Effect of ferric citrate on amyloid-beta peptides behavior.

Amyloid beta (Aß) aggregation and oxidative stress are two of the central events in Alzheimer's Disease (AD). Both these phenomena can be caused by the interaction of Aß with metal ions. In the last years the interaction between ZnII , CuII , and Aß was much studied, but between iron and Aß it is still little known. In this work we determine how three Aß peptides, present in AD, interact with FeIII -citrate. The three Aß peptides are: full length Aß1-42, an isoform truncated at Glutamic acid in position three, Aß3-42, and its pyroglutamated form AßpE3-42. Conformation and morphology of the three peptides, aggregated with and without FeIII -citrate were studied. Besides, we have determined the strength of the interactions Aß/FeIII -citrate studying the effect of ethylenediaminetetraacetic acid as chelator. Results reported here demonstrate that FeIII -citrate promotes the aggregation in all the three peptides. Moreover, Aspartic acid 1, Glutamic acid 3, and Tyrosine 10 have an important role in the coordination with iron, generating a more stable complex for Aß1-42 compared to that for the truncated peptides.

[Biogenic amines in epigeal spontaneous mushrooms: indicators of quality and freshness?]. / Ammine biogene in funghi epigei spontanei: possibili indi- catori di qualità e freschezza?

Purpose of the present study is to determine the qualitative and quantitative composition of nine biogenic amines (BA) in 153 samples of epigeal spontaneous mushrooms (Boletus edulis and Amanita Caesarea), sampled at large and small food retail shops in the Florence area. Each BA has been identified and quantified using a liquid chromatography system with a reversed phase, C18 column and post-column derivatization with o-oftalaldeide. The results have shown a widespread presence of each amine, although with varying frequency and concentration, and an increasing concentration of tyramine and putrescine in samples with poor organoleptic quality than those in good conditions.

Reference values of urinary trans,trans-muconic acid: Italian Multicentric Study.

This article reports the results of a study, conducted in the framework of the scientific activities of the Italian Society for Reference Values, aimed at defining reference values of urinary trans,trans-muconic acid (t,t-MA) in the general population not occupationally exposed to benzene. t,t-MA concentrations detected in 376 subjects of the resident population in three areas of Italy, two in central (Florence and southern Tuscany) and one in northern Italy (Padua), by three laboratories, compared by repeated interlaboratory controls, showed an interval of 14.4-225.0 microg/L (5th-95th percentile) and a geometric mean of 52.5 microg/L. The concentrations measured were influenced by tobacco smoking in a statistically significant way: Geometric mean concentrations were 44.8 microg/L and 76.1 microg/Ll in nonsmokers (264 subjects) and smokers (112 subjects), respectively. In the nonsmoking population, a significant influence of gender was found when concentrations were corrected for urinary creatinine, geometric mean concentrations being 36.7 microg/g creatinine in males (128 subjects) and 44.7 microg/g creatinine in females (136 subjects). The place of residence of subjects did not seem to influence urinary excretion of the metabolite, although personal inhalation exposure to benzene over a 24-h period showed slightly higher concentrations in Padua and Florence (geometric means of 6.5 microg/m(3) and 6.6 microg/m(3), respectively) than in southern Tuscany (geometric mean of 3.9 microg/m(3)). Concentration of t,t-MA in urine samples collected at the end of personal air sampling showed little relationship to personal inhalation exposure to benzene, confirming the importance of other factors in determining excretion of t,t-MA when concentrations in personal air samples are very low.

Most sensitive urinary cotinine cut-off level for environmental tobacco smoke exposure assessment: a pilot study.

The aim of this pilot study was to determine the most sensitive urinary cotinine level able to assess environmental tobacco smoke (ETS) exposure. 54 Florentine subjects (29 males and 25 females), reporting to be nonsmokers and exposed (E) or not exposed (NE) to ETS at home, at work or in places of recreation, were examined. The urinary cotinine concentration was determined using gaschromatographic analysis in samples collected on three consecutive days. 18 subjects (33.3%) reported to be exposed to ETS had a greater median cotinine concentration than 36 ETS-NE subjects (E = 3.3 pg/L vs NE = 2.2 microg/L, median values), with borderline statistical significance (P = 0.05). The 2.5 microg/L cotinine concentration was the only statistically significant cut-off (P = 0.04) discriminating between ETS-E to ETS-NE subjects, identifying 51.9% of the subjects examined as exposed (E). Considering the expanded uncertainty of measurement of the method used (20%), urinary cotinine concentrations higher than 3.1 microg/L, a value whose confidence interval is higher than our proposed cut-off of 2.5 microg/L, mean that to be sure that a subject is exposed to ETS.

Mode-coupling smoluchowski dynamics of a double-stranded DNA oligomer

The local dynamics of a double-stranded DNA d(TpCpGpCpG)(2) is obtained to second order in the mode-coupling expansion of the Smoluchowski diffusion theory. The time correlation functions of bond variables are derived and the (13)C-nmr spin-lattice relaxation times T(1) of different (13)C along the chains are calculated and compared to experimental data from the literature at three frequencies. The DNA is considered as a fluctuating three-dimensional structure undergoing rotational diffusion. The fluctuations are evaluated using molecular dynamics simulations, with the ensemble averages approximated by time averages along a trajectory of length 1 ns. Any technique for sampling the configurational space can be used as an alternative. For a fluctuating three-dimensional (3D) structure using the three first-order vector modes of lower rates, higher order basis sets of second-rank tensor are built to give the required mode coupling dynamics. Second- and even first-order theories are found to be in close agreement with the experimental results, especially at high frequency, where the differences in T(1) for (13)C in the base pairs, sugar, and backbone are well described. These atomistic calculations are of general application for studying, on a molecular basis, the local dynamics of fluctuating 3D structures such as double-helix DNA fragments, proteins, and protein-DNA complexes. Copyright 1999 John Wiley & Sons, Inc.

Dynamics of a double stranded DNA oligomer: mode-coupling diffusion approach and reduced rigid fragment models.

The local dynamics of a double stranded DNA fragment [d(CpGpCpApApApTpTpTpGpCpG)]2 of twelve base pairs is obtained to second order in the mode-coupling expansion of the Smoluchowski diffusion theory. The DNA is considered a fluctuating three-dimensional (3D) structure undergoing rotational diffusion. The starting structure for the calculations is the B canonical structure of the fragment, while the fluctuations are evaluated using molecular dynamics simulations, with the ensemble averages approximated by time averages along a trajectory of length 1.5 ns. The rotational dynamics of the bonds along the double strands are calculated and compared to experimental NMR relaxation rates of different 13C along the sequence: R(Cz), R(Cxy) and R(Hz-->Cz). For a fluctuating 3D structure the mode-coupling diffusion theory is found to be in good agreement with several relative characteristics of the experimental relaxation parameters, while motivations are given for the few differences which are due mainly to poor statistics or to inaccuracies in the diffusion model. With a view to application to larger DNA fragments, discussion is dedicated to the validity of reducing the number of degrees of freedom in the double helix statistics by grouping the atoms in rigid fragments (e.g. the backbone atoms, the sugar atoms and the base atoms of each nucleotide). Consideration is given to the effect on local dynamics properties of reduced descriptions that include only three or four rigid bodies per nucleotide as well as five rigid bodies per base pair. It is found that in general these approximations almost uniformly produce slight increase in the correlation time pattern, which grows as the rigidity in the model increases. The relative effects on the dynamic pattern for the most accurate rigid body models are modest. The errors in C1' and C5' mobilities are more significant if C5' is included in the backbone rigid body. These results offer new tools to analyse NMR relaxation behaviour and new perspectives in studying the role of dynamics in biological macromolecules.

High-performance liquid chromatographic method for determination of ronactolol in urine and plasma: evaluation of pharmacokinetic parameters in healthy humans.

Ronactolol [(+/-)-4'-[2-hydroxy-3-(isopropylamino)propoxy]-p-anisanilide], a new aminopropanol derivative showing beta-adrenoreceptor blocking activity, was administered orally as capsules to healthy humans at three single doses (30, 60, and 120 mg). Two HPLC methods were developed separately for determination of drug levels in urine and plasma. For plasma samples, after addition of internal standard (IS), a single-step extraction of alkalinized plasma was performed with methylene chloride. The organic layer was evaporated to dryness under reduced pressure, and the residue was taken up and chromatographed on a microbore silica column. Ronactolol and IS were detected by a UV detector at a wavelength of 278 nm. Excellent linearity was observed between the peak height ratios (ronactolol:IS) and concentrations in plasma. The lowest limit of detection (signal:noise, 3:1) was 1.5 ng/mL of plasma. Urine samples were directly injected and chromatographed on a microbore C18 column with an ion-pairing mobile phase. Excellent linearity was observed between the peak areas and concentrations in urine. The lowest limit of detection (signal:noise, 3:1) was 75 ng/mL of urine. The assay was used to determine the main pharmacokinetic parameters in healthy humans.

Rapid simultaneous determination of tryptophan and tyrosine in synthetic peptides by derivative spectroscopy.

A method for the simultaneous quantitative determination of tryptophan and tyrosine in synthetic peptides by second-order derivative diode-array spectroscopy is reported. The method does not require hydrolysis of the peptides or a derivatization reaction; the sample is dissolved in 0.1 N NaOH and directly scanned between 262 and 264 nm to detect tyrosine and between 304 and 306 nm to detect tryptophan. From these results the peptide content of the synthetic sample can be easily calculated in a very simple and fast way. The results obtained by the described method with several peptides are compared with those obtained by classical high-performance liquid chromatographic analysis of amino acids after peptide hydrolysis. A very good correlation was found both for the tryptophan/tyrosine molar ratio and the peptide content.

High-performance liquid chromatographic method for assay of otilonium bromide, diazepam, and related compounds in finished pharmaceutical forms.

A rapid, simple, stability-indicating assay procedure for otilonium bromide, a smooth muscle relaxant agent, and diazepam in composite tablet analysis was developed with high-performance liquid chromatography. The tablet matrix was dissolved with water, and drugs were extracted with acetonitrile containing an internal standard. An aliquot was centrifuged and chromatographed on a 5-microns, reversed-phase column with 0.5 M sodium acetate trihydrate buffer containing 5 mM 1-heptanesulfonic acid monohydrate sodium salt:methanol (30:70; v/v; adjusted to pH 6.0 with glacial acetic acid) as the mobile phase. The selectivity of the chromatographic system was demonstrated by resolving both compounds from various potential degradation products of each compound. The method is linear, quantitative, and reproducible.

Simultaneous determination of otilonium bromide and diazepam by first-derivative spectroscopy.

A rapid, simple assay procedure was developed for simultaneous analysis of otilonium bromide, a smooth-muscle relaxant, and diazepam in tablets containing 20 mg of otilonium bromide and 2 mg of diazepam (20:2 tablets) or 40 mg of otilonium bromide and 2 mg of diazepam (40:2 tablets) by "zero-crossing" first-derivative spectroscopy. The tablets were dissolved in 0.01 N HCl, mixtures were centrifuged at 3500 rpm (2472 x g) for 5 min, and first-derivative spectra were recorded. The absolute values of the derivative were measured at 264 nm for determination of otilonium bromide and between 406 and 408 nm (380 nm for analysis of 40:2 tablets) for determination of diazepam. The method is linear, quantitative, and reproducible and can also be used for the tablet dissolution test. Ten tablets of the same batch were analyzed by the described method and by a high-performance liquid chromatographic method, and the results were in good agreement.

Synthesis, characterization, and analytical studies of adosupine, a potential new drug for urinary incontinence.

The synthesis and the spectroscopic characterization of a new potential drug for urinary incontinence, adosupine, is described. Adosupine and its potential synthesis impurities were analyzed by a new HPLC method that was developed with a C18 reversed-phase column. The analysis was made under isocratic conditions, with a mobile phase of acetonitrile:water (15:85, v/v). Resolution of all synthesis impurities was allowed. The method was also applied to stability studies of adosupine in solid state and in solution under different conditions. With the conditions used, only one degradation product was shown by HPLC analysis; it was isolated, characterized, and identified as the hydrolysis product of the lactam ring present in the adosupine structure.

Simultaneous quantitative determination of calcium leucovorin and parabens in lyophilized ampoules by derivative spectroscopy.

A simple and rapid procedure for quantitation of Calcium Leucovorin and parabens simultaneously in lyophilized ampoules formulation by "zero crossing" first-order derivative spectroscopy was developed. The ampoules content was dissolved in a mixture water/ethanol (50/50) and first-derivative spectra were recorded. The absolute values of the derivative at 312 nm for the determination of Calcium Leucovorin and between 244 and 246 nm for the determination of parabens were measured. The method is linear, quantitative and reproducible.

Assessment of exposure to polycyclic aromatic hydrocarbons in police in Florence, Italy, through personal air sampling and biological monitoring of the urinary metabolite 1-hydroxypyrene.

In this study, the authors evaluated exposure to airborne polycyclic aromatic hydrocarbons (PAHs) in workers exposed to exhaust gas from cars, and they assessed the efficiency of urinary 1-hydroxypyrene as an indicator of exposure to pyrene and PAHs. The authors selected 2 groups of police who worked in 2 areas in the city of Florence: 1 group was highly exposed to high-density traffic emissions during the winter and summer of 1997, and the 2nd group experienced low exposure to traffic emissions during the same period. Ambient monitoring was achieved with personal sampling of airborne PAHs during each workshift. Eight hydrocarbons were used as indicators of pollution caused by PAHs (e.g., pyrene, benzo[a]pyrene, benzo[a]anthracene, dibenzo[a,h]anthracene). Biological monitoring was performed through dosing of 1-hydroxypyrene (pyrene metabolite) in urine samples taken at the end of each workshift. The ambient monitoring revealed that PAH concentrations were influenced by both season of sampling and varying intensities of traffic in the different areas. The median concentration of benzo[a]pyrene in winter was twice as high in the high-density traffic area as in the low-density traffic area (i.e., 4.1 ng/m3 versus 1.8 ng/m3). In summer, the high-density traffic area experienced benzo[a]pyrene concentrations that were 6 times higher than in the low-density traffic area (i.e., 1.2 ng/m3 versus 0.2 ng/m3). Benzo[a]pyrene was also correlated highly (r(s) = .92, p < .0001) with the mixture of total PAHs analyzed, thus confirming its function as a good indicator of exposure to PAHs in an urban environment. Levels of urinary 1-hydroxypyrene appeared to be generally influenced by the intensity of traffic, especially during the winter (i.e., median value in winter was 199.2 ng/gm creatinine in the high-density traffic area and 120.5 ng/gm creatinine in the low-density traffic area). An analysis of the general data revealed that 1-hydroxypyrene was, to some degree, related to pyrene, benzo[a]pyrene, and airborne total PAHs, whereas analysis of separate data for the area and the season revealed an emergence of a closer correlation during the winter in the high-traffic area. Therefore, 1 -hydroxypyrene can be considered a good biological indicator of exposure to airborne PAHs in the urban environment, especially in winter and in high-density traffic areas.

[Reference values of solvents and metabolites in biological samples]. / Valori di riferimento di solventi e metaboliti in campioni biologici.

This article presents a review of reference values for organic solvent biomarkers. Some of these results were obtained in the research activities of the Italian Society Reference Values (SIVR). Most experiences show data obtained from control groups during occupational exposures assessment investigations. We considered only data related to the following biomarkers: immodified solvents in blood and urine, their main urinary metabolites. The reference values of the following solvents are reported: benzene, toluene, xylene, nhexane, cyclohexane, trichloroethylene, perchloroethylene, methanol, acetone, N,N-dimethylformamide, carbon disulphide. In the text also the influence of some confounding factors is discussed.

[Evaluation of professional exposure to antiblastic chemotherapeutic agents in a Tuscan hospital environment]. / Valutazione dell'esposizione professionale a chemioterapici antiblastici in ambiente ospedaliero in Toscana.

The aim of this study, carried out from 1997 to 2000, was to evaluate the exposure to antiblastic drugs of hospital staff (38 nurses of 7 oncological Day Hospital care in Tuscany). To evaluate the internal exposure was used biological monitoring, in particular was determined the concentration of urinary cyclophosfamide (CF); pad was made to evaluate the skin contamination. The contamination of working surfaces was determinate, by wipe-test, to verify the decontamination procedures; were used, as markers, CF and 5-fluorouracil (5-FU). We assessed the permeability thorough gloves usually used by the staff and the degradative activity to these drugs by agents used to decontaminate the working surfaces. The results shows, in urine, value lower than the detection limits, although was found a diffused contamination of the working environment. The results confirmed that NaClO is the best product to decontaminate working surfaces and nitrile gloves the ones with less permeability.

[Acute dimethylformamide (DMF) poisoning: a case report]. / Intossicazione acuta da dimetilformamide (DMF): descrizione di un caso.

This paper describes a case of acute occupational intoxication by dimethylformamide in a worker assigned to polyurethanic resin preparation in a simulated leather factory. The peculiarity of this case is constituted by the association of a dimethylformamide classic clinical syndrome, frequently described in the scientific literature (alcohol intolerance, gastroenteric manifestations with liver injury), with coagulation alterations and thrombocytopenia. Measurement of environmental concentrations of the solvents and biological monitoring revealed high levels of exposure to dimethylformamide at the workplace. Our observations confirm the effects of dimethylformamide on hemostasis reported by other authors in previous studies. It is possible to speculate that the effects of dimethylformamide on coagulation and platelets strictly depend on the amount of solvent accumulated in the body.

Local dynamics in biological macromolecules.

The diffusive approach in the optimized Rouse-Zimm approximation to segment relaxation in the nanosecond time domain (ORZLD) is extended to consider chains of nonequivalent units as occurring in biological macromolecules. The correlation times for second-order time correlation functions of each virtual bond on the chain are calculated for some homopolypeptides, and random and regular copolypeptides. The expected correlation times for biological macromolecules organized in multiple domains are discussed via a simple model of the ORZLD hierarchy. Dynamic bond correlation times are compared with static local persistence lengths.

Gas chromatographic-mass spectrometric determination of two new antimycotic agents, 1-[(5-chloro-2-benzofuranyl)(2-chlorophenyl)methyl]-1H-imidazole and 1-[(5-bromo-2-benzofuranyl)phenylmethyl]-1H-imidazole, in rabbit plasma following topical administration: a preliminary comparison with bifonazole.

A method for the analysis of the antimycotic drugs 1-[(5-chloro-2-benzofuranyl)(2-chlorophenyl)methyl]-1H-imidazole, 1-[(5-bromo-2-benzofuranyl)phenylmethyl]-1H-imidazole and bifonazole in rabbit plasma, employing gas chromatography-mass spectrometry with selected-ion monitoring, was developed. The procedure involved single-step purification of the biological matrix via liquid-liquid extraction on Extrelut columns and use of a carrier substance to minimize the negative effects of adsorption sites during the gas chromatographic process. The limits of detection ranged from 0.1 to 1.4 ng/ml, starting from a 200-microliter sample. The method was applied to a preliminary evaluation of percutaneous absorption of both drugs in the rabbit after a single administration, in comparison with bifonazole.

Determination of N-acetyl-S-(N-methylcarbamoyl)cysteine (AMCC) in urine by high-performance liquid chromatography combined with mass spectrometry.

A simple, sensitive and specific method for the quantification of N-acetyl-S-(N-methylcarbamoyl)cysteine (AMCC) in urines of the general population and of exposed workers has been developed. Samples were first diluted with phosphate buffer followed by purification by solid-phase extraction (SPE) with SAX columns prior to analysis by liquid chromatography/mass spectrometry (LC/MS). An external standard was used for the quantification together with selected ion monitoring of m/z 219, [M-H]-. The linear calibration curve showed a good correlation (r2 = 0.98, p < 0.001) between 0.1-110 mg/L of AMCC, and the detection limit was calculated to be 2 microg/L. The within-day and between-day precision, calculated for exposed workers and general population AMCC levels in urine samples, were determined as 1.2 and 3.6%, respectively (as relative standard deviation (RSD)).

[Occupational exposure and individual factors influencing urinary elimination of hippuric acid]. / Esposizione professionale e fattori individuali che condizionano l'eliminazione urinaria di acido ippurico.

The influence of not occupational factors (smoking, alcohol, coffee, drugs, sport, sex, age and body weight) on hippuric acid excretion was analyzed in two groups of healthy male subjects. A group was constituted of 710 painters occupied in wood and coach workings and the other one of 109 not occupational. The 5 degrees, 50 degrees, 95 degrees percentiles of the two distribution were 208, 605, 1784 and 153, 538, 1700 mg/g creatinine respectively. The analysis of variance undertaken on exposed subjects showed that there was a significative difference between urinary hippuric acid levels of subjects consuming alcohol and the not ones. Multiple regression analysis on hippuric acid excretion values distribution in not exposed group showed that only the variable "coffee consumption" resulted statistically significative (Ln hippuric acid = 5.0287 + 0.8062; R2 = 01221). Coffee consumption increase excretion rate (mg/hour) of hippuric acid likely an exposure to a toluene air concentration of 50 mg/m3. The authors suggest an action of caffeine on endogenous amount of benzoic acid.