Biocatalytic membrane reactor development for organophosphates degradation.

Organophosphates (OPs) are highly toxic compounds used as pesticides and nerve agents. The devastating effects, reported in different studies, on the environment and human health indicate a serious scenario for both instantaneous and long terms effects. Bio-based strategies for OPs degradation seem the most promising solutions, particularly when extremophiles enzymes are used. These systems permit OPs degradation with high efficiency and specificity under mild conditions. However, as frequently observed, enzymes can easily lose activity in batch systems, so that a strategy to improve biocatalyst stability is highly needed, in order to develop continuous systems. In this work, for the first time, a continuous biocatalytic system for organophosphates (OPs) detoxification has been proposed by using a triple mutant of the thermostable phosphotriesterase (named SsoPox) isolated from the hyperthermophilic archaeon Sulfolobus solfataricus. The enzyme was covalently immobilized on polymeric membranes to develop a biocatalytic membrane reactor (BMR) able to hydrolyse a pesticide (paraoxon) contained in water. High paraoxon degradation (about 90%) and long term stability (1 year) were obtained when the enzyme was covalently immobilized on hydrophilic membranes. On the contrary, the enzyme in batch system completely loses its activity within few months after its solubilisation in buffer.

A novel KIF5A mutation in an Italian family marked by spastic paraparesis and congenital deafness.

Hereditary spastic paraplegia (HSP) includes a group of diseases characterized by progressive spastic weakness of the lower limbs (pure forms) with possible additional signs (complicated forms). The SPG10 form is due to alteration in the kinesin1A gene (KIF5A) that encodes the neuronal kinesin heavy chain, a protein required for the anterograde axonal transport. We performed clinical, neurophysiological and molecular studies in two siblings affected by AD-HSP complicated by deafness. The screening of the KIF5A gene revealed the novel mutation p.Leu259Gln in two affected siblings and in their father with a pure form of HSP.

Intensification of gas-phase photoxidative dehydrogenation of ethanol to acetaldehyde by using phosphors as light carriers.

In this work a significant improvement of VO(x)/TiO(2) photocatalytic activity in the selective partial oxidation of ethanol to acetaldehyde was achieved by the simultaneous irradiation with light emitting phosphorescent particles and UVA-LEDs as external light source. Photocatalytic tests were carried out in a gas-solid photocatalytic fluidized bed reactor at high illumination efficiency, in which the bed is constituted by VO(x)/TiO(2) photocatalyst at nominal V(2)O(5) content of 5 wt% and suitable selected phosphors, diluted with glass spheres. In this way, phosphors were fluidized together with the catalyst, excited by external UVA-LEDs, emitting their stored energy in close proximity to the catalyst. In the absence of phosphors the ethanol consumption rate initially grows linearly with initial alcohol concentration, then bends towards an asymptotic value for initial ethanol concentration higher than 0.5 vol%. By contrast, when phosphors are present, the ethanol consumption rate increased linearly in the overall range. In all cases acetaldehyde was the main product detected in gas phase with a selectivity of about 97%, ethylene and carbon dioxide the by-products. The results evidenced that the presence of phosphors allowed improved photon transfer, increasing the apparent quantum yield from 2 to 30% together with a high photoreactivity.

District-based abdominal aortic aneurysm screening in population aged 65 years and older.

AIM: Screening for abdominal aortic aneurysms (AAAs) has been carried out in an area of Genoa (Italy) for subjects aged 65 years or more to evaluate prevalence of this disease. METHODS: Between March 2007 and September 2009 8234 subjects were screened. Ultrasound examination of the abdominal aorta and the iliac arterial segments was carried out on each subject and all data related to risk factors were collected. RESULTS: Five hundreds-twelve (6.2%) subjects were found to have an AAA: 469 (10.8%) males and 43 (1.1%) females (significant difference, P < 0.01). Based on the aortic diameter, 403 (4.9%), 80 (1.0%) and 29 (0.3%) had an AAA of 3.0-3.9 cm, 4.0-4.9 cm and ≥ 5.0 cm diameter, respectively. With regards to risk factors, family history of cardiovascular disease only resulted more frequent in subjects with AAA than in those without AAA. CONCLUSION: The prevalence of patients with AAA (6.2%) was similar to previously published estimates. Nevertheless, AAA resulted very high in males. This observation is likely due to screening in a city with a very high percentage of elderly subjects. Family predisposition to cardiovascular disease resulted significant risk factor for AAA. Results of our epidemiological study provide evidence of the usefulness of AAA screening thanks to early diagnosis and appropriate treatment of AAA.

A step forwards in ethanol selective photo-oxidation.

This work focuses on the optimization of a photocatalyst formulation for the selective oxidation of ethanol to acetaldehyde. VO(x)/TiO(2) catalysts at different vanadium loading were studied in a gas-solid photocatalytic fluidized bed reactor at high illumination efficiency, in which the bed is constituted by photocatalyst diluted with alpha-Al(2)O(3) or silica gel. Photocatalytic tests showed the selective formation of acetaldehyde, with ethylene and carbon dioxide as by-products. Selectivity is influenced by the vanadium loading. For ethanol inlet concentration of 0.2 vol%, maximum conversion and acetaldehyde selectivity of 73% and 97%, respectively, were obtained at 100 degrees C on catalyst at nominal 5 wt% V(2)O(5) content (53% of surface monolayer) mixed with alpha-Al(2)O(3). Selective sites were related to surface polymeric vanadates possessing Ti-O-V and V-O-V functionalities while the photoactivity appeared correlated with the catalyst equivalent band gap energy. Increasing the ethanol inlet concentration to 1 vol% and diluting the catalyst with silica gel, total ethanol conversion with about 97% selectivity to acetaldehyde was achieved with a photoreactivity of 0.34 mol m(irradiated)(-3) s(-1). This is three times higher than reported for other photoreactors.

A novel mutation in the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene associated with a severe Rett phenotype.

Mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene have recently been reported in patients with severe neurodevelopmental disorder characterized by early-onset seizures, infantile spasms, severe psychomotor impairment and very recently, in patients with Rett syndrome (RTT)-like phenotype. Although the involvement of CDKL5 in specific biological pathways and its neurodevelopmental role have not been completely elucidated, the CDKL5 appears to be physiologically related to the MECP2 gene. Here we report on the clinical and CDKL5 molecular investigation in a very unusual RTT case, with severe, early-neurological involvement in which we have shown in a previous report, a novel P388S MECP2 mutation [Conforti et al. (2003); Am J Med Genet A 117A: 184-187]. The patient has had severe psychomotor delay since the first month of life and infantile spasms since age 5 months. Moreover, at age 5 years the patient suddenly presented with renal failure. The severe pattern of symptoms in our patient, similar to a CDKL5 phenotype, prompted us to perform an analysis of the CDKL5, which revealed a novel missense mutation never previously described. The X-inactivation assay was non-informative. In conclusion, this report reinforces the observation that the CDKL5 phenotype overlaps with RTT and that CDKL5 analysis is recommended in patients with a seizure disorder commencing during the first months of life.

CADASIL: extended polymorphisms and mutational analysis of the NOTCH3 gene.

CADASIL is a cerebrovascular disease caused by mutations in the NOTCH3 gene. Most mutations result in a gain or loss of cysteine residue in one of the 34 epidermal growth factor-like repeats in the extracellular domain of the Notch3 protein, thus sparing the number of cysteine residues. To date, more than 130 different mutations in the NOTCH3 gene have been reported in CADASIL patients, of which 95% are missense point mutations. Many polymorphisms have also been identified in the NOTCH3 coding sequence, some of them leading to amino acid substitutions. The aim of the present study was to analyze the NOTCH3 gene in a large group of patients affected by leukoencephalopathy and to investigate the presence of genetic variants. The molecular analysis revealed several nucleotide alterations. In particular, we identified 20 different mutations, 22 polymorphisms, and 8 genetic variants of unknown pathological significance never reported previously. We hope that this NOTCH3 gene mutational analysis, performed in such a significant number of unrelated and related patients affected by leukoencephalopathy, will help in molecular screening for the NOTCH3 gene, thus contributing to enlargement of the NOTCH3 gene variation database.

A novel locus for dHMN with pyramidal features maps to chromosome 4q34.3-q35.2.

The distal hereditary motor neuropathy (dHMN) is a rare genetically and clinically heterogeneous disorder characterized by weakness and wasting of distal limb muscles in absence of overt sensory abnormalities. Recently, pyramidal signs have been also described in some patients with dominant or recessive dHMN, and two different loci have been identified in families affected by dHMN complicated with pyramidal dysfunction. We investigated an Italian family affected by an autosomal dominant dHMN complicated by pyramidal signs in order to map a new gene locus. The disease maps to a novel locus in a 26-cM region flanked by D4S1552 and D4S2930 on chromosome 4q34.3-35.2. Three candidate genes (SNX25, CASP3 and TUBB4Q) located in the critical region were screened for the presence of mutations by heteroduplex analysis. No mutations have been detected in the analyzed genes. In conclusion, the new private genetic locus we reported further confirms the wide heterogeneity of dHMN.

A novel Angiogenin gene mutation in a sporadic patient with amyotrophic lateral sclerosis from southern Italy.

Mutations in the Angiogenin gene (ANG) linked to 14q11.2 have been recently discovered to be associated with Amyotrophic Lateral Sclerosis (ALS) in Irish and Scottish populations. In our study we investigated the role of ANG gene in ALS patients from southern Italy. We found a novel mutation in the signal peptide of the ANG gene in a sporadic patient with ALS (SALS). The molecular analysis of the ANG gene also demonstrated an allelic association with the rs11701 single nucleotide polymorphism (SNP) in familial ALS (FALS) but not in SALS patients. Our finding supports the evidence that the ANG gene is involved in ALS.