The advanced therapy classification procedure. Overview of experience gained so far.

The classification procedure, introduced by the European Regulation on advanced therapy medicinal products (ATMPs), has received a tremendous interest from companies, academic and public sponsors developing ATMPs. This procedure gives companies the opportunity to verify whether or not the product they are developing can be considered an ATMP and can therefore benefit from the new regulatory pathway introduced in the European Union for these types of medicinal products. This procedure is optional, free of charge and may take place at any stage of the development of an ATMP in advance of applying for a marketing authorisation. In case of doubt, briefing meetings organised by the European Medicines Agency Innovation Task Force may help preparing for an ATMP classification and are a starting point for the interactions between the Agency and the developers of ATMPs. This article reviews the advantages of the classification procedure for both the developers of ATMPs and the European regulatory network. Since the introduction of this procedure and up to 10 November 2010, the Committee for Advanced Therapies (CAT) has finalised 38 applications for classification.

Myotonic dystrophy CTG repeats in an Italian population sample: evaluation of the polymorphism for forensic applications.

The myotonic dystrophy (DM) CTG repeat polymorphism has been studied in an Italian population sample. Polymerase chain reaction (PCR) amplification, manual polyacrylamide gel electrophoresis (PAGE), and silver staining were employed. Alleles were typed by comparison with a sequenced allelic ladder. A total of 25 different alleles, spanning the range from 5 to 31 CTG triplets, was observed. The heterozygosity was 79%, and no significant deviation from Hardy-Weinberg equilibrium was found. Eighty-one meioses from parentage testing were also analyzed, and a Mendelian pattern of inheritance was observed in all cases. In addition, we could successfully type the DM locus in 20 laboratory-prepared bloodstains, with 1 ng of DNA allowing clear definition of alleles. We conclude that the CTG repeats at the DM locus may be useful for forensic applications.

Comparative in-vitro activity of moxifloxacin, minocycline and azithromycin against Chlamydia spp.

The in-vitro activity of moxifloxacin, a new 8-methoxyquinolone, was compared with minocycline and azithromycin against 40 strains of Chlamydia trachomatis, Chlamydia pneumoniae and Chlamydia psittaci. Both the MIC and the MBC of moxifloxacin ranged from 0.03 to 0.125 mg/L. MICs of minocycline ranged from 0.015 to 0.06 mg/L and MBCs between 0.03 and 0.25 mg/L. MICs of azithromycin ranged from 0.03 to 0.125 mg/L and the MBCs between 0.06 and 0.5 mg/L. MBC values of moxifloxacin were the same as MICs in 32 (80%) of 40 strains tested, whereas those of minocycline and azithromycin were two to four times higher than their MICs. These data confirm those previously obtained indicating that quinolones kill chlamydial strains at concentrations equivalent to their MICs.

PCR-based restriction pattern typing of the vacA gene provides evidence for a homogeneous group among Helicobacter pylori strains associated with peptic ulcer disease.

The results of PCR-based molecular typing of Helicobacter pylori strains by restriction fragment length polymorphism analysis of a 1, 161-bp nucleotide sequence of the midregion of the vacA gene are reported. A total of 48 H. pylori strains isolated from gastric biopsy specimens obtained from 18 patients with peptic ulcer dyspepsia, 15 patients with nonulcer dyspepsia, and 15 asymptomatic H. pylori-infected subjects were studied. Highly heterogeneous restriction patterns were obtained by digestion of PCR products with SauII, BglII, and HhaI, whereas HaeIII digestion resulted in a strictly homogeneous profile for H. pylori strains isolated from 14 of 18 (77.7%) patients with peptic ulcer dyspepsia, but a strictly homogeneous profile was found for strains from only 8 of 15 (53.3%) patients with nonulcer dyspepsia (P = 0.163) and 5 of 15 (33.3%) asymptomatic H. pylori-infected subjects (P = 0.014). A potentially important aspect of the results obtained is the clinical relevance, since a single restriction pattern seems to be able to identify the majority of H. pylori strains associated with peptic ulcer disease.

The DRPLA CAG repeats in an Italian population sample: evaluation of the polymorphism for forensic applications.

The DRPLA CAG repeats polymorphism has been studied in an Italian population sample. PCR amplification, manual PAGE and silver staining were employed. A total of 16 different alleles, spanning the range from 5 to 21 CAG triplettes, was observed. The heterozygosity was 0.81 and no significant deviation from Hardy-Weinberg equilibrium was found 81 meioses from parentage testing were also analyzed and a Mendelian pattern of inheritance was observed in all cases. In addition, we could successfully type DRPLA locus in some forensic specimens, 1 ng of DNA allowing clear definition of alleles. The authors conclude that the DRPLA CAG repeats analysis may be useful for forensic applications.

Detection of serum antibodies against Chlamydia pneumoniae by in vitro neutralization and microimmunofluorescence assays.

Antibodies to Chlamydia pneumoniae (IOL-207) were studied by the microimmunofluorescence (MIF) assay and in vitro neutralization in serum samples from 230 healthy adults and 332 patients at risk of C. pneumoniae infection. In the MIF test, the prevalence of immunoglobulin G (IgG) antibody was 53.9% among healthy subjects and 40.7% in the patients. An MIF antibody titre of > or = 1 : 512, consistent with acute infection by C. pneumoniae was documented in 23 out of 332 patients, and in none of the healthy adults. C. pneumoniae complement-dependent neutralizing antibody was detected in 25.8% and 31.2%, respectively of MIF-positive sera from healthy subjects and patients. The neutralizing antibody detection rate was 52.2% among the 23 patients with MIF titres of > or = 1 : 512. Complement-independent neutralization was observed in only 5 sera from healthy subjects and in 3 sera from patients. The complement-dependent neutralizing ability of sera significantly (p < 0.001) correlated with MIF titres.

The use of surface immunofluorescence assay (SIFA) in the microbiological diagnosis of Lyme borreliosis: a case report.

In this report we describe the use of a newly developed immunofluorescence technique performed with living spirochetes to detect serum antibody to B. burgdorferi s.l. in a case of early Lyme borreliosis. The immunofluorescence method used (surface immunofluorescence assay: SIFA) proved useful in the serological evaluation of suspected cases of Lyme disease.