Dioxin-like Compounds in Lake Fish Species: Evaluation by DR-CALUX Bioassay.

Fish consumption is the principal source of intake of organochlorinated compounds in humans. Compared with other types of foods of animal origin, fish contain the highest levels of polychlorinated biphenyls (PCBs), polychlorinated dibenzo- p-dioxins, and polychlorinated dibenzofurans, all of which are classified as highly toxic organochlorine compounds. Currently, lakes and fish farms in northern Italy are not regularly monitored for PCBs and dioxins in areas contaminated by industrial sources, partially because of the high costs of traditional analytical methods that limit the number of samples to be analyzed. The DR-CALUX cell bioassay is based on the uptake of the cellular aryl hydrocarbon receptor (AhR) for dioxins and dioxin-like compounds. The aim of this study was to assess the levels of dioxins and dioxin-like PCB contamination in Lake Maggiore and Lake Como, two lakes in northwestern Italy, and in nearby areas. The levels were quantified using the cell bioassay DR-CALUX and reference controls in two wild fish species, perch ( Perca fluviatilis) and roach ( Rutilus rutilus), and in a farmed species, rainbow trout ( Oncorhynchus mykiss). Tissue samples collected from the farmed rainbow trout were also submitted to immunohistochemical analysis of CYP1A expression as a marker for environmental pollutant-induced liver damage. The levels of dioxins, furans, and dioxin-like PCBs were all below the maximum levels and action limits set by European Union Regulation, suggesting no risk for human health associated with the consumption of the fish species caught or farmed in these areas.

Innovative oral spray-dried Idebenone systems to improve patient compliance.

Idebenone is a high permeable drug with very slight water solubility that affects the dissolution rate in the biological fluids, causing an irregular and limited in vivo absorption after oral administration. Moreover, it is marketed in Europe as tablets equivalent to 150 mg, with the consequent administration of multiple dose of solid unit to obtain the correct dose, a deterrent for the patients' compliance. According to these considerations, our goal was to develop spray-dried microparticles using a soluble ß-cyclodextrin (CD) polymer and an enhancer of dissolution rate, such as carboxymethyl cellulose, to obtain a formulation easily dosable and soluble in water. The complex in solution was evaluated by phase solubility studies and the Idebenone/CD molar ratio selected was 1:1. According to Higuchi and Connors, adding carboxymethyl cellulose, a Bs-type profile was obtained. This result was due to the presence of carboxymethyl cellulose that competes with the CD in forming Idebenone microsystems, reducing of 10-fold the formulation bulk. UV-Vis absorption, (1)H nuclear magnetic resonance and circular dichroism showed the formation of the CD/Idebenone inclusion complex confirmed also by differential scanning calorimetry, Fourier transform infrared spectroscopy and fluorescence microscope (FM). The water solubility data and the in vitro dissolution tests performed in simulated gastric fluid, showed an increase of the drug water interaction due to the presence of the CD and carboxymethyl cellulose, both able to improve drug wettability, water solubility and dissolution rate. This approach seems to be suitable to produce microsystems which are able to enhance the in vivo absorption of Idebenone after oral administration and to increase the patient compliance.

A Comparative Study on the Influence of Probe Placement on Quality Assurance Measurements in B-mode Ultrasound by Means of Ultrasound Phantoms.

To check or to prevent failures in ultrasound medical systems, some tests should be scheduled for both clinical suitability and technical functionality evaluation: among them, image quality assurance tests performed by technicians through ultrasound phantoms are widespread today and their results depend on issues related to scanner settings as well as phantom features and operator experience. In the present study variations on some features of the B-mode image were measured when the ultrasound probe is handled by the technician in a routine image quality test: ultrasound phantom images from two array transducers are processed to evaluate measurement dispersion in distance accuracy, high contrast spatial resolution and penetration depth when probe is handled by the operator. All measurements are done by means of an in-house image analysis software that minimizes errors due to operator's visual acuity and subjective judgment while influences of ultrasound transducer position on quality assurance test results are estimated as expanded uncertainties on parameters above (measurement reproducibility at 95 percent confidence level): depending on the probe model, they ranged from ±0.1 to ±1.9 mm in high contrast spatial resolution, from ±0.1 to ±5.5 percent in distance measurements error and from ±1 to ±10 mm in maximum depth of signal visualization. Although numerical results are limited to the two examined probes, they confirm some predictions based on general working principles of diagnostic ultrasound systems: (a) measurements strongly depend on settings as well on phantoms features, probes and parameters investigated; (b) relative uncertainty due to probe manipulation on spatial resolution can be very high, i.e. from 10 to more than 30 percent;

Design and development of a rheometer for biological fluids of limited availability.

From studies on the dynamic characterization of human bones, it is noticed that reference data on the viscous behavior of the bone marrow are quite poor. Dependently from marrow limited availability and its opacity, we have not been able to retrieve a tool of appropriate characteristics able to measure bone marrow viscosity. Therefore, principal techniques for the viscosity measurement have been preliminarily examined, and a device suitable for viscosity measurements of biological fluids has been realized. In particular, a rotational rheometer has been developed: it is a coaxial cylinders system, where the fluid flows dragged by the inner cylinder. The device is an absolute rheometer, that is, particularly useful as nowadays it is not known the classification of the bone as far as it concerns its viscous behavior. In this work a preliminary evaluation of the metrological characteristics of the measurement system has been carried out and its main metrological performances have been evaluated.

Identification by a proteomic approach of a plasma protein as a possible biomarker of illicit dexamethasone treatment in veal calves.

Corticosteroids have become the most widespread illegal growth promoters in veal calves and beef cattle. Testing for corticosteroids relies on either direct detection of compounds or their metabolites or indirect detection to identify changes in biological pathways. We used a comparative proteomic approach, based on two-dimensional electrophoresis (2DE), to identify plasma protein markers after short-term dexamethasone administration in veal calves. Twenty-three male Friesian veal calves were treated experimentally with dexamethasone sodium phosphate: 10 received low-dose administration of the drug (0.4 mg day⁻¹ per os) for 20 consecutive days (treatment group); 10 received the drug at therapeutic dosage (2-4 mg kg⁻¹ i.m.) for 3 consecutive days (comparison group). Three animals were not treated (control group). Plasma samples were collected from each animal at six time points (T1-T6; treatment and control group) and at four time points (T1-T4; comparison group) and stored at -80°C until analysis. Plasma proteins were quantified and analysed in triplicate by 2DE. The images were analysed with Bionumerics® software. Comparison of 2DE maps obtained from blood samples at T1 (before treatment) and at T6 (final sampling) showed a significant disappearance (p < 0.001) of two protein spots at T6 in the treatment group. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis and immunoblotting identified these isoforms as serum paraoxonase/arylesterase 1 precursor (PON1). Synthesised in the liver and released into the blood, PON1 has an important role in lipid metabolism. The absence of variation of this protein in the comparison group suggests that the marker has good specificity for detecting illicit corticosteroid treatment.

Influence of FBG sensors length on temperature measures in laser-irradiated pancreas: theoretical and experimental evaluation.

Temperature distribution T(x,y,z,t) in tissue undergoing Laser-induced Interstitial Thermotherapy (LITT) plays a crucial role on treatment outcome. Theoretical and experimental assessment of temperature on ex vivo laser-irradiated pancreas is presented. The aim of this work is to assess the influence of thermometers dimensions on temperature measures during LITT. T(x,y,z,t) inside tissue is monitored by optical sensors, i.e., Fiber Bragg Gratings (FBGs): three FBGs with lengths of 10 mm and nine FBGs of 1 mm, at different distances (2 mm, 5 mm and 10 mm) and different quotes (0 mm, 2 mm and 4 mm) from the laser fiber tip are used. Theoretical punctual T(x,y,z,t) is averaged out on both 10 mm and 1 mm in order to compare numerical predictions with experimental data. Results demonstrate the influence of FBG length on T(x,y,z,t) measures. This phenomenon depends on the distance between sensor and applicator: it is particularly significant close to the applicator tip (2 mm) because of the high spatial T(x,y,z,t) gradient within the tissue. Both theoretical results and experimental ones show that just at a distance of 10 mm from the tip, differences between T(x,y,z,t) provided by FBGs of 10 mm and 1 mm are negligible.

Surgical correction of "rhinoplastic look".

A pointed, narrow and exaggeratedly upturned nasal tip and concave dorsal profile can give the nose an unnatural and artificial appearance that is the unmistakable hallmark of plastic surgery. As a result of changes in social attitudes, noses that have evidently been operated on are no longer acceptable and requests are made for correction. While a more natural dorsal profile can be obtained with camouflage grafts of autologous cartilage or alloplastic material (EPTFE), autologous grafts alone are used to reconstruct the tip. The most complex correction regards an obtuse nasolabial angle, for which the extension graft technique is used. Particular care must be taken during preoperative planning, where the aid of computer simulation and agreement with the patient are essential.

Rib grafts in septorhinoplasty.

Autogenous cartilage has generally been considered the gold standard grafting material in reconstructive septorhinoplasty for volume filling and structural support. In the restructuring of the nasal skeleton, autogenous cartilage can be harvested from the nasal septum, the auricle or the rib, but costal cartilage is considered the best graft material in patients requiring major reconstruction. Rib cartilage is an outstanding material in reconstructive septorhinoplasty, especially in revision surgery and when large amounts of tissue are required. This autologous material has a low rate of complications such as resorption, infection and extrusion compared to homografts and alloplastic implants. In the present study, the authors analyze and discuss the use of autogenous rib cartilage in 54 patients who underwent primary and revision septorhinoplasty. Its use is also suggested in cases in which there is a need to have a fair amount of cartilaginous tissue to be grafted for nasal framework reconstruction and respiratory function improvement.

An air flow sensor for neonatal mechanical ventilation applications based on a novel fiber-optic sensing technique.

In this work, a simple and low-cost air flow sensor, based on a novel fiber-optic sensing technique has been developed for monitoring air flows rates supplied by a neonatal ventilator to support infants in intensive care units. The device is based on a fiber optic sensing technique allowing (a) the immunity to light intensity variations independent by measurand and (b) the reduction of typical shortcomings affecting all biomedical fields (electromagnetic interference and patient electrical safety). The sensing principle is based on the measurement of transversal displacement of an emitting fiber-optic cantilever due to action of air flow acting on it; the fiber tip displacement is measured by means of a photodiode linear array, placed in front of the entrance face of the emitting optical fiber in order to detect its light intensity profile. As the measurement system is based on a detection of the illumination pattern, and not on an intensity modulation technique, it results less sensitive to light intensity fluctuation independent by measurand than intensity-based sensors. The considered technique is here adopted in order to develop two different configurations for an air flow sensor suitable for the measurement of air flow rates typically occurring during mechanical ventilation of newborns: a mono-directional and a bi-directional transducer have been proposed. A mathematical model for the air flow sensor is here proposed and a static calibration of two different arrangements has been performed: a measurement range up to 3.00 × 10(-4) m(3)∕s (18.0 l∕min) for the mono-directional sensor and a measurement range of ±3.00 × 10(-4) m(3)∕s (±18.0 l∕min) for the bi-directional sensor are experimentally evaluated, according to the air flow rates normally encountered during tidal breathing of infants with a mass lower than 10 kg. Experimental data of static calibration result in accordance with the proposed theoretical model: for the mono-directional configuration, the coefficient of determination r(2) is equal to 0.997; for the bi-directional configuration, the coefficient of determination r(2) is equal to 0.990 for positive flows (inspiration) and 0.988 for negative flows (expiration). Measurement uncertainty δQ of air flow rate has been evaluated by means of the propagation of distributions and the percentage error in the arrangement of bi-directional sensor ranges from a minimum of about 0.5% at -18.0 l∕min to a maximum of about 9% at -12.0 l∕min.

Comparative evaluation of ultrasound scanner accuracy in distance measurement.

The aim of the present study is to develop and compare two different automatic methods for accuracy evaluation in ultrasound phantom measurements on B-mode images: both of them give as a result the relative error e between measured distances, performed by 14 brand new ultrasound medical scanners, and nominal distances, among nylon wires embedded in a reference test object. The first method is based on a least squares estimation, while the second one applies the mean value of the same distance evaluated at different locations in ultrasound image (same distance method). Results for both of them are proposed and explained.

Experimental evaluation of errors in the measurement of respiratory parameters of the newborn performed by a continuous flow neonatal ventilator.

Pulmonary ventilators for intensive care provide information on, among many other patient respiratory parameters, patient resistance, compliance and 'work of breathing' values calculated from pressure and flow data patterns according to a widely utilized algorithm. The effects induced by the breathing circuit and analogue filtering of the ventilator measuring system are experimentally investigated during controlled ventilation. Three main phenomena are observed: (a) errors in calculation of resistance and compliance due to filtering of pressure and flow waveforms; (b) the presence of pressure oscillations at the beginning of inspiration and expiration phases; and (c) the phase shift between pressure and flow waveforms. The experimental evaluation of the measuring system of a neonatal ventilator is then conducted and: (a) a delay in pressure and flow measurement synchronization equal to 22 +/- 2 ms is evaluated; moreover, (b) a difference between the values provided by the ventilator and those measured by the reference experimental setup on respiratory parameters such as the compliance, resistance and work of breathing that lies in the range of 7-16% of reading is observed.

The photochemistry of flutamide and its inclusion complex with beta-cyclodextrin. Dramatic effect of the microenvironment on the nature and on the efficiency of the photodegradation pathways.

The photochemistry of the anticancer drug flutamide (FM), 2-methyl-N-[4-nitro-3-(trifluoromethyl)phenyl]propanamide, in homogeneous media and in the beta-cyclodextrin (beta-CD) cavity has been investigated. The photoreactivity of the free molecule has been rationalized on the basis of an intramolecular nitro to nitrite rearrangement followed by cleavage of the nitrite intermediate. The twisted geometry of the nitro group with respect to the aromatic plane plays a key role in triggering such a photoprocess. Incorporation of FM in the beta-CD cavity leads to dramatic effects on both the efficiency and the nature of the photochemical deactivation pathways of the guest molecule. A 20-fold increase in the FM photodecomposition quantum yield and the formation of photoproducts originated by both reduction of the nitro group and cleavage of the amide bond were observed in the presence of the macrocycle. Such a behavior cannot be attributed exclusively to the micropolarity of beta-CD and/or to its role as a reactant. The induced circular dichroism spectra and the nature of the photoproducts formed in these experimental conditions provide indications that the photoreactivity in the beta-CD microenvironment could likely be mediated by structural changes of FM upon complexation.

Tensile experiments and SEM fractography on bovine subchondral bone.

Subchondral bone undecalcified samples, extracted from bovine femoral heads, are subjected to a direct tensile load. The Young's modulus of each sample is determined from repeated tests within the elastic limit. In a last test, the tensile load is increased up to the specimen failure, determining the ultimate tensile strength. The investigation is performed on both dry and wet specimens. The measured Young's modulus for dry samples is 10.3+/-2.5GPa, while that of wet samples is 3.5+/-1.2GPa. The ultimate tensile strengths are 36+/-10 and 30+/-7.5MPa for dry and wet specimens, respectively. SEM micrographs of failure surfaces show characteristic lamellar bone structures, with lamellae composed of calcified collagen fibers. Rudimentary osteon-like structures are also observed. Failure surfaces of wet samples show a marked fiber pull-out, while delamination predominates in dry samples. The obtained results are interpreted on the basis of the deformation mechanisms typical of fiber-reinforced laminated composite materials.

Molecular mechanisms of photosensitization induced by drugs. XII. Photochemistry and photosensitization of rufloxacin: an unusual photodegradation path for the antibacterials containing a fluoroquinolone-like chromophore.

The UVA irradiation of 9-fluoro-2,3-dihydro-10-4'-methyl-1' -piperazinyl-7-oxo-7H-pyrido[1,2,3-de]-1,4-benzo-thiazine-6-carboxylic acid, rufloxacin, a fluoroquinolone antibacterial that shows photosensitizing properties toward biological substrates, leads to formation of two main steady photoproducts characterized by a decarboxylation process and an opening of the piperazinyl ring, respectively. The deprotonation of the 10-piperazinyl group and the dissociation of the 6-carboxyl group of rufloxacin are strictly pH dependent. The photosensitizing activity was tested toward membranes as biological targets. Red blood cell hemolysis and lipid peroxidation were considered as markers of photosensitization. Ultraviolet A-induced damage is strongly influenced by the presence of oxygen, it is triggered by transient species, such as singlet oxygen and free radicals, photogenerated via rufloxacin irradiation, whereas no drug photoproduct is involved in the photosensitization process.

Excision and replacement of nasal septum in aesthetic and functional nose surgery: setting criteria and establishing indications.

Despite the technical details of the excision and replacement of the nasal septum both in aesthetic and functional nasal surgery have been extensively reviewed, in the opinion of the authors a clear and precise definition of the indications of this technique is still lacking. A simplified classification of the nasal septum deformities, based on the site and the direction of the fracture or bending axis, is proposed to establish reproducible guidelines to nasal septum surgery. On the basis of this classification the post-operative results of 227 patients affected by obstructive nasal septum deviation were evaluated. The surgical treatment consisted of conservative septoplasty in 173 cases, while in 54 cases excision and replacement of the nasal septum were performed. A conservative tension release septoplasty was performed for horizontal fracture or angulation of the septum. The more radical excision/replacement surgical approach was preferred when a vertical angulation or bending axis was observed (vertical = normal to the maxillary ridge). Twenty persisting septal deviations were found at the one year post-operative follow-up. Nineteen of these were the outcome of 173 conservative septoplasty, while only one case with unsatisfactory results was the outcome of 54 excision/replacement procedures. Seventeen out of 19 cases originally classified as horizontal deformity who presented at follow up with persisting septum deviation were reclassified as vertical. The reason for surgical failure must be probably identified in a preoperatory classification mistake where vertical deformities were erroneously evaluated horizontal and operated accordingly. The authors suggest excision/replacement of the nasal septum whenever its fracture or major bending axis is vertical.

Upper lateral cartilage suspension over dorsal grafts: a treatment for internal nasal valve dynamic incompetence.

The internal nasal valve incompetence (INVI) can be divided into (1) static: when the articulation between the dorsal edge of the upper lateral cartilages and the dorsal edge of the septum forms an angle less than 10 to 15 degrees and (2) dynamic: when the upper lateral cartilages collapse attracted by the negative pressure during inspiration. To correct both the static and dynamic components in severe cases of INVI, the authors propose a surgical technique consisting of upper lateral cartilages suspension over dorsal grafts; the dorsal margins of the two upper lateral cartilages are pulled dorsally and sutured together over the dorsal edge of the septum and over the dorsal-spreader grafts. The post-operative results of 12 patients affected by INVI with severe nasal obstruction were evaluated. Such a technique, which utilizes both the widening and the suspension effects, is particularly useful in the most severe cases of dynamic INVI.

Liposomal delivery of a 30-mer antisense oligodeoxynucleotide to inhibit proopiomelanocortin expression.

An oligodeoxynucleic sequence of 30 bases (30-mer ODN), complementary to a region of beta-endorphin mRNA, was synthesized to have an antisense effect with regard to the expression of this oligopeptide. Following the solid-phase synthesis of the oligodeoxynucleotide, the 30-mer ODN was encapsulated within liposomes to provide a higher resistance against DNases and an improved entrance into cells. The most suitable liposome formulation as a 30-mer ODN carrier consisted of small unilamellar vesicles (50 nm) with an encapsulation capacity of 4.76 microL/micromol. The liposomal formulations containing dipalmitoyl-DL-alpha-phosphatidyl-L-serine presented fusogenic properties, which are of great importance for the delivery of antisense compounds. The antisense activity of 30-mer ODN-loaded liposomes was evaluated by the determination of beta-endorphin levels in AtT-20 cells. The free 30-mer ODN did not provide any lowering of the beta-endorphin production, whereas the liposomally entrapped compound elicited a concentration-dependent inhibition. The inhibition was determined by a sequence-specific binding of the 30-mer ODN with the target mRNA.

Cytotoxic and cytostatic activity of copper(II) complexes. Importance of the speciation for the correct interpretation of the in vitro biological results.

The cytotoxicity of some copper(II) compounds against the mouse cancer cell line B16, murine L929, human KB cells, and fibroblasts was investigated. All the copper(II) systems tested were shown to have pronounced toxicity against transformed cells and a cytostatic effect against untransformed cells, i.e., human fibroblasts. A careful speciation of the actual in vitro conditions reveals that copper(II) is essentially present as mixed complexes formed with the amino acids of the culture medium, [Cu(glutamine)(histidine)] being the main species. It was found that the cytotoxic activity is related to the amount of copper(II) contained in the tested compounds.

Melanosomes from liver and skin of Rana esculenta L. A comparative chemical study.

1. Melanosomes from skin and liver of Rana esculenta L. have been isolated and some chemical properties of the relevant melanin and protein components were compared. 2. In both cases the pigments show spectroscopic (ESR) and chemical characteristics similar to those of eumelanins. The melanin content in skin melanosomes is higher than in the liver counterparts. 3. Amino acid patterns of the two protein components are different in their quantitative composition and both are characterized by high levels of glycine and proline. 4. The results as a whole indicate that skin and liver melanosomes from the same animal markedly differ in their chemical composition.