Subchronic oral exposure of tungsten induces myofibroblast transformation and various markers of kidney fibrosis.

Tungsten is a naturally occurring transition element used in a broad range of applications. As a result of its extensive use, we are increasingly exposed to tungsten from our environment, including potable water, since tungsten can become bioaccessible in ground sources. The kidneys are particularly susceptible to tungsten exposure as this is the main site for tungsten excretion. In this study, we investigated the prolonged effects of tungsten on the kidneys and how this may impact injury and function. When mice were exposed to tungsten in their drinking water for 1 mo, kidney function had not significantly changed. Following 3-mo exposure, mice were presented with deterioration in kidney function as determined by serum and urine creatinine levels. During 3 mo of tungsten exposure, murine kidneys demonstrated significant increases in the myofibroblast marker α-smooth muscle actin (αSMA) and extracellular matrix products: fibronectin, collagen, and matricellular proteins. In addition, Masson's trichrome and hematoxylin-eosin (H&E) staining revealed an increase in fibrotic tissue and vacuolization of tubular epithelial cells, respectively, from kidneys of tungsten-treated mice, indicative of renal injury. In vitro treatment of kidney fibroblasts with tungsten led to increased proliferation and upregulation of transforming growth factor ß1 (TGFß1), which was consistent with the appearance of fibroblast-to-myofibroblast transition (FMT) markers. Our data suggest that continuous exposure to tungsten impairs kidney function that may lead to the development of chronic kidney disease (CKD).

Operando characterization of chemical reactions in single living cells using SERS.

Detection of chemical reactions in living cells is critical in understanding physiological metabolic processes in the context of nanomedicine. Carbon monoxide (CO) is one of the important gaseous signaling molecules. Surface-enhanced Raman spectroscopy (SERS)-based CO-releasing nanoparticles (CORN) is utilized to investigate the chemical reaction of CO delivery in live cells. Using SERS CORN, carbonyl dissociation from CORN-Ag-CpW(CO)3 to CORN-Ag-CpW(CO)2 in live cells is observed. The subsequent irreversible degradation to CO-free CORN is a consequence of oxidative stress in cells. This observation affirms the step transition of CORN-Ag-CpW(CO)3 in cellular: CORN-Ag-CpW(CO)3 first proceeds via a direct loss of one CO followed by a oxidative decomposition giving rise to CORN-Ag-WO3 and as well as the release of one equivalents of CO. Importantly, the decarbonylation process can be correlated with the level of inflammatory biomarkers. For the first time, we provide unambiguous evidence for the steps transition of CO-release mechanism in cellular.

Tuning the NIR photoabsorption of CuWO4-x nanodots with oxygen vacancies for CT imaging guided photothermal therapy of tumors.

A traditional CuWO4 semiconductor (Eg = 2.25 eV) shows photoabsorption in the visible range with an edge at ∼550 nm, limiting its application in near-infrared (NIR) laser-induced photothermal ablation therapy (PAT). To tune the NIR photoabsorption of CuWO4, we report a trisodium citrate-induced strategy for generating oxygen vacancies. CuWO4 and CuWO4-x nanoparticles are prepared by a facile coprecipitation-solvothermal method in the absence or presence of trisodium citrate (0.2-0.5 g) as the reducing agent. Without trisodium citrate, CuWO4 consists of aggregated particles, and its dispersion has a typical yellow-green color without NIR photoabsorption. With the addition of trisodium citrate from 0.2 to 0.5 g, CuWO4-x samples are composed of monodisperse nanodots with increased sizes from ∼2 to ∼6 nm, and the color of these dispersions becomes darker with increased NIR photoabsorption, that is, from 0.178 to 0.685 at 808 nm. As a result, the aqueous dispersion of CuWO4-x-0.5 nanodots prepared with 0.5 g trisodium citrate exhibits a high photothermal efficiency of 47.6% under 808 nm laser irradiation. Simultaneously, CuWO4-x-0.5 nanodots have high X-ray attenuation as a CT imaging agent due to the presence of a heavy metal element (W). When the CuWO4-x-0.5 dispersion is injected into the tumors of mice, the tumors can be observed by CT and thermal imaging. After 808 nm laser irradiation (1.0 W cm-2, 10 min), cancer cells in vivo can be efficiently ablated by the photothermal effects of CuWO4-x, without obvious toxicity and side effects. Therefore, CuWO4-x can act as a novel all-in-one agent for CT imaging-guided photothermal therapy of tumors.

Separating High-Z Oral Contrast From Intravascular Iodine Contrast in an Animal Model Using Dual-Layer Spectral CT.

RATIONALE AND OBJECTIVES: To show that water and iodine two-material decomposition images from dual-layer dual-energy spectral X-ray computed tomography (DECT) can be used to separate intravascular iodine contrast from simultaneously administered oral tantalum, tungsten, or rhenium contrast in an animal model. MATERIALS AND METHODS: In this Institutional Animal Care and Use Committee approved study, four female Fischer rats were given simultaneous intravenous and oral X-ray computed tomography contrast. Intravenous iodine contrast was administered via tail vein injection. Oral barium, tantalum, tungsten, or rhenium contrast was administered via gavage. The animals were imaged on a dual-layer DECT system at 120 kVp. Water and iodine two-material decomposition images (water equivalent and iodine equivalent images) were used for qualitative analysis. Computer simulations were performed using a customized DECT simulator to better understand why certain high-Z elements disappear in the iodine equivalent images and what is the theoretical range of elements with this property. RESULTS: The iodine and barium contrast appeared only in the iodine equivalent images and could not be differentiated from each other. However, the tantalum, tungsten, and rhenium contrast only appeared in the water equivalent images. This allowed iodine contrast in the bowel wall to be easily segmented from tantalum, tungsten, and rhenium contrast in the bowel lumen. Simulations confirmed that certain high-Z elements will have pixel values of ≤0 mg iodine/mL in the iodine equivalent images due to a K-edge effect associated with DECT systems. CONCLUSIONS: Dual-layer DECT can separate iodine from certain high-Z elements using water equivalent and iodine equivalent images with an increased element range compared to other DECT systems. This K-edge effect could promote the development and approval of new high-Z contrast agents for DECT.

Targeted delivery of tungsten oxide nanoparticles for multifunctional anti-tumor therapy via macrophages.

Tumor-associated macrophages are highly versatile effector cells that have been used to kill tumor cells. Herein, the macrophages as cell-based biocarriers are used for the targeted delivery of photothermal reagents for promoting the efficiency of killing tumor cells by activating the anti-tumor immune response and photothermal therapy (PTT). In this design, macrophages cause the phagocytosis of tumor cells and activate the anti-tumor immune response by secreting plenty of cytokines. Meanwhile, to improve the tumor-killing effect and track the collaborative therapy system in vivo, a novel nanoplatform based on tungsten oxide (W18O49, WO) nanoparticles and fluorescent dyes loaded in polylactic-co-glycolic acid (PLGA) for PTT has been successfully constructed. Subsequently, the nanoparticles are swallowed by macrophages acting as cell-based biocarriers to target the tumor and promote solid tumor ablation in vivo in animal experiments. This system is expected to bring a huge application potential in the visually guided dual-modal therapeutic platform for tumor targeting therapy in vivo.

The use of tungsten as a chronically implanted material.

This review paper shows that tungsten should not generally be used as a chronically implanted material. The metal has a long implant history, from neuroscience, vascular medicine, radiography, orthopaedics, prosthodontics, and various other fields, primarily as a result of its high density, radiopacity, tensile strength, and yield point. However, a crucial material criterion for chronically implanted metals is their long-term resistance to corrosion in body fluids, either by inherently noble metallic surfaces, or by protective passivation layers of metal oxide. The latter is often assumed for elemental tungsten, with references to its 'inertness' and 'stability' common in the literature. This review argues that in the body, metallic tungsten fails this criterion, and will eventually dissolve into the soluble hexavalent form W6+, typically represented by the orthotungstate [Formula: see text] (monomeric tungstate) anion. This paper outlines the metal's unfavourable corrosion thermodynamics in the human physiological environment, the chemical pathways to either metallic or metal oxide dissolution, the rate-limiting steps, and the corrosion-accelerating effects of reactive oxidising species that the immune system produces post-implantation. Multiple examples of implant corrosion have been reported, with failure by dissolution to varying extents up to total loss, with associated emission of tungstate ions and elevated blood serum levels measured. The possible toxicity of these corrosion products has also been explored. As the field of medical implants grows and designers explore novel solutions to medical implant problems, the authors recommend the use of alternative materials.

Functionalized biocompatible WO3 nanoparticles for triggered and targeted in vitro and in vivo photothermal therapy.

We report on dopamine-conjugated hyaluronic acid (HA-D), a mussel-inspired facile capping material that can modify tungsten oxide (WO3) nanoparticles to be both biocompatible and targetable, allowing precise delivery (WO3-HA) to a tumor site. Near-infrared (NIR) irradiated WO3-HA showed a rapid and substantial rise in photothermal heat to complete in vitro thermolysis of malignant MDAMB and A549 cancer cellsbut was found to be relatively less sensitive to normal MDCK cells. A long-term in vivo investigation of ~10 nm HA thickness on WO3 (WO3-HA) nanoparticles demonstrated efficient photo-thermal conversion with time-dependent tumor target accumulation. This long-termin vivo survival study ofWO3-HA showed promising biocompatibility, with a complete recovery from malignant tumor. Due to the importance of keeping simplicity in the design of therapeutic nanoparticles, we therefore expect that this facile scheme (HA-D) would contribute to the biocompatible development of versatile metallic nanoparticles for photothermal applications.

Single-layer tungsten oxide as intelligent photo-responsive nanoagents for permanent male sterilization.

Permanent male sterilization has been recognized as useful tools for the development of neuter experimental animals and fattening livestock, as well as efficient control of pet overpopulation. Traditional routes such as surgical ways, chemical injections, and anti-fertility vaccines have addressed these crucial problems with idea outcomes. However, these routes usually bring out serious pain and infection towards animals, as well as induce long-term adverse reaction and immune suppression. Thus, a convenient, but non-surgical strategy for male sterilization under a mild manner is highly desirable. Here, for the first time, we demonstrate a novel platform for male sterilization by using single-layer WO2.72 nanosheets as smart photo-responsive sterilants. Upon a 980 nm irradiation, these nanoagents can possess intrinsic NIR-induced hyperthermia and sensitize the formation of singlet oxygen due to the cooperation of photothermal and photodynamic effects. Mechanism of cellular injury can be attributed to the denaturation of protein and apoptosis-related death. Moreover, long-term toxicity and possible metabolism route after testicular injection are discussed, indicating the neglectable systemic toxicity and high bio-compatibility of our nanoagents. Overall, our strategy can extremely overcome the shortcomings in various routine routes and suggest the new biological application of nanomaterials.

Low molybdenum state induced by tungsten as a model of molybdenum deficiency in rats.

Organ molybdenum (Mo) concentration and the activity of hepatic sulfite oxidase and xanthine oxidase were compared in tungsten-administered rats as well as rats fed with a low Mo diet to evaluate the use of tungsten-administered rats as a model of Mo deficiency. Twenty-four male 6-week-old Wistar rats were divided into four groups according to diet (AIN93G diet (control diet) or the control diet minus ammonium molybdate (low Mo diet)) and drinking water (deionized water or deionized water containing 200 µg/mL tungsten in the form of sodium tungstate). Mo content in the control and low Mo diets were 196 and 42 ng/g, respectively. Intake of the low Mo diet significantly reduced the Mo content of several organs and serum. Decrease in hepatic sulfite oxidase activity was also induced by the low Mo diet. The administration of tungsten induced marked decreases in organ Mo content and the activity of hepatic sulfite oxidase and xanthine oxidase. These decreases induced by tungsten administration were more pronounced than those induced by just a low Mo diet. Serum uric acid was also reduced by tungsten administration irrespective of Mo intake. Although a comparatively high accumulation of tungsten (3 to 9 µg/g) was observed in the kidneys and liver, adverse effects of tungsten accumulation on liver and kidney function were not observed in serum biochemical tests. These results indicate that tungsten-administered animals may be used as a model of Mo deficiency.

Microneedle-assisted microparticle delivery by gene guns: experiments and modeling on the effects of particle characteristics.

Microneedles (MNs) have been shown to enhance the penetration depths of microparticles delivered by gene gun. This study aims to investigate the penetration of model microparticle materials, namely, tungsten (<1 µm diameter) and stainless steel (18 and 30 µm diameters) into a skin mimicking agarose gel to determine the effects of particle characteristics (mainly particle size). A number of experiments have been processed to analyze the passage percentage and the penetration depth of these microparticles in relation to the operating pressures and MN lengths. A comparison between the stainless steel and tungsten microparticles has been discussed, e.g. passage percentage, penetration depth. The passage percentage of tungsten microparticles is found to be less than the stainless steel. It is worth mentioning that the tungsten microparticles present unfavourable results which show that they cannot penetrate into the skin mimicking agarose gel without the help of MN due to insufficient momentum due to the smaller particle size. This condition does not occur for stainless steel microparticles. In order to further understand the penetration of the microparticles, a mathematical model has been built based on the experimental set up. The penetration depth of the microparticles is analyzed in relation to the size, operating pressure and MN length for conditions that cannot be obtained in the experiments. In addition, the penetration depth difference between stainless steel and tungsten microparticles is studied using the developed model to further understand the effect of an increased particle density and size on the penetration depth.

[Endovascular management of cavernous sinus dural fistulas]. / Manejo endovascular de las fistulas durales al seno cavernoso.

OBJECTIVE: Describe the outcomes of patients diagnosed with indirect carotid-cavernous fistula treated by endovascular methods. DESIGN: A retrospective case series. PARTICIPANTS: Twelve patients with dural cavernous sinus fistula with important ophthalmologic involvement admitted and treated at the National Institute of Neurology and Neurosurgery between February 1990 and January 2005. INTERVENTION: Patients were managed by endovascular embolization for all fistulas. OUTCOME MEASURES: Angiographic controls to 24 hours and at 6 and 12 months were performed. RESULTS: 67 % were female and 33 % male. The mean age was 44 years. 67 % were spontaneous and 33% of traumatic origin. All patients had eye involvement with proptosis (92%) and involvement of the oculomotor nerve (67%). Headache and pulsatile tinnitus were not frequent ophthalmologic data. All were diagnosed by cerebral angiography, 33 % were type C, type D 67 %, and none of the type B classification Barrow. In 17 % of cases the distal arterial robbery showed severe. Predominance of anterior and superior venous drainage in 83 % and 42 % of cases occurred respectively. The surgical approach was arterial in 84% of cases, while in 17 % venous through the superior ophthalmic vein. Cyanoacrylate embolization material was used in 58 % of the cases, as it was associated with the use of removable ball with polyvinyl alcohol particles in 16 % in of venous approach cases. 17% detachable coils were utilized. There were no complications. After angiographic controls at 24 hours 100% occlusion was seen in patients treated with cyanoacrylate (58%) (p = 0.03). The remaining 42% were prescribed maneuver of manual compression. At 12-months angiography all patients had 100% occlusion of the carotid-cavernous fistula. CONCLSUIONS: This is the world's second largest series with indirect carotid-cavernous fistulas treated after trauma. 100 % of cases were cured with the use of a transarterial-controlled approach and N-butyl-cyanoacrylate after long-term observation.

Antimicrobial activity of nanoparticulate metal oxides against peri-implantitis pathogens.

Dental plaque accumulation may result in peri-implantitis, an inflammatory process causing loss of supporting bone that may lead to dental implant failure. The antimicrobial activities of six metal and metal oxide nanoparticles and two of their composites against bacterial pathogens associated with peri-implantitis were examined under anaerobic conditions. The activities of nanoparticles of silver (Ag), cuprous oxide (Cu(2)O), cupric oxide (CuO), zinc oxide (ZnO), titanium dioxide (TiO(2)), tungsten oxide (WO(3)), Ag+CuO composite and Ag+ZnO composite were assessed by minimum inhibitory (bacteriostatic) concentration (MIC) and minimum bactericidal concentration (MBC) determination against Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans. Time-kill assays were carried out to examine the dynamics of the antimicrobial activity with ZnO nanoparticles. MIC and MBC values were in the range of <100 µg/mL to 2500 µg/mL and <100 µg/mL to >2500 µg/mL, respectively. The activity of the nanoparticles tested in descending order was Ag>Ag+CuO>Cu(2)O>CuO>Ag+ZnO>ZnO>TiO(2)>WO(3). Time-kill assays with ZnO demonstrated a significant decrease in growth of all species tested within 4h, reaching 100% within 2h for P. gingivalis and within 3h for F. nucleatum and P. intermedia. Coating titanium surfaces of dental and orthopaedic implants with antimicrobial nanoparticles should lead to an increased rate of implant success.

[The effect of different chemoembolization materials on CT-based attenuation correction in PET/CT]. / Effekt verschiedener Chemoembolisationsmaterialien auf die CT-basierte Schwächungskorrektur der PET/CT.

PURPOSE: Primary and secondary hypervascularized liver tumors may be treated with transarterial chemoembolization (TACE). The purpose of this study was to experimentally quantify the effect of different chemoembolization materials on the PET activity concentration in PET/CT. MATERIALS AND METHODS: Different concentrations of lipiodol, tungsten, tantalum, and a different number of platinum coils embedded in a carrier substance were placed in a liver phantom. An insert filled with only the carrier substance served as the negative control. The liver phantom was placed in a body phantom. The liver phantom was filled with 63.3 KBq [18-F]-Fluor-2-deoxy-D-glucose (FDG)/ml water, the body phantom was filled with 19.7 KBq FDG/ml water. PET/CT was performed and PET attenuation correction was performed based on the CT data. We defined: Activity concentration over embolization material (kBq/ml) approximately measured activity concentration; activity concentration over negative control (kBq/ml) approximately real activity concentration. An overestimation of the activity concentration was quantified by the following ratio: Activity concentration overestimation = activity concentration over embolization material (kBq/ml)/activity concentration over negative control (kBq/ml). RESULTS: All chemoembolization materials led to an overestimation of the PET activity concentration when using CT information for PET attenuation correction. The extent of overestimation is dependent on the concentration and the density of the chemoembolizing agent. PET activity overestimation was 11-151% with lipiodol, 34-1827% with tungsten, 16-1205% with tantalum, and 4-29% with platinum coils. CONCLUSION: Conventional chemoembolization materials cause an overestimation of the PET activity concentration in CT-based attenuation-corrected PET/CT images. This is of importance for the clinical routine since activity concentration quantification may not be used in the presence of chemoembolizing agents for imaging follow-up. If an increased FDG uptake is detected after transarterial chemoembolization, non-attenuation-corrected PET images must be assessed in addition to the attenuation-corrected images in order to differentiate artificially increased tracer uptake from a true increase in activity concentration of the tracer. The use of non-attenuating chemoembolizing materials (e.g. drug-eluting beads) for TACE may serve as an alternative to avoid embolization-associated PET artifacts.

Disposition and clearance of tungsten after single-dose oral and intravenous exposure in rodents.

Tungsten (W) has been nominated for study to the National Toxicology Program (NTP) because of reported associations between concentrations of W in drinking water and childhood leukemia. The disposition of W (administered as sodium tungstate dihydrate in water) in plasma, liver, kidneys, uterus, femur, and intestine of rodents (Sprague-Dawley rats and C57BL/6N mice) was characterized after exposures by oral gavage (1, 10, or 100 mg/kg) or intravenous (1 mg/kg) administration. Each tissue (or plasma) was collected and analyzed by inductively coupled plasma mass spectrometry at 1, 2, 4, or 24 h after dose administration. W was observed in plasma and all tissues after both gavage and i.v. administration. In rats, concentrations in plasma and most tissues peaked at 4 h. In mice, concentrations in plasma and most tissues peaked at 1 h. Although the amount of W in each matrix decreased significantly by 24 h, there was W remaining in several tissues, especially at the higher doses.

Health effects following chronic dosing with tungsten-iron and tungsten-polymer shot in adult game-farm mallards.

Permanent approval of shot composed of tungsten-iron and tungsten-polymer for waterfowl hunting by the U.S. Fish and Wildlife Service was pending the results of the present study that examined the health and reproductive effects of the two shot types on mallards (Anas platyrhynchos) over a 150-day period. We collected data pertaining to the effects of tungsten-iron and tungsten-polymer shot on mortality, body weight, organ weight, tissue pathology, and shot erosion. Thirty-two bird groups (sexes equal) of adult mallards were dosed orally with eight #4 steel shot (control), eight #4 tungsten-iron shot, or eight #4 tungsten-polymer shot on days 0, 30, 60, 90, and 120 of a 150-day trial (26 January 1998 to 25 June 1998). An additional 12 mallards (sexes equal) were dosed orally with eight #4 lead shot (positive control) on day 0 of the study. All lead-dosed ducks died by day 25, whereas no ducks died in the other treatment groups. Significant liver hemosiderosis was present in all control and tungsten-iron-dosed males, in five of eight control and three of eight tungsten-iron-dosed females, and in one tungsten-polymer-dosed male examined. The rate of shot erosion was highest for tungsten-polymer shot (99%), followed by tungsten-iron (72%), and steel (55%) shot. Tungsten-iron or tungsten-polymer shot repeatedly administered to adult mallards did not have deleterious health effects during the 150-day trial based on mortality, body weights, organ weights, and histology of the liver and kidneys.

Reproductive effects and duckling survivability following chronic dosing with tungsten-iron and tungsten-polymer shot in adult game-farm mallards.

Tungsten-iron and tungsten-polymer shot were given conditional approval for waterfowl hunting by the U.S. Fish and Wildlife Service based partly on the results of a 30-day acute toxicity trial utilizing mallards (Anas platyrhynchos). Final approval of the two tungsten-containing shot was contingent on the results of a 150-day study that assessed the health and reproductive effects of tungsten-iron and tungsten-polymer shot in adult mallards. Reproductive data are presented in this paper. Sixteen male and 16 female adult mallards were dosed orally with eight #4 steel shot (control), eight #4 tungsten-iron shot, or eight #4 tungsten-polymer shot on days 0, 30, 60, 90, and 120 of a 150-day trial (26 January 1998 to 25 June 1998). Reproductive performance was assessed during the last 90 days (day 61 to day 150) of the trial. There were no significant differences in egg production and fertility and hatchability of eggs from tungsten-iron- and tungsten-polymer-dosed ducks compared to control ducks. There was no evidence of differences in percent survivability and body weight of ducklings from tungsten-iron and tungsten-polymer mallards compared to ducklings from control ducks. Tungsten-iron or tungsten-polymer shot repeatedly administered to adult mallards during the 150 day trial did not adversely affect reproduction or their offspring.

Activation of extracellular signal-regulated kinases, NF-kappa B, and cyclic adenosine 5'-monophosphate response element-binding protein in lung neutrophils occurs by differing mechanisms after hemorrhage or endotoxemia.

Acute lung injury is frequently associated with sepsis or blood loss and is characterized by a proinflammatory response and infiltration of activated neutrophils into the lungs. Hemorrhage or endotoxemia result in activation of cAMP response element-binding protein (CREB) and NF-kappa B in lung neutrophils as well as increased expression of proinflammatory cytokines, such as TNF-alpha and macrophage-inflammatory peptide-2, by these cells. Activation of the extracellular regulated kinase (ERK) pathway occurs in stress responses and is involved in CREB activation. In the present experiments, hemorrhage or endotoxemia produced increased activation of mitogen-activated protein kinase kinase (MEK)1/2 and ERK2 (p42), but not of ERK1 (p44), in lung neutrophils. ERK1, ERK2, and MEK1/2 were not activated in peripheral blood neutrophils after hemorrhage or endotoxemia. Inhibition of xanthine oxidase led to further increase in the activation of MEK1/2 and ERK2 in lung neutrophils after hemorrhage, but not after endotoxemia. Alpha-adrenergic blockade before hemorrhage resulted in increased activation in lung neutrophils of MEK1/2, ERK1, ERK2, and CREB, but decreased activation of NF-kappa B. In contrast, alpha-adrenergic blockade before endotoxemia was associated with decreased activation of MEK1/2, ERK2, and CREB, but increased activation of NF-kappa B. Beta-adrenergic blockade before hemorrhage did not alter MEK1/2 or ERK1 activation in lung neutrophils, but decreased activation of ERK2 and CREB, while increasing activation of NF-kappa B. Beta-adrenergic inhibition before endotoxemia did not affect activation of MEK1/2, ERK1, ERK2, CREB, or NF-kappa B. These data indicate that the pathways leading to lung neutrophil activation after hemorrhage are different from those induced by endotoxemia.

Xanthine oxidase activity associated with arterial blood pressure in spontaneously hypertensive rats.

Recent evidence in vivo indicates that spontaneously hypertensive rats (SHR) exhibit an increase in oxyradical production in and around microvascular endothelium. This study is aimed to examine whether xanthine oxidase plays a role in overproduction of oxidants and thereby may contribute to hypertensive states as a consequence of the increasing microvascular tone. The xanthine oxidase activity in SHR was inhibited by dietary supplement of tungsten (0.7 g/kg) that depletes molybdenum as a cofactor for the enzyme activity as well as by administration of (-)BOF4272 [(-)-8-(3-methoxy-4-phenylsulfinylphenyl)pyrazolo(1,5-alpha)-1,3, 5-triazine-4-monohydrate], a synthetic inhibitor of the enzyme. The characteristic elevation of mean arterial pressure in SHR was normalized by the tungsten diet, whereas Wistar Koto (WKY) rats displayed no significant alteration in the pressure. Multifunctional intravital videomicroscopy in mesentery microvessels with hydroethidine, an oxidant-sensitive fluoroprobe, showed that SHR endothelium exhibited overproduction of oxyradicals that coincided with the elevated arteriolar tone as compared with WKY rats. The tungsten diet significantly repressed these changes toward the levels observed in WKY rats. The activity of oxyradical-producing form of xanthine oxidase in the mesenteric tissue of SHR was approximately 3-fold greater than that of WKY rats, and pretreatment with the tungsten diet eliminated detectable levels of the enzyme activity. The inhibitory effects of the tungsten diet on the increasing blood pressure and arteriolar tone in SHR were also reproducible by administration of (-)BOF4272. These results suggest that xanthine oxidase accounts for a putative source of oxyradical generation that is associated with an increasing arteriolar tone in this form of hypertension.

Role of reactive oxygen intermediates in lipopolysaccharide-mediated hepatic injury in the rat.

Because reactive oxygen intermediates derived from xanthine oxidase may have an important role in the pathophysiology of lipopolysaccharide-mediated tissue injury, we studied hydrogen peroxide generation using 3-amino-1,2,4-triazole inactivation of hepatic catalase and the ratio of xanthine oxidase to xanthine dehydrogenase activity in rat livers after in vivo lipopolysaccharide administration. We also studied the effect of tungsten, a potent inhibitor of xanthine oxidase, on the toxicity of lipopolysaccharide. There was increased hydrogen peroxide production and enhanced proteolytic conversion from xanthine dehydrogenase to xanthine oxidase in rat livers after lipopolysaccharide administration. Feeding rats a tungsten-rich diet for 4 weeks greatly diminished hepatic xanthine oxidase activity and lessened the rise in intracellular hydrogen peroxide production after lipopolysaccharide treatment. Liver damage, as assessed by the serum transaminase levels and mortality, was also ameliorated by the tungsten-rich diet. These findings suggest that hydrogen peroxide derived from xanthine oxidase contributes to the development of systemic toxicity and liver damage after lipopolysaccharide administration.

Cobalt bioavailability from hard metal particles. Further evidence that cobalt alone is not responsible for the toxicity of hard metal particles.

Hard metal is an alloy of tungsten carbide (WC) in a matrix of cobalt metal (Co). The inhalation of hard metal dust can cause an alveolitis which may progress to interstitial fibrosis. This study was undertaken to compare, both in vivo and in vitro, the bioavailability of cobalt metal when mixed or not with WC and to assess whether this factor had any influence on the cellular toxicity of hard metal particles. In vivo, non-toxic doses of cobalt metal were administered intratracheally in the rat, alone (Co, 0.03 mg/100 g) or mixed with tungsten carbide (WC-Co, 0.5 mg/100 g containing 6.3% of cobalt metal particles). Sequential measurements of cobalt in the lung and in urine demonstrated that the retention time of the metal in the lung was longer in Co- than in WC-Co-treated animals. In vitro, the cellular cobalt uptake was higher when the metal was presented to the macrophages as WC-Co. However, there was no relationship between the cellular uptake of cobalt and the occurrence of toxicity, since the intracellular concentration of cobalt associated with the occurrence of a cytotoxic effect of WC-Co particles was insufficient to exert the same effect when resulting from exposure to Co alone. This clearly indicates that increased bioavailability of cobalt is not the mechanism by which hard metal particles exhibit their cellular toxicity. These observations confirm and extend our previous findings supporting the view that cobalt is not the only component responsible for the toxicity of hard metal particles which should be considered as a specific toxic entity.