The oxidative stress induced in vivo by Shiga toxin-2 contributes to the pathogenicity of haemolytic uraemic syndrome.

Typical haemolytic uraemic syndrome (HUS) is caused by Shiga toxin (Stx)-producing Escherichia coli infections and is characterized by thrombotic microangiopathy that leads to haemolytic anaemia, thrombocytopenia and acute renal failure. Renal or neurological sequelae are consequences of irreversible tissue damage during the acute phase. Stx toxicity and the acute inflammatory response raised by the host determine the development of HUS. At present there is no specific therapy to control Stx damage. The pathogenic role of reactive oxygen species (ROS) on endothelial injury has been largely documented. In this study, we investigated the in-vivo effects of Stx on the oxidative balance and its contribution to the development of HUS in mice. In addition, we analysed the effect of anti-oxidant agents as therapeutic tools to counteract Stx toxicity. We demonstrated that Stx induced an oxidative imbalance, evidenced by renal glutathione depletion and increased lipid membrane peroxidation. The increased ROS production by neutrophils may be one of the major sources of oxidative stress during Stx intoxication. All these parameters were ameliorated by anti-oxidants reducing platelet activation, renal damage and increasing survival. To conclude, Stx generates a pro-oxidative state that contributes to kidney failure, and exogenous anti-oxidants could be beneficial to counteract this pathogenic pathway.

Tolerance to lipopolysaccharide promotes an enhanced neutrophil extracellular traps formation leading to a more efficient bacterial clearance in mice.

Tolerance to lipopolysaccharide (LPS) constitutes a stress adaptation, in which a primary contact with LPS results in a minimal response when a second exposure with the same stimulus occurs. However, active important defence mechanisms are mounted during the tolerant state. Our aim was to assess the contribution of polymorphonuclear neutrophils (PMN) in the clearance of bacterial infection in a mouse model of tolerance to LPS. After tolerance was developed, we investigated in vivo different mechanisms of bacterial clearance. The elimination of a locally induced polymicrobial challenge was more efficient in tolerant mice both in the presence or absence of local macrophages. This was related to a higher number of PMN migrating to the infectious site as a result of an increased number of PMN from the marginal pool with higher chemotactic capacity, not because of differences in their phagocytic activity or reactive species production. In vivo, neutrophils extracellular trap (NET) destruction by nuclease treatment abolished the observed increased clearance in tolerant but not in control mice. In line with this finding, in vitro NETs formation was higher in PMN from tolerant animals. These results indicate that the higher chemotactic response from an increased PMN marginal pool and the NETs enhanced forming capacity are the main mechanisms mediating bacterial clearance in tolerant mice. To sum up, far from being a lack of response, tolerance to LPS causes PMN priming effects which favour distant and local anti-infectious responses.

[Radiological diagnostic approach to idiopathic interstitial pneumonias: findings in high resolution computed tomography]. / Aproximación al diagnóstico radiológico de las neumonías intersticiales idiopáticas. Hallazgos en tomografía computarizada de alta resolución.

A review is presented on the histological and radiological findings in idiopathic interstitial pneumonias, which are included among the diffuse parenchymal lung diseases. Although they may affect other compartments, the lung interstitium is the initial substrate of the parenchymal lesion due to different patterns of inflammation and fibrosis. The current classification, proposed in 2002 as an international multidisciplinary consensus document promoted by the American Thoracic Society and the European Respiratory Society, includes 7 conditions. Based on histological criteria, each histological pattern is associated with an image pattern. They are a group of conditions of unknown origin with common characteristics and differential features that enable them to be individualised as diseases with a different prognosis and treatment. They are rare as idiopathic forms, but share a morphological substrate with other more common diseases of unknown cause, which means they have to be excluded to reach a definitive diagnosis. For this reason it is important that the radiologist is familiar with their characteristic imaging findings.

Mifepristone (RU486) restores humoral and T cell-mediated immune response in endotoxin immunosuppressed mice.

Sepsis and septic shock can be caused by Gram-positive and -negative bacteria and other microorganisms. In the case of Gram-negative bacteria, endotoxin, a normal constituent of the bacterial wall, also known as lipopolysaccharide (LPS), has been considered as one of the principal agents causing the undesirable effects in this critical illness. The response to LPS involves a rapid secretion of proinflammatory cytokines such as tumour necrosis factor (TNF)-α, interleukin (IL)-1, IL-6, interferon (IFN)-γ and the concomitant induction of anti-inflammatory mediators such as IL-10, transforming growth factor (TGF)-ß or glucocorticoids, which render the host temporarily refractory to subsequent lethal doses of LPS challenge in a process known as LPS or endotoxin tolerance. Although protective from the development of sepsis or systemic inflammation, endotoxin tolerance has also been pointed out as the main cause of the non-specific humoral and cellular immunosuppression described in these patients. In this report we demonstrate, using a mouse model, that mifepristone (RU486), a known glucocorticoid receptor antagonist, could play an important role in the restoration of both adaptive humoral and cellular immune response in LPS immunosuppressed mice, suggesting the involvement of endogenous glucocorticoids in this phenomenon. On the other hand, using cyclophosphamide and gemcitabine, we demonstrated that regulatory/suppressor CD4(+) CD25(+) forkhead boxP3(+) and GR-1(+) CD11b(+) cells do not play a major role in the establishment or the maintenance of endotoxin tolerance, a central mechanism for inducing an immunosuppression state.

Renal damage and death in weaned mice after oral infection with Shiga toxin 2-producing Escherichia coli strains.

Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 infections are considered a public health problem in both developed and developing countries because of their increasing incidence and the severity of clinical presentation. Approximately 10% of infected patients develop complications such as haemolytic uraemic syndrome (HUS) characterized by acute renal failure, thrombocytopenia and haemolytic anaemia. The precise sequence of events leading to HUS is still understood incompletely. Because of the lack of a reproducible small animal model for EHEC infections, in vivo studies examining EHEC-host early interactions are limited and insufficient. The aim of this study was to characterize the weaned BALB/c mouse as a model of E. coli O157:H7 infection. In this paper we report that human Shiga toxin 2 (Stx2)-producing EHEC strains can adhere to the intestinal epithelium of weaned BALB/c mice, and produce local damage which leads to systemic disease and death in a percentage of infected mice. The lethality of the EHEC strain is closely age-dependent, and is related to the bacterial ability to colonize intestine and to produce Stx2. It can be concluded that the weaned BALB/c mouse can be used as a small animal model to study host early responses, and the role of bacterial pathogenic factors in the induction of systemic disease, thus providing a useful tool for the evaluation of therapeutic or vaccine approaches.

Air-soil exchange of mercury from background soils in the United States.

The air-surface exchange of mercury (Hg) was measured, using a dynamic polycarbonate flux chamber, for soils with low or "background" Hg concentrations (<0.1 mg/kg) at eleven locations across the contiguous United States. Sampling locations included agricultural, desert, grassland, mixed and pine forest ecosystems (n=1326 soil flux measurements at 46 individual sites). An overall soil Hg flux of 0.9+/-0.2 ng/m2/h for these background soils was obtained by averaging the means for the different locations. Soil Hg fluxes were significantly lower in dark conditions than in the light for all but the grassland sites. Mean inlet air Hg concentrations were 1.0+/-0.1 ng/m3 in the dark and 1.3+/-0.2 ng/m3 in the light. Soil temperature inside and outside of the chamber, air temperature, relative humidity, and irradiance were measured concurrently with soil Hg flux. Soil-air Hg exchange was weakly predicted by environmental variables (R2 from 0.07 to 0.52). For a single location, flux was better correlated with soil moisture than other measured environmental parameters, suggesting that soil moisture might be an important driver for Hg emissions from background soils. In addition, based on data collected we suggest some quality control measures for use of Tekran 2537A analyzers when measuring low mercury fluxes. Using basic scaling procedures, we roughly estimate that natural emissions from soils in the contiguous U.S. release approximately 100 Mg/yr of Hg to the atmosphere.

[Spermatic cord mass as a manifestation of systemic vasculitis: a case report and review of the literature]. / Masa en el cordón espermático como primera manifestación de una vasculitis sistémica: presentacion de un caso y revisión de la literatura.

We report a case of necrotizing granulomatous vasculitis in the spermatic cord in a 35-year-old man with an antecedent of brain stroke 3 years before. The clinical manifestation was as a painless left scrotal mass. The diagnosis was established by histological examination of the spermatic cord. We discuss the physical findings, radiological features and pathological findings, reviewing the literature for previous similar cases.

Tricuspid regurgitation during pregnancy: two-dimensional and pulsed Doppler echocardiographic observations.

Although precordial murmurs occur frequently during pregnancy, their origin is rarely known. To investigate the nature of these murmurs, 2-dimensional (2-D) and pulsed Doppler echocardiography was performed in 81 asymptomatic pregnant women who had new systolic precordial murmurs. End-systolic right atrial dimensions, early diastolic tricuspid anular diameters, and end-diastolic right ventricular diameters were measured from the apical 4-chamber view and compared with measurements from 26 young nonpregnant women without known heart disease. Tricuspid regurgitation (TR) was detected by pulsed Doppler echocardiography in 35 of 81 pregnant women. Thirty-two pregnant women had normal 2-dimensional and pulsed Doppler echocardiographic findings. Other valvular or congenital lesions were detected in 14 pregnant women. The right-sided intracardiac diameters were larger in all groups of pregnant women than in non-pregnant control subjects (p less than 0.001). The tricuspid anular diameter was larger in the pregnant women with TR than in pregnant women with normal pulsed Doppler echocardiographic findings (24 +/- 2 vs 22 +/- 3 mm, p less than 0.015). No patient had clinical difficulty associated with a murmur. Functional TR, therefore, often occurs in pregnancy and appears to be a result of dilation of the tricuspid anulus. TR may contribute to the development of murmurs in pregnant women.

Case report: MRI in the diagnosis of testicular Leydig cell tumour.

We report the appearance of three cases of Leydig cell tumours on MRI. This imaging method showed well-defined and peripheral intratesticular tumours displaying marked and homogeneous enhancement when contrast medium was used. This latter finding was only observed in Leydig cell tumours when they were compared in a series of 104 patients with different scrotal pathologies.

Effects of cereal grain supplementation on apparent digestibility of nutrients and concentrations of fermentation end-products in the feces and serum of horses consuming alfalfa cubes.

Twenty geldings (five groups; similar age and BW) were used in a completely randomized design experiment to determine effects of grain supplementation of an alfalfa-cube diet on apparent nutrient digestibility and hindgut fermentation. The geldings were housed individually, fed their diets in two equal meals (0600 and 1800), and adapted to five dietary treatments over 6 wk. The treatments were alfalfa cubes (1% of BW; DM basis) without (control) or with one of four rolled cereal grains (i.e., barley, corn, naked oats, or oats) to provide a target level of 0.4% of BW as total nonstructural carbohydrates (TNC). Due to acute laminitis, three geldings (one in the control group and two in the barley group) were excluded. Because of this and multiple incidents of gas colic, TNC level was decreased to 0.2% of BW to ensure the geldings' health throughout the adaptation (7 d) and sample collection (5 d) periods. Grain intakes varied (P < 0.05) and reflected the different TNC concentrations. Apparent digestibilities of DM, OM, CP, NDF, ADF, and cellulose were not affected (P > 0.05) by grain supplementation and averaged 63.2, 63.1, 79.5, 42.7, 39.9, and 50.3%, respectively. Regardless of the source, grain supplementation increased (P < 0.05) apparent digestibility of TNC (from 85.6 to 94.6%) and decreased (P < 0.05) fecal pH (from 7.04 to 6.74). Fecal concentrations of total VFA (mg/g of DM) were greatest for the barley and naked oats diets (averaging 11.73), intermediate for the oats diet (8.00), and least for the control and corn diets (averaging 5.00; P < 0.05). Fecal concentrations of lactate (microg/g of DM) were greatest for the barley diet (254), intermediate for the oats diet (138), and least for the remaining diets (averaging 100; P < 0.05). Fecal concentrations of NH3 N (mg/g of DM) were greatest for the naked oats diet (1.68), intermediate for the barley and oats diets (averaging 0.86), and least for the remaining diets (averaging 0.63; P < 0.05). Serum concentration of lactate was 46% higher (P < 0.05) for the control than for the grain diets (averaging 0.05 mg/100 mL). Feeding barley, corn, naked oats, and oats contributed to 13, 15, 8, and 20% higher (P < 0.05) serum NH3 N concentrations than the control diet (0.25 mg/100 mL). Higher (P < 0.05) serum concentrations of urea N (mg/100 mL) were detected for the control, barley, and naked oats diets (averaging 25.28) than for the corn or oats diets (averaging 22.21). Results suggest that horses consuming alfalfa cubes could be supplemented with rolled barley, corn, naked oats, or oats at levels not exceeding 0.2% of BW as TNC without affecting nutrient digestion or overall health negatively.

[Useful tips for CT-angiography]. / Consejos útiles para la realización de una TC-coronariografía.

Multidetector CT scanners are now available in most hospitals; this technology makes it possible to perform cardiac studies. The technique depends on the technology, and certain points must be taken into consideration to improve the results of cardiac studies. We discuss four key points for ensuring high-quality multidetector CT studies. Under the heading patient preparation, we include deciding whether to use beta blockers, aspects related to the routine use of vasodilators, some guidelines for patient breath holding, etc. The next section discusses imaging acquisition, including when to use multisegmentation or even the possibility of prospective acquisition to reduce the dose of radiation. The third point deals with contrast administration to achieve good enhancement of the vessels. The last section reviews postprocessing techniques and comments on possible errors in 3D reconstructions or the need to render reconstructions of the entire thorax to rule out other diseases.

[MR-arthrography of the shoulder. Description and evaluation of the technique using a US-guided pediatric spinal needle]. / RM-artrografía de hombro. Descripción y valoración de la técnica guiada con ecografía y utilizando una aguja espinal infantil.

OBJECTIVE: To describe the technique of US-guided shoulder arthrography using a pediatric needle (modified Valls- Melloni technique) and to assess its efficacy. MATERIAL AND METHODS: Descriptive study of articular puncture for 48 magnetic resonance imaging arthrographs of the shoulder in 48 consecutive patients. The puncture was performed by a radiologist without prior experience in the technique. We used an anterior approach to the shoulder, guiding the puncture using US according to the Valls-Melloni technique; however, we used a pediatric spinal needle (Yale spinal; 22G: 0.7 x 40 mm). The efficacy of the technique was evaluated using the following variables: time employed, number of attempts, extravasation of contrast outside the joint, pain reported by the patient (on a scale from 0 to 10), and immediate or late complications of the technique. RESULTS: The time required for the procedure was 15.2+/-2.6 min (mean+/-standard deviation). A single puncture sufficed in 45 patients (94%); two attempts were necessary in two patients (4%) and three in one patient (2%). Contrast extravasation outside the joint occurred only in two patients (4%). The mean pain reported was 3.6 points (confidence interval: 3.1-4). Three patients (6%) had a vasovagal reaction. No late complications were observed. CONCLUSION: US-guided shoulder arthrography using a pediatric spinal needle is fast, simple, and safe; it can be performed by any radiologist, even without prior experience in the technique.

Cuticular hydrocarbons of triatomines.

Triatomine insects (Hemiptera) are the vectors of Chagas disease. Their cuticular surface is covered by a thin layer of lipids, mainly hydrocarbons, wax esters, fatty alcohols, and free or esterified fatty acids. These lipids play a major role in preventing a lethal desiccation, altering the absorption of chemicals and microorganism penetration, they also participate in chemical communication events. Lipid components are biosynthetically related, the synthesis of long chain and very long chain fatty acids was first shown in the integument of Triatoma infestans through the concerted action of fatty acid synthases (FAS's) and fatty acyl-CoA elongases. A final decarboxylation step produces the corresponding hydrocarbon. Capillary gas chromatography coupled to mass spectrometry analyses showed that cuticular hydrocarbons of Triatominae comprise saturated straight and methyl-branched chains, from 18 to more than 43 carbon atoms. Odd-chain hydrocarbons, mostly from 27 to 33 carbons, are the major straight chains. Different isomers of mono, di, tri, and tetramethylcomponents, mostly from 29 to 39 atoms in the carbon skeleton, account for the major methyl-branched hydrocarbons. The presence, absence, and relative quantities of these hydrocarbons represent characters for their chemical phenotype, and are useful for differentiating genera, species and populations. In this review, we will discuss the metabolic pathways involved in hydrocarbon formation, and their structure, together with their role in insect survival. We will also review the utility of cuticular hydrocarbon fingerprints in chemotaxonomy.

Relevance of neutrophils in the murine model of haemolytic uraemic syndrome: mechanisms involved in Shiga toxin type 2-induced neutrophilia.

It has been demonstrated that infections due to Shiga toxins (Stx) producing Escherichia coli are the main cause of the haemolytic uraemic syndrome (HUS). However, the contribution of the inflammatory response in the pathogenesis of the disease has also been well established. Neutrophils (PMN) represent a central component of inflammation during infections, and patients with high peripheral PMN counts at presentation have a poor prognosis. The mouse model of HUS, by intravenous injection of pure Stx type 2 (Stx2), reproduces human neutrophilia and allows the study of early events in the course of Stx2-induced pathophysiological mechanisms. The aim of this study was to address the contribution of PMN on Stx2 toxicity in a murine model of HUS, by evaluating the survival and renal damage in mice in which the granulocytic population was depleted. We found that the absence of PMN reduced Stx2-induced lethal effects and renal damage. We also investigated the mechanisms underlying Stx2-induced neutrophilia, studying the influence of Stx2 on myelopoyesis, on the emergence of cells from the bone marrow and on the in vivo migration into tissues. Stx2 administration led to an accelerated release of bone marrow cells, which egress at an earlier stage of maturation, together with an increase in the proliferation of myeloid progenitors. Moreover, Stx2-treated mice exhibited a lower migratory capacity to a local inflammatory site. In conclusion, PMN are essential in the pathogenesis of HUS and neutrophilia is not merely an epiphenomenon, but contributes to Stx2-damaging mechanism by potentiating Stx2 toxicity.

Endogenous glucocorticoids modulate neutrophil function in a murine model of haemolytic uraemic syndrome.

Haemolytic uraemic syndrome (HUS) is caused by Shiga-toxin-producing Escherichia coli (STEC). Although, Shiga toxin type 2 (Stx2) is responsible for the renal pathogenesis observed in patients, the inflammatory response, including cytokines and polymorphonuclear neutrophils (PMN), plays a key role in the development of HUS. Previously, we demonstrated that Stx2 injection generates an anti-inflammatory reaction characterized by endogenous glucocorticoid (GC) secretion, which attenuates HUS severity in mice. Here, we analysed the effects of Stx2 on the pathogenic function of PMN and the potential role of endogenous GC to limit PMN activation during HUS development in a murine model. For this purpose we assessed the functional activity of isolated PMN after in vivo treatment with Stx2 alone or in simultaneous treatment with Ru486 (GC receptor antagonist). We found that Stx2 increased the generation of reactive oxygen intermediates (ROI) under phobol-myristate-acetate (PMA) stimulation and that the simultaneous treatment with Ru486 strengthened this effect. Conversely, both treatments significantly inhibited in vitro phagocytosis. Furthermore, Stx2 augmented in vitro PMN adhesion to fibrinogen (FGN) and bovine serum albumin (BSA) but not to collagen type I (CTI). Stx2 + Ru486 caused enhanced adhesion to BSA and CTI compared to Stx2. Whereas Stx2 significantly increased migration towards N-formyl-methionyl-leucyl-phenylalanine (fMLP), Stx2 + Ru486 treatment enhanced and accelerated this process. The percentage of apoptotic PMN from Stx2-treated mice was higher compared with controls, but equal to Stx2 + Ru486 treated mice. We conclude that Stx2 activates PMN and that the absence of endogenous GC enhances this activation suggesting that endogenous GC can, at least partially, counteract PMN inflammatory functions.

Estimation of heritability by parent-offspring regression.

The implications of bias due to previous inbreeding of parents and genotype x environmental interaction on narrow sense heritability (h(2)) estimates by parent-offspring regression are enumerated. To remove the bias caused by genotype x environment interaction, an analysis of covariance model could be used. In special cases, where phenotypic expression is a result of two organisms interacting, such as in symbiotic N2 fixation, an analysis of covariance model with a test of heterogeneity of slopes is recommended. When host genotype x strain interactions are significant, separate heritability estimates for each strain are suggested to take advantage of genotype x strain interaction, which may be a major factor contributing to the expression of N2 fixation traits.

SAS applications for Tai's stability analysis and AMMI model in genotype x environmental interaction (GEI) effects.

A user-friendly graphical data analysis to perform stability analysis of genotype x environmental interactions, using Tai's stability model and additive main effects and multiplicative interaction (AMMI) biplots, are presented here. This practical approach integrates statistical and graphical analysis tools available in SAS systems and provides user-friendly applications to perform complete stability analyses without writing SAS program statements or using pull-down menu interfaces by running the SAS macros in the background. By using this macro approach, the agronomists and plant breeders can effectively perform stability analysis and spend more time in data exploration, interpretation of graphs, and output, rather than debugging their program errors. The necessary MACRO-CALL files can be downloaded from the author's home page at http://www.ag.unr.edu/gf. The nature and the distinctive features of the graphics produced by these applications are illustrated by using published data.

An accurate and simplified method to calculate angiographic left ventricular ejection fraction.

An accurate and simplified method to calculate left ventricular (LV) ejection fraction (EF0 derived from the ellipsoidal formula for LV volume calculation is described. The LV Minor axis (D) is obtained from the average of three equidistant LV diameters at end-diastole (Ded) and end-systole (Des), and the shortening fraction of D2 (% delta D2) calculated as (D2ed - D2es)/D2ed. EF is calculated as EF = [delta D2 + ([1 - delta D2] X delta L)] X 100, where delta L = the shortening fraction of the long axis. The coefficient of correlation between the EF by this method and the EF derived from measurements of LV volumes with the area-length method was 0.98, SEE = 3.57% (n = 50). No significant over- or underestimation was observed according to the regression equation Y = 0.922X + 0.82. Thus, this simplified method allows accurate LVEF calculation without the need for planimetry of LV area.

Beneficial effects of the alpha-adrenergic antagonist nicergoline during acute myocardial ischemia and reperfusion in the dog.

Recent experimental and clinical data have stimulated interest in the use of alpha-adrenergic antagonists in acute myocardial infarction. We evaluated nicergoline, a new relatively selective alpha 1-antagonist which uniquely lowers heart rate. Open-chest dogs, randomized to control (n = 25) or intravenously treated group (n = 20; 0.5 mg/kg bolus, then 0.10 to 0.15 mg/kg/min), underwent coronary artery occlusion (CAO) followed after 25 minutes by coronary artery reperfusion (CAR). Nicergoline decreased heart rate by 47 +/- 5 bpm and mean aortic pressure by 39 +/- 4 mm Hg. Following CAO, nicergoline reduced total coronary collateral resistance (radiolabeled microspheres; 698 +/- 75 vs 2167 +/- 530 mm Hg/ml/min/gm, p less than 0.05), increased the ischemic zone/nonischemic zone flow ratio (0.14 +/- 0.04 vs 0.06 +/- 0.02, p less than 0.05), and reduced the rise in intramyocardial CO2 tension in the ischemic zone (mass spectrometry, p less than 0.001). Furthermore, the drug decreased the rate of ventricular tachycardia (VT; 191 +/- 13 vs 243 +/- 3 bpm, p less than 0.001) and the incidence of ventricular fibrillation (VF; 1 of 20 [5%] vs 7 of 25 [28%], p less than 0.05). Following CAR, nicergoline did not significantly reduce the incidence of VF but did lower rate (154 +/- 8 vs 212 +/- 10 bpm, p less than 0.001) and incidence (p less than 0.05) of VT. Thus nicergoline reduced severity of ischemia and afforded protection against arrhythmias induced by myocardial ischemia and reperfusion. The observed reduction in heart rate may have contributed importantly to these beneficial effects. Clinical investigation of this potentially useful vasodilator seems warranted.