Search

Coherent structures play a dominant role in affecting the wake features of submarine propellers. Large-Eddy Simulation (LES) is especially well suited for investigating such influence, since the large energy-carrying structures of the flow are fully resolved with no turbulence modeling. However, detailed validations with experiments are rather limited in the literature, especially for the forward mode of operation, due to the challenges associated to both computations and experiments of this class of flows. Here the focus will be especially on the comparison of present LES computations of a notional seven-bladed propeller in open-water conditions with Particle Image Velocimetry (PIV) results, to assess carefully the capability of LES to capture the flow physics. A comparison across three values of advance coefficient will be also presented, based on the full three-dimensional fields from computations, in order to illuminate the influence of this parameter on flow features and wake coherence.

Large-Eddy Simulation, with an Immersed-Boundary method, is adopted to analyze the wake of a notional submarine propeller, using a cylindrical grid composed of about 840 million nodes. The accuracy of the overall approach is validated by comparisons with available measurements. Three different load conditions are investigated, allowing an assessment of the sensitivity to the rotational speed. Results demonstrate that tip and hub vortices are the main coherent structures. However, several additional vortices are produced across the span of the propeller blades. The evolution of turbulent kinetic energy in the propeller wake is substantially affected by coherent structures. The highest values occur at the axis and at outer radii, due to the hub and the tip vortices, respectively. In addition, downstream evolution of turbulence associated to the tip vortices is not monotonic. In contrast, fluctuations at the wake axis, originating mainly from instability of the hub vortex, keep decreasing starting from the near wake.

A new diclofenac salt called diclofenac-choline (DC) has recently been proposed for the symptomatic treatment of oropharyngeal inflammatory processes and pain because its greater water solubility allows the use of high concentrations, which are useful when the contact time between the drug and the oropharyngeal mucosa is brief, as in the case of mouthwashes or spray formulations. The antioxidant activity of DC has not yet been investigated, and so the aim was to use luminol-amplified-chemiluminescence (LACL) to verify whether various concentrations of DC (1.48, 0.74 and 0.37 mg/mL for incubation times of 2, 4 and 8 min) interfere with oxygen and nitrogen radicals during the course of human neutrophils respiratory bursts; electron paramagnetic resonance (EPR) spectroscopy was used to investigate its direct antiradical (scavenger) activity. The EPR findings showed that DC has concentration-dependent scavenging activity against the ABTS, the DPPH, and the hydroxyl radicals, but no activity on superoxide anion, as has been previously reported in the case of other NSAIDs. LACL revealed an inhibitory effect that was statistically significant after only 2 min of incubation, and similar after 4 and 8 min. The effects on the peroxynitrite radical paralleled those observed in the previous test. High concentrations and short incubation times showed that there is no interference on PMN viability, and so the inhibitory findings must be attributed to the effect of the drug. The anti-inflammatory effects of DC cannot be attributed solely to the inhibition of prostaglandin synthesis, but its effects on free radicals and neutrophil bursts suggest that they may contribute to its final therapeutic effect.

Background IgA nephropathy (IgAN) is a common complex disease with a strong genetic involvement. We aimed to identify novel, rare, highly penetrant risk variants combining family‐based linkage analysis with whole‐exome sequencing (WES). Methods Linkage analysis of 16 kindreds of South Italian ancestry was performed using an ‘affected‐only’ strategy. Eight most informative trios composed of two familial cases and an intrafamilial control were selected for WES. High‐priority variants in linked regions were identified and validated using Sanger sequencing. Custom TaqMan assays were designed and carried out in the 16 kindreds and an independent cohort of 240 IgAN patients and 113 control subjects. Results We found suggestive linkage signals in 12 loci. After sequential filtering and validation of WES data, we identified 24 private or extremely rare (MAF

This Chapter deals with the experimental survey of the flow field around a catamaran advancing in steady drift. The survey was aimed at investigating the dynamics of the keel vortices with particular emphasis on the mechanisms underlying their generation, detachment, evolution and destabilization. Detailed flow measurements were undertaken by Stereo-PIV (S-PIV) at selected cross planes along and downstream of the catamaran. The test matrix consists of two Froude numbers (Fr=0.4 and 0.5) and two drift angles (6° and 9°). In the experiments, the model was kept fixed at the dynamical values of trim and sinkage, as measured in a previous Planar Motion Mechanism (PMM) test campaign. The major topological and kinematical features of the keel vortices are highlighted through the analysis of the mean and fluctuating components of the velocity and vorticity field. A conditional average technique was used to investigate the interaction between the keel-vortex and free-surface in the wake of the model.

Resveratrol (trans-3,4',5-trihydroxystilbene, CAS 501-36-0), a natural antioxidant and polyphenol found in grapes and wine, has been found to pharmacologically precondition the heart in nitric oxide (NO)-dependent manner. In the vascular system, NO functions as an endogenous inhibitor of leukocyte chemotaxis, adherence, and activation. The present study was designed to determine if resveratrol, through NO, can block the proadhesive molecules generated in the ischemic reperfused myocardium. Isolated hearts were prepared from properly anesthetized rats, and mounted on a Langendorff apparatus. The hearts were randomly assigned to one of the three groups: (i) control, (ii) resveratrol, and (iii) resveratrol + NG-nitro-L-arginine ethyl ester (L-NAME). The hearts were perfused in the absence (n = 6) or presence of 10 micromol/L resveratrol (n=6) or resveratrol + L-NAME (n = 6) for 15 min. All the hearts were then subjected to 30 min ischemia followed by 2 h of reperfusion. Ventricular function was monitored, infarct size and apoptotic cell death measured, and the proadhesive molecules and malonaldehyde formation determined in the perfusate. Resveratrol significantly improved postischemic ventricular function and reduced myocardial infarct size compared to the non-treated control group. The amount of proadhesive molecules including soluble intracellular adhesion molecule-1 (sICAM-1), endothelial leukocyte adhesion molecule-1 (sE-Selectin) and vascular cell adhesion molecule-1 (sVCAM-1) were each significantly decreased during reperfusion in the resveratrol group. L-NAME, a NO blocker, completely abolished such beneficial effects of resveratrol. The results support an anti-inflammatory action of resveratrol through a NO-dependent mechanism.

This study reports the immunomodulatory activity on human Monocyte Derived Dendritic Cells (MDDCs) of a vaccine preparation shown to be effective against an HPV16-related tumour in an animal model. The vaccine is composed of extract from Nicotiana benthamiana leaves containing HPV16 E7 protein expressed by a potato virus X-derived vector (NbPVX-E7). The effect of the extract was evaluated on MDDC differentiation and maturation by monitoring the phenotypic expression of specific markers. The results show that NbPVX-E7 does not induce monocyte differentiation to dendritic cells, but does induce MDDC maturation. Plant extract does not influence MDDC-uptake of E7-FITC while it significantly improves the Ovalbumin-FITC uptake, considered as a model antigen. Importantly, NbPVX-E7-pulsed MDDCs/PBMCs are able to prime human blood-derived lymphocytes from healthy individuals to induce HPV16 E7-specific cytotoxic activity. This is a propaedeutic study for a possible use of E7-containing plant extract in human immunotherapy of HPV-related lesions.

There is growing interest in natural chemical compounds from aromatic, spicy, medicinal and other plants with antioxidant properties in order to find new sources of compounds inactivating free radicals generated by metabolic pathways within body tissue and cells, mainly polymorphonuclear leukocytes (PMNs) whose overregulated recruitment and activation generate a large amount of reactive oxygen species (ROS) and reactive nitrogen species (RNS), leading to an imbalance of redox homeostasis and oxidative stress. The aim of this study was to examine whether a propylene glycol extract of Calendula officinalis interferes with ROS and RNS during the PMN respiratory bursts, and to establish the lowest concentration at which it still exerts antioxidant activity by means of luminol-amplified chemiluminescence. Electron paramagnetic resonance (EPR) spectroscopy was also used in order to confirm the activity of the C. officinalis extract. The C. officinalis extract exerted its anti-ROS and anti-RNS activity in a concentration-dependent manner, with significant effects being observed at even very low concentrations: 0.20 μg/ml without L-arginine, 0.10 μg/ml when L-arginine was added to the test with phorbol 12-myristate 13-acetate and 0.05 μg/ml when it was added to the test with N-formyl-methionyl-leucyl-phenylalanine. The EPR study confirmed these findings, 0.20 μg/ml being the lowest concentration of C. officinalis extract that significantly reduced 2,2-diphenyl-1-picrylhydrazyl. These findings are interesting for improving the antioxidant network and restoring the redox balance in human cells with plant-derived molecules as well as extending the possibility of antagonizing the oxidative stress generated in living organisms when the balance is in favor of free radicals as a result of the depletion of cell antioxidants.

Adult rats were treated for ten days with capsaicin or with NaCl 0.9% directly injected into the lateral cerebral ventricles through a surgically implanted cannula. A third group of rats was implanted with the same cannula but did not receive any treatment. The food intake and the body weight were recorded for at least six weeks after stopping the treatment. The animals were always kept at constant ambient temperature of 22 °C. The body weight of the capsaicin-treated group was reduced by the treatment, and showed a regular but lower degree of recovery trend than the control groups after the treatment period. In fact the capsaicin treated animals never reached the body weight of the controls. Nevertheless, food intake did not significantly vary after the capsaicin treatment. On the basis of these and previous findings, we can assume that capsaicin injected into the cerebral ventricles to rats kept at constant ambient temperature can acts on hypothalamic neurons, but a permanent action on metabolic pathways can not be excluded.

The expression pattern of the pannexin2 protein (Px2) in healthy and ischemized brains of adult rats was investigated. A polyclonal antibody for rat Px2 was generated in chicken and purified for affinity. This antibody was used to study by Western blot, Enzyme-Linked Immunosorbent Assay, and immunohistochemistry, the expression pattern of Px2 in healthy brain of adult rats and in the hippocampus of rats submitted to bilateral clamping of carotid arteries for 20 min, followed by different times of reperfusion (I/R) (8 h, 24 h, 48 h, 72 h, 14 days and 30 days). Immunohistochemical studies visualized the wide and complex expression pattern of Px2 in the healthy brain. All Px2(+) positive cells were neurons which also showed no puncta on their cellular membranes. Both pyramidal cells and interneurons, the majority of which were positive to parvalbumin, were stained in healthy hippocampus. The number of Px2 interneurons in the hippocampus showed a progressive reduction at successive time intervals after I/R, with a negative peak of about -40% after 72 h from I/R. Interneurons which were positive for both Px2 and parvalbumin, represented about the 85% of all parvalbumin cells stained in the hippocampus. This percentage rested grossly unmodified at different time intervals after I/R in spite of the progressive neuronal depletion. Concomitantly, an intense astrogliosis occurred in the hippocampus. Most of the astroglial cells expressed de novo and for a transient time (from 24 h to 14 days from I/R), Px2. Primary co-cultures of hippocampal neurons and astrocytes were submitted to transient ischemia-like injury. This set of experiments further confirmed the in vivo results by showing that Px2 is de novo and transiently expressed in astroglial cells following a transient ischemia-like injury. These results suggested the expression of Px2 in the astrocytes may be induced either from injured neurons or by biochemical pathways internal to the astrocyte itself. In conclusion, our results showed the transient expression of Px2 in astrocytes of reactive gliosis occurring in the hippocampus following I/R injury. We hypothesize that Px2 expression in astrocytes following an ischemic insult is principally involved in the formation of hemichannels for the release of signaling molecules devoted to influence the cellular metabolism and the redox status of the surrounding environment.

Recent studies have documented that grapes and grape juices are equally cardioprotective as red wine. The existing reports implicate that the skin and seeds of the grapes containing polyphenolic antioxidants are instrumental for the cardioprotective properties of grapes. The present study examines if the flesh of grapes also possesses any cardioprotective abilities. Three groups of randomly selected rats were fed, water only (control), flesh of the grapes (2.5 mg/kg b. wt.) or the skins (2.5 mg/kg b. wt.) for 30 days. At the end of the 30 days, isolated perfused hearts were made ischemic for 30 min followed by 2 h of reperfusion in the working mode. The results demonstrated that both flesh and skin of the grapes could protect the hearts from ischemic reperfusion injury as evidenced by improved postischemic ventricular recovery and reduced myocardial infarct size. High performance liquid chromatography (HPLC) revealed that skin and flesh contained comparative amounts of glucose, fructose, tartaric acid, malic acid, shikimic acid, and trans-caftaric acid. In addition, the flesh contained reduced amounts (compared to skin) of cis-coutaric, trans-coutaric, caffeic, p-coumaric, cinnamics, and catechin/epicatechin. Total polyphenolic index was also lower in flesh compared to skin. The anthocyanins were present exclusively in the skin. Electron paramagnetic resonance (EPR) spectrometry of hydroxy radicals indicated that both flesh and skins possessed equal amount of ROS scavenging activities. Total malonaldehyde content in the heart was reduced comparatively with either flesh or skin. The results indicate for the first time that the flesh of grapes are equally cardioprotective as skin, and antioxidant potential of skin and flesh of grapes are comparable with each other despite of the fact that flesh does not possess any anthocyanin activities.

The present paper reviews two "unconventional" approaches to experimentally address the diagnostics, analysis and solution of hydroacoustic problems in naval engineering. The first approach, based on direct pressure fluctuation measurements combined with detailed flow measurements in the proximity of the noise source, provides a direct estimate of the flow phenomena at the origin of sound generation and emission. The second approach is a relatively recent strategy to investigate the sources of acoustic noise in the aeronautical field and concerns the use of volumetric techniques, such as Tomographic PIV (Elsinga, et al., 2006), in combination with acoustic analogies. The abilities of the two approaches are investigated with reference to the analysis of the noise sources in some test cases consisting of an isolated propeller, a propeller operating in the wake a surface ship, and an open-water propeller-rudder system. Both approaches are shown to enable the physical interpretation of the potential mechanism of noise generation and emission from a naval propeller.

OBJECTIVE: The largest Italian asbestos cement factory had been active in Casale Monferrato until 1986: in previous studies a substantial increase in the incidence of pleural mesothelioma was found among residents without occupational exposure to asbestos. To estimate exposure to asbestos in the population, this study evaluated the presence of histological asbestosis and the lung burden of asbestos fibres (AFs) and asbestos bodies (ABs). METHODS: The study comprises the consecutive series of necropsies performed at the Hospital of Casale Monferrato between 1985 and 1988. A sample of lung parenchima was collected and stored for 48 out of 55 necropsies. The AF concentration was measured with a TEM electron microscope with x ray mineralogical analysis. The ABs were counted and fibrosis evaluated by optical microscopy. The nearest relative of each subject was interviewed on occupational and residential history. Mineralogical and histological analyses and interviews were conducted in 1993-4. RESULTS: Statistical analyses included 41 subjects with AF, AB count, and interview. Subjects without occupational exposure who ever lived in Casale Monferrato had an average concentration of 1500 AB/g dried weight (gdw); Seven of 18 presented with asbestosis or small airway lung disease (SAL). G2 asbestosis was diagnosed in two women with no occupational asbestos exposure. One of them had been teaching at a school close to the factory for 12 years. Ten subjects had experienced occupational asbestos exposure, seven in asbestos cement production: mean concentrations were 1.032 x 10(6) AF/gdw and 96,280 AB/gdw. Eight of the 10 had asbestosis or SAL. CONCLUSION: The high concentration of ABs and the new finding of environmental asbestosis confirm that high asbestos concentration was common in the proximity of the factory. Subjects not occupationally exposed and ever living in Casale Monferrato tended to have higher AB concentration than subjects never living in the town (difference not significant). The concentrations of ABs and AFs were higher than those found in other studies on nonoccupationally exposed subjects.

Oxidative stress is increasingly recognised as a pivotal factor that plays a number of roles in the inflammatory response to environmental signals. It has been claimed that Aesculus hippocastanum extracts have antioxidant and anti-inflammatory activity, but these claims are mainly based on the results of chemical reactions and folk-medicine.The aim of this study was to examine whether a bark extract of Aesculus hippocastanum interferes with reactive oxygen/nitrogen species (ROS/RNS) during the course of human neutrophil respiratory bursts, and to establish the lowest concentration at which it still has antioxidant activity by means of luminol amplified chemiluminescence (LACL). We also studied its ability to counteract lipid peroxidation (LPO) in human cells. Before investigating its antioxidant effects on human cells, we analysed its scavenging activity against ABTS*+, hydroxyl radical, superoxide anion, and Fremy's salt (those last three by means of electron paramagnetic resonance (EPR) spectrometry).The extract of Aesculus hippocastanum exerted its anti-ROS/RNS activity in a concentration-dependent manner with significant effects being observed for even very low concentrations: 10 microg/ml without L-Arg, and 5 microg/ml when L-Arg was added to the fMLP test. The LPO assay confirmed these results, which were paralleled by the EPR study.These findings are interesting for improving the antioxidant network and restoring redox balance in human cells, and extend the possibility of using plant-derived molecules to antagonise the oxidative stress generated in living organisms when the balance is in favour of free radicals as a result of the depletion of cell antioxidants.

The aim of the study was to explore the antiradical activity of sulfurous water, used for inhalatory therapy (characterized by the presence of sulfhydryl [HS]) by means of electron paramagnetic resonance (EPR) spectroscopy. The effects of sulfurous water corresponding to the concentrations from 16 down to 0.25 μg/mL of HS were tested by means of Fenton reaction (HO•), KO2-crown ether system (O2-•), and EPR of Tempol and of Fremy's salt radical. All of these assays were made using natural sulfurous water or degassed sulfurous water (no detectable HS) or reconstituted sulfurous water (degassed plus NaHS). The free radicals were significantly inhibited by natural water with HS concentrations ranging from 16 to 1 μg/mL for HO•, Tempol, and Fremy's salt, and O2-• was significantly inhibited from 16 and 2 μg/mL. The tests of degassed water did not reveal any significant differences from baseline values. The tests of reconstituted water led to significant results overlapping those obtained using natural water, thus confirming the importance of the presence of HS group (reductive activity). The positive effects of the activity of sulfurous thermal water is partially based on the patients' subjective sense of well-being and partially on symptomatic (or general) clinical improvements that are sometimes difficult to quantify. These findings indicate that, in addition to their known mucolytic activity and trophic effects on respiratory mucosa, the HS groups in sulfurous water also have antioxidant activity that contributes to the water's therapeutic effects on upper and lower airway inflammatory diseases.

Platelets are the second most abundant cell type in blood and are essential for maintaining haemostasis. Their count and volume are tightly controlled within narrow physiological ranges, but there is only limited understanding of the molecular processes controlling both traits. Here we carried out a high-powered meta-analysis of genome-wide association studies (GWAS) in up to 66,867 individuals of European ancestry, followed by extensive biological and functional assessment. We identified 68 genomic loci reliably associated with platelet count and volume mapping to established and putative novel regulators of megakaryopoiesis and platelet formation. These genes show megakaryocyte-specific gene expression patterns and extensive network connectivity. Using gene silencing in Danio rerio and Drosophila melanogaster, we identified 11 of the genes as novel regulators of blood cell formation. Taken together, our findings advance understanding of novel gene functions controlling fate-determining events during megakaryopoiesis and platelet formation, providing a new example of successful translation of GWAS to function.

We present correction procedures that compensate for the limited spatial resolution of particle image velocimetry (PIV) and hot wire anemometry (HWA) measurements. Both techniques provide two components of the velocity vector. The correction method, which is based on DNS velocity fields, is then applied to measurements in the far field of two round jets at a turbulent Reynolds number of about 179, for PIV, and 370, for HWA. Both large- and small-scale quantities are discussed. Amongst the latter, the correction of the velocity derivative variance is larger for the PIV data than for the HWA data. However, the opposite is true for the correction of the isotropy ratios of the velocity derivatives. After compensation, the two techniques provide similar values of the mean energy dissipation rate and also of one isotropy ratio.

A group of 18 patients, before undergoing otoplastic surgery, was treated with miocamycin 600 mg in a single dose; middle ear tissue samples were collected at 2, 3, 4 and 6 h. A group of 20 patients suffering from secretory otitis media (SOM) was also treated with 600 mg of miocamycin in a single dose; collections of middle ear fluid (MEF) were performed at 1, 2, 4 and 6 h. Blood withdrawals from all patients occurred at the same time intervals. The miocamycin determination was performed by means of a microbiological method employing Sarcina lutea ATCC 9341. The highest serum levels (1.06 +/- 0.13 mg/l) were found at 2 h in the group from which middle ear mucosa had been withdrawn and at 1 h (2.2 +/- 0.5 mg/l) in the group from which the MEF had been collected; at 6 h miocamycin could be determined in both groups, with values ranging from 0.1 to 0.2 mg/l. In middle ear mucosa the miocamycin concentration was 1.52 +/- 0.27 mg/kg at 2 h and 0.2 +/- 0.09 mg/kg at 6 h. In MEF, the miocamycin concentration was 2.1 +/- 0.27 mg/l at 1 h and 0.24 +/- 0.05 mg/l at 6 h. The miocamycin concentration determined at 1 h in MEF was virtually equal to that in serum at the same time. At the following experimental times of withdrawal, the miocamycin concentration in mucosa specimens, as well as in MEF samples, proved to be markedly higher than values simultaneously found in serum.

Network of Excellence to facilitate the continuation of leadership of the European Hydrodynamic testing facilities. The objective of this alliance is to develop a formal and lasting structure to coordinate the definition and introduction of novel measurement, observation and analysis technologies for hydrodynamic model testing. This benchmark was defined to compare the efficiency of the Stereo PIV systems of the HTA Members and to compare their know-how. A Stereo PIV study on a flat plate wake is presented. The main aim of the present work is the efficiency testing of a Stereo PIV system. In fact the study of a flat plate wake, semi-submerged, near the free surface is a good benchmark for SPIV techniques. To achieve this work a flat plate (80 cm height and 50 cm chord) is vertically submerged to a depth of 30 cm; so the tip is near the free surface and is possible to test the acquisition system in presence of bubbles and other disturbances. Many tests are performed at varying incidence (5, 20 and 40 degrees) and velocity in a towing tank. The used Stereo PIV system is a fully submersible probe (Pereira et al. 2003); it is equipped with two 2048x2048 px2 CCD cameras and a 200 mJ Nd-YAG laser. The system is modular and can be assembled into different configurations. Hollow glass spheres of 30 mm diameter are used as seeding. The investigation zone is about 250 mm x 300 mm, the laser thickness is about 5 mm. The used target for the Soloff et Al.(1997)'s based calibration is a two-dimensional dotted grid, with a centre mark; grid spacing is 20 mm x 20 mm. Instead a pin target and uniform flow acquisitions are used for the flow-based calibration. The measurements give information on instabilities of the tip vortex of the plate. In fact the tip structures varying intensities and geometry with the Reynolds number and with the incidence the test comprises also measurement planes along the chord at defined distances from the leading edge

Twenty patients undergoing tonsillectomy were treated with a peroral administration of 600 mg of miocamycin every 12 hours for 4 days. On the 5th day, after a last administration of a dose of 600 mg the ablation of the tonsils was carried out on groups of 4 subjects, each one at the following times after oral intake of the drug: 2, 4, 6, 8, 12 h. Twenty-nine patients, admitted to hospital to undergo lung resection were treated with peroral administration of miocamycin in accordance with the above mentioned dose scheme. The operation was carried out on groups of 5 subjects, each on the fifth day at the following times after the last administration: 2, 3, 4, 6, 8 and 12 h (4 subjects). Simultaneously blood was withdrawn for the determination of miocamycin in serum. Miocamycin was measured by a microbiological procedure using Sarcina lutea ATCC 9341. The highest levels of miocamycin were observed after 2h in tonsils (3.2 +/- 0.82 mg/kg) and serum (1.3 +/- 0.33 mg/l). After 12h miocamycin proved to be still measurable in the tissue (0.12 +/- 0.05 mg/kg), whereas it was not detected in serum. In pulmonary tissue, the highest levels of miocamycin were likewise identified at the 2nd hour (2.82 +/- 0.59 mg/kg), simultaneously with the highest serum levels (2.3 +/- 0.61 mg/l). At the 12th hour miocamycin could still be dosed in 3 tissue samples, with values between 0.1 and 0.2 mg/kg and was found just at dosing limits in only one serum sample.

Host defense, as it generally applies to humans, refers to the individual's ability to withstand infections. Human host defense mechanisms are numerous, diversified, complex, and often interdependent. The administration of drugs may influence some phases of immunocompetence mechanisms. The effect of several antimicrobial agents on organic defenses has been studied. The parameters considered were chemotaxis, phagocytosis, intracellular killing, superoxide-anion production, antibody production, lymphocyte subset behavior, and natural killer cell activity. Some antibacterial agents inhibit these parameters, whereas others can enhance some of these immune parameters to differing extents. Some antibacterial agents have a neutral effect on these parameters.

Augmentin (875 amoxicillin and 125 mg potassium clavulanate) was administered orally to patients with chronic bronchitis. Concentrations of amoxicillin and clavulanic acid were measured in serum, sputum and urine. Peak serum levels for amoxicillin of 11.23 +/- 2.61 micrograms/ml were observed at 2 hours and for clavulanic acid of 2.55 +/- 0.54 micrograms/ml at 1 hour. After 9 hours, 50% of the amoxicillin and 39% of the clavulanic acid had been renally excreted. The peak sputum concentration of amoxicillin was 1.31 +/- 0.42 micrograms/ml at 4 hours and of clavulanate was 0.79 +/- 0.23 micrograms/ml at 2 hours. Patients awaiting surgery received an oral dose of augmentin as above. Samples of lung, tonsil, middle ear mucosa and prostate were obtained and tissue concentrations of both compounds measured. Peak levels of amoxicillin ranged from 0.87 micrograms/g (tonsil) to 2.56 micrograms/g (lung) and of clavulanic acid from 0.20 micrograms/g (prostate) to 0.56 micrograms/g (lung) between 3 and 4 hours after dosing.

In 265 patients operated for breast carcinoma the monoclonal antibody serum test CA 15.3 was predictive of metastatic diffusion of the disease. Its level increased in cases of distant metastasis with no significant difference between multiple and single sites (p = 0.014). The concentration of the marker was higher in 21 (23.8%) patients without nodal involvement and in 19 (27.5%) with nodal involvement (p = 0.193). Our study suggests that CA 15.3 may be an aid in the follow-up of patients with metastatic diffusion of breast cancer.

MTT (3-(4, 5-dimethyl-2-thiazolyl)-2, 5-dihphenyltetrazolium bromide) assay is a widely used method to assess cell viability and proliferation. MTT is readily taken up by cells and enzymatically reduced to formazan, a dark compound which accumulates in cytoplasmic granules. Formazan is later eliminated by the cell by a mechanisms often indicated as exocytosis, that produces characteristic needle-like aggregates on the cell surface. The shape of formazan aggregates and the rate of exocytosis change in the presence of bioactive amyloid beta peptides (Abeta) and cholesterol. Though the cellular mechanisms involved in MTT reduction have been extensively investigated, the exact nature of formazan granules and the process of exocytosis are still obscure. Using Nile Red, which stains differentially neutral and polar lipids, and a fluorescent analog of cholesterol (NBD-cholesterol), we found that formazan localized in lipid droplets, consistent with the lipophilic nature of formazan. However, formazan granules and aggregates were also found to form after killing cells with paraformaldehyde fixation. Moreover, formazan aggregates were also obtained in cell-free media, using ascorbic acid to reduce MTT. The density and shape of formazan aggregates obtained in cell-free media was sensitive to cholesterol and Abeta. In cells, electron microscopy failed to detect the presence of secretory vesicles, but revealed unusual fibers of 50 nm of diameter extending throughout the cytoplasm. Taken together, these findings suggest that formazan efflux is driven by physico-chemical interactions at molecular level without involving higher cytological mechanisms.

Though the fluid dynamic fields past stationary NACA airfoils have been deeply investigated and understood, open questions still remain when unsteady pitching conditions are considered. This is even of greater relevance in the aero-naval applications where such unsteadiness becomes critical in maneuvering situations. Rotors in helicopters and maneuvering rudders in ships and submarines are clear examples of such cases. Regarding the physics, the variation of the angle of attack generates a modification of the circulation around the airfoil which is likely to be compensated, in the framework of the Kutta theory, by a vortex shedding in order to have a net vorticity balance equal to zero. So when the angle of attack is varied continuously, a street of vortices has to be expected downstream the airfoil. In order to experimentally investigate this problem, the flow past a NACA-0009 airfoil, with span to chord ratio larger than four, has been tested; to simulate the continuous variation of the angle of attack, the airfoil has been put in rotation around its aerodynamic center according to a sinusoidal law. To investigate the influence of the oscillation amplitude and of the pitching frequency, velocity and vorticity fields are presented as well as Jeong's invariant on 2D velocity fields.

The expression pattern of pannexin1, a gene coding for a protein that forms gap junction channels, was studied as both mRNA and protein in the CNS of adult mouse. Pannexin1 was widely expressed in the CNS by neuronal cell types but not glial cells, except for Bergmann glial cells of the cerebellar cortex. Cells positive to Ca-binding proteins, principally parvalbumin, but also calbindin and calretinin, as well as glutamate decarboxylase 67 kDa isoform, were pannexin1-positive. Pannexin1 labeling was found in cells which are known to exhibit spontaneous and synchronous discharge, such as neurons of the inferior olivary complex and the reticular thalamic nucleus, and also in neurons whose electrical activity is not coupled with neighboring cells, such as motoneurons of the spinal cord. The analysis of cellular localization showed puncta that surrounded cell bodies (e.g. the pyramidal cells of hippocampus) or restricted areas inside the cell bodies (e.g. the spinal motoneurons). In Bergmann glial cells the staining was present as fine grains that covered a large part of the cellular surface. Pannexin1 stained cells that previous studies have reported as expressing connexin36, another protein forming gap junction channels. Thus, it was possible that these two proteins could be integrated in the same functions. Since connexin36 expression levels change after seizures, we examined the expression of both pannexin1 and connexin36 in cerebral cortex, hippocampus, cerebellum and brain stem at different time intervals (2, 4 and 8 h) after i.p. injection of 4-aminopyridine, which resulted in systemic seizures. The only modification of the expression levels observed in this study concerned the progressive decrement of the connexin36 in the hippocampus, while pannexin1 expression was unchanged. This finding suggested that pannexin1 and connexin36 are involved in different functional roles or that they are expressed in different cell types and that only those expressing the Cx36 are induced to apoptosis by epileptic seizures.

The pool of different carnitine derivatives is formed by carnitine, acetylcarnitine, propionylcarnitine and isovalerylcarnitine. Isovalerylcarnitine is a compound performing activities that differ from those of the other carnitine esters. Its activity on proteolytic enzymes and on the calpain system has been demonstrated in the past. Both the calpain and the caspase systems belong to the protease family and lead to cytochrome activation and apoptosis. The two systems can interact to promote apoptosis. In view of this proapoptotic activity of isovalerylcarnitine, studies were carried out to ascertain whether this carnitine derivative influences cell-reaction processes associated with apoptosis. U937 leukemic cells were selected for these studies because they are a well-established model for the assessment of cellular immune responses. In addition to nuclear morphologic alterations produced by apoptosis that can be detected by specific histochemical and microscopic methods, we also took other cell functions into consideration, such as phagocytosis, cell killing and cell growth, which are indices of immune function related to apoptosis. Unlike reference carnitine forms, isovalerylcarnitine produced an early and marked increase in phagocytosis and also an increase in cell killing. Cell proliferation was reduced. The hypothesis is set forth that isovalerylcarnitine may be a caspase-activating, proapoptotic factor that resembles various anticancer agents, which induce early apoptosis that coincides with early activation of caspase. This hypothesis is supported by the ability of isovalerylcarnitine to induce early phagocytosis and cell killing.

A new chemical named parrodienes has been extracted from parrots' plumage. From the chemical point of view, parrodienes are polyunsaturated aldehydes similar to carotenoids. On the basis of this similarity we organized some biological experiments to evaluate the inhibition of lipoperoxidation of cell membranes induced by CCl4, protection against ultraviolet rays, anti-inflammatory activity and protection from an increase of ornithine-decarboxylase as marker of tumoral skin alteration. The results of these experiments showed that unsaturated dienes (parrodienes) play an important role on the inhibition and prevention of many biological processes that are at the basis of different pathological disorders.

In view of the high concentration of resveratrol found in a new autochthonous wine (Uvalino) and the notable antioxidant activity of this substance, we decided to assess whether this wine could inhibit the production of free radicals. Nowadays, free radicals are considered to be the most noxious factors for tissues, triggering the development of many diseases. The assessments were carried out using a direct and more precise technique, electronic paramagnetic resonance (EPR), which is able to detect the ability of an antioxidant agent to inhibit the formation of hydroxyl radicals (*OH), which are among the most noxious reactive oxygen species (ROS). The results show that Uvalino wine is able to eliminate ROS production almost completely. Consequently, it has beneficial effects on health in all the diseases in which ROS plays an important pathogenetic role.

The ability of two samples of red wine with different resveratrol concentrations to inhibit hydroxyl radicals (*OH) produced by a Fenton-type reaction was assessed using the method of electronic paramagnetic resonance (EPR). One sample was an autochthonous wine, Uvalino, which has a very high resveratrol concentration; the second was another red wine with a much lower resveratrol concentration. The ability of the sample of Uvalino wine to obstruct hydroxyl radicals was evident, but it wasn't much better than the ability of the sample of wine with a lower resveratrol concentration. The resveratrol concentration of wine is an important factor for the inhibition of the formation of free radicals, especially hydroxyl radicals; however, it is not the only one responsible for this property of wine. Resveratrol concentration can act synergically with other factors, such as polyphenols, which are also contained in wine and have antioxidant properties.

The antioxidant activity of a novel series of derivatives with a carotenoid-like structure was studied. These derivatives have recently been isolated chemically as a result of studies on the pigments present in a particular species of birds, namely parrots. These novel derivatives, which are also called parrodienes, have been proved to possess interesting biological properties that differ from those that carotenoids are known to have. The objective of this study was to demonstrate the ability of these novel compounds to inhibit the formation of reactive oxygen species, especially their ability to block the formation of hydroxyl radicals, which are among the most reactive products of oxygen reactions and which produce the greatest damage to cells and tissues. The technique used to assess this antioxidant capacity of parrodienes was electron paramagnetic resonance, which allows direct assessment of inhibition of hydroxyl radical formation (.OH). The results show that these derivatives, especially octatriene, are able to exert evident antioxidant activity, thus confirming that their antioxidant properties are important for their biological activity.

As a part of Ogliastra Project we present preliminary data about the first phase of an epidemiological survey led in a geographically, culturally and genetically isolated population of Sardinia. In particular, we focus our study in a region of Eastern-central Sardinia called Ogliastra, whose population has been demographically stable without massive immigration because of geographic barriers. Population structure of Ogliastra consists of 23 small isolated villages, presenting high endogamy and inbreeding. Furthermore, centenarian cultural isolation and conservatism has generated a great deal of homogeneity in life style and eating habits. Our survey took place in Perdasdefogu, an Ogliastra village of about 2400 inhabitants. Study design is a cross sectional survey aimed at determining prevalence of diseases presenting familial aggregation. Enrollment in the study is based on voluntary participation of the population who gave informed consent. Almost 1400 people, ranging from age 3 to 97 years, underwent an extensive evaluation comprehensive of blood sample tests, anthropometrical and blood pressure measurements, a structured interview to collect socio-demographic data, living habits, exposure to most common risk factors, medical and medication history, information about familial disorders and a physical examination. Clinics for examining people and collecting and analyzing blood samples are all located at the same center in Perdasdefogu. Our physicians examined all people participating in the study and they entered all data on PCs using an electronic medical record based on the structured interview drawn up by the study group. Such an electronic medical record is made of several modules allowing consultation of a pharmacological archive in order to correctly enter medicines taken by patients , consultation of the ICD IX for the correct pathological classification and of an anatomic and semeiotic archive in order to better define the physical examination. The common purpose of every module is to obtain standardized information. All collected data are then automatically transferred to a central database located in Cagliari where they are added to the genealogical and genetic information in order to be available, to authorized personnel, for statistical analysis. Preliminary results will be presented.

The objective of this study was to assess whether tyrosol and caffeic acid are able to inhibit lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-alpha release. TNF is one of the most important cytokines involved in inflammatory reactions. The results show that both tyrosol and caffeic acid are able to inhibit LPS-induced TNF-alpha release from human monocytes, even at low doses. Their mechanisms of action are discussed and we conclude that high doses of the two compounds are not required to achieve effective inhibition of inflammatory reactions due to TNF-alpha release.

Some well-known antioxidant phenols present in extravirgin olive oil have also been found in white wine. Both tyrosol and caffeic acid are phenols that are present not only in extravirgin olive oil, but also in wine, especially white wine. Their antioxidant properties are well known, but their biological effects have not yet been elucidated. In a previous study we found that these substances were able to inhibit tumor necrosis factor alpha release. The present study was carried out to assess whether these compounds are able to inhibit other inflammatory cytokines, such as interleukin-1 beta and interleukin-6. The results show that low concentrations of these phenols, which can be found in the bloodstream after intake of moderate quantities of white wine, exert significant inhibitory activity on the release of several inflammatory cytokines.

Resveratrol, a phytoalexin found in grapes and wine, has been shown to have anti-inflammatory properties. Since endothelium is activated during inflammation by some cytokines released by macrophages and many other cells, we tested whether resveratrol could modulate endothelial cell activation. We studied the effect of resveratrol treatment in vitro on the expression of vascular cell adhesion molecule-1 by tumour necrosis factor alpha-stimulated human umbilical vein endothelial cells. In addition, we studied the effect of resveratrol treatment in vivo (in a murine experimental model) on the modulation of tumour necrosis factor alpha-induced vascular permeability. Resveratrol, used at the concentrations present in human plasma following moderate wine consumption, was demonstrated to be an inhibitor of the adhesion molecule expression by tumour necrosis factor alpha-stimulated endothelial cells. In addition, resveratrol significantly prevented the cytokine-induced vascular leakage. Our results (both in vitro and in vivo) may explain some aspects of the anti-inflammatory effects of resveratrol.

The present study demonstrates that intrathecal injection of interleukin-1 (IL-1) in rats at a dose of 50 pg or 500 pg induced a significant hyperalgesic effect in the hot-plate test. The hyperalgesia or the altered nociceptive responses were not considered to be the result of sensitization to the hot plate, since the nociceptive alteration caused by intrathecal administration of IL-1 stopped after 20 min. The results suggest that IL-1 can affect pain responsiveness in a dose-dependent manner and that it is able to modulate neuronal functions. Taken together, our findings and the data from the literature suggest that IL-1-induced hyperalgesia is related not only to the route of administration but also to the algesia testing method used.

We have examined the most recent literature dealing with the relationship between tumor growth and neovascularization (tumoral neoangiogenesis). It is quite evident that antiangiogenic therapy against solid tumors has an important theoretical basis. Many antiangiogenic substances are presently under study, and some inhibitors of angiogenesis, such as TNP-740, platelet factor 4, linomide, tecogalan, etc., are currently being tested in phase 1 or 2 clinical trials. These studies have, however, underscored the possibility of numerous side effects due to the lack of specificity of these substances against endothelial cells. The most promising and rational use of antiangiogenic factors in anticancer therapy seems to be their association with traditional chemo- or radiotherapy. Gene therapy also offers considerable theoretical promise as demonstrated by the transfection of thrombospondin 1 complementary DNA into a human breast carcinoma cell line.

Rats were taught to self-administer beta-endorphin (0.1 microgram or 0.5 microgram/microliter) or NaCl 9% by pressing a lever that activated a pump connected with a cannula implanted in the lateral cerebral ventricle (icv). The pump delivered 1 microliter at each lever pressing. After a training period of 2 weeks, the self-injection behaviour was studied before, during and after nociceptive stimulation. In response to a nociceptive stimulation, 4 rats increased their self-injection of beta-endorphin at 0.5 microgram/microliter per injection. This effect is specific for beta-endorphin since under identical conditions 6 rats did not increase the injection of isotonic NaCl saline solution. Another 6 rats did not increase their self-injection during nociceptive electrical stimulation and the post-nociceptive period when the dose of beta-endorphin was 0.1 microgram/microliter per injection. However these animals self-administered significantly higher levels of beta-endorphin during the pre-control period. Also studied were the acute effects of 10 micrograms of icv beta-endorphin on tail-flick latency in sec. The acute administration of 10 micrograms of beta-endorphin induced a long-lasting analgesia. The results show that the rats increased beta-endorphin self-administration during the pre-control period when the dose was 0.1 microgram/microliter and during the nociceptive stimulation period when the dose was 0.5 microgram/microliter. In the former case the self-administration had the profile of a voluntary doping drug intake, in the latter case the profile was that of an antalgic self-medication.

Two or three minutes after intracerebroventricular (i.c.v.) injection of Tween 80 plus ethanol as vehicles for capsaicin, strong convulsive movements occurred in the treated rats; they twisted and turned, dashed themselves against their cage and all died. Tween 80 in saline without ethanol also produced convulsions and death in all the rats treated. Capsaicin dissolved in the vehicle seemed to furnish protection against such death. Moreover, when capsaicin in Tween 80 was administered i.c.v. 10 min before the Tween vehicle it prevented the death which would have been caused by the i.c.v. vehicle. Thus a new effect is added to those already discovered in the numerous pharmacological studies on capsaicin.

The function of alveolar macrophages (AMS) in patients with pneumonia (n = 7) (Group A) and chronic obstructive pulmonary disease (COPD) (n = 11) (Group B) was investigated by evaluating the rate of phagocytosis and of the intracellular killing. A control group of healthy subjects (n = 8) was also included. Phagocytosis frequency (PHF), phagocytosis index (PHI) and intracellular killing towards Candida albicans were then evaluated. PHF and PHI were found to be significantly (p0.05) lower in Group B patients than in the control group, while intracellular killing showed a behaviour similar to controls. PHF and PHI values observed in the patients of Group A and in the control group did not show any significant difference; the intracellular killing rate proved on the contrary to be significantly (p0.05) lower than that observed in the controls.