Flow cytofluorometric analysis of the binding of Vicia villosa lectin to T lymphoblasts: lack of correlation with cytolytic function.

The relationship between the binding of Vicia villosa (VV) lectin and the expression of cytolytic function in T lymphoblasts has been investigated using flow cytofluorometric techniques. Spleen cells activated in vitro in 5-day mixed leukocyte cultures (MLC) were incubated sequentially with VV, rabbit anti-V antiserum, and fluoresceinated sheep anti-rabbit IgG. When these stained MLC cells were passed on a flow cytometer gated to exclude nonviable cells and small lymphocytes, a single heterogeneous peak of fluorescence was seen, as compared to control MLC cells that had not been incubated with VV. Fluorescence of lymphoblasts was dependent upon lectin dose and was eliminated when staining was performed in the presence of N-acetyl-D-galactosamine, the appropriate competitive sugar for VV. T cell blast populations activated against H-2, Mls, or parasite antigens all had comparable levels of fluorescence after staining with VV, although the cytolytic activity of these cells varied widely. Furthermore, when MLC lymphoblasts binding large or small amounts of VV were sorted on the basis of their relative fluorescence intensity and tested for cytolytic function, no appreciable difference in activity between the 2 populations was observed. These results are inconsistent with the hypothesis that VV binds selectively to cytolytic T lymphocytes.

Bystander-Activated Memory CD8 T Cells Control Early Pathogen Load in an Innate-like, NKG2D-Dependent Manner.

During an infection the antigen-nonspecific memory CD8 T cell compartment is not simply an inert pool of cells, but becomes activated and cytotoxic. It is unknown how these cells contribute to the clearance of an infection. We measured the strength of T cell receptor (TCR) signals that bystander-activated, cytotoxic CD8 T cells (BA-CTLs) receive in vivo and found evidence of limited TCR signaling. Given this marginal contribution of the TCR, we asked how BA-CTLs identify infected target cells. We show that target cells express NKG2D ligands following bacterial infection and demonstrate that BA-CTLs directly eliminate these target cells in an innate-like, NKG2D-dependent manner. Selective inhibition of BA-CTL-mediated killing led to a significant defect in pathogen clearance. Together, these data suggest an innate role for memory CD8 T cells in the early immune response before the onset of a de novo generated, antigen-specific CD8 T cell response.

Dose-dependent effect of atrial natriuretic peptide on blood pressure, heart rate, and skin blood flow of normal volunteers.

Synthetic atrial natriuretic peptide, containing 26 amino acids in the rat sequence, L-364, 343 (Ileu-ANP), was infused intravenously at increasing rates (1-40 micrograms/min) into four normal volunteers. Mean intraarterial blood pressure decreased and heart rate increased in cumulative-dose-dependent fashion. Skin blood flow as measured with a laser Doppler device rose already with a cumulative dose of 55 micrograms Ileu-ANP and further rises were directly related to dose. The only side effects observed were those accompanying symptomatic hypotension at higher doses. These findings provide strong evidence that Ileu-ANP acts as a vasodilator in normal volunteers.

Role of ATP-dependent potassium channels in pulmonary vascular tone of fetal lambs with congenital diaphragmatic hernia.

High mortality in newborn babies with congenital diaphragmatic hernia (CDH) is principally due to persistent pulmonary hypertension. ATP-dependent potassium (K(ATP)) channels might modulate pulmonary vascular tone. We have assessed the effects of Pinacidil, a K(ATP) channel opener, and glibenclamide (GLI), a K(ATP) channel blocker, in near full-term lambs with and without CDH. In vivo, pulmonary hemodynamics were assessed by means of pressure and blood flow catheters. In vitro, we used isolated pulmonary vessels and immunohistochemistry to detect the presence of K(ATP) channels in pulmonary tissue. In vivo, pinacidil (2 mg) significantly reduced pulmonary vascular resistance (PVR) in both controls and CDH animals. GLI (30 mg) significantly increased pulmonary arterial pressure (PAP) and PVR in control animals only. In vitro, pinacidil (10 microM) relaxed, precontracted arteries from lambs with and without CDH. GLI (10(-5) microM) did not raise the basal tone of vessels. We conclude that activation of K(ATP) channels could be of interest to reduce pulmonary vascular tone in fetal lambs with CDH, a condition often associated with persistent pulmonary hypertension of the newborn.

Quality of life depends on the drinking pattern in alcohol-dependent patients.

AIMS: In patients with alcohol dependence, health-related quality of life (QOL) is reduced compared with that of a normal healthy population. The objective of the current analysis was to describe the evolution of health-related QOL in adults with alcohol dependence during a 24-month period after initial assessment for alcohol-related treatment in a routine practice setting, and its relation to drinking pattern which was evaluated across clusters based on the predominant pattern of alcohol use, set against the influence of baseline variables METHODS: The Medical Outcomes Study 36-Item Short-Form Survey (MOS-SF-36) was used to measure QOL at baseline and quarterly for 2 years among participants in CONTROL, a prospective observational study of patients initiating treatment for alcohol dependence. The sample consisted of 160 adults with alcohol dependence (65.6% males) with a mean (SD) age of 45.6 (12.0) years. Alcohol use data were collected using TimeLine Follow-Back. Based on the participant's reported alcohol use, three clusters were identified: 52 (32.5%) mostly abstainers, 64 (40.0%) mostly moderate drinkers and 44 (27.5%) mostly heavy drinkers. Mixed-effect linear regression analysis was used to identify factors that were potentially associated with the mental and physical summary MOS-SF-36 scores at each time point. RESULTS: The mean (SD) MOS-SF-36 mental component summary score (range 0-100, norm 50) was 35.7 (13.6) at baseline [mostly abstainers: 40.4 (14.6); mostly moderate drinkers 35.6 (12.4); mostly heavy drinkers 30.1 (12.1)]. The score improved to 43.1 (13.4) at 3 months [mostly abstainers: 47.4 (12.3); mostly moderate drinkers 44.2 (12.7); mostly heavy drinkers 35.1 (12.9)], to 47.3 (11.4) at 12 months [mostly abstainers: 51.7 (9.7); mostly moderate drinkers 44.8 (11.9); mostly heavy drinkers 44.1 (11.3)], and to 46.6 (11.1) at 24 months [mostly abstainers: 49.2 (11.6); mostly moderate drinkers 45.7 (11.9); mostly heavy drinkers 43.7 (8.8)]. Mixed-effect linear regression multivariate analyses indicated that there was a significant association between a lower 2-year follow-up MOS-SF-36 mental score and being a mostly heavy drinker (-6.97, P < 0.001) or mostly moderate drinker (-3.34 points, P = 0.018) [compared to mostly abstainers], being female (-3.73, P = 0.004), and having a Beck Inventory scale score ≥8 (-6.54, P < 0.001), at baseline. The mean (SD) MOS-SF-36 physical component summary score was 48.8 (10.6) at baseline, remained stable over the follow-up and did not differ across the three clusters. Mixed-effect linear regression univariate analyses found that the average 2-year follow-up MOS-SF-36 physical score was increased (compared with mostly abstainers) in mostly heavy drinkers (+4.44, P = 0.007); no other variables tested influenced the MOS-SF-36 physical score. CONCLUSION: Among individuals with alcohol dependence, a rapid improvement was seen in the mental dimension of QOL following treatment initiation, which was maintained during 24 months. Improvement was associated with the pattern of alcohol use, becoming close to the general population norm in patients classified as mostly abstainers, improving substantially in mostly moderate drinkers and improving only slightly in mostly heavy drinkers. The physical dimension of QOL was generally in the normal range but was not associated with drinking patterns.

Submerged Vanes for Flow Control and Bank Protection in Streams, HR-255, 1984

This study was conducted for the purpose of evaluating a new concept for a bank-protection structure: The Iowa Vane . The underlying idea involves countering the torque exerted on the primary flow by its curvature and vertical velocity gradient, thereby eliminating or significantly reducing the secondary flow and thus reducing the undermining of the outer banks and the high-velocity attack on it. The new structure consists of an array of short, vertical, submerged vanes installed with a certain orientation on the channel bed. A relatively small number of vanes can produce bend flows which are practically uniform across the channel. The height of the vanes is less than half the water depth, and their angle with the flow direction is of the order of l0 degrees. In this study, design relations have been established. The relations, and the vanes' overall performance, have been tested in a laboratory model under different flow and sediment conditions. The results are used for the design of an Iowa-Vane bank protection structure for a section of East Nishnabotna River along U.S. Highway 34 at Red Oak, Iowa.

Portable School Stop Signs and Other Non-Uniform School Stop Control Devices, HR-190, 1978

Portable (roll-out) stop signs are used at school crossings in over 300 cities in Iowa. Their use conforms to the Code of Iowa, although it is not consistent with the provisions of the Manual on Uniform Traffic Control Devices adopted for nationwide application. A survey indicated that most users in Iowa believe that portable stop signs provide effective protection at school crossings, and favor their continued use. Other non-uniform signs that fold or rotate to display a STOP message only during certain hours are used at school crossings in over 60 cities in Iowa. Their use does not conform to either the Code of Iowa or the Manual on Uniform Traffic Control Devices. Users of these devices also tend to favor their continued use. A survey of other states indicated that use of temporary devices similar to those used in Iowa is not generally sanctioned. Some unsanctioned use apparently occurs in several states, however. A different type of portable stop sign for school crossings is authorized and widely used in one state. Portable stop signs similar to those used in Iowa are authorized in another state, although their use is quite limited. A few reports in the literature reviewed for this research discussed the use of portable stop signs. The authors of these reports uniformly recommended against the use of portable or temporary traffic control devices. Various reasons for this recommendation were given, although data to support the recommendation were not offered. As part of this research, field surveys were conducted at 54 locations in 33 communities where temporary stop control devices were in use at school crossings. Research personnel observed the obedience to stop control and measured the vehicular delay incurred. Stopped delay averaged 1.89 seconds/entering vehicle. Only 36.6 percent of the vehicles were observed to come to a complete stop at the study locations controlled by temporary stop control devices. However, this level of obedience does not differ from that observed at intersections controlled by permanent stop signs. Accident experience was compiled for 76 intersections in 33 communities in Iowa where temporary stop signs were used and, for comparative purposes, at 76 comparable intersections having other forms of control or operating without stop control. There were no significant differences in accident experience An economic analysis of vehicle operating costs, delay costs, and other costs indicated that temporary stop control generated costs only about 12 percent as great as permanent stop control for a street having a school crossing. Midblock pedestrian-actuated signals were shown to be cost effective in comparison with temporary stop signs under the conditions of use assumed. Such signals could be used effectively at a number of locations where temporary stop signs are being used. The results of this research do not provide a basis for recommending that use of portable stop signs be prohibited. However, erratic patterns of use of these devices and inadequate designs suggest that improved standards for their use are needed. Accordingly, nine recommendations are presented to enhance the efficiency of vehicular flow at school crossings, without causing a decline in the level of pedestrian protection being afforded.

Glucose-dependent insulinotropic polypeptide receptor knockout mice are impaired in learning, synaptic plasticity, and neurogenesis.

Glucose-dependent insulinotropic polypeptide (GIP) is a key incretin hormone, released from intestine after a meal, producing a glucose-dependent insulin secretion. The GIP receptor (GIPR) is expressed on pyramidal neurons in the cortex and hippocampus, and GIP is synthesized in a subset of neurons in the brain. However, the role of the GIPR in neuronal signaling is not clear. In this study, we used a mouse strain with GIPR gene deletion (GIPR KO) to elucidate the role of the GIPR in neuronal communication and brain function. Compared with C57BL/6 control mice, GIPR KO mice displayed higher locomotor activity in an open-field task. Impairment of recognition and spatial learning and memory of GIPR KO mice were found in the object recognition task and a spatial water maze task, respectively. In an object location task, no impairment was found. GIPR KO mice also showed impaired synaptic plasticity in paired-pulse facilitation and a block of long-term potentiation in area CA1 of the hippocampus. Moreover, a large decrease in the number of neuronal progenitor cells was found in the dentate gyrus of transgenic mice, although the numbers of young neurons was not changed. Together the results suggest that GIP receptors play an important role in cognition, neurotransmission, and cell proliferation.

Circadian clock-dependent and -independent rhythmic proteomes implement distinct diurnal functions in mouse liver.

Diurnal oscillations of gene expression controlled by the circadian clock underlie rhythmic physiology across most living organisms. Although such rhythms have been extensively studied at the level of transcription and mRNA accumulation, little is known about the accumulation patterns of proteins. Here, we quantified temporal profiles in the murine hepatic proteome under physiological light-dark conditions using stable isotope labeling by amino acids quantitative MS. Our analysis identified over 5,000 proteins, of which several hundred showed robust diurnal oscillations with peak phases enriched in the morning and during the night and related to core hepatic physiological functions. Combined mathematical modeling of temporal protein and mRNA profiles indicated that proteins accumulate with reduced amplitudes and significant delays, consistent with protein half-life data. Moreover, a group comprising about one-half of the rhythmic proteins showed no corresponding rhythmic mRNAs, indicating significant translational or posttranslational diurnal control. Such rhythms were highly enriched in secreted proteins accumulating tightly during the night. Also, these rhythms persisted in clock-deficient animals subjected to rhythmic feeding, suggesting that food-related entrainment signals influence rhythms in circulating plasma factors.

Haemodialysis acutely reduces the plasma levels of ADMA without reversing impaired NO-dependent vasodilation.

End-stage renal disease patients have endothelial dysfunction and high plasma levels of ADMA (asymmetric omega-NG,NG-dimethylarginine), an endogenous inhibitor of NOS (NO synthase). The actual link between these abnormalities is controversial. Therefore, in the present study, we investigated whether HD (haemodialysis) has an acute impact on NO-dependent vasodilation and plasma ADMA in these patients. A total of 24 patients undergoing maintenance HD (HD group) and 24 age- and gender-matched healthy controls (Control group) were enrolled. The increase in forearm SkBF (skin blood flow) caused by local heating to 41 degrees C (SkBF41), known to depend on endothelial NO production, was determined with laser Doppler imaging. SkBF41 was expressed as a percentage of the vasodilatory reserve obtained from the maximal SkBF induced by local heating to 43 degrees C (independent of NO). In HD patients, SkBF41 was assessed on two successive HD sessions, once immediately before and once immediately after HD. Plasma ADMA was assayed simultaneously with MS/MS (tandem MS). In the Control group, SkBF41 was determined twice, on two different days, and plasma ADMA was assayed once. In HD patients, SkBF41 was identical before (82.2+/-13.1%) and after (82.7+/-12.4%) HD, but was lower than in controls (day 1, 89.6+/-6.1; day 2, 89.2+/-6.9%; P<0.01 compared with the HD group). In contrast, plasma ADMA was higher before (0.98+/-0.17 micromol/l) than after (0.58+/-0.10 micromol/l; P<0.01) HD. ADMA levels after HD did not differ from those obtained in controls (0.56+/-0.11 micromol/l). These findings show that HD patients have impaired NO-dependent vasodilation in forearm skin, an abnormality not acutely reversed by HD and not explained by ADMA accumulation.

Guidelines for Removal of Traffic Control Devices in Rural Areas, TR-527, 2005

It is commonly regarded that the overuse of traffic control devices desensitizes drivers and leads to disrespect, especially for low-volume secondary roads with limited enforcement. The maintenance of traffic signs is also a tort liability concern, exacerbated by unnecessary signs. The Federal Highway Administration’s (FHWA) Manual on Uniform Traffic Control Devices (MUTCD) and the Institute of Transportation Engineer’s (ITE) Traffic Control Devices Handbook provide guidance for the implementation of STOP signs based on expected compliance with right-of-way rules, provision of through traffic flow, context (proximity to other controlled intersections), speed, sight distance, and crash history. The approach(es) to stop is left to engineering judgment and is usually dependent on traffic volume or functional class/continuity of system. Although presently being considered by the National Committee on Traffic Control Devices, traffic volume itself is not given as a criterion for implementation in the MUTCD. STOP signs have been installed at many locations for various reasons which no longer (or perhaps never) met engineering needs. If in fact the presence of STOP signs does not increase safety, removal should be considered. To date, however, no guidance exists for the removal of STOP signs at two-way stop-controlled intersections. The scope of this research is ultra-low-volume (< 150 daily entering vehicles) unpaved intersections in rural agricultural areas of Iowa, where each of the 99 counties may have as many as 300 or more STOP sign pairs. Overall safety performance is examined as a function of a county excessive use factor, developed specifically for this study and based on various volume ranges and terrain as a proxy for sight distance. Four conclusions are supported: (1) there is no statistical difference in the safety performance of ultra-low-volume stop-controlled and uncontrolled intersections for all drivers or for younger and older drivers (although interestingly, older drivers are underrepresented at both types of intersections); (2) compliance with stop control (as indicated by crash performance) does not appear to be affected by the use or excessive use of STOP signs, even when adjusted for volume and a sight distance proxy; (3) crash performance does not appear to be improved by the liberal use of stop control; (4) safety performance of uncontrolled intersections appears to decline relative to stop-controlled intersections above about 150 daily entering vehicles. Subject to adequate sight distance, traffic professionals may wish to consider removal of control below this threshold. The report concludes with a section on methods and legal considerations for safe removal of stop control.

Spatiotemporal adaptive multiscale multiphysics simulations of two-phase flow

We present a spatiotemporal adaptive multiscale algorithm, which is based on the Multiscale Finite Volume method. The algorithm offers a very efficient framework to deal with multiphysics problems and to couple regions with different spatial resolution. We employ the method to simulate two-phase flow through porous media. At the fine scale, we consider a pore-scale description of the flow based on the Volume Of Fluid method. In order to construct a global problem that describes the coarse-scale behavior, the equations are averaged numerically with respect to auxiliary control volumes, and a Darcy-like coarse-scale model is obtained. The space adaptivity is based on the idea that a fine-scale description is only required in the front region, whereas the resolution can be coarsened elsewhere. Temporal adaptivity relies on the fact that the fine-scale and the coarse-scale problems can be solved with different temporal resolution (longer time steps can be used at the coarse scale). By simulating drainage under unstable flow conditions, we show that the method is able to capture the coarse-scale behavior outside the front region and to reproduce complex fluid patterns in the front region.

Fluorescence flow cytometer to determine urine particle reference intervals in doping control samples.

Background: Urine is still the matrix of choice to fight against doping, because it can be collected non-invasively during anti-doping tests. Most of the World Anti-Doping Agency's accredited laboratories have more than 20 years experience in analyzing this biological fluid and the majority of the compounds listed in the 2010 Prohibited List - International Standard are eliminated through the urinary apparatus. Storing and transporting urine samples for doping analyses does not include a specific protocol to prevent microbial and thermal degradation. The use of a rapid and reliable screening method could enable determine reference intervals for urine specimens in doping control samples and evaluate notably the prevalence of microbial contamination known to be responsible for the degradation of chemical substances in urine.Methods: The Sysmex(R) UF-500i is a recent urine flow cytometer analyzer capable of quantifying BACT and other urinary particles such as RBC, WBC, EC, DEBRIS, CAST, PATH. CAST, YLC, SRC as well as measuring urine conductivity. To determine urine anti-doping reference intervals, 501 samples received in our laboratory over a period of two months were submitted to an immediate examination. All samples were collected and then transported at room temperature. Analysis of variance was performed to test the effects of factors such as gender, test type [in-competition, out-of-competition] and delivery time.Results: The data obtained showed that most of the urine samples were highly contaminated with bacteria. The other urine particles were also very different according to the factors.Conclusions: The Sysmex(R) UF-500i was capable of providing a snapshot of urine particles present in the samples at the time of the delivery to the laboratory. These particles, BACT in particular, gave a good idea of the possible microbial degradation which had and/or could have occurred in the sample. This information could be used as the first quality control set up in WADA (World Anti-Doping Agency) accredited laboratories to determine if steroid profiles, endogenous and prohibited substances have possibly been altered. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

Logistics Development in Finnish and Swedish Companies with Respect of Russia and Four Asian Countries: Traffic Flow and Warehousing Analysis from Current Situation and Likely Development Trends

On selvää, että tänä päivänä maailmankaupan painopiste on hiljalleen siirtymässä Aasiaan ja varsinkin Kiina on ollut huomion keskipisteessä. Erityisesti valmistavien yritysten perspektiivistä muutos on ollut merkittävä ja tämä tosiasia kasvattaa yrityksissä paineita luoda kustannustehokkaita toimitusketjuratkaisuja,joiden vasteaika on mahdollisimman lyhyt. Samaan aikaan kun tarkastellaan kuljetusvirtoja, huomattaan että maanosien välillä on suuri epätasapaino. Tämä on enimmäkseen seurausta suurten globaalisti toimivien yritysten toimitusketjustrategioista. Useimmat näistä toimijoista optimoivat verkostonsa turvautumalla 'paikalliseen hankintaan', jotta he voisivat paremmin hallita toimitusketjujaan ja saada näitä reagointiherkimmiksi. Valmistusyksiköillä onkin monesti Euroopassa pakko käyttää kalliita raaka-aineita ja puolivalmisteita. Kriittisiksi tekijöiksi osoittautuvat kuljetus- ja varastointikustannukset sekä näiden seurauksena hukka-aika, joka aiheutuu viivästyksistä. Voidakseen saavuttaa optimiratkaisun, on tehtävä päätös miten tuotteet varastoidaan: keskitetysti tai hajautetusti ja integroida tämä valinta sopivien kuljetusmuotojen kanssa. Aasiasta Pohjois-Eurooppaan on halpaa käyttää merikuljetusta, mutta operaatio kestää hyvin pitkään - joissain tapauksessa jopa kahdeksan viikkoa. Toisaalta lentokuljetus on sekä kallis että rajoittaa siirrettävien tuotteiden eräkokoa.On olemassa kolmaskin vaihtoehto, josta voisi olla ratkaisuksi: rautatiekuljetus on halvempi kuin lentokuljetus ja vasteajat ovat lyhyemmät kuin merikuljetuksissa. Tässä tutkimuksessa tilannetta selvitetään kyselyllä, joka suunnattiin Suomessa ja Ruotsissa toimiville yrityksille. Tuloksien perusteella teemme johtopäätökset siitä, mitkä kuljetusmuotojen markkinaosuudet tulevat olemaan tulevaisuudessa sekä luomme kuvan kuljetusvirroista Euroopan, Venäjän, Etelä-Korea, Intian, Kiinan ja Japanin välillä. Samalla on tarkoitus ennakoida sitä, miten tarkastelun kohteena olevat yritykset aikovat kehittää kuljetuksiaan ja varastointiaan tulevien vuosien aikana. Tulosten perusteella näyttää siltä, että seuraavan viiden vuoden kuluessa kuljetuskustannukset eivät merkittävissä määrin tule muuttuman ja meri- sekä kumipyöräkuljetukset pysyvät suosituimpina vaihtoehtoina.Kuitenkin lentokuljetusten osuus laskee hiukan, kun taas rautatiekuljetusten painotus kasvaa. Tulokset paljastavat, että Kiinassa ja Venäjällä kuljetettava konttimäärä kasvaa; Intiassa tulos on saman suuntainen, joskaan ei niin voimakas. Analyysimme mukaan kuljetusvirtoihin liittyvä epätasapaino säilyy Venäjän kuljetusten suhteen: yritykset jatkavat tulevaisuudessakin vientiperusteista strategiaansa. Varastoinnin puolella tunnistamme pienemmän muutoksen, jonka mukaan pienikokoisten varastojen määrät todennäköisesti vähenevät tulevaisuudessa ja kiinnostus isoja varastoja kohtaan lisääntyy. Tässä kohtaa on mainittava, että suomalaisilla yrityksillä on enemmän varastoja Keski- ja Itä-Euroopassa verrattuna ruotsalaisiin toimijoihin, jotka keskittyvät selkeämmin Länsi-Euroopan maihin. Varastoja yrityksillä on molemmissa tapaukissa paljolti kotimaassaan. Valitessaan varastojensa sijoituskohteita yritykset painottavat seuraavia kriteereitä: alhaiset jakelukustannukset, kokoamispaikan/valmistustehtaan läheisyys, saapuvan logistiikan integroitavuus ja saatavilla olevat logistiikkapalvelut. Tutkimuksemme lopussa päädymme siihen, että varastojen sijoituspaikat eivät muutu satamien rakenteen ja liikenneyhteyksien takia kovinkaan nopeasti.