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Introduction and objectives The particle size distribution and the median mass aerodynamic diameter (MMAD) of an inhaled product are important characteristics which may influence the deposition of the drug in the lung and as a consequence may also affect its therapeutic index. We evaluated the relationship between in vitro MMAD and in vivo lung deposition (LD, expressed as fraction of the delivered dose) and the exhaled fraction (EF), using relevant literature from scintigraphic studies conducted in healthy volunteers and asthmatic patients. Moreover the relationship between the ratio of EF to LD and MMAD has been further assessed. Methods Data from 21 studies in healthy volunteers (15 with pressurised metered dose inhalers (pMDI) and 6 with dry powder inhalers (DPI)) and 11 studies in asthmatic patients (8 pMDI and 3 DPI) have been evaluated. Asthmatic patients had a FEV1 >70% predicted, supporting pooling of the data with that of healthy volunteers. Results LD increased when MMAD decreased, so that when MMAD is about 1 µm the LD is more than 50%of the delivered dose and becomes markedly lower when MMAD approach 4 µm. EF is low and did not change markedly. On the contrary, the ratio EF/LD is independent of the MMAD, suggesting that the EF is proportional to the LD and not affected by MMAD. Conclusion These data demonstrate that the EF/LD ratio is independent from the MMAD providing reassurance that a smaller particle size will not be associated with a higher exhaled fraction.

Rat intoxication with a single dose of the hepatotoxin carbon tetrachloride induces a significant modification of liver protein kinase C total activity which depends on the degree of the intrahepatocyte oxidatiye unbalance provoked by various concentrations of the haloalkane. Low carbon tetrachloride amounts stimulate total protein kinase C activity, while one order of magnitude higher amounts exert strong enzyme inhibition. The latter effect is due to an early inactivation followed with progress of time by a proteolytic degradation of the enzyme. A pathological recruitment of the calcium-dependent protein kinase C regulatory enzymes calpain and calpastatin appears responsible for protein kinase C loss. The prolonged excess of cytosolic calcium which characterizes the single high dose carbon tetrachloride poisoning also leads to inactivation of calpain II and calpastatin in a time-dependent manner.

4-Hydroxynonenal (4-HNE), produced during the oxidative lipid breakdown of biological membranes, modulates various biochemical processes in normal liver and in hepatoma cells. It is very probable that the effects of 4-HNE are related to the quantity formed in the cells and the cells' ability to metabolize it. Aldehyde catabolism takes place within the cells through oxidative and reductive enzymes, and through conjugation with intracellular glutathione. In this paper, the various enzymatic pathways involved in the metabolism of 4-HNE were studied in normal hepatocytes and in hepatoma cells. The hepatocyte pathway undergoes a complex variety of change during neoplastic transformation. In hepatoma cells, generally, 4-HNE metabolism was due mainly to aldehyde dehydrogenases, whereas in normal hepatocytes 4-HNE metabolism was mainly due to alcohol dehydrogenase and glutathione-S-transferase. The increase in oxidative enzymes compared to normal tissue was not the same in all types of hepatoma: in HTC hepatoma cells, the enzyme levels were considerably higher; in AH-130 hepatoma cells of Yoshida, they were lower in subcellular particles and similar in the cytosol. Indeed, consumption of externally-added 4-HNE in hepatoma cells was proportional to their content of 4-HNE metabolizing enzymes.

Protective Ni-Aluminide coatings produced by microwave assisted SHS Synthesis of Ni, Al, Cr and Si powders on Ti substrate.

Increasing of oxidation resistance of aluminide coatings obtained by Microwave ignition temperature in SHS Synthesis.

To date more than 4000 compounds are recognized to belong to the class of flavonoids. These natural phenolic drugs are poorly soluble in water and are rapidly degraded and metabolized in the human body, but nevertheless are very promising for their potential contribution to the prevention and therapy of major chronic diseases, including cardiovascular and neurodegenerative diseases and cancer. In recent years a number of flavanols (e.g. catechins), flavonols (e.g. quercetin, myricetin) and isoflavones (e.g. genistein, daidzein) have been confirmed to possess strong antioxidant, anti-inflammatory, anti-proliferative and anti-aging activities. Incorporation into lipidic or polymer-based nanoparticles appears to markedly help the oral delivery of flavonoids, as these particles can protect the drug from degradation in the gastrointestinal tract and, by virtue of their unique absorption mechanism through the lymphatic system, also from first-pass metabolism in the liver. In addition, both oral and parenteral administration of flavonoids exploits a pharmacologic delivery route that guarantees sustained release of the active principle at the desired site of action. A comprehensive review of studies currently available on the in vitro and in vivo experimental administration of flavonoids by means of nanovectors may be of use as a foundation for the development of advanced delivery systems for these powerful compounds, in view of their adoption in primary and secondary disease prevention.

The effect of recombinant protein from the envelope (gp120) of the HIV on B lymphocytes purified from either HIV-infected individuals or healthy seronegative controls was examined. B cells from peripheral blood and lymph nodes of HIV-infected individuals spontaneously secreted TNF-alpha; this secretion was augmented by the presence of gp120, whereas B cells from healthy seronegative donors failed to secrete significant levels of TNF-alpha in the presence or absence of gp120. In a coculture system of B cells and chronically HIV-infected T cells (ACH-2), where viral expression is largely mediated by TNF-alpha, gp120 increased virus expression only if the B cells were obtained from HIV-infected individuals. The effects of gp120 on viral expression in this system were not mediated via CD4 receptor binding or FcR binding of anti gp120-gp120 immune complexes. Besides its effect on cytokine production, gp120 also stimulated Ig secretion in B cells from HIV-infected individuals, but not from normal donors. Finally, it was demonstrated by in situ hybridization that germinal centers of lymph nodes from HIV-infected individuals contain large amounts of HIV RNA that is in close proximity to germinal center B cells. These findings suggest that the hyperplastic germinal centers of lymph nodes provide an unique environment for virus expression and accumulation where gp120 stimulates B cells to secrete HIV inductive cytokines, such as IL-6 and TNF-alpha, and thereby further enhances virus expression in infected cells in a paracrine manner.

This study was carried out on 33 patients who were sero-positive for liver-kidney microsomal antibodies (LKM) in order to examine clinical features and the presence of underlying hepatitis C virus infection. Twenty-four sera were positive for antibodies against HCV (anti-HCV) as detected by enzyme immunoassay and confirmed by recombinant immunoblot assay. These patients had chronic liver disease and the majority of those treated with interferon responded favourably. Three of the nine anti-HCV-negative patients had idiopathic chronic hepatitis and two responded favourably to steroids. Two patients were diagnosed as having toxic hepatitis and the other four had various extrahepatic disorders without evidence of liver involvement. The immunoblotting analysis showed reactivity with a 50 kDa microsomal protein which presumably corresponded to cytochrome P-450 db1 both in anti-HCV-positive and -negative sera. In addition a few anti-HCV-positive sera also reacted with a 35 kDa microsomal antigen. Autoimmune markers different from LKM were absent in both groups. The high prevalence of antibodies to the hepatitis C virus among LKM-positive sera confirms that this infection plays a role in forms of chronic hepatitis that had previously been labelled autoimmune. In patients with LKM the presence of anti-HCV may help to forecast a therapeutic response to interferon, while its absence may forecast response to steroid therapy.

This work presents an implementation of neocognitron neural network, using a high performance computing architecture based on GPU (graphics processing unit). Neocognitron is an artificial neural network, proposed by Fukushima and collaborators, constituted of several hierarchical stages of neuron layers, organized in two-dimensional matrices called cellular planes. For the high performance computation of face recognition application using neocognitron it was used CUDA (compute unified device architecture) as API (application programming interface) between the CPU and the GPU, from GeForce 8800 GTX of NVIDIA company, with 128 ALU's. As face image databases it was used a face database created at UFSCar, and the CMU-PIE (Carnegie Mellon University pose, illumination and expression) database. The load balancing was achieved through the use of cellular connections as threads organized in blocks, following the CUDA philosophy of development. The results showed the feasibility of this type of device as a massively parallel data processing tool, and that smaller the granularity and the data dependency of the parallel processing, better is its performance.

The reconstruction of a three-dimensional surface from a set of unorganized points is a fundamental process in a lot of applications, including laser range scanners, medical imaging, and others. This work presents a modified version of the Neural Meshes algorithm for surface reconstruction from point cloud, called Neural Meshes with Edge Swap, or Neural Meshes ES. From the original basic Neural Meshes algorithm we developed a new heuristic that include an edge-swap operation, making the algorithm less sensible to parameters variation, improving the quality of the generated surfaces. The results show that the edge-swap operation is able to avoid a series of incorrect reconstructed areas in the mesh, mainly in the concave structures.

Microwaves at 2.45 GHz used to ignite the combustion synthesis of Ni and Al powder mixtures to produce NiAl and a complex Ni-Al-Ti intermediate layer on titanium. Wear resistance of the complex Ni-Al-Ti layer were measured in dry-sliding against alumina using a tribometer. Other mechanical properties, as scratch resistance and elastic modulus, were measured. These showed that Ni-Al-Ti layer presents increased performances with respect to polycrystalline NiAl.

The involvement of CCl4 biotrasformation mechanism in decreasing the Protein Kinase C activity has been analyzed in hepatocytes isolated from phenobarbital-pretreated rats. A significant inhibition (55%) and an almost total disappearance (87%) of the enzyme activity were observed at 15 min and at 30 min incubation with CCl4, respectively. Cell preincubation with Trolox or desferrioxamine allowed a marked whilst not complete protection of both cytosolic and particulate Protein Kinase C activity. These results show that the CCl4 reactive metabolites play a primary role in hepatocyte Protein Kinase C impairment and suggest that besides lipid peroxidation other mechanisms-possibly a derangement of Ca2+ homesostasis-may be involved in this process.

1. The effects of heavy metals on lipid peroxidation in the gills and digestive gland of mussels exposed for six days to Cu 2+ , Cd 2+ or Zn 2+ (40 μg/l/animal) were investigated. 2. In the tissues of Cu-exposed mussels a significant increase of the level of malondialdehyde (MDA), which is indicative of the peroxidative process, and a decrease of the concentration of glutathione were observed. 3. Moreover, in the digestive gland of mussels, copper exposure results in an increase of other carbonyl compounds and in the lysosomal accumulation of lipofuscin granules. 4. The exposure of mussels to Zn or to Cd did not elicit any of the above effects. 5. The results are discussed in relation to the possible role that Cu-induced lysosomal lipofuscin accumulation may play in heavy metal detoxification.

The onset of the lipoprotein secretory block provoked by CCl4 in the whole animal was monitored after purification of liver Golgi membranes. Both lipid transit through the apparatus and hexosylation of the lipoprotein are markedly inhibited 5-15 min after poisoning. Pre-treating the animal with alpha-tocopherol, shown to prevent lipid peroxidation without modifying the covalent binding due to CCl4 metabolites, affords little protection against lipid accumulation in the Golgi, but total preservation of galactosyl transferase activity. While haloalkylation therefore appears to be the major mechanism of damage in the early phases of CCl4-induced derangement of lipid secretion, lipid peroxidation is probably more involved later; this is indicated by the marked, though never complete, protection against fatty liver afforded at 24 h after CCl4 poisoning by supplementation of the membrane with alpha-tocopherol.

Background Ectopic pregnancy (EP) is a major reproductive health issue, whose underlying causes remain largely unknown. The unusual macrophage presence in the oviduct affected by EP could indicate macrophage contribution to the pathology. Methods Macrophages have important functions in reproduction that are reviewed in this work. They are needed for tissue remodelling and immune-regulatory roles, and are present both in the ovary and uterus. Numerous cytokines regulate monocytes recruitment, differentiation and function in the reproductive tract, among them leukaemia inhibitory factor (LIF), colony-stimulating factor 1 and transforming growth factor-beta are indispensable and non-redundant for reproductive outcome. Cytokine types and levels are modulated by estrogen, progesterone and seminal plasma, which drive the differentiation of monocytes to immunity cells or to immunosuppressed trophic and scavenging macrophages. Results Many risk factors for EP involve an inflammatory reaction that can induce the release of mononuclear phagocytes from the bone marrow and/or favour immunosuppressed trophic differentiation of newly recruited mononuclear phagocytes in the reproductive tract. These observations strengthen the hypothesis that immunosuppressed trophic and scavenging macrophages may have a role in EP onset. Conclusions Macrophages may contribute to the regulation of tubal motility through prostaglandin production and induction of progesterone secretion. Considerations about LIF also suggest that macrophages may have a central role in ectopic receptivity.

Microwave assisted SHS of NiAl compounds on Ti substrate in order to promote the formation of a complex Ni-Al-Ti interface which enhances the NiAl coating adhesion. This interlayer presents higer hardness, toughness and capability of stopping the fracture propagation.

Despite an increasingly understanding of the pathogenetic mechanisms of sepsis, its mortality remains extremely high, caused mainly by hemodynamic impairment-related alterations frequently present in severe sepsis. Currently, treatment of sepsis is based on hemodynamic support, antibiotic therapy, surgical excision of infectious foci and immunomodulatory therapy. In fact, a massive host inflammatory infection response has recently emerged to substantially contribute to the development of septic shock and multiple organ dysfunction. Many clinical trials on various pharmacological agents have been conducted: glucocorticoids, nonsteroidal anti-inflammatory drugs (NSAIDs), antithrombin III (AT III), anti-endotoxin monoclonal antibodies, nitric oxide inhibitors, interleukin-1 receptor antagonist, anti-tumor necrosis factor (TNF) antibodies. Apart from some likely favourable findings connected to low doses of glucocorticoids, most studies yielded disappointing results. Nevertheless, the use of recombinant human activated protein C (drotrecogin-alpha) has recently proven to have a mortality reduction effect particularly in patients with severe sepsis and dysfunction of at least two organs. Furthermore, the early treatment of hemodynamic instability with volume expanders and vasopressors (early goal-directed therapy), and a strict glycemic control represent important measures in order to significantly reduce mortality from severe sepsis and septic shock, and are fundamental guidelines recommended by most scientific societies (Surviving Sepsis Campaign).

Oxygen supply to all tissues is possible only in a condition of adequate blood circulation. Oxygen demand is the driving force that is responsive of hemodynamic adjustment. The human body acts on four modulators (intravascular volume, inotropy, vasoactivity, chrono-tropy) in order to adjust the hemodynamic state. Hemodynamic monitoring consists of techniques able to assess the hemodynamic status and to understand the mechanism of patient decompensation; its goal is to maintain adequate tissue perfusion through appropriate therapeutic interventions. An early diagnosis of hemodynamic alteration is crucial for an early treatment; several reports have explored the effectiveness of hemodynamic manipulations and results are conflicting: too many variables can, in fact, modify the results: timing and lenght of the treatment, drugs used, etc. However, at least, in some specific settings, as sepsis, early intervention has a positive impact on mortality. In this presentation it will be briefly analyzed the most common parameters used in the ICU. Arterial pressure, central venous pressure, pulmonary artery catheter derived parameters, SvO2 and their relation with organ perfusion are considered and positive and negative aspects of this type of monitoring is reviewed. Starting from these considerations we would like to underline the importance of understanding the physiological basis of monitoring and the correct interpretation of data in order to have improvement on patient outcome.

OBJECTIVE Oxidative stress may be one of the important complex pathogenetic mechanisms that lead to damage in scleroderma; free radicals may provoke endothelial injury, fibroblast proliferation and fragmentation of autoantigens favouring induction of autoantibodies. The present study investigates the oxidant status in scleroderma patients by measuring the urinary concentration of 8-isoprostaglandin-F2alpha, an F2-isoprostane, and a product of free radical-mediated peroxidation of arachidonic acid. METHODS Forty-three scleroderma patients (42 women and 1 man, mean age 54.1 yr, mean disease duration 9.0 yr) underwent clinical evaluation and instrumental investigations in order to assess skin, vascular, lung and heart involvement. Von Willebrand factor was evaluated as marker of vascular dysfunction in 36 out of the 43 cases. The urinary level of 8-isoprostaglandin-F2alpha was measured in all scleroderma patients and in the 43 age- and sex-matched healthy controls. RESULTS Urinary levels of 8-isoprostaglandin-F2alpha were higher in scleroderma patients than in the healthy control group (341.7 vs 147.6 pg/mg creatinine; P < 0.001). Values of 8-isoprostaglandin-F2alpha were strongly correlated with the nailfold videocapillaroscopy pattern and lung involvement (P = 0.002 and 0.003, respectively), showing increasing levels with the progression of pulmonary severity. Correlation between 8-isoprostaglandin-F2alpha level and von Willebrand factor narrowly failed to reach statistical significance (P = 0.05). There was no correlation between 8-isoprostaglandin-F2alpha concentration and disease activity, vascular, skin and heart involvement, disease pattern or autoantibody profile. CONCLUSIONS Our study further supports the role of oxidant stress in the pathogenesis of scleroderma, showing a strong correlation between a marker of free radical damage with both the severity of lung involvement and the videocapillaroscopic patterns.

Sickle cell disease has been frequently associated with sensorineuronal hearing loss. Several studies have demonstrated a significant prevalence rate of sensorineuronal hearing loss in black patients reporting a range of 11-41%, while few data are reported for white people with Hb S/beta thalassemia. In this paper we evaluated the prevalence of sensorineuronal hearing loss in a Sicilian population affected by sickle cell disease.Otologic and audiologic examinations were performed in 50 patients with S-beta thalassemia (37 with the beta(s)beta(0th) and 13 with the beta(s)beta(+th)) and 23 patients with sickle cell anemia (beta(s)beta(s)) observed at the Department of Pediatric Hematology and Oncology, University of Catania.A sensorial hearing loss of more than 25 dB was recorded in 24% of subjects with Hb S/beta0-thalassemia, in 23% of subjects with Hb S/beta+-thalassemia and in 30% of subjects with sickle cell anemia. We found an increase in the frequency of hearing loss with increasing age.Our data showed that sensorineuronal hearing loss is a common complication in white patients with sickle cell anemia, and in patients with severe forms of Hb S/beta-thalassemias.

The Eighth Conference on Retroviruses and Opportunistic Infections (CROI) marked a moment of great uncertainty among physicians on how to manage antiretrovirals in the long term. Several protocols of structured therapy interruption (STI) suggest that a better, less toxic utilization of these drugs can be achieved, with similar or even greater efficacy than standard highly active antiretroviral therapy (HAART). A new frontier in the pharmacological control of HIV infection is the development of drugs capable of interfering with the attachment and entry of HIV; different molecules can prevent the in vitro fusion of viral and cellular membrane induced by activation of gp41 Env, whereas others target the interaction of gp120 Env with chemokine receptors. Drug transporters are becoming an important element in the pharmacological control of HIV infection, particularly for protease inhibitors.

A hyperdynamic circulatory pattern in advanced liver disease is known since a long time. The first studies evaluating cardiac function in cirrhosis were performed in patients with alcoholic liver disease and thus this condition was attributed to the toxic effects of ethanol. A reduced performance of the left ventricle after physical and pharmacological strains along with an altered diastolic function has been demonstrated also in postviral cirrhosis. Many factors are involved in advanced cirrhosis whereas little is known in the earlier stages of disease.To this aim we have investigated patients with different stages of hepatitis C virus (HCV)-related liver disease to detect the time-course of diastolic dysfunction. An impaired relaxation and increased thickness of left ventricular walls along with an altered pattern of transmitral flow can be easily detected by means of echocardiography.In chronic hepatitis diastolic function is preserved but increased thickness of left ventricle parietal walls can be detected in patients with fibrosis on liver biopsy. The typical pattern of diastolic dysfunction is observed in Child A cirrhotic patients and in Child C ascitic patients but thickness of parietal walls is more relevant in the former group. Chronic aldosterone blockade could exert favourable effects in heart remodeling suggesting a potential role of these drugs in cirrhotic cardiomyopathy.The presence of increased thickness of left ventricle parietal walls in chronic hepatitis C in the precirrhotic stage point to a putative role of HCV in this heart structural abnormality that can become a co-factor in the more advanced stages of cirrhosis when portal hypertension and its deleterious effects on systemic hemodynamics, cardiac function and structure become manifest.

PRO-140, a monoclonal antibody against the HIV coreceptor CCR5, is under investigation by Progenics and the Aaron Diamond AIDS Research Center (ADARC) as a potential treatment for HIV infection [211441], [286246], [286247]. Phase I/II trials were expected to commence during 2001 [395621], [409142], despite being initially planned for 2000 [322637], [361819], [365216], [375598], [408483]. In January 1998, ADARC and Progenics reported that the HIV binding site on the CCR5 coreceptor is distinct from betachemokine binding domains, which they claimed may allow for the development of therapeutics with fewer side effects [273391], 421256]. In vitro studies have shown PRO-140 potently blocked all of 17 primary HIV isolates that use CCR5 as a fusion coreceptor [342173]. In October 2000, Progenics was awarded an SBIR grant to fund a 2-year project exploring the breadth, potency and durability of PRO-140 therapy in laboratory and animal models of HIV infection. This project was a collaboration between Progenics, Weill Medical College of Cornell University and the Scripps Research Institute [385982]. In May 1999, the company entered into an agreement with Protein Design Labs (PDL) for the humanization by PDL of PRO-140 [325445]. In November 1997, Progenics was awarded a 600,000 dollars grant from the NIAID for the examination of new approaches to HIV vaccine design based on CCR5 [268407].

Following an outbreak of bovine spongiform encephalopathy (BSE) in dairy cows in the United Kingdom (UK), 153 definite and probable human cases of new variant Creutzfeldt-Jakob disease (nvCJD) have been reported, almost exclusively in the UK. Although exposure to the BSE agent is the most plausible interpretation for the occurrence of nvCJD, the causal link between the BSE prion and nvCJD is still debated. This review discusses the pros and cons of nvCJD as a separate nosographic entity, the scientific basis for a correlation between BSE and nvCJD, the validity of the current diagnostic criteria for CJD and nvCJD, the contribution of epidemiology to the detection of a causal relation between BSE and nvCJD, and the present and future directions of the epidemiological research on BSE, CJD and nvCJD.

An increasing body of evidence from animal models, human specimens and cell lines points to reactive oxygen species as likely involved in the pathways, which convey both extracellular and intracellular signals to the nucleus, under a variety of pathophysiological conditions. Indeed, reactive oxygen species (ROS), in a concentration compatible with that detectable in human pathophysiology, appear able to modulate a number of kinases and phosphatases, redox sensitive transcription factors and genes. This type of cell signalling consistently implies the additional involvement of other bioactive molecules that stem from ROS reaction with cell membrane lipids. The present review aims to comprehensively report on the most recent knowledge about the potential role of ROS and oxidised lipids in signal transduction processes in the major events of cell and tissue pathophysiology. Among the lipid oxidation products of ROS-dependent reactivity, which appear as candidates for a signalling role, there are molecules generated by oxidation of cholesterol, polyunsaturated fatty acids and phospholipids, as well as lysophosphatidic acid and lysophospholipids, platelet activating factor-like lipids, isoprostanes, sphingolipids and ceramide.

In vivo studies on human colon adenocarcinoma showed decreased transforming growth factor-beta1 (TGF-beta1) antiproliferative cytokine content in tumour tissue related to malignancy progression, with a corresponding decrease in lipid peroxidation aldehydic end-product, 4-hydroxynonenal (HNE). The tumour mechanism to escape TGF-beta1-mediated growth inhibition may be due to an altered TGF-beta1 receptor system. Subsequent in vitro analyses showed a differential distribution of TGF-beta1 receptors depending on the human colon cancer cell line considered (CaCo-2 or HT-29): compared to HT-29 cells, CaCo-2 cells showed a decrease of the two main TGF-beta1 receptors, RI and RII. Notwithstanding their partial TGF-beta1 RI and RII deficiency, treatment of CaCo-2 cells with adequate doses of the cytokine (10 ng/ml) was able to induce apoptosis. Of note, co-treatment of these cells with 1 microM HNE increased the apoptotic effect. The constant low concentration of TGF-beta1 in the tumour mass may be related to the low content of antiproliferative HNE observed in colon cancer: the latter phenomenon, which reduces TGF-beta1 production in the tumour area, may represent a favourable condition for neoplastic progression. The enhancement of TGF-beta1-induced apoptosis by HNE in CaCo-2 cells supports this hypothesis. The different transcriptional components regulated by the distinct signaling pathways of these two molecules might be proposed; in particular, crosstalk between the MAPK and the Smad pathway could modulate and co-operate in the transcription of target genes involved in regulation of cell proliferation.

Among the oxidative breakdown products of omega-6 unsaturated fatty acids, the aldehyde 4-hydroxy-2,3-nonenal (HNE) is receiving increasing attention for its potential pathophysiological implication, which at least partly lies on the demonstrated ability to modulate gene expression of a number of genes. Here we show that a marked down-modulation of HNE nuclear localisation in cells of a macrophage line (J774-A1) can be afforded by treatment with sulfydryl and carbonyl reagents without significantly interfering with cell viability. As regards the addition of thiol-group reagents to the cell suspension, N-ethylmaleimide (NEM) led to a sustained decrease of HNE nuclear localisation, while 4-(chloromercuri)-benzene-sulfonic acid (PCMBS) gave a similar but more transient effect. Hydroxylamine (HYD), a carbonyl-group reagent, was also able to inhibit HNE nuclear localisation. The actual efficacy of the inhibitors used was then tested on the HNE-induced stimulation of transforming growth factor beta1 (TGFbeta1) production by J774-A1 cells. Indeed, the thiol reagents NEM and PCMBS, both markedly down-modulating HNE nuclear localisation, were able to inhibit HNE-induced increase of TGFbeta1 protein synthesis. The carbonyl reagent HYD was less effective on this respect, producing strong but incomplete protection against HNE-induced TGFbeta1 increase. Taken together, the results indicate that sulfydryl groups are involved in the process of HNE cellular internalisation, while both sulfydryl and carbonyl groups are involved in the process of HNE nuclear translocation, and consequently in the modulation of gene expression by the aldehyde. Further, an actual demonstration is provided that HNE-induced effect on gene regulation can be efficiently counteracted by suitable interference with HNE biochemistry.

4-Hydroxynonenal (HNE) in the concentration range detectable in many pathophysiologic conditions is able to modulate signal transduction cascades and gene expression. Here, we report the stimulating effect of 1 μM HNE on the release of the monocyte chemotactic protein-1 (MCP-1) by murine macrophages. MCP-1-increased export following 1-h cell treatment with HNE proved to be comparable to that exerted by standard amounts of bacterial lipopolysaccharide (LPS). However, the key molecular event in HNE-induced secretion of MCP-1 appeared to be the increased activity of β-PKC isoforms, which are recognized as playing a role in the regulation of cell protein transport and secretion. On the other hand, in LPS-stimulated cells, the δ isoform was seen to be involved and was probably related to LPS-mediated effects on MCP-1 expression and synthesis. In conclusion, HNE might interact with other pro-inflammatory stimuli, like LPS, in a concerted amplification of MCP-1 production and secretion.

The investigation of toxic compounds which need a previous metabolic activation often gets advantage by the employment of cell suspensions, in most of the cases rat hepatocytes. In particular, this model system allows the characterization of lipid peroxidation kinetics, the evaluation of its pathogenetic role in the induction of cell death and the analysis of the interaction among different prooxidant compounds in bringing about both oxidative damage and cytolysis.

Objective To assess the influence of hepatitis C virus (HCV) genotypes on the clinical outcome of liver disease, we analysed 2307 patients. Results The most frequently represented genotypes were 1 b (40%) and 2 (28.1%). Patients with these genotypes had a median age higher than patients with other genotypes (P < 0.01). The overall survival of subjects with genotype 1 b was poorer than the survival of patients with other genotypes (P< 0.01). Liver cirrhosis was found in 280 patients (12.1%), and type 1 b was the most represented isolate among them (P < 0.01). Sixty-two patients (22%) developed hepatocellular carcinoma (HCC) during a follow-up of 1481.8 cumulative years (estimated crude incidence rate, 4.1 cases per 100 person-years for all cirrhotics; 5.9 cases for genotype 1 a; 4.5 cases for genotype 1 b; and 2.8 cases for genotypes non-1). Considering the whole population of 2307 patients, only genotype 1 b was associated significantly with both cirrhosis and the development of HCC. One hundred and nineteen cirrhotic patients underwent treatment with interferon in uncontrolled studies. Interferon therapy was associated with both better survival (P

Because of its high radiation hardness, diamond can be used better than other materials in the intense radiation field characterizing the interior region of a particle beam in an accelerator. In effect, the measurements reported here were carried out by placing diamond detectors under continuous irradiation in the 26-MeV proton beam of the 15-MV TANDEM accelerator of Southern National Laboratory (LNS) of INFN in Catania. The diamond detectors were built in the Rome Tor Vergata Laboratory. Diamond films were deposited by microwave plasma enhanced chemical vapor deposition on silicon substrates. A four-pixel beam monitor prototype was then realized by depositing four titanium gold contacts on the diamond surface. The signal was found to be stable and reproducible. The collected charge in DC mode was more than 20 000 electrons/protons for a diamond thickness of 65 μm, thus exhibiting a gain of approximately 10 4 with respect to the Faraday cup. For the measured samples, both response and release times of approximately 1 s were observed in the above experimental conditions. An analysis of the relative sensitivity between pixels was also performed. No differences of the detector current and Faraday cup current ratio were observed for different pixels, indicating the homogeneity of the beam monitor response.

CD30 is a member of the TNF receptor superfamily, previously shown to be expressed on Hodgkin's lymphoma cells and on normal activated lymphocytes. We here show that CD30 is highly expressed on recently activated human gamma delta T cells. Elevated surface levels of this molecule persisted in long-term cultures of gamma delta cells, without further cell stimulation. CD30 acted as a co-stimulus in gamma delta T cells by potentiating the intracellular Ca(2+) fluxes induced by CD3 cross-linking. The engagement of CD30 enhanced the expression of several cytokines induced upon CD3 stimulation such as IL-4 and IFN-gamma but not IL-10. The CC chemokines RANTES and macrophage inflammatory protein-1beta were constitutively expressed and not affected by stimulation. The inducible expression of the neutrophil chemoattractant IL-8 was enhanced by CD30 co-stimulation, as well as that of the CC chemokines I-309 and MDC, whereas the secretion of the monocyte chemotactic protein-1 was not detected. Triggering of CD30 may therefore modulate the expression of several cytokines released by gamma delta cells; the expression of its physiologic ligand by APC and neutrophils at the site of infection may contribute to determine the outcome of an immune response.

Liver ischemia-reperfusion is characterized by an increased oxygen-dependent free radical chain-reaction rate and an increased steady-state concentration of reactive oxygen species. The aim of this study was to evaluate the in situ generation of reactive oxygen species and its relationship with phagocyte activation and recruitment in reperfused rat liver. Rat livers were subjected to 2 hours of selective lobular ischemia and reperfusion for up to 12 hours. The following parameters were determined: in situ liver chemiluminescence, understood to reflect the tissue steady-state concentration of singlet oxygen ((1)O(2)); myeloperoxidase tissue activity; the number of neutrophils; and the degree of necrosis. An early chemiluminescence burst was measured after 30 minutes of blood reflow (early phase of oxidative stress), followed by a relapse and a further increase after 4 to 12 hours of reperfusion (late phase of oxidative stress). Both early and late phases were modified by pretreatment with gadolinium chloride (GdCl(3)), pointing to a key role of the Kupffer cells. Neutrophils infiltrated into the liver, myeloperoxidase activity, in situ chemiluminescence, and necrosis were found to be strongly correlated over the 4- to 12-hour reperfusion period (r =.960; average of the 4 correlation coefficients). Together with resident phagocytes, neutrophil recruitment and activation appear to provide a major contribution to the increase of oxygen-dependent free-radical reactions and amplification of liver reperfusion damage. Surface chemiluminescence appears to properly describe the in situ and in vivo progressive organization of the acute inflammatory response with phagocyte-mediated liver injury.

Replication of the human immunodeficiency virus (HIV), the causative agent of the acquired immunodeficiency syndrome (AIDS), is under the control of both viral and host factors. Among the latter, the regulatory network of cytokines has been shown to affect virtually every step of the virus life cycle, from cell entry to budding of new progeny virions. Proinflammatory cytokines, such as tumour necrosis factor alpha, can either trigger or potentiate HIV expression via activation of the cellular transcription factor NF-kappaB. Other molecules, including interleukin 6 (IL-6) and the interferons, can up-regulate HIV expression by acting predominantly at post-transcriptional and/or post-translational levels. Anti-inflammatory cytokines, including transforming growth factor beta, IL-4 and IL-10, counteract these effects but can also potentiate viral replication under different experimental conditions. Chemotactic cytokines (chemokines) have recently entered the arena of host factors controlling viral spreading as potent inhibitors competing with the virus for cell-surface 7-transmembrane domain receptors also acting, together with CD4, as entry co-receptors for HIV. The cytokine network is constitutively activated in most HIV-infected individuals, as demonstrated by recent analysis of intracellular signalling molecules such as the Janus kinase/signal transducer and activator of transcription pathway. Finally, cytokines have already shown their potential use as pharmacological agents able to restore at least some of the compromised immune functions in infected individuals, as exemplified by the potent enhancing effect of IL-2 on the number of circulating CD4+ T lymphocytes.

This study examines the impact of direct-to-consumer (DTC) pharmaceutical advertising on prescription drug knowledge and the requesting behavior of consumers. The authors developed and tested a conceptual model of prescription drug knowledge and requests. Consumers' belief that drug advertising can educate them was associated with a greater amount of drug knowledge, and the belief they would upset physicians by asking for specific drugs was associated with less knowledge. The belief that drug advertising reduces prices was associated with greater probability of drug requests, and the belief that physicians should be the sole source of drug information was associated with lesser probability of request. Preference for generic drugs was associated with a lesser likelihood of requesting a specific drug. Media exposure and drug advertising awareness were associated with higher drug knowledge and a greater probability of drug requesting.

The U937-derived chronically HIV-infected U1 cell line and uninfected U937 cell clones were efficiently lysed by both unstimulated (NK) and IL-2-stimulated (lymphokine-activated killer; LAK) peripheral blood mononuclear cells (PBMC) of healthy HIV-seronegative donors. Pretreatment of target cells with IFN-gamma down-modulated killing of both U1 cells and two U937 cell clones, and up-regulated MHC class I expression. In contrast, TNF-alpha enhanced the sensitivity of infected U1 cells, but not of U937 cell clones to NK / LAK cell lysis. Co-cultivation of IL-2-stimulated PBMC with U1 cells triggered expression and replication of HIV by cell-cell contact, and this effect was inhibited by anti-TNF-alpha antibodies (Ab); virus production was partially inhibited by zidovudine. Of interest, anti-TNF-alpha Ab protected U1 cells from LAK cell activity. Thus, TNF-alpha can induce HIV expression from chronically infected U1 cells, but also plays an important role in sensitizing these cells to lysis.

Journal de Gynecologie Obstetrique et Biologie de la Reproduction - Vol. 36 - N° 4 - p. 327-328

The effect of acute glutathione (GSH) depletion induced by GSH-depleting agent L-buthionine-(S,R)-sulfoximine (BSO) on hepatic microsomal glucose-6-phosphatase (G6Pase) activity in male Wistar rats was investigated. Liver GSH evaluated in high-performance liquid chromatography after administration of 4 mmol.Kg-1 BSO i.p. was decreased by 19% and 50% at the time-points of 1.5 h and 3 h, respectively. In these conditions, a significant decrease in Vmax and an increasing trend in K(m) of hepatic G6Pase activity were observed, especially in 3 h BSO-rats. Alterations in kinetic parameters of G6Pase were calculated in both intact and detergent-treated microsomes, using glucose-6-phosphate and pyrophosphate as substrate. A little increase in thiobarbituric acid-reactive substances and a limited decrease in 5,5'-dithiobis(2-nitrobenzoate)-reactive protein thiols were also noted. The results of this study show that acute GSH depletion induced by BSO is able to affect hepatic microsomal G6Pase activity. A possible explanation to account for the effect of BSO-induced GSH depletion on hepatic G6Pase system is discussed.

Abstract Xanthan Gum is the most popular biopolymer used today to provide hole cleaning and suspension capabilities to water based drilling fluids but it presents some limitations as regards temperature sensitivity and tolerance to field contaminants. In this paper a Scleroglucan is proposed as a better alternative to Xanthan Gum for drilling fluid compositions. Superior benefits offered by Scleroglucan on hole cleaning, rate of penetration, temperature sensitivity and mud tolerance to shale have been demonstrated by successful field applications and by extensive laboratory studies. Introduction Scleroglucan is a biopolymer produced by fermentation of a plant pathogen fungus of genus Sclerotium that has already found wide applications for enhanced oil recovery such as for polymer flooding and water shutoff. Owing to its excellent suspending and viscosifying power, its high resistance to shear rate and its unique thermal stability it is presently considered one of the best mobility control agent in high salinity and high temperature reservoirs. So far, only a few authors have considered the potential advantages of using Scleroglucan also for drilling applications. In the patent literature Scleroglucan has been recently proposed for drilling fluids, spacers, and cement slurries. The Scleroglucan non ionic nature and major thermal stability suggest that it should be able to perform better than Xanthan Gum for many drilling applications. No published data have never been reported, however, regarding drilling applications of Scleroglucan. By the authors the performances of Xanthan Gum and Scleroglucan based muds have been directly compared by drilling nearby wells with carefully designed formulations. Substitution of Xanthan Gum with Scleroglucan in a potassium based mud resulted in a reduction of dilution rates (-20%) and time spent in mud conditioning or bit balling (-60%). Besides, improved mud rheology also contributed to the realization of much faster drilling rates through the water sensitive shales of the 12 1/4" section (+40%). In this paper field observations are discussed and extended with the results of laboratory studies in which fundamental rheology plays a major role. Rheological measurements are performed both in stationary and oscillatory fields. Low shear rate viscosities at 0.06 sec and Herschel & Bulkley yield stresses are used to quantify hole cleaning capabilities and critical MBT of lab formulations and field muds. Evaluation of cleaning efficiency and cutting concentration is performed with an in house developed program using the Walker and Mayes theory modified with the Herschel & Bulkley rheological approach. The oscillation parameters Storage Modulus (G'), Loss Modulus (G") and Loss Tangent (G"/G') are used to characterize the "undisturbed" structure and thixotropy of the systems. According to existing theories and to the results of the authors field experience and laboratory studies with analogous formulations, Loss Tangent and Storage Modulus are related to the degree of structuration and to the "strength" of the structure that develops in biopolymer solutions and muds under static conditions. As a consequence, these data are analyzed to determine the suspension capability and resistance to shale contamination of the systems. On the basis of field experience and laboratory studies the main advantages offered by Scleroglucan for drilling muds compositions are confirmed and underlined. The reported field data refer to the first application of Scleroglucan in a Potassium Formate based mud. In the same field other wells were drilled using Scleroglucan in different operative conditions and mud formulations and always with good results. The discussion of these data will be reported elsewhere. P. 105