Ion concentrations in nasal airway surface liquid: a prediction model for the identification of cystic fibrosis carriers.

BACKGROUND: Cystic fibrosis (CF) carriers seem to have a higher risk to develop chronic rhino-sinusitis (CRS), although the full underlying mechanisms are unknown. Ion concentrations in nasal airway surface liquid (ASL) may be influenced by the heterozygosity for CF gene mutation, with possible impacts on the development of CRS. METHODS: A cheap and feasible standardized technique was designed to measure the ion levels in nasal ASL. With this purpose we collected, under basal conditions, samples from the nasal cavity of 165 adults: 14 homozygous for CF, 83 carriers and 68 healthy controls. Sodium (Na) and Chlorine (Cl) concentrations were then evaluated among different groups. RESULTS: Statistical analysis revealed a significant difference of Na and Cl values between controls and carriers and between controls and homozygotes. Receiver operating characteristic (ROC) curves and derived indicators (Youden's index and Area Under the Curve, AUC) were used to further evaluate the diagnostic capability of Na and Cl concentrations to differentiate heterozygotes from controls. ROC curves demonstrated that the optimal diagnostic cut-off value of Na is at 124, and the optimal cut-off value of Cl is at 103,2. CONCLUSION: ASL sampling can be considered a new diagnostic tool for providing quantitative information on nasal ion composition. According to our findings, Na and Cl concentrations of nasal ASL could represent a useful tool to assess heterozygotes and healthy controls.

Multiscale Modeling of Thiol Overoxidation in Peroxiredoxins by Hydrogen Peroxide.

In this work, we employ a multiscale quantum-classical mechanics (QM/MM) scheme to investigate the chemical reactivity of sulfenic acids toward hydrogen peroxide, both in aqueous solution and in the protein environment of the peroxiredoxin alkyl hydroperoxide reductase E from Mycobacterium tuberculosis (MtAhpE). The reaction of oxidation of cysteine with hydrogen peroxides, catalyzed by peroxiredoxins, is usually accelerated several orders of magnitude in comparison with the analogous reaction in solution. The resulting cysteine sulfenic acid is then reduced in other steps of the catalytic cycle, recovering the original thiol. However, under some conditions, the sulfenic acid can react with another equivalent of oxidant to form a sulfinic acid. This process is called overoxidation and has been associated with redox signaling. Herein, we employed a multiscale scheme based on density function theory calculations coupled to the classical AMBER force field, developed in our group, to establish the molecular basis of thiol overoxidation by hydrogen peroxide. Our results suggest that residues that play key catalytic roles in the oxidation of MtAhpE are not relevant in the overoxidation process. Indeed, the calculations propose that the process is unfavored by this particular enzyme microenvironment.

Urinary excretion of ciprofloxacin after administration of extended release tablets in healthy volunteers. Swellable drug-polyelectrolyte matrix versus bilayer tablets.

This paper builds on a previous paper in which new ciprofloxacin extended-release tablets were developed based on a ciprofloxacin-based swellable drug polyelectrolyte matrix (SDPM-CIP). The matrix contains a molecular dispersion of ciprofloxacin ionically bonded to the acidic groups of carbomer, forming the polyelectrolyte-drug complex CB-CIP. This formulation showed that the release profile of the ciprofloxacin bilayer tablets currently commercialised can be achieved with a simpler strategy. Thus, since ciprofloxacin urine concentrations are associated with the clinical cure of urinary tract infections, the goal of this work was to compare the urinary excretion of SDPM-CIP tablets with those of the CIPRO XR® bilayer tablets. A batch of SDPM-CIP tablets was manufactured by the wet granulation method and the CB-CIP ionic complex was obtained in situ. Fasted healthy volunteers received a single oral dose of 500 mg ciprofloxacin of either formulation in a randomised crossover study. Urinary concentrations were assessed by HPLC at intervals up to 36 h. Pharmacokinetic parameters (rate of urinary excretion, maximum urine excretion rate, tmax, area under the curve, amount and percentage of the ciprofloxacin dose excreted in urine) showed no statistical differences between both formulations at any of the time intervals of collection. The processing conditions to obtain SDPM-CIP tablets are easy to scale up since they involve technology currently employed in the pharmaceutical industry and the process is less challenging to implement. In addition, SDPM-CIP tablets met pharmacopoeial quality specifications.

The role of hyaluronan as a drug carrier to enhance the bioavailability of extended release ophthalmic formulations. Hyaluronan-timolol ionic complexes as a model case.

The aim of this work was to obtain information concerning the properties of ophthalmic formulations based on hyaluronic-drug ionic complexes, to identify the factors that determine the onset, intensity and duration of the pharmacotherapeutic effect. Dispersions of a complex of 0.5% w/v of sodium hyaluronate (HyNa) loaded with 0.5% w/v of timolol maleate (TM) were obtained and presented a counterionic condensation higher than 75%. For comparison a similar complex obtained with hyaluronic acid (HyH) was also prepared. Although the viscosity of HyNa-TM was significantly higher than that of HyH-TM, in vitro release of TM from both complexes showed a similar extended drug release profile (20-31% over 5h) controlled by diffusion and ionic exchange. Ocular pharmacokinetic study performed in normotensive rabbits showed that HyNa-TM complex exhibited attractive bioavailability properties in the aqueous humor (AUC and Cmax significantly higher and later Tmax) compared to commercial TM eye-drops. Moreover, a more prolonged period of lowered intra-ocular pressure (10h) and a more intense hypotensive activity was observed after instillation of a drop of HyNa-TM as compared to the eye-drops. Such behavior was related to the longer pre-corneal residence times (400%) observed with HyNa-TM complex. No significant changes in rabbit transcorneal permeation were detected upon complexation. These results demonstrate that the ability of HyNa to modulate TM release, together with its mucoadhesiveness related to the viscosity, affected both the pharmacokinetic and pharmacodynamic parameters. The HyNa-TM complex is a potentially useful carrier for ocular drug delivery, which could improve the TM efficacy and reduce the frequency of administration to improve patient compliance.

Hyaluronan as drug carrier. The in vitro efficacy and selectivity of Hyaluronan-Doxorubicin complexes to affect the viability of overexpressing CD44 receptor cells.

We report Doxorubicin ionic complexes with hyaluronic acid (HiH-Dx) or its sodium salt (HiNa-Dx) as tumor-targeting delivery system. The complexes were prepared in situ by mixing aqueous solutions of Dx.HCl with HiH or HiNa. Clear colloidal dispersions with a high degree of counterionic condensation (cc) were obtained. Affinity constants (logKcc) of HiNa-Dx and HiH-Dx were 7.96 and 8.08, respectively. Delivery rates of Dx from the complexes were measured in a Franz-type bicompartimental device. In line with the high affinity constants, loaded Dx was slowly released from the complexes. To test the targeting potential of the complexes, carcinogenic A549 cells overexpressing the CD44 receptors were used. HTR8/SVneo cells without overexpression of CD44 were used as control. In A549 cells, cytotoxicity of both HiH-Dx and HiNa-Dx complexes was 3-fold higher than that of the reference solution. However, no differences were observed between the complexes and free Dx solution in HTR8/SVneo cells. Flow cytometry data suggested successful uptake of Dx in cells, with a greater internalization of Dx in A549 cells than in HTR8/SVneo cells when the complexes were used. Similarly, microscopy imagines revealed a higher concentration of Dx in A549 cells with the complexes. This work provides more detailed information in order to contribute to more solid bases to evaluate the potentiality of Hi as an antineoplastic drug carrier convenient for being used in specific therapeutic indications with minimal side effects.

Phenotype/genotype correlation and cystic fibrosis related diabetes mellitus (Italian Multicenter Study).

BACKGROUND: A genotype/phenotype correlation between early onset cystic fibrosis related diabetes (CFRD) and the N1303K mutation of the CF gene was previously identified in a small series of 28 CFRD patients, out of 313 CF patients. PATIENTS AND METHODS: In order to confirm the observation, data of 141 CFRD patients out of 1,229 CF patients attending 14 Italian CF centers were collected. All patients were older than 10 years and had been genotyped. RESULTS: DeltaF508 was the most frequent mutation (147/282 alleles: 52%) and N1303K the second most frequent mutation (18/282 alleles: 6.3%) in CFRD patients, without significant difference as compared with CF patients without DM (52% vs 48.6% and 6.3% vs 5.1%, respectively). W1282X was the third most frequent mutation in CFRD patients, more frequent than in CF patients without DM (5.3% vs 2%; p<0.001). CONCLUSIONS: Unlike the previous study, we did not find a higher frequency of the N1303K mutation in CFRD patients; moreover, data from this large CF series showed a significant correlation between the W1282X mutation and CFRD.

[Italian Cystic Fibrosis Registry: 10 years of activity]. / Registro Italiano Fibrosi Cistica: 10 anni di attività.

Cystic Fibrosis (CF) is a recessive autosomic genetic disease with an incidence in mediterranean countries of about 1:3500 born alive. In Italy the considerable genetic variability makes it difficult to identify all the homozygous subjects and, consequently, to estimate the incidence of the disease in healthy carriers. The disease is evolutive and affects various systems, most of all the respiratory and gastrointestinal systems. Not many years ago, when the clinical definition of CF was first introduced, average survival did not exceed the pediatric age. Nowadays with ever advancing diagnostic and therapeutical techniques many CF patients survive until an adult age. It is therefore necessary to plan adequate health service interventions so as to satisfy as much as possible the needs of both the patients and their families. To this end data collected since 1.1.1988 by the Italian registry for CF (year of birth, sex, region of birth and residence, diagnosis procedures, results of sweat test, pancreatic insufficiency, DNA analysis, status: alive, dead, lost to follow up) of all the patients, diagnosed in the 18 Reference Centres and the 3 local Centres for CF, have proved to be extremely useful. Since the birth of the Registry on 31.12.1997, data relating to 2458 patients alive on 1.1.1988 and 1159 born during the last ten years, for a total of 3617 subjects (1756 females and 1861 males), have been recorded. As already mentioned a considerable increase in life expectancy of CF patients (from 1988 to 1990 the average age of death was 14 years, from 1994 to 1997 it was 19) and a consequent increase in the percentage of adult patients have been observed.

[Prevalence of Chlamydia pneumoniae in respiratory infections in children: an ambulatory diagnostic problem]. / Prevalenza di Chlamydia pneumoniae nelle infezioni respiratorie del bambino: un problema diagnostico ambulatoriale.

It has been recently suggested that Chlamydia Pneumoniae infection is a common finding among children with acute respiratory diseases. Chlamydia cell culture is difficult and time-consuming to perform. Polymerase chain reaction (PCR) is a more rapid but also more expensive technique used to identify Chlamydia in pharyngeal swab, but it can be performed only in few specialized laboratories. We tested a rapid enzyme immuno-assay to detect Chlamydia in 20 children with respiratory infections (mean age 3.29 years; male:female ratio = 12:8) and in 21 healthy children (mean age 4.70 years male:female ratio = 15:6). Prevalence of Chlamydia isolation from pharyngeal swab was very high in both patients and healthy children without a significative difference in the two considered groups (45% vs 42%, p = 0.8). Specific Chlamydia IgG antibodies were undetectable in all patients and healthy children. Nine out of 20 patients affected by acute respiratory disease were Chlamydia-positive and 11 out 20 were Chlamydia-negative: these two groups didn't differ in regard to clinical and laboratory features, whereas duration of symptoms was significantly longer in Chlamydia-positive patients (9.3 vs 5.5 days, p = 0.014). Our study suggests a high prevalence of Chlamydia pharyngeal swab positivity in both healthy and sick children. Diagnosis of Chlamydia infection was not feasible on the basis of the considered clinical and laboratory findings.

Genetic history of cystic fibrosis mutations in Italy. I. Regional distribution.

Earlier analysis of the Italian population showed patterns of genetic differentiation that were interpreted as being the result of population settlements going back to pre-Roman times. DNA disease mutations may be a powerful tool in further testing this hypothesis since the analysis of diseased individuals can detect variants too rare to be resolved in normal individuals. We present data on the relative frequencies of 60 cystic fibrosis (CF) mutations in Italy and the geographical distribution of the 12 most frequent CF mutations screened in 3492 CF chromosomes originating in 13 Italian regions. The 12 most frequent mutations characterize about 73% of the Italian CF chromosomes. The most common mutation, delta F508, has an average frequency of 51%, followed by N1303K and G542X, both with average frequencies around 5%. Multivariate analyses show that the relative frequencies of CF mutations are heterogeneous among Italian regions, and that this heterogeneity is weakly correlated with the geographical pattern of non-DNA 'classical' genetic markers. The northern regions are well differentiated from the central-southern regions and within the former group the western and eastern regions are remarkably distinct. Moreover, Sardinia shows the presence of mutation T338I, which seems absent in any other European CF chromosome. The north-western regions of Italy, characterized by the mutation 1717-1G-->A, were under Celtic influence, while the north-east regions, characterized by the mutations R1162X, 2183AA-->G and 711 + 5G-->A, were under the influence of the Venetic culture.

[Hospital admissions in a pediatric department]. / Percorso dei ricoveri ospedalieri in una divisione pediatrica.

Admission of 429 children in Pediatric Department were examined. 249 children had been taken to emergency ward by parents, 131 had been hospitalized by other physician. 175 children had been examined before admission by panel doctor who decided admission of 114 in Pediatric Department. Parents decided admission of 30 children. The necessity of admission in Department was only in 33 per cent of children admitted.

The treatment of dermatophytoses of the glabrous skin: a comparison of undecylenic acid and its salt versus tolnaftate.

In a double-blind study, the efficacy of 1% tolnaftate cream, 3% undecylenic acid and its zinc salt, and a placebo cream were tested in dermatophytosis of the glaborous skin and groin. Ninety-seven subjects completed the study: 33 received tolnaftate, 23 of these subjects were cured clinically and mycologically. Thirty-two subjects received 3% undecylenic acid and 20% zinc undecylenate as a cream. Of these, 21 were cured clinically and mycologically. Only three of the 32 subjects receiving placebo were cured clinically and mycologically. Both tolnaftate and undecylenic acid and its zinc salt are effective in this condition.

[The vaccination against measles: effectiveness and care in use (author's transl)]. / Validità e limiti della vaccinazione antimorbillosa.

The A. deals with three clinic aspects of outstanding importance of the measles and its relevant vaccination; that is: the complications of the disease, the contraindications to live virus vaccine and the possible reactions to vaccine. It is particularly stressed the possibility of neurological complications due either to disease or to vaccination posthumous.

[Case of L. monocytogenes meningitis in the non-neonatal period]. / Su un caso di meningite da L. monocytogenes in età non neonatale.

A case of Listeria monocytogenes meningitidis in a little girl of Envie (Cuneo) is reported. The epidemiologic study has shown the presence of significatively high antibodies titers also in the sera of other members of the family.

Treatment of "tinea pedis" with griseofulvin and topical antifungal cream.

A protocol for determining the antifungal efficacy of systemic or topical drugs in tinea pedis is presented. In this study, (1) no patient had concomitant onychomycosis; (2) the clinical types were separated into (a) plantar scaling, (b) intertriginous, and (c) vesicular instep; (3) the soles were treated for three months (time related to the shedding of all stratum corneum); (4) the follow-up period for soles was three months (related to characteristics of the drug and its depot effect on the target area, the horny layer); (5) the final evaluation related to the percentage of patients "clinically and mycologically cured" at the end of the follow-up period. With this protocol, ultramicrosize griseofulvin (Gris-PEG) alone, topical clotrimazole (Lotrimin) alone, and a combination of the two were tested in seventy-three patients with tinea pedis. The results were as follows: for plantar scaling type of tinea pedis, the combination was not better than griseofulvin alone; for intertriginous tinea pedis, the combination was definitely better than griseofulvin alone; and topical 1 percent clotrimazole was much less effective than griseofulvin.

Superficial mycoses: treatment with a new, broad-spectrum antifungal agent: 1% clotrimazole solution.

A 1% clotrimazole solution was used as an antifungal for Tinea versicolor, Cutaneous Candidiasis and Dermatophytosis. All studies were double-blind and controlled, active vs vehicle. The final evaluation was made at a two-week follow-up after the treatment was stopped, which for most of the clinical conditions studied, was 28 days. The 1% clotrimazole solution was found to be effective with no local irritation noted.

Fruiting organs of Cladosporium werneckii.

Submerged mycelia of a strain of Cladosporium werneckii isolated from tinea nigra palmaris, when cultured on enriched corn-meal agar media, developed fruiting bodies resembling perithecia.