowels. This report consists of an update on the testing and performance of the various shapes and sizes of dowels. It also documents the results of the first series of performance surveys and draws interim conclusions about the performance of various bar shapes, sizes, spacings, and basket configurations. In addition to the study of elliptical steel dowel performance, fiber reinforced polymers (FRP) are also tested as elliptical dowel material (in contrast to steel) on a section of the highway construction north of the elliptical steel test sections.

09032-017 Miller Creek Watershed Final Report

Miller Creek is on the 2006 Section 303d Impaired Waters List and has a 19,926 acre watershed. All indicators, as reported in the Miller Creek assessment, show that the impairment is due to sediment and nutrient delivery from upland runoff which contributes to elevated water temperatures, excessive algae, and low dissolved oxygen levels within the stream. In an effort to control these problems, the Miller Creek Water Quality Project will target areas of 5 tons per acre or greater soil loss or with 0.5 tons per acre or greater sediment delivery rates. The assessment revealed these targeted priority lands make up 32% or 6,395 acres of the Miller Creek watershed. Priority lands include cropland, pasture land, timber, and sensitive riparian areas. It is the goal of this project to reduce sediment delivery by 70% on 60% or 3,837 acres of these priority lands. This will be accomplished through installation of strategically placed structural practices, rotational grazing systems, and buffer strips. These practices will reduce soil loss, reduce sediment delivery, improve water quality, and improve wildlife habitat in the watershed. Utilizing partnerships with NRCS and IDALS-DSC will be important in making this project successful. In addition to using matching funds from EQIP, WHIP, and CRP, the Monroe SWCD is committed to prioritizing local cost share funds through IFIP and REAP for use in the Miller Creek Watershed.

Water-filtered infrared-A radiation (wIRA) is not implicated in cellular degeneration of human skin.

BACKGROUND: Excessive exposure to solar ultraviolet radiation is involved in the complex biologic process of cutaneous aging. Wavelengths in the ultraviolet-A and -B range (UV-A and UV-B) have been shown to be responsible for the induction of proteases, e. g. the collagenase matrix metalloproteinase 1 (MMP-1), which are related to cell aging. As devices emitting longer wavelengths are widely used in therapeutic and cosmetic interventions and as the induction of MMP-1 by water-filtered infrared-A (wIRA) had been discussed, it was of interest to assess effects of wIRA on the cellular and molecular level known to be possibly involved in cutaneous degeneration. OBJECTIVES: Investigation of the biological implications of widely used water-filtered infrared-A (wIRA) radiators for clinical use on human skin fibroblasts assessed by MMP-1 gene expression (MMP-1 messenger ribonucleic acid (mRNA) expression).Methods: Human skin fibroblasts were irradiated with approximately 88% wIRA (780-1400 nm) and 12% red light (RL, 665-780 nm) with 380 mW/cm(2) wIRA(+RL) (333 mW/cm(2) wIRA) on the one hand and for comparison with UV-A (330-400 nm, mainly UV-A1) and a small amount of blue light (BL, 400-450 nm) with 28 mW/cm(2) UV-A(+BL) on the other hand. Survival curves were established by colony forming ability after single exposures between 15 minutes and 8 hours to wIRA(+RL) (340-10880 J/cm(2) wIRA(+RL), 300-9600 J/cm(2) wIRA) or 15-45 minutes to UV-A(+BL) (25-75 J/cm(2) UV-A(+BL)). Both conventional Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and quantitative real-time RT-PCR techniques were used to determine the induction of MMP-1 mRNA at two physiologic temperatures for skin fibroblasts (30 degrees C and 37 degrees C) in single exposure regimens (15-60 minutes wIRA(+RL), 340-1360 J/cm(2) wIRA(+RL), 300-1200 J/cm(2) wIRA; 30 minutes UV-A(+BL), 50 J/cm(2) UV-A(+BL)) and in addition at 30 degrees C in a repeated exposure protocol (up to 10 times 15 minutes wIRA(+RL) with 340 J/cm(2) wIRA(+RL), 300 J/cm(2) wIRA at each time). RESULTS: Single exposure of cultured human dermal fibroblasts to UV-A(+BL) radiation yielded a very high increase in MMP-1 mRNA expression (11 +/-1 fold expression for RT-PCR and 76 +/-2 fold expression for real-time RT-PCR both at 30 degrees C, 75 +/-1 fold expression for real-time RT-PCR at 37 degrees C) and a dose-dependent decrease in cell survival. In contrast, wIRA(+RL) did not produce cell death and did not induce a systematic increase in MMP-1 mRNA expression (less than twofold expression, within the laboratory range of fluctuation) detectable with the sensitive methods applied. Additionally, repeated exposure of human skin fibroblasts to wIRA(+RL) did not induce MMP-1 mRNA expression systematically (less than twofold expression by up to 10 consecutive wIRA(+RL) exposures and analysis with real-time RT-PCR). CONCLUSIONS: wIRA(+RL) even at the investigated disproportionally high irradiances does not induce cell death or a systematic increase of MMP-1 mRNA expression, both of which can be easily induced by UV-A radiation. Furthermore, these results support previous findings of in vivo investigations on collagenase induction by UV-A but not wIRA and show that infrared-A with appropriate irradiances does not seem to be involved in MMP-1 mediated photoaging of the skin. As suggested by previously published studies wIRA could even be implicated in a protective manner.

1012-007 Yellow River Headwaters Final Report

The Yellow River Headwaters Watershed (YRHW) drains 26,730 acres of rural land within Winneshiek and Allamakee Counties. While portions of the river have been designated as a High Quality Resource by the State of Iowa, other portions appear on the State's 303(d) List of Impaired Waters due to excessive nutrients, sediment and other water quality issues. The Winneshiek SWCD was fortunate enough to secure WSPF/WPF funds for FY2009 to begin addressing many of the sources of the identified problems, especially along the all-to-critical stream corridor. Initial landowner I producer interest has exceeded expectations and several key BMPs have been installed within the identified critical areas. Yet due to the current budget constraints in the WSPF/WPF programs, we currently have greater landowner I producer interest than we do funds, which is why the District is applying for WIRB funding, to provide supplemental incentives to continue the installation of needed Grade Stabilization Structures, Terraces and Manure Management Systems in identified critical areas. Other funding, currently available to the District, will cover the remaining portions of the project's budget, including staff and our outreach efforts.

1233-015IJ Miller Creek Watershed Final Report

Miller Creek, a 19,926 acre watershed, is listed on the 2008 Section 303d Impaired Waters List. All indicators, as reported in the Miller Creek assessment, show that the impairment is due to nutrient and sediment delivery from upland runoff which contributes to elevated water temperatures, excessive algae, and low dissolved oxygen levels within the stream. The WIRB board provided implementation grant funds in 2010 for a three year project to treat targeted areas of 5 tons per acre or greater soil loss with an estimated reduction of 2,547 tons. As of December 1, 2012, with 95% of the funds allocated, the final results are estimated to provide a sediment delivery reduction of 4,500 tons and an estimated phosphorus reduction of 5,700 lbs per year. These accomplishments and the completion of the three year Miller Creek WIRB project represent "Phase I" of the SWCD's goals to treat the Miller Creek watershed. This application represents "Phase II" or the final phase of the Miller Creek water quality project. The Monroe SWCD plans to reduce sediment delivery by 70% on an additional 245 acres of priority land. This goal will be accomplished through installation of strategically placed structural practices, BMPs, and grazing systems. These practices will reduce soil loss, nutrient runoff, and sediment delivery as well as improve water quality and wildlife habitat in the watershed. Utilization of partnerships with NRCS and IDALS-DSC will continue to be an important part to the success of the project. Project goals will be achieved by utilizing matching funds from EQIP, and the Monroe SWCD has approved the use of District IFIP cost share funds specifically for use in the Miller Creek Watershed.

1011-006 Competine Creek Watershed Final Report

The Competine Creek watershed is a 24,956 acre sub-watershed of Cedar Creek. The creek traverses portions of three counties, slicing through rich and highly productive Southern lowa Drift Plain soils. The watershed is suffering from excessive sediment delivery and frequent flash floods that have been exacerbated by recent high rainfall events. Assessment data reveals soil erosion estimated to be 38,435 tons/year and sediment delivery to the creek at 15,847 tons/year. The Competine Creek Partnership Project is seeking WIRB funds to merge with IDALS-DSC funds and local funds, all targeted for structural Best Management Practices (BMPs) within the 2,760 acres of High Priority Areas (HPAs) identified by the assessment process. The BMPs will include grade stabilization structures, water and sediment basins, tile-outlet terraces, CRP, and urban storm water conservation practices. In addition, Iowa State University Extension-Iowa Learning Farm is investing in the project by facilitating a crop sampling program utilizing fall stalk nitrate, phosphorous index, and soil conditioning index testing. These tests will be used by producers as measures of performance to refine nutrient and soil loss management and to determine effective alternatives to reduce sediment and nutrient delivery to Competine Creek.

07003-001 Clear Lake Watershed Final Report

Clear Lake, Iowa's third largest natural lake, is a premier natural resource and popular recreational destination in north central Iowa. Despite the lake's already strong recreational use, water quality concerns have not allowed the lake to reach its full potential. Clear Lake is listed on Iowa's 2004 303(d) Impaired Waters List due to excessive levels of phosphorus, bacteria, and turbidity. Urban storm water runoff from the 8,600 acre watershed is a significant contributor to Clear Lake's impairment. Local communities have been working towards the goal of making improvements at all 30 storm water outlets that have a drainage area of five acres or more and have a cost effective solution. Many improvements have already been made, and now there are only seven storm water outlet sites remaining that still need protection in order to meet the goal. The storm water improvements have been very effective in reducing contaminants in urban runoff, achieving reduction levels in the 50-80% range. The proposed Clear Lake Storm Water Improvement Project will address the remaining seven outlet sites and take place over three years. The first year will consist of performing engineering and design of storm water best management practices (BMPs) at the seven outlet sites to determine if a cost effective solution exists for each. Years two and three will consist of installing two storm water improvements each year to implement the most cost effective BMPs at a minimum of four of the seven sites. The grant request addresses one of the main priorities of the Iowa Watershed Improvement Grant: storm water runoff.

Increased male offspring's risk of metabolic-neuroendocrine dysfunction and overweight after fructose-rich diet intake by the lactating mother.

An adverse endogenous environment during early life predisposes the organism to develop metabolic disorders. We evaluated the impact of intake of an iso-caloric fructose rich diet (FRD) by lactating mothers (LM) on several metabolic functions of their male offspring. On postnatal d 1, ad libitum eating, lactating Sprague-Dawley rats received either 10% F (wt/vol; FRD-LM) or tap water (controls, CTR-LM) to drink throughout lactation. Weaned male offspring were fed ad libitum a normal diet, and body weight (BW) and food intake were registered until experimentation (60 d of age). Basal circulating levels of metabolic markers were evaluated. Both iv glucose tolerance and hypothalamic leptin sensitivity tests were performed. The hypothalamus was dissected for isolation of total RNA and Western blot analysis. Retroperitoneal (RP) adipose tissue was dissected and either kept frozen for gene analysis or digested to isolate adipocytes or for histological studies. FRD rats showed increased BW and decreased hypothalamic sensitivity to exogenous leptin, enhanced food intake (between 49-60 d), and decreased hypothalamic expression of several anorexigenic signals. FRD rats developed increased insulin and leptin peripheral levels and decreased adiponectinemia; although FRD rats normally tolerated glucose excess, it was associated with enhanced insulin secretion. FRD RP adipocytes were enlarged and spontaneously released high leptin, although they were less sensitive to insulin-induced leptin release. Accordingly, RP fat leptin gene expression was high in FRD rats. Excessive fructose consumption by lactating mothers resulted in deep neuroendocrine-metabolic disorders of their male offspring, probably enhancing the susceptibility to develop overweight/obesity during adult life.

Lipoprotein distribution and biological variation of 24S- and 27-hydroxycholesterol in healthy volunteers.

24S- and 27-hydroxycholesterol are obligatory intermediates of cholesterol catabolism and play an important role in the maintenance of whole-body cholesterol homeostasis. Using an HPLC-MS method for oxysterol quantification, the distribution of esterified and unesterified oxysterols in lipoprotein subfractions as well as the influence of daytime, food intake and menstrual cycle on oxysterol concentrations were investigated in healthy volunteers. Moreover, reference intervals for 24S- and 27-hydroxycholesterol in plasma as well as the corresponding levels for 27-hydroxycholesterol in the HDL subfraction were established in 100 healthy volunteers. Both circulating oxysterols are mainly transported in association with HDL and LDL--primarily in the esterified form. No significant diurnal changes and no variations during menstrual cycle of either absolute or cholesterol-related plasma levels were detected. In contrast to 24S-hydroxycholesterol in plasma and 27-hydroxycholesterol in the HDL subfraction, the 95% reference intervals of 27-hydroxycholesterol both in plasma and the non-HDL subfraction were higher in males than in females. The concentrations of 27-hydroxycholesterol in plasma and the non-HDL subfraction showed strong positive correlations with the concentrations of cholesterol, non-HDL cholesterol and triglycerides. Our data on the lipoprotein distribution of oxysterols as well as on their intra- and inter-individual variation set the stage for future clinical studies.

07032-012 Lake MacBride Watershed Final Report

Lake Macbride is considered to be one of the top four lakes for fishing in the state of Iowa. It is widely used by the public and contributes significant economic benefits to the county. Lake Macbride is situated in the North Corridor which is one of the fastest growing areas in the state. The lake has a surface area of 940 acres and drains 16,205 acres. Lake Macbride is on the Iowa 303(d) list for excessive sediments and nutrients. In 2001, Lake Macbride State Park received over 2.5 million dollars from the Marine Fuel Tax and Fish and Wildlife Trust to install 2 silt basins and stabilize over 12 miles of shoreline in the lake. Also in 2001, the Johnson County SWCD received a WSPF allocation from DSC to address agriculture and urban runoff issues in the watershed. Section 319 funding was received in 2002 to continue watershed efforts to the present. A watershed assessment was completed in 2003 to guide watershed activities. In 2005, a TMDL was completed for the lake. Since 2001, over $645,000 dollars has been spent by landowners and funding partners to install conservation practices in the watershed. Watershed efforts have resulted in the reduction of over 4200 tons of soils from being delivered into Lake Macbride. Nutrient reductions have also occurred from the development of nutrient management plans on 2000 acres. The District is in the process of wrapping up watershed efforts on private land. A series of 13 structures is planned to be installed in the State Park over the next several years. One of the last remaining items that still needs addressed is 1,400 feet of eroding shoreline adjacent to Lake Macbride along Cottage Reserve Road. The road is under the jurisdiction of the Johnson County Board of Supervisors. Both the Board of Supervisors and the IDNR are willing to contribute substantial dollars to address the 250 tons of soil that are being directly delivered to Lake Macbride each year.