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Biomedical subjects

A Triolo

Publications and source records attributed to A Triolo.

28 records · Page 2Linked to original sources

Analysis of platelet-activating factor by gas chromatography-mass spectrometry: low-energy electron impact of the corresponding 3-acetyl-2-tert.-butyldimethylsilyl derivative.

A method for the analysis of platelet-activating factor in platelets employing gas chromatography and selected-ion monitoring mass spectrometry with low-energy electron impact and stable isotope dilution was developed. The procedure involved Bligh and Dyer extraction of the sample followed by thin-layer chromatographic purification. Platelet-activating factor is successively hydrolysed to the corresponding 2-acetyl-1-O-alkylglycerol by digestion with phospholipase C, and the product is allowed to isomerize to the more thermodynamically stable 3-acetyl-1-O-alkylglycerol before column purification and derivatization of the free OH with tert.-butyldimethylchlorosaline-imidazole. This reagent is of common use in platelet-activating factor derivatization, but is made to react with 2-acetyl instead of 3-acetyl isomer. The advantages of using the latter for the final derivatization are discussed and this method is compared with others currently available for gas chromatographic-mass spectrometric analysis of platelet-activating factor.

Blood Platelets↗

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.

Administration, Topical↗

A new glucose clamp algorithm: clinical validation.

A new glucose clamp technique for in vivo studies of insulin sensitivity was validated clinically. Eighteen patients (10 males, 8 females, age 35-80 years, body mass index 34.6-17.04) were connected to a computer-assisted artificial pancreas "Betalike R", using a new algorithm based on a "minimal model", to carry out the glucose clamp technique automatically and especially to overcome the well-known problems of its priming phase. We performed the euglycemic hyperinsulinemic clamp in four patients and the hyperglycemic hyperinsulinemic clamp in 14. In one patient both clamps were done. The mean priming time to reach steady-state glycemia was 20 min. Plasma insulin concentrations were measured every 20 min. This new automatic glucose clamp technique enables the priming phase to be run without any significant overshoot, and accidental variations of glycemia in steady state were reduced to a minimum. The system showed satisfactory safety and stability in controlling the patient's glycemia and assured high speed of the priming phase.

Adult↗

[Evaluation of a new immunoturbidimetry technique for measuring microalbuminuria].

Subclinical elevation of urinary albumin excretion is a good predictor of later clinical proteinuria. A simple, sensitive and rapid immunoturbidimetric method was developed to quantify urinary albumin excretion (URIN-PAK ImmunoMICRO LAB, Miles Italia Spa). In the presence of polyethylene glycol 6000, immunocomplex between human albumin and its specific antibody are rapidly formed (5-50 min, at room temperature). Absorbance reading are mode U 340 nm (Automatic Analyzer RA 1000, Technicon). The test is specific for albumin failing to cross react with other plasma proteins present in urine, as well as with glibenclamide, chlorpropamide, phenformin, hemoglobin, glucose, urea and thymol. The present method correlates with SCLAVO H-ALBUMIN RIA Kit (r = 0.9917). The test is suitable for clinical use.

Albuminuria↗

Focal cerebral ischemia: reduction in size of infarcts by ventriculo-subarachnoid perfusion with fluorocarbon emulsion.

A new method for brain resuscitation following acute focal ischemic insult has been developed in this laboratory. The technique utilizes a surrogate route to supply cerebral metabolites and employs highly oxygenated fluorocarbons (OFNS), which are efficient gas transport and exchange agents, perfused through the ventriculo-subarachnoid spaces. We previously described a return of aerobic metabolism and EEG after severe global ischemia by oxygenated perfusions and now report treatment-induced reduction in the size of experienced cerebral infarction. Twenty-eight cats were anesthetized (choralose and urethane), tracheotomized and placed in a stereotactic frame. Physiologic adjustments assured arterial blood pCO2 28-35 Torr, pO2 100-150 Torr pH 7.4 and glucose less than 200 mg%. The left middle cerebral artery was exposed transorbitally and temporarily clipped along with both common carotids for 2 h. One hour later (3 h after ischemic onset) the treated group were perfused by the ventriculo-cisternal route either with OFNS [pO2 = 600 Torr; 3 ml/min 6 h, 2 ml/min 2 h, 1 ml/min 2 h, 0.5 ml/min 2 h at 10 mm Hg intracranial pressure (ICP)] or with the vehicle perfusate. Eighteen to twenty hours after the ischemic insult the animals were sacrificed. Sections of fresh brain of 0.5 mm thickness were incubated in 1% triphenyl tetrazolium chloride. The infarcted areas were confirmed with classic neuropathologic techniques. Areas of infarction (expressed in cm3 and as % of the brain) were measured using a planimeter. OFNS-treated brains contained 80% less infarcted tissue than the vehicle-perfused or untreated stroked animals. The infarcted areas were significantly treatment reduced (P less than 0.05 ANOVA and Bonferroni tests).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Theophylline in the treatment of the elderly with chronic obstructive pulmonary disease.

The authors describe the pharmacological effects of theophylline and elucidate its mechanisms of action according to latest hypothesis. The narrow therapeutic range of the drug and the existence of factors capable of modifying its distribution and clearance in the elderly, are taken into consideration, as well. The behaviour of theophylline concentrations in two groups of elderly subjects affected by chronic bronchitis, treated respectively with intravenous aminophylline and anhydrous theophylline administered orally are assessed. Usefulness of associating the two routes of administration is underlined. In fact, the delay in the achievement of the therapeutic range observed after oral intake can be made up for by the initial peak induced by intravenous injection. Our observations in the elderly are consistent with the necessity of therapeutic monitoring. In addition, a careful dosage of the drug can avoid the occurrence of untoward effects.

Aged↗

Metabolism of isbufylline in humans. Isolation, identification, and synthesis of plasma and urine metabolites.

Isbufylline metabolism after oral administration to humans was studied. The main metabolites detected by the HPLC method, in plasma, were 1-methyl-7-(2-hydroxy-2-methyl-propyl) xanthine (I), 1,3-dimethyl-7-(2-hydroxy-2-methyl-propyl) xanthine (II), and 1-methyl-7-(2-methyl-propyl) xanthine (III). The main metabolites detected in urine were 1-methyl-7-(2-hydroxy-2-methyl-propyl) xanthine (I), 1,3-dimethyl-7-(2-carboxy-propyl) xanthine (IV), and 1,3-dimethyl-7-(2-hydroxymethyl-propyl) xanthine glucuronic acid (V)-Gluc. They were isolated by HPLC, identified by GC/MS, HPLC/MS, or HPLC/MS/MS, and finally synthesized. Recovery of these metabolites, along with the absence of unmetabolized isbufylline in the urine, indicated biotransformation and renal excretion as the main routes of isbufylline elimination in humans. HPLC quantitation of the characterized urine metabolites revealed that 49% of the drug was eliminated as (I), 9% as (V)-Gluc, and 5% as (IV).

1-Methyl-3-isobutylxanthine↗

Separation and characterization of a yeast alcohol dehydrogenase conjugate with theophylline.

The purpose of this work is to characterize the structural difference of conjugates in order to condition the immunoreactivity of the enzymatic tracer in the homogeneous immunoassays. Conjugates between yeast alcohol dehydrogenase (ADH) and 7-theophyllincarboxyalkyl acids were obtained by the mixed-anhydride method, and characterized with kinetic, electrophoretic and chromatographic techniques (IEF, MCC, CF). The enzyme activities were found to be inversely proportional to the substitution grade but not correlated to the number of carbon atoms of the carboxyalkyl spacer arms. Only the oxidative form (ADH2) of the enzyme was found capable of reacting with theophylline derivatives in the experimental conditions. Enzyme conjugates were resolved into three components, due to the different distribution of theophylline moieties on the surface of the protein, as confirmed by examination of the tryptic patterns of the single components.

Alcohol Dehydrogenase↗