Unlabeled Semen Analysis by Means of the Holographic Imaging

The morphology, the motility, and the biochemical structure of the spermatozoon have often been correlated with the outcome of in vitro fertilization and have been shown to be the sole parameters of the semen analysis in predicting the success of intracytoplasmic sperm injection and intracytoplasmic morphologically selected sperm injection. In this context, digital holography has demonstrated to be an attractive technique to perform a label-free, noninvasive, and high-resolution technique for characterization of live spermatozoa. The aim of this chapter is to summarize the recent achievements of digital holography in order to show its high potentiality as an efficient method for healthy and fertile sperm cell selection, without injuring the specimen and to explore new possible applications of digital holography in this field

Fishery reserves in the Mediterranean Sea: the Gulf of Castellammare case study

The effects of fisheries management based on artificial reefs and on trawl banning are explored in the Gulf of Castellammare fishery reserve by means of biological (from trammel and trawl survey) data collected during several research programs between 1990 and 2001. The artificial reefs have caused an increase of diversity but not of biomass, as suggested by the comparison between the associated fish assemblage and that of nearby sandy bottoms. The associated species however do not have any trophic relation to the boulders, except the twobanded seabream, Diplodus vulgaris. An overall increase of experimental trammel net yields in the artificial reef area was observed from 1990 to 1998, due mainly to pelagic species associated with the boulders. The trawl ban caused a dramatic increase of groundfish biomass in the protected area (+711% after four years, total species). Different species had different increase rates, from 2-fold for the musky octopus, Eledone moschata to 127-fold for the gurnard, Lepidotrigla cavillone. Eight and ten years after the ban started, the yields did not vary significantly in the overall area, but decreased near to (both outside and inside) the protected area, probably due to increased legal and illegal trawling. The mean size did not increase in three studied species, except for the monkfish, Lophius budegassa. In conclusion the Gulf of Castellammare fishery reserve is considered a positive example of marine coastal fisheries management, especially considering the effects of the trawl ban on the abundance of groundfish stocks, although the cooperation between scientists and administrative bodies is still far from optimal

Innovative Caciocavallo cheeses made from a mixture of cow milk with ewe or goat milk.

This study assessed and compared the physicochemical, microbiological, and sensorial characteristics of Caciocavallo cheeses, made from cow milk and a mixture of cow with ewe or goat milk, during ripening. Different cheese-making trials were carried out on an industrial scale following the standard procedure of pasta filata cheeses, with some modifications. The percentage of the different added milk to cow milk influenced compositional and nutritional characteristics of the innovative products, leading to a satisfactory microbiological and sensorial quality

MEASUREMENT OF MACRO-SCALE INDENTATION MODULUS USING THE PRIMARY HARDNESS STANDARD MACHINES AT INRIM

In this paper it is described the experimental procedure and the statistical method for the measurement of indentation modulus by using the primary hardness standard machine at INRIM, in the macro-scale range. Indentation modulus is calculated on the basis of Doerner-Nix linear model and from accurate measurements of indentation load, displacement, contact stiffness and Vickers hardness impression imaging. Load is provided by dead-weight masses and displacement is measured by a laser-interferometric system, perpendicular with respect to the Vickers pyramid vertex. The geometrical dimension of the Diamond Pyramid Hardness (DPH) impression is measured by means of a micro-mechanical system and optical microscopy imaging technique. Applied force and indentation depth are measured simultaneously, 16 Hz of sampling rate, and the resulting indentation curve is obtained. Preliminary tests are performed on metals and alloys samples. Considerations and comments on the accuracy of the proposed method and analysis are discussed

Homology-based modeling of universal stress protein from listeria innocua up-regulated under acid stress conditions

An Universal Stress Protein (USP) expressed under acid stress condition by Listeria innocua ATCC 33090 was investigated. The USP was up-regulated not only in the stationary phase but also during the exponential growth phase. The three dimensional (3D) structure of USP was predicted using a combined proteomic and bioinformatics approach. Phylogenetic analysis showed that the USP from Listeria detected in our study was distant from the USPs of other bacteria (such as Pseudomonas spp., Escherichia coli, Salmonella spp.) and clustered in a separate and heterogeneous class including several USPs from Listeria spp. and Lactobacillus spp. An important information on the studied USP was obtained from the 3D-structure established through the homology modelling procedure. In detail, the Model_USP-691 suggested that the investigated USP had a homo-tetrameric quaternary structure. Each monomer presented an architecture analogous to the Rossmann-like α/β fold with 5 parallel β-strands and 4 α-helices. The analysis of monomer-monomer interfaces and quality of the structure alignments confirmed the model reliability. In fact, the structurally and sequentially conserved hydrophobic residues of the β-strand 5 (in particular the residues V146 and V148) were involved in the inter-chains contact. Moreover, the highly conserved residues I139 and H141 in the region α 4 were involved in the dimer association and functioned as hot spots into monomer-monomer interface assembly. The hypothetical assembly of dimers was also supported by the large interface area and by the negative value of solvation free energy gain upon interface interaction. Finally, the structurally conserved ATP-binding motif G-2X-G-9X-G(S/T-N) suggested for a putative role of ATP in stabilizing the tetrameric assembly of the USP. Therefore, the results obtained from a multiple approach, consisting in the application of kinetic, proteomic, phylogenetic and modelling analyses, suggest that Listeria USP could be considered a new type of ATP-binding USP involved in the response to acid stress condition during the exponential growth phase

Raw milk from vending machines: Effects of boiling, microwave treatment, and refrigeration on microbiological quality

Abstract In Italy, the sale of raw milk from vending machines has been allowed since 2004. Boiling treatment before its use is mandatory for the consumer, because the raw milk could be an important source of foodborne pathogens. This study fits into this context with the aim to evaluate the microbiological quality of 30 raw milk samples periodically collected (March 2013 to July 2013) from 3 vending machines located in Molise, a region of southern Italy. Milk samples were stored for 72h at 4°C and then subjected to different treatments, such as boiling and microwaving, to simulate domestic handling. The results show that all the raw milk samples examined immediately after their collection were affected by high microbial loads, with values very close to or even greater than those acceptable by Italian law. The microbial populations increased during refrigeration, reaching after 72h values of about 8.0 log cfu/mL for Pseudomonas spp., 6.5 log cfu/mL for yeasts, and up to 4.0 log cfu/mL for Enterobacteriaceae . Boiling treatment, applied after 72h to refrigerated milk samples, caused complete decontamination, but negatively affected the nutritional quality of the milk, as demonstrated by a drastic reduction of whey proteins. The microwave treatment at 900 W for 75s produced microbiological decontamination similar to that of boiling, preserving the content in whey proteins of milk. The microbiological characteristics of raw milk observed in this study fully justify the obligation to boil the raw milk from vending machines before consumption. However, this study also showed that domestic boiling causes a drastic reduction in the nutritional value of milk. Microwave treatment could represent a good alternative to boiling, on the condition that the process variables are standardized for safe domestic application

Biodiversity of Lactobacillus plantarum from traditional Italian wines

In this study, 23 samples of traditional wines produced in Southern Italy were subjected to microbiological analyses with the aim to identify and biotype the predominant species of lactic acid bacilli. For this purpose, a multiple approach, consisting in the application of both phenotypic (API 50CHL test) and biomolecular methods (polymerase chain reaction-denaturing gradient gel electrophoresis and 16S rRNA gene sequencing) was used. The results showed that Lactobacillus plantarum was the predominant species, whereas Lb. brevis was detected in lower amount. In detail, out of 80 isolates 58 were ascribable to Lb. plantarum and 22 to Lb. brevis. Randomly amplified polymorphic DNA-polymerase chain reaction was used to highlight intraspecific variability among Lb. plantarum strains. Interestingly, the cluster analysis evidenced a relationship between different biotypes of Lb. plantarum and their origin, in terms of wine variety. Data acquired in this work show the possibility to obtain several malolactic fermentation starter cultures, composed by different Lb. plantarum biotypes, for their proper use in winemaking processes which are distinctive for each wine

Transcriptome and metabolome reprogramming in tomato plants by Trichoderma harzianum strain T22 primes and enhances defence responses against aphids

Beneficial fungi in the genus Trichoderma are among the most widespread biocontrol agents of plant pathogens. Their role in triggering plant defences against pathogens has been intensely investigated, while, in contrast, very limited information is available on induced barriers active against insects. The growing experimental evidence on this latter topic looks promising, and paves the way towards the development of Trichoderma strains and/or consortia active against multiple targets. However, the predictability and reproducibility of the effects that these beneficial fungi is still somewhat limited by the lack of an in-depth understanding of the molecular mechanisms underlying the specificity of their interaction with different crop varieties, and on how the environmental factors modulate this interaction. To fill this research gap, here we studied the transcriptome changes in tomato plants (cultivar “Dwarf San Marzano”) induced by Trichoderma harzianum (strain T22) colonization and subsequent infestation by the aphid Macrosiphum euphorbiae. A wide transcriptome reprogramming, related to metabolic processes, regulation of gene expression and defence responses, was induced both by separate experimental treatments, which showed a synergistic interaction when concurrently applied. The most evident expression changes of defence genes were associated with the multitrophic interaction Trichoderma-tomato-aphid. Early and late genes involved in direct defence against insects wereinduced (i.e. peroxidase, GST, kinases and polyphenol oxidase, miraculin, chitinase), along with indirect defence genes, such as sesquiterpene synthase and geranylgeranyl phosphate synthase. Targeted and untargeted semi-polar metabolome analysis revealed a wide metabolome alteration showing an increased accumulation of isoprenoids in Trichodermatreated plants. The wide array of transcriptomic and metabolomics changes nicely fit with the higher mortality of aphids when feeding on Trichoderma treated plants,herein reported,and with the previously observed attractiveness of these latter towards the aphid parasitoid Aphidius ervi.Moreover, Trichoderma treated plants showed the over-expression of transcripts coding for several families of defence-related transcription factors (bZIP, MYB, NAC, AP2-ERF, WRKY), suggesting that the fungus contributes to the priming of plant responses against pest insects. Collectively, our data indicate that Trichoderma treatment of tomato plants induces transcriptomic and metabolomic changes, which underpin both direct and indirect defence responses

Management of a patient with multiple device replacements and extractions: When the leadless pacemaker is a viable solution

Leadless pacemaker (LPs) is a safe device and the implantation rates of this device is increasing. The device extraction and replacement are today a challenging procedures especially in case of infections, fragile and older patients or in unfavorable venous anatomy; LPs can be a valid alternative strategy in these cases. We report a case of management of a patient with multiple previous device replacements and extractions, with malfunction of transvenous pacemaker and with a fibrous membrane between the walls of the ventricular lead and the superior vena cava (SVC), who underwent a successful LP implantation