Real-life experience on effectiveness and safety of dupilumab in adult patients with moderate-to-severe atopic dermatitis.

BACKGROUND: Dupilumab, a fully human monoclonal antibody targeting the alpha subunit of IL-4 was recently approved for the treatment of moderate-to-severe atopic dermatitis (AD) in adult patients. OBJECTIVE: To assess dupilumab effectiveness and safety in adults with moderate-to-severe AD in a real-life Italian multicentre retrospective cohort. METHODS: Adult moderate-to-severe AD patients, referring to 39 Italian centers, received dupilumab in the context of a national patient access program. Disease assessment was performed at baseline, after 4 and 16 weeks of treatment using Eczema-Area-and-Severity-Index (EASI) score, itch and sleep numerical-rating-score (itch-NRS, sleep-NRS) and Dermatology-Life-Quality-Index (DLQI). RESULTS: A total of 109 (71 M/38F) patients was studied. There was a significant reduction in EASI score, itch-NRS, sleep-NRS and DLQI from baseline to week 4 and a further significant decline to week 16. EASI 50, EASI75 and EASI90 were achieved by 59.6%, 28.4% and 9.3% of patients at 4 weeks and by 87.2%, 60.6% and 32.4% of them at 16 weeks, respectively. Adverse events were experienced by 19.2% (21/109) of the patients and they were all mild in intensity, being conjunctivitis the most common side effect. CONCLUSIONS: Dupilumab significantly improved disease severity, pruritus, sleep loss and quality of life with an acceptable safety profile.

Acute juvenile pityriasis rubra pilaris: a case report after mononucleosis infection.

A 24-year-old male boy presented dermatosis which first appeared acutely after an infection at age 17. Clinical and histopathologic examinations were consistent with a diagnosis of juvenile pityriasis rubra pilaris type III. Treatment with UVB narrow-band led to complete resolution of the dermatitis within 1 year. Pityriasis rubra pilaris is a papulosquamous disorder of unknown etiology, which can be treated with retinoids, methotrexate, cyclosporine, and narrow-band phototherapy.

Age-related alteration of the adenosine/dopamine balance in the rat striatum.

An antagonistic interaction between adenosine A2A- and dopamine D2-receptors has been described. Radioligand binding experiments showed a predominant reduction in the number of D2 vs. A2A-receptors in the striatum of aged compared to young rats. The A2A-receptor-mediated antagonistic modulation of D2-receptor binding remained unchanged in aged animals. In striatal homogenates a significant increase in adenosine and no change in dopamine content was found in aged vs. young rats. These results reveal the existence of an age-dependent imbalance of adenosine vs. dopamine in favor of adenosine, which involves both presynaptic and postsynaptic mechanisms.

Adenosine A2A receptor stimulation enhances striatal extracellular glutamate levels in rats.

The influence of CGS 21680, an adenosine A2A receptor agonist, on striatal glutamate extracellular levels was tested in a microdialysis study in rats. CGS 21680 (10 mu M), infused intrastriatally through the microdialysis probe, greatly enhanced glutamate extracellular levels. These results show that striatal adenosine A2A receptors are involved in the regulation of striatal glutamate extracellular levels. They also suggest that adenosine A2A receptor antagonists may possess neuroprotective effects in models of striatal excitotoxicity.

Simultaneous high-performance liquid chromatographic determination of adenosine and dopamine in rat striatal tissue with combined ultraviolet absorbance and electrochemical detection.

A method is described for the simultaneous determination of biogenic amines, adenosine and their metabolites in rat striatal tissue using high-performance liquid chromatography with ultraviolet spectrophotometric and electrochemical detection. Peaks in the chromatograms of striatal tissue extracts were identified by retention times and by on-line analysis of peak spectra for adenosine and its metabolites, and by comparing current ratios of the dual-electrode coulometric detector for monoamines and metabolites. The assay gives a linear response over the concentration range of 0.15-0.60 micrograms/ml for biogenic amines, 0.5-2.0 micrograms/ml for serotonin, 5-20 micrograms/ml for hypoxanthine, adenosine and N-methyladenosine, and 10-40 micrograms/ml for inosine. The limit of detection for striatal homogenates was 3.5 ng/g for monoamines, 9 ng/g for serotonine, 140 ng/g for hypoxanthine, 290 ng/g for inosine and 80 ng/g for adenosine. The recovery ranged from 88.5% for vanillylmandelic acid to 110.3% for dopamine. The method was used to measure biogenic amines, adenosine and related metabolites in rat striatal tissues.

Different effects of harmine on plasma concentrations of L-dopa and on cerebral dopamine metabolism in rabbits and rats.

The effects of short-term intravenous administration of harmine, a monoamine oxidase inhibitor, on the plasma concentrations of L-Dopa and on dopamine levels in the brain striata of rats and rabbits after L-Dopa administration were studied. Harmine affects the L-Dopa plasma concentrations in rabbits but not in rats: in fact in the former species the area under the concentration-time curve observed after administration of L-Dopa alone increased significantly when animals were pretreated with harmine. Dopamine striatal levels increased in concert with plasma L-Dopa concentrations after administration of L-Dopa in rats and rabbits. However pretreatment with harmine resulted in a significant increase of dopamine levels in the brain striata of rabbits only. These results suggest that harmine or one of its metabolites affect the brain dopamine system not merely as a type A monoamine oxidase inhibitor but with a modulatory effect, without a precise indication of site or mode of action of harmine.

Determination of thyreostatic residues in cattle plasma by high-performance liquid chromatography with ultraviolet detection.

A procedure is described for the concurrent assay of thiouracil, methylthiouracil, propylthiouracil, phenylthiouracil and methimazole in bovine plasma. In this procedure, reversed-phase high-performance liquid chromatography is performed after liquid-liquid extraction of plasma with ethyl acetate. Compounds are quantified by ultraviolet detection using a wavelength of 276 nm for thiouracil, methylthiouracil, propylthiouracil and phenylthiouracil and 258 nm for methimazole. The linearity range, precision, recovery and detection limits were determined and the method was shown to be applicable to samples of plasma from young bulls experimentally treated with methylthiouracil.

Activity of liposomal amphotericin B (AmBisome) against Leishmania infantum and tissue distribution in mice.

Preliminary observations have shown that AmBisome, a liposomal formulation of amphotericin B (Vestar Inc.), is effective and non-toxic in animal and human visceral leishmaniasis. The activity of multiple doses of this drug on Leishmania infantum, in BALB/c mice was investigated, and amphotericin B concentration in liver and spleen was determined. Groups of infected mice were treated intravenously with 3, 5, or 7 doses of AmBisome (3 mg/kg) over 3, 10 and 25 days, respectively. The antileishmanial activity of the drug was compared with that of meglumine antimoniate (28 mg Sbv/kg per day over 21 days). Three consecutive daily doses of AmBisome were sufficient to clear all parasites from the liver of mice, while antimony did so only after 21 doses. Twenty-four-48 h after their last dose all the AmBisome-treated mice showed very high amphotericin B concentrations in liver (61.2-76.2 micrograms/g) and spleen (39.8-72.1 micrograms/g) with no overt signs of toxicity. Mice that received 2 or 4 doses at intervals of 5 to 8 days, maintained drug levels as high as those detected after 3 consecutive doses over 11 and 26 days, respectively. This should enable visceral leishmaniasis treatment on an intermittent or outpatient basis, thereby reducing overall treatment costs.

High-performance liquid chromatography-electrochemical detection of 3-methylhistidine in human urine.

A reversed-phase high-performance liquid chromatographic method is described for the determination of 3-methylhistidine content in human urine using pre-column derivatization with phenylisothiocyanate, isocratic elution with 15 mM sodium acetate-acetonitrile (92:8, v/v) and electrochemical detection. The limit of quantitation was 0.1 pmol. The method has been applied in routine analyses of 3-methylhistidine in both clinical and research work.

Simultaneous high-performance liquid chromatographic analysis of buspirone and its metabolite 1-(2-pyrimidinyl)-piperazine in plasma using electrochemical detection.

A selective and sensitive high-performance liquid chromatographic method with coulometric detection is described for the quantitation of buspirone and its active metabolite, 1-(2-pyrimidinyl)piperazine, in plasma samples of mice treated orally with buspirone (10 mg/kg body weight). The analytes are extracted with a carboxylic acid solid-phase extraction column before chromatography. A dual-electrode electrochemical detector is used. The limit of detection is 50 pg for buspirone and 35 pg for 1-(2-pyrimidinyl)piperazine.

Determination of cimetidine and related impurities in pharmaceutical formulations by high-performance liquid chromatography.

The analytical characteristics of cimetidine tablets were studied. A high-performance liquid chromatographic method was developed in order to assay cimetidine and its related impurities simultaneously. A reversed-phase system and diode-array detector were used.

Determination of naltrexone and 6 beta-naltrexol in plasma by high-performance liquid chromatography with coulometric detection.

A simple and reliable reversed-phase high-performance liquid chromatographic method with coulometric detection is described for the quantitation of naltrexone and its metabolite, 6 beta-naltrexol, in plasma samples of healthy volunteers who received orally 50 mg of naltrexone. The analytes and the internal standard, naloxone, are extracted with an octadecyl solid-phase extraction column before chromatography. The mobile phase is 0.01 M potassium phosphate (pH 3)-acetonitrile (85:15, v/v) and it is pumped at 0.8 ml/min. The coulometric detector is formed by two electrodes set at +0.20 V and +0.70 V, with a palladium reference electrode. The limit of quantitation observed was 5 ng/ml for both naltrexone and 6 beta-naltrexol. This method can be used to investigate pharmacokinetic parameters of different pharmaceutical preparations of this opioid antagonist.

Plasma catecholamine responses during a personalized physical stress as a dynamic characterization of essential hypertension.

High performance liquid chromatography (HPLC) with electrochemical detection proves to be a reliable method for determination of plasma catecholamines (CA) to assess the possible role of the sympathetic nervous system (SNS) in essential hypertension (EH). The present investigation in a group of 15 normotensive (N) and 13 stable EH patients, homogeneous for age and duration of hypertension, was carried out without treatment in the supine position, up-right position and during a personalized bicycle exercise. Mean blood pressure, mean heart rate, plasma renin activity and plasma aldosterone were also evaluated at the various exertion phases. Norepinephrine (NE) and epinephrine (E) showed a progressive increase in N and in EH patients, reaching the highest values at maximum effort. However, EH patients showed higher E plasma levels than N before maximum effort. Dopamine (DA) reached the highest values in N at maximum effort and in EH patients at recovery time. These findings allow us to foresee the possibility of a better characterization of the SNS role in EH.

High-performance liquid chromatographic determination of L-3-(3,4-dihydroxyphenyl)-2-methylalanine (alpha-methyldopa) in human urine and plasma.

A procedure is described for the determination of alpha-methyldopa (MD) [L-3-(3,4-dihydroxyphenyl)-2-methylalanine], its metabolite and catecholamines in the urine and plasma of patients undergoing MD therapy, by high-performance liquid chromatography with dual working electrode coulometric detection. An efficient sample preparation procedure is presented for the isolation of endogenous MD, its metabolite and catecholamines from plasma or urine. After deproteinization of a plasma sample with methanol containing 2% of 0.5 M perchloric acid and dilution of a urine sample (1:200), MD, dihydroxyphenylacetic acid (DOPAC), 3-O-methylmethyldopa (3-OMMD) and homovanillic acid (HVA) were separated with a Supelcosil LC-18 column. Catecholamines were extracted from the supernatant of deproteinized plasma or from urine by ion exchange on a Sephadex CM-25 column and subsequent adsorption on alumina. The use of the same mobile phase for the concurrent assay of MD, its metabolite and catecholamines increased considerably the efficiency of sample separation. Recoveries were close to 100% for MD, DOPAC, 3-OMMD and HVA and 70% for catecholamines. The effects of various experimental parameters related to mobile phase composition on chromatographic performance are reported. The purity of the eluted compounds was tested by recording both the first detector response (oxidation current) and the second detector response (reduction current). The ratio of the detector responses yielded a chemical reversibility ratio for the detected compound. A number of applications such as monitoring data from patients under MD therapy are presented.

Application of a high-performance liquid chromatography coulometric method for the estimation of mebendazole and its metabolites in human sera.

A novel, sensitive high-performance liquid chromatographic method, making use of coulometric detection, for the estimation of mebendazole and its metabolites in the sera of eight hydatidosis patients was devised. Recovery rates, precision, accuracy and sensitivity for each compound are reported and compared with those of the previously published methods.