WES in a family trio suggests involvement of TECPR2 in a complex form of progressive motor neuron disease.

We have performed whole-exome sequencing in a family trio with a 16-year-old girl suffering of progressive motor neuron disease. There was no family history of the disease and no parental consanguinity. Our exome analysis indicated the proband as a compound heterozygote for two missense variants in the TECPR2 gene according to a recessive mode of inheritance. The TECPR2 gene has been reported as a positive regulator of autophagy which is an essential mechanism for maintaining neuron homeostasis and survival and plays a key role in major adult and pediatric neurodegenerative diseases. Variants in this gene have been found responsible for a recently described form of hereditary spastic paraplegia called SPG49 in two previous reports. We propose that both variants causing amino acid substitution, p.Leu684Val and p.Thr903Met, inherited in trans-phase compound heterozygote form, can be responsible for the phenotype observed in our patient. We also consider the possible contribution of a heterozygous variant in the SPG7 gene. Sanger sequencing confirmed the segregation of variants within the family tree including the patient's unaffected brother.

Biotechnological potential of the seaweed Cladophora rupestris (Chlorophyta, Cladophorales) lipidic extract.

Recently, with the advent of modern technologies, various marine organisms including algae are being studied as sources of natural substances effective on classical microorganisms and able to also combat the new trend of acquired resistance in microbes. In the present study the antimicrobial activity of the lipidic extract of the green seaweed Cladophora rupestris collected in a Mediterranean area, in two sampling periods (January and April), was assayed. The chemical characterization of the lipidic fractions was performed by gas-chromatography and multinuclear and multidimensional NMR spectroscopy. In the lipidic extract of C. rupestris collected in January an antibacterial activity against Enterococcus sp., Streptococcus agalactiae and Vibrio cholerae non-O1 was recorded; by contrast, bacterial inhibition was measured on several Vibrio species only in April. The fatty acid profile of C. rupestris lipidic extract, analyzed by gas chromatography, resulted mainly composed of palmitic, myristic, oleic, α linolenic, palmitoleic and linoleic acids. Moreover, since α-linolenic acid was the predominant ω3 fatty acid in April, we suggest its involvement in the antibacterial activity observed in this month, taking also into account that pure α-linolenic acid resulted effective towards some vibrios strains. C. rupestris fatty acid profile revealed also an interesting composition in polyunsaturated fatty acids in both the considered periods with the ω6/ω3 ratio lower than 1, leading to conclude that this macroalga may be employed as a natural source of ω3. Finally, the (1)H NMR spectrum in CDCl3 of algal lipid fractions showed the characteristic signals of saturated (SAFAs) and unsaturated fatty acids (UFAs) as well as other metabolites and a marked difference in free fatty acids (FFAs) content for the two examined algal lipid fractions. It is noteworthy that C. rupestris lipidic extracts show, by NMR spectroscopy, the signal pattern of polyhydroxybutyrate, a natural biocompatible and biodegradable polymer. In conclusion, on account of its antimicrobial activity, nutritional value and bioplastic content, C. rupestris lipidic extract can be considered a promising source for future biotechnological applications.

Mar Piccolo of Taranto: Vibrio biodiversity in ecotoxicology approach.

Microorganisms play an indispensable role in the ecological functioning of marine environment. Some species are sensitive while others are insensitive for a specific pollutant. The aim of this work is a preliminary study of the quantitative and qualitative distribution of cultivable vibrios in sediments and water samples characterized by different toxicity levels. For 1 year, in three suitably selected sampling stations of Mar Piccolo in Taranto (Ionian Sea, Italy), we have evaluated the toxicity level by Microtox® system, vibrios, total, and fecal coliform densities. The results of the Microtox® tests showed sediments characterized by an elevated level of toxicity, while the interstitial water of the same sites always showed biostimulatory phenomenon. The quantitative results show that vibrios and coliforms are more abundant in water than in sediment samples. The most often isolated strains were: Vibrio alginolyticus, Vibrio mediterranei, Vibrio metschinkovii, and Vibrio splendidus II. This work is the first example of study on the distribution of Vibrio species related to toxicity evaluation conducted by the Microtox® bioassay. The results show the different distribution of Vibrionaceae in two environmental matrices analyzed and characterized by different levels of toxicity.

The lipidic extract of the seaweed Gracilariopsis longissima (Rhodophyta, Gracilariales): a potential resource for biotechnological purposes?

In recent years seaweeds increasingly attracted interest in the search for new drugs and have been shown to be a primary source of bioactive natural products including antibiotics. In the present investigation the antimicrobial activity of Gracilariopsis longissima lipidic extract was assayed and its chemical characterization was carried out by means of advanced analytical techniques such as gas-chromatography and multinuclear and multidimensional NMR spectroscopy. G. longissima lipidic extract showed an antibacterial activity against several Vibrio species. These results are interesting considering both the resistance against antibiotics developed by vibrios and the need to control fish and shellfish diseases due to vibriosis. Analysis of fatty acid methyl esters performed by gas-chromatography showed that palmitic acid methyl ester (16:0) was the predominant saturated fatty acid (42%), while, among monounsaturated fatty acids, oleic acid methyl ester (18:1) prevailed (8.5%). Because the palmitic acid represents the main component of fatty acids we hypothesized its involvement in the antibacterial activity observed. However, a pure sample of palmitic acid did not show an antibacterial activity. The fatty acid profile of G. longissima revealed also an interesting composition in polyunsaturated fatty acids and in particular the ratio of ω-3 to ω-6 fatty acids was >1 thus suggesting that this macroalga may be used as a natural source of ω3. Moreover, the (1)H NMR spectrum in CDCl(3) of algal lipid fraction shows the characteristic signals of saturated and unsaturated fatty acids as well as other metabolites. Interestingly, in the lipid extract the presence of polyhydroxybutyrate, a linear biodegradable and biocompatible polyester, was clearly identified by NMR spectroscopy. In conclusion, the lipidic extract of G. longissima on account of its antimicrobial activity, nutritional value and content in biodegradable and biocompatible polyester represents an interesting potential biotechnological resource.

Isolation of alkane-degrading bacteria from deep-sea Mediterranean sediments.

AIMS: To isolate and identify alkane-degrading bacteria from deep-sea superficial sediments sampled at a north-western Mediterranean station. METHODS AND RESULTS: Sediments from the water/sediment interface at a 2400 m depth were sampled with a multicorer at the ANTARES site off the French Mediterranean coast and were promptly enriched with Maya crude oil as the sole source of carbon and energy. Alkane-degrading bacteria belonging to the genera Alcanivorax, Pseudomonas, Marinobacter, Rhodococcus and Clavibacter-like were isolated, indicating that the same groups were potentially involved in hydrocarbon biodegradation in deep sea as in coastal waters. CONCLUSIONS: These results confirm that members of Alcanivorax are important obligate alkane degraders in deep-sea environments and coexist with other degrading bacteria inhabiting the deep-subsurface sediment of the Mediterranean. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that the isolates obtained have potential applications in bioremediation strategies in deep-sea environments and highlight the need to identify specific piezophilic hydrocarbon-degrading bacteria (HCB) from these environments.

A test battery approach for ecotoxicological characterization of Mar Piccolo sediments in Taranto (Ionian Sea, Southern Italy).

The eco-toxicological approach is based on the determination of the toxic effects on organisms pertaining to various ecosystems and supplies information about the contaminants mixture bioavailability, in complex matrices as sediments. The use of a single species for a correct evaluation of the toxicity levels can be reductive, concerning the complexity of the ecosystem. In this work we have used species with various evolutionary levels and habitats; in particular, three different organisms: two amphipods species (Corophium insidiosum and Gammarus aequicauda) and one bacterium Vibrio fischeri. We have compared these organisms for the evaluation of sediments toxicity in four sites along the Ionian coast (Taranto, Italy); in particular, three sites in Mar Piccolo and one site in Mar Grande. The toxicity of sediments measured using Vibrio fischeri (Microtox Solid Phase Test protocol) has been compared with the mortality of the two amphipods. Both in polluted (Mar Piccolo sites) and in non-polluted environments (Mar Grande), the results of the three biological tests carried out converge into the evaluation of sediments quality monitored. In conclusion, these preliminary results show the potential use of Corophium insidiosum and Gammarus aequicauda as test species for a correct evaluation of sediments quality, together with Vibrio fischeri.

Culturable heterotrophic bacteria in seawater and Mytilus galloprovincialis from a Mediterranean area (Northern Ionian Sea-Italy).

Although Mar Piccolo of Taranto (Ionian Sea, Italy) is one of the most important Mytilus galloprovincialis farming areas, data concerning the natural bacterial microbiota of these mussels and their surrounding environment are still scant. This study was carried out seasonally, throughout a year, to determine culturable heterotrophic bacteria both in the water and mussels samples collected at three sampling sites in the Northern Ionian Sea: S. Vito, Lido Gandoli and Lido Silvana. Culturable heterotrophic bacteria abundance was determined by spread plate on Marine Agar. Heterotrophic bacteria were identified by several morphological, culture and biochemical methods. Bacterial concentrations were higher in the mussel samples compared to the corresponding seawater throughout the year. Among Gram negative heterotrophic bacteria, Aeromonas prevailed both in the water (18%) and mussel samples (40%). Other genera such as Moraxella, Pseudomonas, Alcaligenes, Acinetobacter, Flavobacterium, Chromobacterium, Photobacterium and Flexibacter were present with different percentages of isolation. Bacilli were predominant among Gram positive bacteria. Some genera (Lucibacterium and Vibrio) were present only in mussel samples. The results obtained contribute to improve the knowledge on both the bacterial abundance and diversity in mussels and the surrounding seawater in the Northern Ionian Sea.

Comparison of n-eicosane and phenanthrene removal by pure and mixed cultures of two marine bacteria.

The biotransformation activities of two hydrocarbonoclastic marine bacteria, Corynebacterium sp. and Sphingomonas sp. 2MPII, on n-eicosane and phenanthrene were investigated. During a 56-day experiment, in pure and mixed cultures, Corynebacterium sp. and Sphingomonas sp. 2MPII removed about 70% of the initial n-eicosane and phenanthrene concentrations (1 and 0.4 g L(-1), respectively). In pure cultures, culturable cell abundances increased over time, from 0.8 to 8.6 x 10(-11) CFU L(-1) (Corynebacterium sp.) and from 2.1 to 16 x 10(-11) CFU L(-1) (Sphingomonas sp. 2MPII ) but remained barely constant in mixed cultures. We defined a biotransformation index based on the number of culturable cells rather than the culture protein content, with the biotransformation cell yield (BCY) expressed in grams hydrocarbon CFU(-1) per day to better characterize hydrocarbon removal in pure and mixed cultures. The BCY was markedly higher in mixed than in pure cultures, increasing by a factor of 2-10.7 and 2.3-4.7 for n-eicosane and phenanthrene removal, respectively.

Effects of heavy ions on visual function and electrophysiology of rodents: the ALTEA-MICE project.

ALTEA-MICE will supplement the ALTEA project on astronauts and provide information on the functional visual impairment possibly induced by heavy ions during prolonged operations in microgravity. Goals of ALTEA-MICE are: (1) to investigate the effects of heavy ions on the visual system of normal and mutant mice with retinal defects; (2) to define reliable experimental conditions for space research; and (3) to develop animal models to study the physiological consequences of space travels on humans. Remotely controlled mouse setup, applied electrophysiological recording methods, remote particle monitoring, and experimental procedures were developed and tested. The project has proved feasible under laboratory-controlled conditions comparable in important aspects to those of astronauts' exposure to particle in space. Experiments are performed at the Brookhaven National Laboratories [BNL] (Upton, NY, USA) and the Gesellschaft für Schwerionenforschung mbH [GSI]/Biophysik (Darmstadt, FRG) to identify possible electrophysiological changes and/or activation of protective mechanisms in response to pulsed radiation. Offline data analyses are in progress and observations are still anecdotal. Electrophysiological changes after pulsed radiation are within the limits of spontaneous variability under anesthesia, with only indirect evidence of possible retinal/cortical responses. Immunostaining showed changes (e.g. increased expression of FGF2 protein in the outer nuclear layer) suggesting a retinal stress reaction to high-energy particles of potential relevance in space.

[The experience of the Operative Unit SETIL in Puglia]. / L'esperienza dell'Unità Operativa SETIL in Puglia.

(Epidemiological study on childhood tumors and leukaemia) study is an epidemiological case-control study, conducted in 15 Italian regions, including Apulia, to assess the possible risk of childhood cancer (leukaemia, LNH, and Neuroblastomes) resulting from exposures to electromagnetic fields (EMFs).

Influence of n-alkanes and petroleum on fatty acid composition of a hydrocarbonoclastic bacterium: Marinobacter hydrocarbonoclasticus strain 617.

This study concerns the effects of various long-chain n-alkanes, n-alkane mixtures and Arabian Light crude oil on the fatty acid (FA) composition of a sedimentary marine bacteria (Marinobacter hydrocarbonoclasticus strain 617), growing under aerobic conditions. The cultures with n-alkanes, as compared with soluble carbon sources, led to greater amounts of saturated and methyl branched FA (mainly belonging to a delta10 series). We observed the appearance or increase of saturated and unsaturated FA with the same carbon chain length (CCL) as the n-alkane carbon source (maximum for n-alkane CCL corresponding to the 'range' of the de novo synthesized fatty acids). We also observed a strong control of the oddness/evenness of the CCL of the FA by the oddness/evenness of the n-alkane. A n-alkane utilization index, (saturated + branched)/monounsaturated fatty acids (SAFA + BFA/MUFA) enabled discriminating between soluble carbon sources and hydrocarbons.

Enhanced biodegradation of phenanthrene by a marine bacterium in presence of a synthetic surfactant.

The biodegradation of phenanthrene by the marine strain Sphingomonas sp. 2MPII (DSMZ 11572) was enhanced by the solubilizating properties of the nonionic surfactant Tween 80. After 197 h of incubation, 85 +/- 4% of the initial amount of phenanthrene (0.4 g l-1) was biodegraded in presence of Tween 80 (0.5 g l-1) as opposed to 52 +/- 5% without this synthetic surfactant. These results confirm that the activity of the strain 2MPII is limited by the bioavailability of the polycyclic aromatic hydrocarbon (PAH) substrate in the aqueous phase. Tween 80 appears to be efficient in increasing the bioavailability of hydrophobic compounds such as PAHs.

[Measurement of radon concentrations in schools at 2 communities of Puglia and oncologic risk]. / Indagine sulle concentrazioni di radon nelle scuole di due comuni pugliesi e rischio oncologico.

Measurements of radon's concentration have been effected in the schools of two communes of Puglia: Polignano and Valenzano. The average concentration of radon, based on the prospecting effected in the research period (spring/summer 1997), appeared, for both communes, lower than national average (75 Bq/m3). However, these average concentrations is below estimated caused of the limitation of the research period, owing to organization only at the period spring/summer. In fact, by the extrapolation applying a right factor of conversion, of the "winter" average concentrations from that average calculated during the research period (considering the evident limits of such procedure) we obtain, respectively, for both schools of Polignano and Valenzano, an average concentrations about of 75 Bq/m3 e 103 Bq/m3. The research shows the existence of a correlation among the obtained results and two important factors: the buildings materials utilized and the geological conformation of the subsoil; in particular, the buildings having main wall (tufa) or built by mixed materials (cement, bricks and tufa) measured the highest radon concentrations. Nevertheless, the absolute risk to develop a pulmonary tumour due to the radon exposition, limitedly for the population of the common of Valenzano, appears lightly superior in respect of the national risk (6 cases on 100,000 fellows) resulted about 8.5 case on 100,000 fellows.

Marinobacter hydrocarbonoclasticus gen. nov., sp. nov., a new, extremely halotolerant, hydrocarbon-degrading marine bacterium.

On the basis of phenotypical characteristics and analysis of 16S rRNA sequence, a new species belonging to a new genus is described, and the name Marinobacter hydrocarbonoclasticus is proposed. This organism, isolated from Mediterranean seawater near a petroleum refinery, is a gram-negative, aerobic, rod-shaped bacterium. It grows at NaCl concentrations of 0.08 to 3.5 M and uses various hydrocarbons as the sole source of carbon and energy. Its DNA has a guanine-plus-cytosine content of 52.7 mol%. The 16S rRNA analysis shows a clear affiliation between M. hydrocarbonoclasticus and the gamma group of the phylum Proteobacteria. A close phylogenetic relationship appears among the species Marinomonas vaga, Oceanospirillum linum, Halomonas elongata, and Pseudomonas aeruginosa. Because of the impossibility of finding a single most closely related species, we suggest that this bacterium be assigned to a new genus, at least temporarily. The possibility of a revision of this status when new data appear is, however, not excluded. The type strain is M. hydrocarbonoclasticus SP.17 (= ATCC 49840).

[Effects of isometheptene on gallbladder contraction]. / Effetti dell' isometeptene sulla contrazione della colecisti.

In order to verify the influence of the adrenergic system on the gallbladder contraction the Authors studied the effect of isometeptene, a sympathomimetic drug active on beta receptors, on gallbladder emptying. The gallbladder was studied on 6 subjects using a real-time (linear array) equipment with a 3,5 MHz transducer. In fasting subjects the gallbladder emptying was obtained by a fatty meal. In the next day the fatty meal stimulation was associated with the administration of isometeptene. The mean values +/- S.D. of the follow parameters were evaluated: -gallbladder basal volume -gallbladder residual volume both after fatty meal and after the association with isometeptene -gallbladder maximum emptying percentage in two experiences. The results show that the isometeptene doesn't inhibit significantly the gallbladder emptying induced by a fatty meal.

[Evaluation of gallbladder volume and motility with echotomography]. / Valutazione del volume e della motilita' colecistica con l'ecotomografia.

The gallbladder volume has been evaluated in 22 normal volunteer subjects (12 men and 10 women) by the real time ultrasound B scanning, using the formula V = 1/6 abc (a = length; b = width, c = antero-posterior diameter). The fasting GB volume, the residual GB volume after Bronner's meal (every 15 min. for 90 min.) and the maximum emptying (%) were evaluated in each subject. It was not observed any statistically significative difference between the two groups (fasting volume: 18.5 +/- 6.0 ml in men and 15.2 +/- 5.1 ml in women; residual volume: 4.3 +/- 1.5 ml in men and 3.5 +/- 1.3 ml in women). The maximum GB emptying in women occurs later. In order to verify its reproducibility, the experience has been repeated in two different days in seven men without any statistically significative difference.

[Echotomographic determination of gallbladder diameters]. / Determinazione ecotomografica dei diametri della colecisti.

The investigation methods of gallbladder physiology in normal man are usually limited because often they are invasives and hazardous (radiations). The gallbladder size was accurately evaluated by US with study "in vitro". The variations in gallbladder-size measurements in the same observer and among many observers by US, demonstrate that Real-Time ul-ultrasonography is a simple, accurate, non-invasive and potentially valuable mean in the study of gallbladder motility.