SLC6A15, a novel stress vulnerability candidate, modulates anxiety and depressive-like behavior: involvement of the glutamatergic system.

Major depression is a multifactorial disease, involving both environmental and genetic risk factors. Recently, SLC6A15 - a neutral amino acid transporter mainly expressed in neurons - was proposed as a new candidate gene for major depression and stress vulnerability. Risk allele carriers for a single nucleotide polymorphism (SNP) in a SLC6A15 regulatory region display altered hippocampal volume, glutamate levels, and hypothalamus-pituitary-adrenal axis activity, all markers associated with major depression. Despite this genetic link between SLC6A15 and depression, its functional role with regard to the development and maintenance of depressive disorder is still unclear. The aim of the current study was therefore to characterize the role of mouse slc6a15 in modulating brain function and behavior, especially in relation to stress as a key risk factor for the development of mood disorders. We investigated the effects of slc6a15 manipulation using two mouse models, a conventional slc6a15 knock-out mouse line (SLC-KO) and a virus-mediated hippocampal slc6a15 overexpression (SLC-OE) model. Mice were tested under basal conditions and following chronic social stress. We found that SLC-KO animals displayed a similar behavioral profile to wild-type littermates (SLC-WT) under basal conditions. Interestingly, following chronic social stress SLC-KO animals showed lower levels of anxiety- and depressive-like behavior compared to stressed WT littermates. In support of these findings, SLC-OE animals displayed increased anxiety-like behavior already under basal condition. We also provide evidence that GluR1 expression in the dentate gyrus, but not GluR2 or NR1, are regulated by slc6a15 expression, and may contribute to the difference in stress responsiveness observed between SLC-KO and SLC-WT animals. Taken together, our data demonstrate that slc6a15 plays a role in modulating emotional behavior, possibly mediated by its impact on glutamatergic neurotransmission.

Bacterial dynamics in a raw cow's milk Caciotta cheese manufactured with aqueous extract of Cynara cardunculus dried flowers.

AIMS: To investigate the bacterial dynamics of a Caciotta cheese traditionally manufactured in the Montefeltro area (Central Italy) with raw cow's milk and an aqueous extract of dried flowers from Cynara cardunculus as a coagulating agent. METHODS AND RESULTS: Conventional methods and a combined PCR-DGGE approach, relying on culture-dependent and -independent analyses, were used to investigate the cheese bacterial community, with a special focus on lactic acid bacteria. A heterogeneous population, including enterococci, lactococci, lactobacilli, food spoilage and other banal micro-organisms, was found. SIGNIFICANCE AND IMPACT OF THE STUDY: The study contributed to highlighting the influence of different technological parameters on bacterial dynamics of a raw milk Caciotta cheese coagulated with vegetable rennet. CONCLUSIONS: None of the species found in the vegetable rennet became dominant during the cheese-making and a prevailing role of the adventitions microbita coming from the raw milk and the dairy environment was highlighted.

Characterisation of Lactobacillus helveticus strains producing antihypertensive peptides by RAPD and inverse-PCR of IS elements.

Lactobacillus helveticus is used for the manufacture of cheeses and milk-based products. Although it is not considered a probiotic microorganism, some strains demonstrated beneficial effects through the production of antihypertensive peptides from the hydrolysis of casein during milk fermentation. Strain-specificity of bioactive peptide production by L. helveticus makes the availability of reliable typing methods essential for both legal and good manufacturing processes. Accordingly, RAPD and inverse-PCR of five insertion sequence elements were comparatively evaluated for the molecular characterisation of four L. helveticus dairy cultures producing antihypertensive peptides and fourteen reference strains. Calculation of discriminatory indices and cluster analysis of the DNA fingerprints confirmed the suitability of both approaches for acceptable strain differentiation. Although RAPD was more discriminating, for a few test strains a neat discrimination was only achieved through multiplex inverse-PCR, thus suggesting the suitability of a combined analytical approach for a finer strain discrimination.

Phenotypic, genotypic and technological characterization of predominant lactic acid bacteria in Pecorino cheese from central Italy.

AIMS: Identification and biotyping of lactic acid bacteria (LAB) isolated from raw-milk Pecorino cheese manufactured in the Marche region (central Italy) for selection of suitable starter cultures or adjuncts. METHODS AND RESULTS: Preliminary characterization with morphological and biochemical assays were undertaken for 112 Gram-positive and catalase-negative isolates. Unequivocal identification of the isolates was obtained through restriction analysis of the amplified 16S rRNA gene and sequencing of 360-380 bp amplicons. Fifty-nine isolates belonging to LAB species generally recognized as safe and potentially utilized as starters or flavour-producing adjuncts were preselected and tested for their acidifying, proteolitic and autolytic activities. Fifty-five of these isolates were also subject to RAPD (randomly amplified polymorphic DNA) fingerprinting and unweighted pair-group method with arithmetic averages (UPGMA) cluster analysis for the estimation of genotypic intra-species variation. As a result, in Pecorino cheese, a heterogeneous lactic acid bacteria population, which includes strains with metabolic characteristics of technological interest, was characterized. CONCLUSIONS: The polyphasic approach proposed allows the bacterial ecology of Pecorino cheese to be investigated and allows to assess the potential role of autochthonous LAB strains for the dairy industry. SIGNIFICANCE AND IMPACT OF THE STUDY: The great economic importance of Pecorino cheese encouraged a deeper knowledge of its microbiota, which is known to influence the peculiar sensory properties of this cheese, also in view of its exploitation.

Occurrence and characterization of Aeromonas spp. in mussels from the Adriatic Sea.

Samples of mussels (Mytilus galloprovincialis) were collected from natural beds on the Adriatic Sea in front of Ancona Province (Marches, Central Italy) officially recognized for human consumption. The mussels were analyzed to evaluate the prevalence of Aeromonas spp. pathogenic strains. Out of 144 samples, 32 Aeromonas strains were isolated and 12 showed virulence and enterophatogenicity on mice. Isolates were also examined to check their sensitivity to different antimicrobial agents. All isolates were sensitive to imepenem, neomycin, ciprofloxacin, tetracycline, oxytetracycline, doxycicline, chloramphenicol, polymyxin B, and colistin sulfate, but resistant to penicillin, carbenicillin, cephalotin, sulfamethoxazole, lincomycin, novobiocin, and rifampicin. Multiple resistance to antimicrobial agents with different mechanism of action was observed. Owing to the high occurrence of pathogenic Aeromonas strains, the risk of developing human infections by eating raw or undercooked mussels is envisaged.

Tori of confusion: binaural localization cues for sources within reach of a listener.

To a first-order approximation, binaural localization cues are ambiguous: many source locations give rise to nearly the same interaural differences. For sources more than a meter away, binaural localization cues are approximately equal for any source on a cone centered on the interaural axis (i.e., the well-known "cone of confusion"). The current paper analyzes simple geometric approximations of a head to gain insight into localization performance for nearby sources. If the head is treated as a rigid, perfect sphere, interaural intensity differences (IIDs) can be broken down into two main components. One component depends on the head shadow and is constant along the cone of confusion (and covaries with the interaural time difference, or ITD). The other component depends only on the relative path lengths from the source to the two ears and is roughly constant for a sphere centered on the interaural axis. This second factor is large enough to be perceptible only when sources are within one or two meters of the listener. Results are not dramatically different if one assumes that the ears are separated by 160 deg along the surface of the sphere (rather than diametrically opposite one another). Thus for nearby sources, binaural information should allow listeners to locate sources within a volume around a circle centered on the interaural axis on a "torus of confusion." The volume of the torus of confusion increases as the source approaches the median plane, degenerating to a volume around the median plane in the limit.

Normal tension glaucoma: a ten-year follow-up.

The authors describe the results obtained in 10 years of observation of a group of 41 eyes affected by normal tension glaucoma. An evolution of perimetric defects was found in 29.26% of the eyes examined.