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Ragusano is a raw milk pasta-filata cheese produced in the southeastern region of Sicily at farmhouse level using natural rennet and traditional wooden tools. The cheese manufacturing process involves two rounds of cooking, stretching of the overnight-ripened curd, moulding, brining and aging. Slight differences in the manufacturing process among producers lead to different flavor and texture of the final product. In an attempt to define the key steps of the process that lead to a safe cheese, we monitored the production of PDO Ragusano cheese in five farms. During each of three subsequent days, two blocks of Ragusano were produced per farm, for an overall count of thirty blocks of cheese. Chemical and microbiological analysis were performed on milk, production intermediates and different sections (sub-rind, intermediate, and core) of cheese aged for 2, 3 and 4 months. Results point to the importance of the second cooking of the curd as a key step in establishing the necessary conditions for a fast acidification. In fact, in the only farm where this step takes place for 10 minutes only (as opposed to an average length of 150 minutes), acidification of the curd appears to be noticeably slower, with a concomitant higher susceptibility to contamination. Manufacturing conditions proved to be very effective in causing death of the small number of Staphylococcus aureus cells found as contaminants of raw milk. No contaminations by Listeria monocytogenes, Salmonellae and Escherichia coli were detected at any stage of the process. On the other hand, after 2 months from manufacturing already, cheeses showed water activity levels which are hardly compatible with growth of the above mentioned pathogens, and which decreased further with subsequent aging. Our results show that the cheese-making technology of Ragusano guarantees high levels of microbiological safety as long as good practices are mantained throughout the entire process. The high temperatures used for cooking and stretching the curd, fast acidification, low aw values and high salt content contribute to make Ragusano a hostile environment for the growth of pathogens. [ABSTRACT FROM AUTHOR]

The advantages and applications of the videolaparoscopic technique (VL) versus open surgery in the treatment of acute and complicated appendicitis are not well defined. The aim of this study was to identify which of the two procedures is more suitable. The study examined 124 patients, 73 females (57.5%) and 51 males (42.5%). We compared 62 patients in the laparoscopic group with 62 open surgery patients. We analysed the results of the two groups (VL, open) for age, gender, operative time, hospital stay, complications and costs. The mean patient age was 24.1 years (range: 4-70). The conversion rate was 1.6% (1 case/62). The patients in the laparoscopic group were predominantly female (p < 0.0001). The average age of VL patients compared to open surgery was significantly higher (p < 0.01). The mean operative time was not significantly different between the two groups. The hospitalisation time was shorter for the laparoscopic group (4.77 vs. 6.39 days, p < 0.01). Complications were 0% for VL and 4.81% for the open group. The average operation cost in the open group was 8070.00 euros (+/- 4267) and 6818.00 euros (+/- 1446,00) for VL (p < 0.05). Laparoscopic appendectomy has significant advantages over traditional open surgery and should be the first choice in cases of acute but uncomplicated appendicitis.

Accurate segregation of chromosomes, essential for the stability of the genome, depends on 'bi-orientation'-simultaneous attachment of each individual chromosome to both poles of the mitotic spindle. On bi-oriented chromosomes, kinetochores (macromolecular complexes that attach the chromosome to the spindle) reside on the opposite sides of the chromosome's centromere. In contrast, sister kinetochores shift towards one side of the centromere on 'syntelic' chromosomes that erroneously attach to one spindle pole with both sister kinetochores. Syntelic attachments often arise during spindle assembly and must be corrected to prevent chromosome loss. It is assumed that restoration of proper centromere architecture occurs automatically owing to elastic properties of the centromere. Here we test this assumption by combining laser microsurgery and chemical biology assays in cultured mammalian cells. We find that kinetochores of syntelic chromosomes remain juxtaposed on detachment from spindle microtubules. These findings reveal that correction of syntelic attachments involves an extra step that has previously been overlooked: external forces must be applied to move sister kinetochores to the opposite sides of the centromere. Furthermore, we demonstrate that the shape of the centromere is important for spindle assembly, because bipolar spindles do not form in cells lacking centrosomes when multiple chromosomes with juxtaposed kinetochores are present. Thus, proper architecture of the centromere makes an important contribution to achieving high fidelity of chromosome segregation.