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Photocatalytic technique had already been employed in the treatment of olive mill wastewater (OMW) using the photocatalysis in suspension. The coupling of photocatalytic and membrane techniques should result in a very powerful process bringing great innovation to OMW depollution. Despite the potential advantages using these hybrid photoreactors, research on the combined use of photocatalysis and membranes has so far not been sufficiently developed. The present paper describes a study to assess the photocatalytic efficacy of a new ceramic membrane containing titanium dioxide, irradiated by UV light, used to abate the pollutant load of OMW. Good results were obtained (more than 90% of the phenol content was removed and the COD decrease was of the order of 46-51% in 24 h) particularly using the ceramic membrane compared with those offered by analogous catalytic membranes made of metallic or polymeric materials.

The present research was devoted to studying the kinetics of the artificial rancidification of peanut oil (PO) when a sample of this oil was isothermally heated at 180°C in an air stream. The formation of radical species due to heating was evaluated using a radical index whose value was determined using a biosensor method based on a superoxide dismutase (SOD), while the increasing toxicity was monitored using a suitable toxicity measuring probe based on the Clark electrode and immobilized yeast cells. An extra virgin olive oil was isothermally rancidified under the same experimental conditions and the corresponding data were used for the purpose of comparison. Both the so-called "model-fitting" and the classical kinetic methods were applied to the isothermal process biosensor data in order to obtain the kinetic constant rate value at 180°C., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Purpose: We measured premature ejaculation related female sexual distress using a new diagnostic tool, the Female Sexual Distress Scale-Revised-Premature Ejaculation questionnaire., Materials and Methods: In this large-scale, Internet based population study we evaluated 2,109 women in a stable relationship during the last 6 months. The 1,361 women in the premature ejaculation group had no female sexual disorder but the partner had premature ejaculation alone. The 748 controls had no female sexual disorder and a partner without premature ejaculation. We determined questionnaire content and discriminant validity, internal consistency and test-retest reliability. Multivariate logistic regression with propensity score reweighting was done to determine the clinical impact of demographics on the perception of sexual distress., Results: The questionnaire was well understood. Internal consistency was greater than 0.90 and 0.84 in the premature ejaculation and control groups, respectively. Test-retest reliability was 0.82 (95% CI 0.72-0.87) and 0.85 (95% CI 0.79-0.92) in the premature ejaculation and control groups, respectively. The questionnaire had a high AUC of 0.90 (95% CI 0.89-0.91). The new cutoff score of 12 or greater had 79.1% sensitivity (95% CI 73.8-82.5), 99.5% specificity (95% CI 98.0-100.0), 99.3% positive predictive value (95% CI 98.7-100.0) and 67.9% negative predictive value (95% CI 64.2-73.2). Median questionnaire scores were significantly higher in the premature ejaculation group than in controls (20, 95% CI 19-21 vs 6, 95% CI 6-7, p <0.0001). Logistic regression adjusted and unadjusted by propensity score indicated that women in the premature ejaculation group had a 7.12 (95% CI 5.98-10.14, p <0.0001) to 9.83 (95% CI 7.94-12.15) greater probability of sexual distress than controls., Conclusions: The Female Sexual Distress Scale-Revised-Premature Ejaculation questionnaire fulfills psychometric requirements for measuring sexual distress related to partner sexual dysfunction., (Copyright © 2013 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.)

Within the framework of research carried out by our team aimed at developing new immunological methods to determine proteins such as immunoglobulins G in different biological matrixes, for instance, serum and milk, tests were performed on several immunosensors based on different transducer types, i.e. amperometric (classical or screen-printed) electrodes for hydrogen peroxide. Lastly the feasibility of constructing immunosensors based on surface plasmon resonance (SPR) was investigated. "Competitive" immunological procedures were used in the first two cases. Conversely, the surface plasmon resonance transduction technique allowed a "direct" measurement. Applications were performed on human serum, powdered milks for babies and particularly on several animal milks, in the case of buffalo milk seeking a routine control method to identify possible inflammatory affections in the animals., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Background and Methods: Several standard powdered black pigments were characterized by means of thermogravimetry TG-DTG and allied techniques. These pigments were used to make standard plaster frescoes at this purpose prepared. The latter ones were subjected to Raman and reflectance analysis. The results obtained, together with TG data, were chemometrically processed and used to identify an analogous standard fresco fabricated by an unknown commercial black pigment, obtaining excellent results., Results: The same colorimetric and reflectometric techniques, coupled with suitable chemometric techniques, were then successfully used to identify the type of black pigment present in an ancient roman fresco of the Imperial Age (30 B.C.)., Conclusion: TG-DTG resulted useful techniques to autenticate powdered black pigments.Colorimetry and Raman, but also the only colorimetry, were useful to identify an ancient black pigment in situ.

Background: The thermal oxidative degradation of aged and non aged cellulose samples of commercial paper was studied using thermogravimetry and derivative thermogravimetry under a forced air flow up to 800°C., Results: TG and DTG data were processed using two non-isothermal-based model-fitting methods and one based on linear least squares to calculate Ea trend values, measured as a function of artificially induced sample age. The Ea trends thus obtained were compared in order to assess their potential for yielding archaeometric curves. As the trends of first two methods show an inversion of the direction between non aged cellulose samples and artificially aged samples, while the third method does not, an in-depth study was carried out using a multilinearity assumption., Conclusions: The results are discussed and the outcomes indicate that the above cited inversion is real and not linked to the method. Additionally, it was evidenced that the number of points used for the estimation of linear least squares model parameters is of capital importance.

The last year we fabricated some new immunosensors for the analysis of lactoferrin protein. In the present research the immunological and analytical characteristics of the immunosensor method have been extensively investigated. The study was therefore extended to cover the ability of the analyte and the corresponding antibody to produce the immunocomplex. A rough estimation of the K(aff) value was obtained at the midpoint of the Langmuir curve, where K(aff)=1/IC(50). The K(aff) value was found to be of the order of 10(6)M(-1). In addition we attempted in the present study to reduce the excessively long time required for each measurement, due to the fact that, in the previous researches the measurement procedure used was the classic "competition" ELISA type, employing an ad hoc enzymatic marker. One possible way of reducing measurement time was found to be to perform a kind of completely innovative "direct" measurement developed by us, which still uses an enzymatic marker and an amperometric transducer, as in the previous competition method.

Two different immunosensors, recently developed for the determination of antibacterial proteins (lactoferrin and immunoglobulin G) in buffalo milk and in other commercial animal milks samples, were used in the present study. The aim was to propose these immunosensor methods for routine control of important diet products, such as cow and goat milks, and in particular buffalo milk. To this end we employed two different kinds of immunosensors: one for the analysis of immunoglobulin G (IgG), the other was a new amperometric immunosensor for lactoferrin analysis. Lactoferrin and IgG immunosensors were also used for the determination of lactoferrin and immunoglobulin G in buffalo milk on different days of lactation.

To completely overcome the problem of the presence of urea in the serum, which can be the cause (especially at low immunoglobulin G concentrations) of a small but non negligible interference in the enzyme reaction of the enzymatic marker, when the measurement was performed by a potentiometric immunosensor that we constructed and characterized in previous work, and which used urease as marker, we have now constructed an entirely different and highly innovative immunosensor. This new device uses the enzyme alkaline phosphatase as marker, sodium phenylphosphate as substrate but above all, a tyrosinase biosensor obtained by coupling a Clark type gas diffusion amperometric electrode and the tyrosinase enzyme, immobilized in a cellulose triacetate membrane, as transducer. After optimizing the 'competitive' measurement procedures, the new immunosensor was used to determine both HIgG and the anti-HIgG, with a limit of detection (LOD) of the order of 3x10 -11 M. Clearly this highly innovative construction geometry makes the immunosensor extremely selective. This makes it possible to determine immunoglobulin G both in human serum and milk without the slightest interference by any urea present in these biological matrixes.

Thorough research was carried out on Lactoferrin immunosensor development. Furthermore, two different competitive procedures were used for Lactoferrin determination, in which either the antigen (Lactoferrin) or the antibody (anti-Lactoferrin) was, respectively, conjugated with horseradish peroxidase enzyme using a biotinylation process. The biotinylation of Lactoferrin and the subsequently used competition procedure for the immunosensor measurement were to get ready. Three different kinds of immunosensors were implemented, in all cases using the peroxidase enzyme as marker and hydrogen peroxide as substrate, but alternatively using as transducers one of the following sensors: (i) an amperometric electrode for H2O2, (ii) a Clark electrode and (iii) an iodide electrode. After optimizing the "competitive" measurement procedures and the transducer, the new Lactoferrin immunosensor was used for the determination of Lactoferrin content in human milk and in different types of dried milks or other dairy products, specifically produced and sold in chemist's shops to feed unweaned children in the first few months of life.

As a follow-up of the research programs carried out by our group concerning the artificial isothermal rancidification process in extra virgin olive oil (EVOO), in the present work the trends of both the total antioxidant capacity and the total polyphenols concentration as well as the main kinetic parameters of the process during the thermal oxidation of EVOO were studied and compared. In addition, the possibility of evaluating the increase in radicals concentration during the thermal oxidation process using a superoxide dismutase biosensor was also studied. The present investigation concerning this important food product is highly topical as it refers to the state of alteration of the EVOO used for cooking or frying, as a function of the temperature reached.

The process of artificial rancidification of extra-virgin olive oil due to heating in an oxidizing atmosphere was studied by testing an actual kinetic model of the process and monitoring the thermal oxidative degradation of the polyphenols contained in it. To this end, a series of oxidative degradation experiments were carried out on extra-virgin olive oil samples under isothermal conditions at 98, 120, 140, 160, and 180 degrees C using a thermostatic silicon oil bath. The experimental procedure used in this study carefully followed the recommendations regarding the study of olive oil rancidification set out in the AOM procedure. The change in polyphenol concentration with time was monitored at selected temperatures using a tyrosinase biosensor operating in an organic phase (n-hexane). The activation energy for the polyphenol degradation process determined using the MacCallum method was found to be practically constant throughout most of the process. Furthermore, the application of the so-called "model-fitting" method to this process enabled the specific constant rates to be determined at the above-mentioned selected temperatures. In addition, a confirmation of the activation energy value was obtained by the "model-fitting" method and the algorithm of the kinetic model equation best-fitting the experimental curve representing the whole process was checked. Finally, further very interesting observations were made, for instance, the half-life concentration values of polyphenols at selected temperatures between 98 and 180 degrees C.

In this paper our previous researches dealing with compatibility, thermoanalytical characterization, the kinetics of thermal degradation of acetaminophen, either pure or contained in some commercial pharmaceutical formulations, have found applications outlets. In a previous investigation the possible interactions between acetaminophen and four excipients contained in the commercial pharmaceutical formulations were tested. As a continuation of this research in the present study an analytical method based on differential scanning calorimetry (DSC) was applied to determine the acetaminophen content of four commercial pharmaceutical formulations. For a fifth drug it was shown that the method is not applicable owing to observed incompatibility with one of the excipients. Finally, the analytical results obtained were compared with those derived from two UV spectrophotometric methods (one, i.e., "direct method," recommended by the Pharmacopeia and the other based on the first-order derivative UV spectra).

Fabriano paper was aged by irradiation with ultraviolet light (k=310) in a veterometer for 300 hours. At fixed time intervals, samples of the paper under test were analysed by titanium dioxide photosensor to determine electrochemically the "environmental persistence" index, by a suitable conductimeter method, to determine the specific conductivity variation and by thermogravimetry to determine the moisture content, the onset temperature of the cellulose degradation process and the value of the activation energy of the same process. The behaviour of these different types of indicators displayed approximately monotonous trends as a function of time.

Quercetin is a widely distributed plant flavonoid possessing a variety of chemical and biological activities, including chelation, free-radical scavenging, and antioxidant activity. Atrazine is a selective triazine herbicide that has been the subject of an international revision program for human and ecological health risks because of its persistence in the environment. In a previous study, we demonstrated that atrazine was clastogenic in the Allium cepa test. In this present study, we investigated whether quercetin affords protection from the chromosome breaks induced by atrazine. In a preliminary assay, 0.1-20 microg/ml quercetin produced no toxicity or clastogenic activity in the Allium cepa test. Subsequently, we evaluated the effects of 0.5 and 5 microg/ml quercetin on the clastogenicity of 2.5, 5.0, and 7.5 microg/l atrazine. Quercetin (0.5 microg/ml) significantly reduced the frequency of total aberrations induced by 7.5 microg/l atrazine, while both concentrations of quercetin significantly decreased the frequency of fragments induced by 7.5 microg/l atrazine. The results of this study indicate that plant flavonoids such as quercetin may protect against the genotoxic effects of atrazine. Efforts to understand the extent to which plant flavonoids influence the biological activities of genotoxicants and the mechanisms involved in the interactions could help to better discern the advantages and disadvantages of their use and to clarify their possible protective role against pollutants., (Copyright (c) 2006 Wiley-Liss, Inc.)

Thermogravimetry (TG) and differential scanning calorimetry (DSC) were used to assess the compatibility between acetaminophen (Ac) and some excipients (polyvinylpyrrolidone (P), magnesium stearate (M), citric acid (C), aspartame (As), mannitol (Mn), cellulose (Cll) and starch (S)) in several of the more commercially available pharmaceutical formulations and in solid binary mixtures. The present study compared thermodynamic data on acetaminophen melting and vaporization processes of pure acetaminophen with those found for several solid mixtures and in some commercially available acetaminophen-based dosage forms. Appreciable modifications occur only for solid mixtures with high content of excipient. Acetaminophen-based dosage forms and its solid binary mixtures usually show "additivity" of calorimetric peaks number of pure components in their calorimetric curve profiles, thus revealing a good thermoanalytical compatibility between acetaminophen and the excipients examined, except for samples containing appreciable content of mannitol.

The class II products of the major histocompatibility complex have a distribution restricted to certain tissues and cells. For instance, they are constitutively expressed by B lymphocytes, but not by resting T lymphocytes. In this study, we report the identification of a novel DNase I hypersensitive site within a putative regulatory region of the human HLA-DRA gene, the so-called far upstream region. This hypersensitive site was present in the genome of the DRalpha-positive human B-lymphoid Raji cell line, and absent in the DRalpha-negative T-lymphoid Jurkat cell line. In addition, this hypersensitive site was also present in transgenic B lymphocytes isolated from the murine transgenic line TG 53, carrying a single integrated copy of the human HLA-DRA gene per haploid genome. The correlation between DRA expression and the presence of this far upstream hypersensitive site suggests novel long distance chromatin remodeling mechanisms possibly shared by human and murine class II genes.

Fresh (larch and fir, in its white and red varieties) and ancient wood samples (dating respectively to the 13th, 15th and 17th centuries) were subjected to thermogravimetric analysis (TG and DTG). The resulting thermogravimetric data were then used to construct archeometric curves for the wood varieties tested. In a preliminary approach, it was attempted to correlate the onset temperature of the thermogravimetric step corresponding to cellulose decomposition with the age (expressed in centuries) of the samples, although the results obtained were anything but brilliant. More encouraging results were obtained by examining the relationship between wood sample age and the value of the (percent cellulose/percent lignin) ratio computed from the thermogravimetric data. Lastly, a procedure for processing data obtained from the TG curves was applied to a kinetic analysis of the processes that take place when wood samples are subjected to a temperature regime with a constant heating rate, obtaining values for the activation energy of the TG step corresponding to the decomposition of cellulose. Also using these data it was attempted to construct archeometric curves, obtaining results that varied quite significantly according to the wood species tested.

An immobilised Pseudomonas putida cell based biosensor, able to work directly in organic solvent (n-hexane), was designed and built. The response, in n-hexane, to benzene and some of its derivatives was studied. The proposed biosensor was found to be suitable for determining benzene in hydrophobic matrices.

A thermal and a kinetic analysis on the decomposition processes of a commercial drug named diamplicil (AD), obtained by an antibiotic combination of ampicillin (A) and dicloxacillin (D), have been carried out to find their thermal stability. The DSC/TG curves of this commercial drug were compared with those of its active components and an excipient, the magnesium stearate (M). Kinetic study was carried out using both isothermal and dynamic TG curves. Decomposition mechanisms for both active components and commercial drug tested were not found. The kinetic data obtained by the non-isothermal isoconversional method showed that D component causes a decrease of the kinetic stability of the active A component. Additive magnesium stearate does not decrease the stability of the two components. Moreover, storage time values at room temperature were calculated.

An enzyme inhibition biosensor, developed in our laboratory and previously used for the analysis of compounds with anticholinesterase activity (e.g. physostigmine, neostigmine, pyridostigmine nicotine and organophosphorus compounds) has now been tested for the analysis of another recently synthesized cholinesterase inhibitor, i.e. eptastigmine. In addition nicotinic acid and nicotinamide, although displaying weaker inhibition properties, were also tested in pharmaceutical products using the same inhibition enzyme sensor. The biosensor consisted of a hydrogen peroxide amperometric electrode coupled to a functionalised nylon membrane chemically bonding both the enzymes butyrylcholinesterase and choline oxidase; a butyrylcholine standard solution in glycine buffer acted as substrate. The response of the system to all the inhibitors considered was characterised completely and the analysis of several pharmaceutical formulations containing nicotinamide or nicotinic acid was also performed.

A physical-chemical characterisation of several marbles frequently used in ancient times for artistic or decorative purposes was performed in support the work of historians and restorers. The data were obtained using several different types of instrumental chemical methods (Thermogravimetry, Differential Thermal Analysis, X-ray Diffractometry and ICP Plasma Emission Spectroscopy) and have been summarised in short tables. The data have already proved useful in the identification of a small number of finds (statues or architectonic elements) from Ancient Rome (Imperial Age, 2nd-3nd cent. A.D.) for the purpose of which also a well-known pattern recognition analysis software package was used for data processing. In practice, the research showed that an organised set of chemical data obtained using several modern instrumental methods can provide a valid basis for the reasonably rapid and reliable identification of the type of marble used to make artistic artifacts that have not yet been subjected to typological study.

Enzymatic electrodes based on superoxide dismutase enzyme were developed. Using the superoxide dismutase enzyme sensor assembled according to the classical model, poor results were obtained. Results were improved by adopting a new way of assembling the biosensor using a cellulose triacetate layer in which the SOD enzyme is entrapped and sandwiched between two gas-permeable membranes, or using a kappa-carrageenan gel layer entrapping the enzyme, sandwiched between an external gas permeable membrane and an internal cellulose acetate membrane, coupled in each case to the oxygen amperometric transducer. Results obtained by applying the newly developed biosensor to assaying hydrophobic compounds showing radical scavenging properties, operating in dimethylsulfoxide, were also satisfactory.

The experimental evaluation of properties of defence against free radicals represents an extremely interesting heuristic and applicational objective. Research was carried out to experimentally evaluate the scavenging properties of several fruits and plants, fresh or from a herbalist's using an amperometric superoxide dismutase (SOD) biosensor recently developed by the present authors. The superoxide radical was produced in solution using the xanthine/xanthine oxidase system; measurements were carried out by comparing biosensor response to superoxide radical both in the presence and absence of the sample considered.

In the present study, Thermogravimetry (TG) and Differential Scanning Calorimetry (DSC) are simultaneously applied to determine the thermal properties of two antibiotic salts, disodium and calcium phosphomycin, used either pure or in association with several excipients. This study was carried out kinetically as well by mathematical elaboration of the TG curves performed according to an isothermal procedure applied at different fixed temperatures. Kinetic parameters showing agreement with those produced by the isothermal method were also obtained by means of a non-isothermal method using a dynamic TG curve alone. The main aims of the work were to provide reliable kinetic parameters (kinetic constant k, activation energy E(a) and pre-exponential factor A) to evaluate the thermal stability of phosphomycin salts in the presence and absence of the excipients generally contained in the phosphomycin-based pharmaceutical forms available on the market, and to obtain information concerning compatibility towards the active components. These kinetic parameters were then used to extrapolate shelf-life and half-life values at room temperature for pure active components in the solid state and for their pharmaceutical derivatives.

The results reported in this paper regard the setting up of a polymeric membrane ISE that is selective for cholic acids (CA) and able to work in a flow system, especially in flow injection analysis (FIA), based on the exchanger (tetrakisdecylammoniumcholate, TDACh), which has proved effective, is of very simple but suitable structure and is above all easy to synthesise starting from commercially available chemicals. A complete analytical characterisation of the sensor was performed working both in batch conditions and in FIA, using in the latter case a 'wall jet' type of flow cell. The response toward different bile acid sodium salts such as the CA, deoxycholic (DCA), chenodeoxycholic (CDCA), ursodeoxycholic (UDCA), taurocholic (TCA) sodium salts was checked. The application to the analysis of different commercial drugs by FIA was also performed to determine the UDCA or CDCA acid content of several pharmaceutical formulations. Lastly, a preliminary study is presented concerning the use of the investigated electrochemical sensor as high performance liquid chromatography (HPLC) detector.

A superoxide dismutase biosensor was used to determine the antioxidant properties of scavenger molecules and the antiradical activity of healthy and diseased human kidney tissues; this biosensor is based on the use of the enzyme superoxide dismutase (SOD), which is physically entrapped in a kappa-carrageenan gel membrane, and of a transducer consisting of an amperometric hydrogen peroxide electrode. Several compounds with scavenging properties were tested, including some commercial drugs.

The human leukaemic K562 cell line can be induced in vitro to undergo erythroid differentiation by a variety of chemical compounds, including haemin, butyric acid, 5-azacytidine and cytosine arabinoside. Differentiation of K562 cells is associated with an increased expression of embryo-fetal globin genes, such as the zeta, epsilon and gamma globin genes. Therefore the K562 cell line has been proposed as a useful in vitro model system to determine the therapeutic potential of new differentiating compounds as well as to study the molecular mechanism(s) regulating changes in the expression of embryonic and fetal human globin genes. Inducers of erythroid differentiation which stimulate gamma-globin synthesis could be considered for possible use in the experimental therapy of those haematological diseases associated with a failure in the expression of adult beta-globin genes. In this paper we demonstrated that the G + C selective DNA-binding drugs chromomycin and mithramycin were powerful inducers of erythroid differentiation of K562 cells. Erythroid differentiation was associated with an increase in the accumulation of (a) Hb Gower 1 and Hb Portland and (b) gamma-globin mRNA.

To investigate the cytokines involved in the interaction between circulating (B and T lymphocytes) and non-circulating (stromal cells) elements present in lymphoid tissue, highly purified populations were isolated from human tonsils and the cytokine production and mRNA expression (interleukin-1 alpha, -2, -4, -5, -6, -8, -10, leukocyte inhibitory factor, granulocyte-macrophage colony-stimulating factor, and interferon-gamma) were assessed both by immunoassay and reverse transcriptase polymerase chain reaction under resting conditions and after activation with tumor necrosis factor-alpha. Under basal conditions most cytokines were not detected, except for interleukin-8 which was produced by T lymphocytes and lymphoid cells. Activation by tumor necrosis factor-alpha induced interleukin-8 production by B lymphocytes. Tonsillar T lymphocytes expressed mRNA for interleukin-1 alpha, -8, -10, -4, leukocyte inhibitory factor, and interferon-gamma, only interleukin-4 was expressed by resting peripheral blood T lymphocytes. Tonsillar B lymphocytes were mRNA positive for interleukin-1 alpha, -8, -10, leukocyte inhibitory factor, and interferon-gamma, these were not expressed by peripheral blood B lymphocytes. Stromal cells constitutively produce interleukin-6 whose levels increased 5 times upon tumor necrosis factor-alpha activation Granulocyte-macrophage colony-stimulating factor and interleukin-8 were detected only after tumor necrosis factor-alpha activation. Only stromal cells constitutively express interleukin-6 and granulocyte-macrophage colony-stimulating factor and show a cytokine pattern different from that described for other non-lymphoid cells, such as follicular dendritic cells. These data indicate that in the human tonsil population, lymphoid and non-lymphoid cells can be distinguished by different patterns of cytokine expression.

Background: The recent development of biosensor technologies for biospecific interaction analysis enables the monitoring of a variety of molecular reactions in real time by surface plasmon resonance (SPR). If the ligand is a biotinylated single stranded DNA, this technology could monitor DNA-DNA hybridization. This approach could be of great interest in virology, since the hybridization step is oftenly required to confirm specificity of molecular diagnosis., Objectives: To determine whether real-time molecular diagnosis of human immunodeficiency virus type I (HIV-1) could be performed using biosensors and SPR technology., Study Design: Specific hybridization of a biotinylated HIV-1 oligonucleotide probe immobilized on a sensor chip to single stranded DNA obtained by asymmetric polymerase-chain reaction (PCR) was determined using the BIAcore biosensor., Results: Direct injection of asymmetric PCR to a sensor chip carrying an internal HIV-1 oligonucleotide probe allows detection of hybridization by SPR using biosensor technology. This enabled us to apply a real-time, one-step, non-radioactive protocol to demonstrate the specificity of amplification of HIV-1 genomic sequences by PCR., Conclusion: The procedure described in this study for HIV-1 detection is simple, fast (PCR and SPR analyses take 30 min), reproducible and could be proposed as an integral part of automated diagnostic systems based on the use of laboratory workstations and biosensors for DNA isolation, preparation of PCR reactions and analysis of PCR products.

We have studied the effects of chromomycin and of a triple-helix-forming oligonucleotide (TFO) that recognizes Sp1 binding sites on protein-DNA interactions and HIV-1 transcription. Molecular interactions between chromomycin, the Sp1 TFO and target DNA sequences were studied by gel retardation, triplex affinity capture using streptavidin-coated magnetic beads and biosensor technology. We also determined whether chromomycin and a TFO recognizing the Sp1 binding sites of the HIV-1 long terminal repeat (LTR) inhibit the activity of restriction enzyme HaeIII, which recognizes a sequence (5'-GGCC-3') located within these Sp1 binding sites. The effects of chromomycin and the TFO on the interaction between nuclear proteins or purified Sp1 and a double-stranded oligonucleotide containing the Sp1 binding sites of the HIV-1 LTR were studied by gel retardation. The effects of both chromomycin and TFO on transcription were studied by using an HIV-1 LTR-directed in vitro transcription system. Our results indicate that low concentrations of chromomycin potentiate the effects of the Sp1 TFO in inhibiting protein-DNA interactions and HIV-1-LTR-directed transcription. In addition, low concentrations of chromomycin do not affect binding of the TFO to target DNA molecules. The results presented here support the hypothesis that both DNA binding drugs and TFOs can be considered as sequence-selective modifiers of DNA-protein interactions, possibly leading to specific alterations of biological functions. In particular, the combined use of chromomycin and TFOs recognizing Sp1 binding sites could be employed in order to abolish the biological functions of promoters (such as the HIV-1 LTR) whose activity is potentiated by interactions with the promoter-specific transcription factor Sp1.

We investigated whether stromal cells obtained from human tonsils could interact and modulate the proliferation of the osteosarcoma cell in order to determine why lymph node metastases usually have a low incidence and remain occult using routine examinations. The effects of the supernatant of resting or activated stromal cells were analysed on osteoblastic cell proliferation of three different cell lines (HOS, U2, OS, MG-63). Only the proliferation of MG-63 was significantly inhibited. The direct adhesion of stromal cells to the osteosarcoma cell lines caused a greater inhibition of the proliferation of all three lines tested.

Study Objective: To determine the clinical effects of the alkalinization of 2% mepivacaine with epinephrine used for epidural block during cesarean section., Design: Randomized, double-blind, placebo-controlled (standard commercial preparation of 2% mepivacaine with epinephrine) study., Setting: Inpatient obstetric department at a general hospital., Patients: Seventy patients scheduled for elective cesarean section under epidural anesthesia., Interventions: Two groups of 35 patients each receiving either the standard commercial preparation of mepivacaine or the pH-adjusted solution (prepared with the addition of 0.1 meq/ml of sodium bicarbonate to the standard commercial solution)., Measurements and Main Results: Measurements of sensory (pinprick) and motor (Bromage's criteria) block were taken at 1- to 2-minute intervals beginning after the completion of the epidural injection. Increasing the pH of the mepivacaine resulted in a significant shortening of the time of analgesia onset (9.3 minutes compared with 16.01 minutes, p less than 0.01) and of peak effect (11.1 minutes compared with 21.2 minutes, p less than 0.01). The alkalinization did not affect duration of the block, intensity of motor block, or mean dose of local anesthetic used., Conclusion: The alkalinization allowed the surgery to proceed more rapidly, significantly decreasing the time interval between epidural block and delivery of the infant.

Enzyme sensors for urea and creatinine were developed by coupling an ammonia gas-diffusion electrode with triacetate cellulose membranes entrapping urease or creatinine deiminase enzymes. Satisfactory results were obtained by using these sensors both in standard solutions and in authentic biological matrices.

The two most commonly used spectrometric methods for the determination of the phosphorus content of human bile are compared. The optimum experimental conditions are studied, and the analytical characteristics of the two methods, using both standard samples and human bile, are evaluated. The methods are compared on the basis of their sensitivity, precision and accuracy, and the correlation between the two techniques demonstrated using fifteen samples of human bile. Data obtained by both methods have been used to calculate lithogenic index values.

Two new polymeric membrane electrodes selective to cholate and to benzylpenicillinate, based respectively on PVC membranes containing polybenzyl-propargilamine as conductor polymer and benzyldimethylcethylammonium-cholate, or -benzylpenicillinate as exchangers, have been prepared, and applied to the determination of cholic acids, anionic surfactants and antibiotics. Results are compared with those ones obtained by analogous PVC sensors and liquid membrane sensors, previously described.

This study was designed to evaluate the maternal intraoperative and postoperative analgesia and neonatal effects of adding sufentanil to epidural anesthesia for cesarean section before the skin incision. Forty-five multipara were randomized in three equal groups to receive sufentanil 80 micrograms, 50 micrograms, or saline with the epidural lidocaine. Intraoperative and postoperative analgesia and side effects were recorded. Infants were evaluated by Apgar score at one and five minutes and by Neurologic and Adaptive Capacity Score and Early Neonatal Neurobehavioral Scale at one, four and 24 hours of life. Superior analgesia, with no intraoperative requirements for supplemental narcotics, was obtained when either 80 micrograms or 50 micrograms sufentanil were added to lidocaine. Postoperative analgesia was prolonged after sufentanil, but side effects increased with the greater dose. The infants whose mothers received 80 micrograms sufentanil showed a mild neurobehavioral depression. It is therefore concluded that the addition of 50 micrograms of sufentanil improves both intraoperative and postoperative analgesia without significant neonatal effects.

A potentiometric method based on phosphate precipitation with Pb2+ and on a lead ion selective electrode is applied to the determination of bile phosphates. The method is rapid and simple and does not require extensive sample pretreatment.

A new type of 10% lipidic emulsion based on safflower oil and soya (Liposyn II, Abbott, Latina) characterised by a high linoleic/linolenic acid ratio was administered in random, double blind fashion with 10% Intralipid to 20 subjects submitted to major surgery of the digestive system. Dosage was 1.2 g/kg at an infusion rate of 2.6 ml/kg/h. In order to evidence possible in vivo inter-reactions between proteins of the acute post-aggressive phase and these emulsions, serial samples were used to measure haemogasanalytic parameters, plasma levels of triglycerides and serum levels of protein C-reactive and fourth complement factor. Both emulsions proved well tolerated and there was no evidence of general or administrative site reactions. The plasma levels of triglycerides underwent an increase during the test, with no significant differences between the two groups of patients. Notwithstanding the high serum levels of protein C-reactive of the subjects in question, there was no evidence of activation of the complement system nor alteration in haemogasanalytic parameters with the two lipidic emulsions adopted.

A cholate-selective liquid membrane electrode has been applied to the determination of the pool of cholic acids in human bile. Pretreatment of the sample was necessary for reliable potentiometric determination; different procedures were tried for pretreatment of human bile.

A simple method for direct determination both of choline and lecithin (phosphatidylcholine) in human bile and blood sera was developed. An enzyme electrode, based on immobilized choline oxidase on nylon net and an oxygen Clark electrode was assembled. Phosphatidylcholine can be determined by use of phospholipase D as hydrolyzing agent. Reliable results were obtained in the case of determination of lecithin and choline in bile samples.

A reliable cholesterol electrode was assembled with a Clark type oxygen electrode and cholesterol oxidase immobilised on a nylon net fixed on the electrode surface. Determination of free (unesterified) and total (esterified and unesterified) cholesterol in human bile samples was performed. Results were compared with the enzymatic-spectrophotometric procedure based on the Roeschlau method. The electrode has a usable life of more than two weeks, and more than 300 determinations could be performed. The CV values for free and total cholesterol in bile samples is 5-6%. The comparison between the proposed procedure and the spectrophotometric procedure is fair for the half of the results (delta less than 5%) and acceptable in the other half (delta less than 25%).

The values of conditional formation constants for the cobalt(II) chloride-1-nitroso-2-naphthol system were determined at 25 degrees in ethanol-benzene mixtures of different compositions by spectrophotometry in the visible region. Comparisons were made with the results previously obtained with the isomeric ligand 2-nitroso-1-naphthol. In both cases the L/M complexation ratio of the species found increases with increasing ethanol content of the medium.