Risk associations for intestinal parasites in symptomatic and asymptomatic schoolchildren in central Mozambique.

OBJECTIVES: Chronic infections by enteric parasites including protist and helminthic species produce long-term sequelae on the health status of infected children. This study assesses potential associations linked with enteric parasite infections in symptomatic and asymptomatic children in Zambézia province, Mozambique. METHODS: In this prospective cross-sectional study, stool samples and epidemiological questionnaires on demographics and risk associations were collected from symptomatic children (n = 286) from clinical settings and asymptomatic (n = 807) children from 17 schools and creches aged 3‒14 years. We detected enteric parasites using PCR-based methods. We calculated prevalence (adjusted for age, sex, house construction, drinking water, and latrine use) and odds ratios (ORs) for risk associations with logistic regression, after adjusting for district, neighbourhood and symptoms. RESULTS: Numbers and adjusted prevalence (95% confidence intervals in parentheses) for the symptomatic and asymptomatic populations were Giardia duodenalis 120, 52% (22-82), 339, 42% (25-59); followed by Strongyloides stercoralis 52, 14% (9‒20), 180, 20% (15-25). Risk associations for G. duodenalis included drinking untreated river/spring water, OR 2.91 (1.80-4.70); contact with ducks, OR 14.96 (2.93‒76.31); dogs, OR 1.92 (1.04-3.52); cats, OR 1.73 (1.16-2.59), and a relative with diarrhoea, OR 2.59 (1.54‒4.37). Risk associations for S. stercoralis included having no latrine, OR 2.41 (1.44-4.02); drinking well water, OR 1.82 (1.02-3.25), and increasing age, OR 1.11 (1.04-1.20). CONCLUSIONS: We found a high prevalence of intestinal parasites regardless of the children's symptoms. Drinking well or river water, domestic animals, and latrine absence were contributing factors of human infections.

The battle of two ions: Ca2+ signalling against Na+ stress.

Soil salinity adversely affects plant growth, crop yield and the composition of ecosystems. Salinity stress impacts plants by combined effects of Na+ toxicity and osmotic perturbation. Plants have evolved elaborate mechanisms to counteract the detrimental consequences of salinity. Here we reflect on recent advances in our understanding of plant salt tolerance mechanisms. We discuss the embedding of the salt tolerance-mediating SOS pathway in plant hormonal and developmental adaptation. Moreover, we review newly accumulating evidence indicating a crucial role of a transpiration-dependent salinity tolerance pathway, that is centred around the function of the NADPH oxidase RBOHF and its role in endodermal and Casparian strip differentiation. Together, these data suggest a unifying and coordinating role for Ca2+ signalling in combating salinity stress at the cellular and organismal level.

Calcium Signaling during Salt Stress and in the Regulation of Ion Homeostasis.

Soil composition largely defines the living conditions of plants and represents one of their most relevant, dynamic and complex environmental cues. The effective concentrations of many either tolerated or essential ions and compounds in the soil usually differ from the optimum that would be most suitable for plants. In this regard, salinity - caused by excess of NaCl - represents a widespread adverse growth condition but also shortage of ions like K+, NO3- and Fe2+ restrains plant growth. During the past years many components and mechanisms that function in the sensing and establishment of ion homeostasis have been identified and characterized. Here, we reflect on recent insights that extended our understanding of components and mechanisms, which govern and fine-tune plant salt stress tolerance and ion homeostasis. We put special emphasis on mechanisms that allow for interconnection of the salt overly sensitivity pathway with plant development and discuss newly emerging functions of Ca2+ signaling in salinity tolerance. Moreover, we review and discuss accumulating evidence for a central and unifying role of Ca2+ signaling and Ca2+ dependent protein phosphorylation in regulating sensing, uptake, transport and storage processes of various ions. Finally, based on this cross-field inventory, we deduce emerging concepts and arising questions for future research.

A novel technique for single-shot energy-resolved 2D x-ray imaging of plasmas relevant for the inertial confinement fusion.

A novel x-ray diagnostic of laser-fusion plasmas is described, allowing 2D monochromatic images of hot, dense plasmas to be obtained in any x-ray photon energy range, over a large domain, on a single-shot basis. The device (named energy-encoded pinhole camera) is based upon the use of an array of many pinholes coupled to a large area CCD camera operating in the single-photon mode. The available x-ray spectral domain is only limited by the quantum efficiency of scientific-grade x-ray CCD cameras, thus extending from a few keV up to a few tens of keV. Spectral 2D images of the emitting plasma can be obtained at any x-ray photon energy provided that a sufficient number of photons had been collected at the desired energy. Results from recent inertial confinement fusion related experiments will be reported in order to detail the new diagnostic.

Directional bremsstrahlung from a Ti laser-produced x-ray source at relativistic intensities in the 3-12 keV range.

Front and rear side x-ray emission from thin titanium foils irradiated by ultraintense laser pulses at intensities up to ≈5 × 10(19) W/cm2 was measured using a high-resolution imaging system. Significant differences in intensity, dimension, and spectrum between front and rear side emission intensity in the 3-12 keV photon energy range was found even for 5 µm thin Ti foils. Simulations and analysis of space-resolved spectra explain this behavior in terms of directional bremsstrahlung emission from fast electrons generated during the interaction process.

Novel x-ray multispectral imaging of ultraintense laser plasmas by a single-photon charge coupled device based pinhole camera.

Spectrally resolved two-dimensional imaging of ultrashort laser-produced plasmas is described, obtained by means of an advanced technique. The technique has been tested with microplasmas produced by ultrashort relativistic laser pulses. The technique is based on the use of a pinhole camera equipped with a charge coupled device detector operating in the single-photon regime. The spectral resolution is about 150 eV in the 4-10 keV range, and images in any selected photon energy range have a spatial resolution of 5 microm. The potential of the technique to study fast electron propagation in ultraintense laser interaction with multilayer targets is discussed and some preliminary results are shown.

A comparison of the beta-D-xyloside, odiparcil, to warfarin in a rat model of venous thrombosis.

BACKGROUND: A significant need exists for new chronic oral anticoagulation therapies to replace warfarin. Previous studies have shown that beta-D-xylosides, which prime glycosaminoglycan (GAG) synthesis, have antithrombin and antithrombotic activity. In the following report, a new orally active beta-D-xyloside (odiparcil) has been characterized in a rat model of venous thrombosis and its efficacy and bleeding liability compared to warfarin. Additionally, studies were conducted to investigate odiparcil's ex vivo antithrombin and antiplatelet activity, and also to explore the potential utility of protamine sulfate as a neutralizing agent. METHODS AND RESULTS: In vivo thrombosis studies were conducted in a rat inferior vena cava model, and bleeding studies in a rat tail transection model. Following oral dosing, warfarin and odiparcil produced dose-related suppression of thrombus formation. A therapeutically relevant dose of warfarin in this model (international normalized ratio; INR 3.0) achieved approximately 65% inhibition of thrombus formation. Warfarin caused dose-related significant increases in bleeding indices. Odiparcil antithrombotic activity was limited by its mechanism to a maximum suppression of thrombus formation of 65-70%, and did not prolong bleeding indices. Additionally, odiparcil-induced heparin cofactor II (HCII)-dependent antithrombin activity was shown to be a function of dermatan sulfate-like GAG production. Other than thrombin-related effects, no odiparcil effects on platelet function were observed. In antidote studies, it was demonstrated that odiparcil-induced antithrombotic activity could be partially neutralized by protamine sulfate. CONCLUSIONS: These experiments suggest that an antithrombotic approach based upon xyloside induction of circulating GAGs may have the potential to approximate the efficacy of warfarin and yet with a reduced risk to hemostasis.

Prediction of portwine stain clearance and required number of flashlamp pumped pulsed dye laser treatments.

BACKGROUND AND OBJECTIVE: Portwine stain (PWS) response to flashlamp pumped pulsed dye laser (FPPDL) treatment is variable and unpredictable. Our aim was to develop a model to predict treatment outcome and the required number of treatments. STUDY DESIGN/MATERIALS AND METHODS: We hypothesized that PWS clearance decreases exponentially with the number of treatments, as a consequence of the exponential decrease of laser light fluence in human skin. Of 70 patients with a PWS in the head/neck area, the color difference between normal skin and PWS was measured with a chromameter, prior to, and at several stages during FPPDL treatment. Through the obtained values of each of the four color parameters, DeltaL*, Deltaa*, Deltab*, and DeltaE, mono-exponentially decreasing functions were fitted. RESULTS: All four color parameters showed decreasing exponential functions, with decay rates that were not significantly different, and a mean R(2) of 0.6, indicating a reliable fit-quality. CONCLUSIONS: Our model suggests that individual prediction of treatment outcome and the required number of treatments is possible in an early stage of FPPDL treatment of PWS, in theory already after one single laser treatment.

Small GTP-binding protein Ral modulates regulated exocytosis of von Willebrand factor by endothelial cells.

Weibel-Palade bodies are endothelial cell-specific organelles, which contain von Willebrand factor (vWF), P-selectin, and several other proteins. Recently, we found that the small GTP-binding protein Ral is present in a subcellular fraction containing Weibel-Palade bodies. In the present study, we investigated whether Ral is involved in the regulated exocytosis of Weibel-Palade bodies. Activation of endothelial cells by thrombin resulted in transient cycling of Ral from its inactive GDP-bound to its active GTP-bound state, which coincided with release of vWF. Ral activation and exocytosis of Weibel-Palade bodies were inhibited by incubation with trifluoperazine, an inhibitor of calmodulin, before thrombin stimulation. Functional involvement of Ral in exocytosis was further investigated by the expression of constitutively active and dominant-negative Ral variants in primary endothelial cells. Introduction of active Ral G23V resulted in the disappearance of Weibel-Palade bodies from endothelial cells. In contrast, the expression of the dominant-negative Ral S28N did not affect the amount of Weibel-Palade bodies in transfected cells. These results indicate that Ral is involved in regulated exocytosis of Weibel-Palade bodies by endothelial cells.

Histologic evaluation of skin damage after overlapping and nonoverlapping flashlamp pumped pulsed dye laser pulses: A study on normal human skin as a model for port wine stains.

BACKGROUND AND OBJECTIVE: In the treatment of port wine stains (PWS) with the flashlamp pumped pulsed dye laser (FPPDL), no consensus exists about overlapping of pulses. The advantage of overlapping pulses is homogeneous lightening of the PWS; the risk is redundant tissue damage. The aim of this study was to determine the histopathologic effect on human skin of pulsed dye laser pulses with various degrees of overlap, with normal human skin as a model for PWS. STUDY DESIGN/MATERIALS AND METHODS: Eighteen healthy white volunteers were irradiated with pulsed dye laser pulses with increasing radiant exposure and with different degrees of overlap. Biopsy samples were taken and histologically analysed. RESULTS: Overlapping of pulses on normal human skin enhances depth of vascular damage with approximately 30%. Adjacent pulses also show this effect. We found no histologic signs of serious damage to epidermis or dermal connective tissue by using radiant exposure levels of 6-8 J/cm(2), regardless of pulse application. CONCLUSIONS: Reasoning that the mechanism of tissue injury is comparable for normal and PWS skin, we conclude that it is safe to treat PWS with overlapping FPPDL pulses to achieve homogeneous lightening.

Comparing the antithrombotic efficacy of a humanized anti-factor IX(a) monoclonal antibody (SB 249417) to the low molecular weight heparin enoxaparin in a rat model of arterial thrombosis.

A humanized inhibitory anti-factor IX(a) antibody (SB 249417) has been compared to enoxaparin (Lovenox) in a rat model of arterial thrombosis. Pretreatment of rats with either SB 249417 (3.0 mg/kg, i. v.) or enoxaparin (30.0 mg/kg, i.v. or s.c.) resulted in comparable and significant reductions in thrombus formation. However, the efficacious dose of enoxaparin resulted in >30-fold increase in the aPTT over baseline, while the efficacious dose of SB 249417 prolonged the aPTT by only approximately 3-fold. Additionally, pretreatment with SB 249417 resulted in sustained blood flow and arterial patency throughout the experiment in >80% of rats treated. In contrast, <30% of rats pretreated with enoxaparin remained patent throughout the experiment. The data in this report indicate that the selective inhibition of factor IX(a) with the monoclonal antibody SB 249417 produces a superior antithrombotic profile to that of the low molecular weight heparin enoxaparin.

Monocyte chemoattractant protein-1 is upregulated in rats with volume-overload congestive heart failure.

BACKGROUND: Chemokines are potent proinflammatory and immune modulators. Increased expression of chemokines, eg, monocyte chemoattractant protein-1 (MCP-1), has recently been described in clinical and experimental heart failure. The present report is aimed at exploring the expression, localization, and binding site regulation of MCP-1, a member of the C-C chemokine family, in a rat model of volume-overload congestive heart failure (CHF). METHODS AND RESULTS: An aortocaval fistula was surgically created between the abdominal aorta and inferior vena cava. Rats with CHF were further subdivided into compensated and decompensated subgroups. Northern blot analysis and real-time quantitative polymerase chain reaction demonstrated upregulation of MCP-1 mRNA expression correlating with the severity of CHF (288+/-22, 502+/-62, and 826+/-138 copies/ng total RNA for sham, compensated, and decompensated animals, respectively; n=5, P:<0.05). MCP-1 protein was localized by immunohistochemistry in cardiomyocytes, vascular endothelium and smooth muscle cells, infiltrating leukocytes, and interstitial fibroblasts, and its intensity increased with severity of CHF. In addition, rats with CHF displayed a significant decrease of (125)I-labeled MCP-1 binding sites to myocardium-derived membranes (384.3+/-57.0, 181.3+/-8.8, and 123.3+/-14.1 fmol/mg protein for sham, compensated, and decompensated animals, respectively). CONCLUSIONS: Volume-overload CHF in rats is associated with alterations in the expression, immunohistochemical localization, and receptor binding of the MCP-1 chemokine in the myocardium. These changes were more pronounced in rats with decompensated CHF. The data suggest that activation of the MCP-1 system may contribute to the progressive cardiac decompensation and development of CHF in rats with aortocaval fistula.

An inhibitory anti-factor IX antibody effectively reduces thrombus formation in a rat model of venous thrombosis.

An inhibitory anti-factor IX/IXa antibody (BC2) has been investigated as an anti-thrombotic agent in a rat venous thrombosis model. The treatment of rats post-injury with a single bolus dose of BC2 (3 mg/kg, i.v.) resulted in an approximately 4 fold reduction in venous thrombus mass (P = 0.043). This efficacy was matched by a minimal (<2.5 fold) prolongation of the aPTT and had no effect on the prothrombin time (PT). Heparin by comparison, given as a bolus followed by continuous infusion, at doses comparable in efficacy at reducing thrombus formation, prolonged the aPTT >50 fold. These results demonstrate that the anti-factor IX/IXa antibody (BC2), when compared to heparin, can effectively reduce venous thrombosis with less disruptive consequences on blood clotting.

Antithrombotic efficacy of a novel murine antihuman factor IX antibody in rats.

A murine antihuman factor IX monoclonal antibody (BC2) has been generated and evaluated for its capacity to prolong the activated partial thromboplastin time (aPTT) in vitro and ex vivo and to prevent arterial thrombosis in a rat model in vivo. BC2 extended aPTT to a maximum of 60 to 80 seconds at 100 to 1000 nmol/L in vitro (rat and human plasma, respectively) and ex vivo (rat) after dosing of rats up to 6 mg/kg in vivo. BC2, administered as bolus (1 to 6 mg/kg) followed by infusion (0.3 to 2 mg x kg(-1) x h(-1)), dose-dependently prevented thrombosis of an injured rat carotid artery (FeCl(3)-patch model), increased time to artery occlusion, and reduced incidence of vessel occlusion. BC2 efficacy in preventing arterial thrombosis exceeded that of heparin (bolus 15 to 120 U/kg followed by infusion 0.5 to 4.0 U x kg(-1) x min(-1)), whereas the latter rendered the blood incoagulable (aPTT>1000 seconds). BC2 demonstrated complete antithrombotic efficacy also as a single bolus given either as prevessel or postvessel injury as evidenced by reduction of thrombus mass (from 4.18+/-0.49 to 1.80 +/-0.3 mg, P<0.001), increasing vessel patency time (from 14.9+/-0.9 minutes to 58.3+/-1.7 minutes, P<0.001) and decreasing incidence of vessel occlusion from 100% to 0% in vehicle- versus BC2-treated rats, respectively. BC2 (3 mg/kg, IV) administered in a single bolus resulted in 50% reduction in thrombus mass (P<0.01), extended vessel patency time (P<0.001), extended aPTT only 4-fold, and had no effect on blood loss via a tail surgical wound; heparin, at doses that reduced thrombus mass to a similar extent, extended aPTT beyond 1000 seconds (over 500-fold) and increased blood loss from 1.8+/-0.7 to 3.3 +/-0.6 mL (P<0.001). These data suggest that BC2 may provide enhanced therapeutic efficacy in humans at lesser interference with blood hemostasis than heparin.

Small GTP-binding protein RalA associates with Weibel-Palade bodies in endothelial cells.

In endothelial cells von Willebrand factor (vWF) and P-selectin are stored in dense granules. so-called Weibel-Palade bodies. Upon stimulation of endothelial cells with a variety of agents including thrombin, these organelles fuse with the plasma membrane and release their content. Small GTP-binding proteins have been shown to control release from intracellular storage pools in a number of cells. In this study we have investigated whether small GTP-binding proteins are associated with Weibel-Palade bodies. We isolated Weibel-Palade bodies by centrifugation on two consecutive density gradients of Percoll. The dense fraction in which these subcellular organelles were highly enriched, was analysed by SDS-PAGE followed by GTP overlay. A distinct band with an apparent molecular weight of 28,000 was observed. Two-dimensional gel electrophoresis followed by GTP overlay revealed the presence of a single small GTP-binding protein with an isoelectric point of 7.1. A monoclonal antibody directed against RalA showed reactivity with the small GTP-binding protein present in subcellular fractions that contain Weibel-Palade bodies. The small GTPase RalA was previously identified on dense granules of platelets and on synaptic vesicles in nerve terminals. Our observations suggest that RalA serves a role in regulated exocytosis of Weibel-Palade bodies in endothelial cells.

Effects of the novel antimigraine agent, frovatriptan, on coronary and cardiac function in dogs.

The effects of frovatriptan (VML 251/SB-209509) on coronary artery function were investigated in isolated coronary arteries from beagle dogs. Low concentrations of frovatriptan produced contraction with -logEC50 7.55 +/- 0.08 (n = 11). The maximal observed contraction attained was 56 +/- 7% of the control 5-hydroxytryptamine (5-HT; 10 microM) response. At high concentrations of frovatriptan (>6 microM), reversal of sumatriptan (10 microM)-induced contractions was noted. In arteries precontracted with the thromboxane mimetic, U46619, frovatriptan produced a bell-shaped concentration-response relation with a maximal response at 600 nM. Concentrations of frovatriptan >2 microM produced marked reversal of tone, with full relaxation of precontracted tissues at 200 microM. In anesthetized, open-chest mongrel dogs, intravenous (n = 5) or intracoronary (n = 5) artery administration of frovatriptan (0.0001-1 mg/kg) had no consistent effect on left ventricular end-diastolic pressure, left end-systolic pressure, cardiac contractility, aortic blood flow, systemic peripheral resistance, coronary blood flow, coronary vascular resistance, mean arterial blood pressure, or heart rate when compared with vehicle (n = 3). Intravenous sumatriptan produced minor effects on blood pressure and heart rate. Intracoronary artery administration of sumatriptan (0.0003 mg/kg) produced an increase in systemic peripheral resistance to 120.5 +/- 8.2% compared with vehicle (97.8 +/- 5.4%; p < 0.05). This dose of sumatriptan also produced a significant increase in coronary blood flow and decrease in coronary vascular resistance. Intravenous administration of sumatriptan produced a dose-related reduction in left ventricular diastolic pressure with a reduction to 58.3 +/- 8.3% and 41.7 +/- 25% of control values observed at 0.3 and 1 mg/kg, respectively; however, administration of sumatriptan by an intracoronary route had no effect. In a model of myocardial infarction, comparable doses of sumatriptan (1.0 mg/kg) or frovatriptan (0.1 mg/kg), in terms of their effect on carotid vascular resistance, had no significant effect on infarct size. Frovatriptan had no effect on coronary blood flow after reperfusion; however, sumatriptan produced a significant reduction in coronary blood flow for < or =3 h. These studies show that frovatriptan has the capability of relaxing coronary arteries in vitro, has no overall effect on cardiac function at rest with no effect on coronary hemodynamics after myocardial infarction, and has a profile superior to that of sumatriptan.

Small GTP-binding proteins in human endothelial cells.

Small GTP-binding proteins of the Ras superfamily control an extensive number of intracellular events by alternating between GDP- and GTP-bound conformation. The presence of members of this protein family was examined in human umbilical vein endothelial cells employing RT-PCR. Sequence analysis of 215 cDNA clones revealed the presence of a total of 28 different partial cDNAs encoding small GTP-binding proteins. Two sequences corresponded to novel isoforms of Rab2 and Rab9. In addition, human analogues of Rab4b, Rab7, Rab9, Rab14 and Rab15 were identified. Besides Rab proteins, members of other subfamilies were detected as well. As a first step towards elucidation of the function of the different small GTP-binding proteins identified we have isolated full length cDNA corresponding to Rab30 from a human endothelial cell cDNA library. In order to assess the subcellular localization of Rab30, we expressed epitope-tagged Rab30 cDNA in monkey kidney COS-1 cells. Immunoelectron-microscopy of transfected COS-1 cells indicated that Rab30 is associated with Golgi stacks.

Characterization of portwine stain disfigurement.

Portwine stain disfigurement is caused by several factors. To what extent and in which proportion these factors influence the overall perceived disfigurement is incompletely understood. In this study, the contribution of seven portwine stain characteristics to overall portwine stain disfigurement was assessed. Color slides were taken from 90 patients with untreated portwine stains in the head/neck area. From these slides, overall portwine stain disfigurement was judged by a panel of 16 lay persons. The reliability of the average ratings of this panel was established with weighted kappa analysis (kappa = 0.51) and by calculating the Cronbach alpha coefficient (0.99). Using a previously tested multi-item questionnaire, the following portwine stain characteristics were rated quantitatively by a panel of five professionals: color, patchiness, boundary, size, shape, surface structure, and hypertrophy of the underlying tissue. By means of multiple linear regression analysis, the ratings for overall portwine stain disfigurement (panel of lay persons) were compared with the ratings for the individual portwine stain characteristics (panel of professionals). From the results of this analysis, the percentual contribution of each of the characteristics to overall portwine stain disfigurement was calculated. Size turned out to be the most important portwine stain characteristic, being responsible for almost half of the overall disfigurement. Color and boundary are the next two most important characteristics, contributing 18.7 and 12.4 percent, respectively. The other four characteristics together account for 10 percent. In our model, 13 percent of overall portwine stain disfigurement remains unexplained. We expect patient features to account for this. We feel that these results may have consequences for laser treatment of portwine stains. Reducing the size and fading out the boundary of the stain probably reduce overall portwine stain disfigurement more effectively than primarily trying to lighten the often persistent center of the stain.